Experimental induction of feline viral rhinotracheitis virus re-excretion in FVR-recovered cats. The re-excretion of feline viral rhinotracheitis (FVR) virus (feline herpesvirus I) by FVR-recovered cats is recorded both spontaneously and following a variety of stimuli, namely, corticosteroid administration, change of housing, and parturition and lactation. At least 27 of 33 (82%) FVR-recovered cats studied were shown to be viral carriers. The carrier state was characterised by periods of viral latency interspersed with episodes of viral shedding. Administration of 0-75 mg dexamethasone trimethylacetate and 2-25 mg prednisolone on days 0,2 and 4 resulted in re-excretion after a mean lag period of 7-2 days in 22 of 32 (69%) FVR-recovered cats on a total of 31 of 57 (54%) occasions. Rehousing resulted in virus re-excretion after a mean lag period of 7-2 days in four of 22 (18%) cats tested on a total of six of 40 (15%) occasions. Apparently spontaneous shedding occurred on a total of 10 occasions in nine of 31 (29%) cats during a mean observation period of 8-8 months. Four of six FVR-recovered queens in a total of four of 10 litters (40%) shed virus within two to 10 weeks of parturition. Serum neutralising antibody titres were generally boosted at the time of first re-infection but afterwards remained essentially constant. Although 82% of cats in these studies were shown to be viral carriers, only 45% of cats shed virus spontaneously or as a result of the natural stress situations and it is postulted that these naturally excreting cats are of most significance epidemiologically.


Induction of proliferation in dense non-starved 3T3 cells by Ca++ ionophore A23187. The proliferogenic effect of the Ca++ ionophore A23187 was tested in dense non-starved 3T3 cells. Whereas continuous exposure during 48 h of cells to the ionophore at concentrations is larger than or equal to 0.4 muM cytotoxic, a short exposure for 30 s up to 4 min at 0.2 muM was proliferogenic. It was also found that such short exposures to the ionophore caused a transient increase in the intracellular level of cyclic GMP and a roughly simultaneously appearing decrease in the intracellular level of cyclic AMP.


Alteration of the cytoplasmic endonuclease cleavage pattern of SV-40 deoxyribonucleic acid in the presence of ATP, actinomycin D and ethidium bromide. Endonuclease activities of cytoplasmic extracts of BSC-1 (E1) and BHK-21-cl 13 (E2) cells were assayed with SV-40 as a substrate and analysed by velocity sedimentation in alkaline and neutral sucrose gradients. Endonucleases were found to require Mg2+ ions for their activity. E1 endonuclease generated linear 6 S DNA fragments, pointing to non-random double-strand cleavage of DNA ; the action of E2 endonuclease resulted in double-strand DNA cleavage to fragments heterogeneous in size. Both endonucleases were inhibited by ATP. The inhibitory effect of actinomycin D (AD) was proportional to the AD/DNA molar ratio. AD+ATP association as well as the presence of ethidium bromide altered the cleavage pattern of E1 towards the predominance of single-strand breaks.


[Investigations concerning the evidence of herpes simplex virus type 2 (HSV-2) antibodies in patients with carcinoma of the cervix (author's transl)]. The neutralizing antibodies to HSV-1 and HSV-2 were determined in the sera of 128 patients. Infection was detectable in nearly 100% of the cases in each of the three investigated groups (patients with carcinoma of the cervix, female patients with chronic recurrent HSV infection in the genital area and a control group without and history of HSV infection). The percentage of patients displaying HSV-2 antibodies in the group with carcinoma of the cervix (38%) is significantly higher than in the control group (12%). The results are compared with the findings of other authors and the possible causal significance of HSV in carcinogenesis is discussed.


Fluorometric determination of the quality of FITC conjugates. A fluorometric method for the evaluation of FITC conjugates is described. By this method, the titer and the dilution required for complete antigen detection can be determined and an objective comparison of the various conjugate specimens is possible.


[Identification of the optimal requirement of growing animals for polyunsaturated fatty acids]. Data obtained from studying eight groups of animals of a growing age showed their food rations to ensure proper growth and development when they contain optimal amounts of all food ingredients, including polyunsaturated fatty acids. Therefore, the fatty component in the food rations of test animals should contain both animal and vegetable fats, the latter comprising quantitatively 15 to 25 per cent of diurnal fat allowance. Such an amount of vegetable oil fully provides the rattlings with polyunsaturated fatt acids in a proportion comprising 4.3-5.8 per cent of the diurnal calorific value of the ration. For this reason, in investigations involving animal tests, or experimental substantiation of various problems pertaining to baby alimentation the authors recommend to use in feeding rattlings rations balanced in all the ingredients, including also the fatty acids composition.


[Effect of the nature of the nutrition and of the functional state of the hypophyseal-adrenal system on the "convulsion readiness" of the body]. The effect of some amino acids, succinate and different doses of ACTH on the convulsion response readiness of the organism under the action of a strong sound stimulant was studied. With enrichment of the ration with glutamic acid and also following a parenteral introduction of large doses of succinate a definite protective effect was revealed and, conversely, administration of ACTH (intramuscularly, for 7 day in a dose of 5 Units per 100 g of body weight) increased the convulsion response readiness of the organism. A possible mechanism underlying the detected changes in the convulsion response readiness of the organism under the active factors under study are discussed.


[Contribution to retrospective roentgenographic follow-up of solitary pulmonary nodules (author's transl)]. The diagnostic problems of malignant solitary pulmonary nodules are demonstrated in 5 cases with a roentgenographic history of many years. The slow growth rate of adenocarcinoma and alveolar cell carcinoma--mainly in woman--and the possibility of a scar tissue carcinoma has to be considered. Peripheral pulmonary nodules in adult persons suspicious of carcinoma must be verified histologically, even if their growth rate is very slow and their roentgenographic history lasts many years. If this is not possible a thoracotomy should be performed.


[Main signs of neurologic disease during pregnancy (author's transl)]. The most important neurologic diseases which may start during pregnancy are described. On the basis of clinical, easily discovered, signs the essential differential diagnosis and specific diagnostic techniques (electromyography, electroneurography, angiography, computotumography) and their indications are outlined. In some detail prognosis and indications for common diseases (epilepsy, M.S.) are discussed. Treatment of those diseases is especially considered in which emergency drug therapy may be needed.


Metabolic fate of N-n-butyl-N-(4-hydroxybutyl)-nitrosamine and its analogues. Selective induction of urinary bladder tumours in the rat. 1. The metabolic fates of N-n-butyl-N-(4-hydroxybutyl)nitrosamine (BBN) and N,N-di-n-butylnitrosamine (DBN) were investigated in the rat and other animal species, to elucidate a possible relationship between metabolism and organotropic carcinogenicity to the urinary bladder of these N-nitrosamines. 2. The principal urinary metabolite of BBN as well as of DBN in the rat was N-n-butyl-N-(3-carboxypropyl)nitrosamine (BCPN), which was demonstrated to be the active form of these compounds as bladder carcinogen. The species difference in response to BBN or DBN is discussed on the basis of the urinary excretion rate of BCPN. 3. Metabolism in vivo and carcinogenicity of a number of BBN analogues were investigated in the rat and a general scheme for biotransformation of N-alkyl-N-(omega-hydroxyalkyl)nitrosamines is given. 4. A possible correlation of structure and metabolism with organotropic carcinogenicity of BBN analogues is discussed, with special reference to selective induction of bladder tumours. 5. A clear demonstration of overlapping carcinogenic and mutagenic activities is presented for BBN, DBN and related compounds.


Endocrine dysfunction and slipped captial femoral epiphysis. Five patients with concomitant endocrinopathy and slipped capital femoral epiphysis were studied in detail. One had diabetes and hypothyroidism, one had hypothyroidism, one had hypergonadotropic hypogonadism and two had a craniopharyngioma (one of whom had severe panhypopituitarism post-operatively). An additional seven patients with cranio-pharyngioma revealed marked delay in closure of epiphyses and an additional undiagnosed case of slipped capital femoral epiphysis. Of the six patients with slipped capital femoral epiphysis, three had bilateral and three unilateral involvement. Of the five patients undergoing surgical stabilization, there was significant delay of epiphyseodesis, prompting us to recommend concomitant bone grafting. Histological examination of the femoral head from a three year old child with panhypopituitarism showed marked irregularity of the growth plate and loss of columnar integrity, which may be a predisposing factor to slipping in older children with endocrinopathies. The effects of various hormones on the physis are specifically discussed, especially as they relate to the possible etiology of slipped capital femoral epiphysis.


FORMATION OF H-addition radicals in adenine derivatives: part II. The formation of H-addition radicals in monocrystals of adenosine, adenosine-HCl, adenine-HCl-1/2H2O, and adenine-2 HCl by X-irradiation has been st-died by using electron spin resonance spectroscopy at 9.5 GHz and at 35 GHz. In all crystals, both H-addition radicals at position C2 and at position C8 were observed. The coupling constants of these two H-addition radicals are different and depend strongly on the protonation state of the adenine base. INDO calculations reproduce well the observed trends of the coupling constants. It is shown that the C2-addition radical is transformed into the C8-addition radical by heat and vice versa the C8-addition into the C2-addition by light of lambda greater than 360 nm.


Prophage induction by alkylating ethyl methylaminosulfonate. Treatment of the lysogenic strain M. lysodeikticus 53-40 (N5) with ethyl methylaminosulfonate results in a slight increase in infective center concentration of N5 phages which is only detectable once disturbing cells had been eliminated by lysozyme. After conversion of ethyl methylaminosulfonate with radioactive labelled guanosine at 37 degrees C, N7-ethyl-guanine could be ascertained. Therefore the biological activity of the drug may be due to an ethylation of DNA.


[Modification of glyceraldehyde-3-phosphate dehydrogenase from rabbit skeletal muscle by [3-(3-bromoacetylpyridinio)-propyl]-adenosine pyrophosphate (author's transl)]. The NAD analogue [3-(3-acetylpyridinio)-propyl]adenosine pyrophosphate forms enzymically inactive complexes with glyceraldehyde-3-phosphate dehydrogemase from yeast and rabbit skeletal muscle. In the latter enzyme four mol of the analogue are bound with equal affinity inhibiting the enzyme in a competitive way: K1 = 0.0 mM as compared to the dissociation constant KD = 0.6 mM. The brominated derivative [3-(3-bromoacetylpyridinio)-propyl]adenosine pyrophosphate is covalently bound to both enzymes causing irreversible loss of enzymic activity. Complete inactivation of the enzyme from muscle required two moles of the analogue per mol of tetramer. The remaining two sites are still able to bind two mol of NAD+ without regain of enzymic activity. In the case of the yeast enzyme four mol of the analogue are bound. Inactivation of the rabbit muscle enzyme is accompanied by the disappearance of two out of four highly reactive sulfhydryl groups; in the yeast enzyme the four active site cysteine residues are still able to react with DTNB, the reactivity being diminished significantly. Hybrid formation between the native enzymes from yeast and skeletal muscle is not affected by the modification of the enzyme. Similarly the sedimentation properties of the covalently modified enzyme are indistinguishable from those of the native molecule. This indicates that both the native and the irreversibly inhibited enzyme are identical regarding their quaternary structure.


[Paraneoplastic hypoglycemia due to malignant histiocytoma (Doege-Potter syndrome) (author's transl)]. A case report is given on a male patient who was 52 years old when he died. 9 years ago a nephrectomy was performed because of a densely-packed mesenchymal tumor of the adipose tissue of the pelvis renalis. 7 years after the surgical intervention he suffered from several hypoglycemic crises. An operation was performed, and 2 large tumor nodes were removed from the greater omentum. A retroperitoneal relapse was inoperable. Nevertheless, the disorder of the carbohydrate metabolism disappeared after the operation. At the autopsy, one year later, a large retroperitoneal tumor with metastases in the lymph nodes was found. The neoplastic growth was classified as a malignant histiocytoma, and the hyperglycemia was taken as a paraneoplastic syndrome (Doege-Potter).


[The biological behaviour of experimental tumours of the central nervous system (author's transl)]. In 31 experimental groups, 1955 rats were exposed to methyl-, dimethyl-, trimethyl-, and ethylnitrosourea or dimethylphenyltriazene. Altogether, 3102 tumours were registered. 1930 of them were located in the central nervous system. Derived from neurooncological classifications in man, these tumours were subdivided corresponding to a three-step grading scheme. In contrast to brain tumours in man, in rats the degree of dedifferentiation is of minor importance with regard to the clinical course of the cancer disease. In rats CNS tumours lead to death of the animals within a few hours or days after the onset of clinical symptomatology and should be regarded as biologically highly malignant. The grading allows a more exact valuation of the frequency of distinct tumour types in different experimental groups. Following transplacental application, isomorphous neoplasms occur significantly more frequent as compared with anisomorphous ones (grad II and III) which dominate when the carcinogen is given repeatedly to adult animals. Parallels are drawn to human neurooncology.


[Transplacental carcinogenic action of diethylnitrosamine in the Dzungarian hamster (author's transl)]. After transplacental application (30 mg/kg i.p., 3 days until 6 hours a. p.), diethylnitrosamine induces a high rate (76.2 per cent) of bronchiolar tumours in the lung of the Dzungarian hamster. These tumours were classified as papillary or tubulous adenomas and carcinomas. Furthermore, eome tumours were observed in other organs. The problem of organotropism of diethylnitrosamine, especially among different hamster hamster species, is discussed.


[Histological diagnosis and differential diagnosis of localized alveolar cell carcinoma (author's transl)]. The diagnosis of the alveolar cell carcinoma is based on the fact that it is a cytological and histological highly differentiated tumour. It consists thoroughly out of cylindrical cells. Furthermore, a diffuse carcinosis of the alveoli has to exist, i. e. lining of the intact alveolar walls by cancer cells. The adenocarcinoma is the only alternative in differential diagnosis to the alveolar cell carcinoma in view of the cytological and histological criteria. Pulmonary metastases of extrapulmonary adenocarcinomas can exactly imitate the histologic pattern of the alveolar cell carcinoma and can be diagnosed only by the clinician.


[Disturbances of sexual function following rectal excision and sphincter preserving resections of the rectum (author's transl)]. Male sexual function represents a many-sided effect. The disturbances aredeveloped in variable degress. Impotency after excision of malignant tumours occurs in each second patient and after anterior resection is in a third of patients observed. A modest chance for recovery is possible. Careful anatomic dissection is of utmost importance to safe guard the sexual function following curative excision of cancer. This way is less likely to cause injury to the nerves in the critical areas.


Pig kidney phosphofructokinase. Purification to homogeneity and qualitative basic characterization. Phosphofructokinase has been purified from pig kidney by extraction with phosphate buffer at pH 8, followed by alcohol treatment, affinity chromatography on matrix-bound Cibacron blue F3G-A, and gel chromatography on Sepharose 6B. Using sodium dodecyl sulphate electrophoresis the enzyme was found to be homogeneous and to have a specific activity of about 80 units/mg protein. Like other phosphofructokinases, at pH 7.0 the enzyme exhibits a sigmoidal dependence in its activity on the fructose 6-phosphate concentration and is strongly inhibited by ATP. The degree of citrate inhibition is influenced by the concentration of the two substrates. ATP strengthens and fructose 6-phosphate relieves the inhibition by citrate. AMP and cAMP are able to overcome the ATP inhibition. The ADP activation curve is biphasic. The molecular weight of the subunit of pig kidney phosphofructokinase was determined to be 88 000 by means of sodium dodecyl sulphate electrophoresis.


Mitochondrial redox components of human adrenocortical steroid hydroxylases under physiological conditions and in focal hyperplasia of the zone fasciculata. Cytochrome P450, aa3 and adrenodoxin were characterized in human adrenocortical mitochondria. Human adrenodoxin demonstrated a distinctive electron paramagnetic spectrum exhibiting anisotropic g values indicative of axial symmetry. The observed g values were (formula: see text). Adrenodoxin content of crude human adrenal mitochondria was 0.65 +/- 0.01 nmol/mg of mitochondrial protein and was in a stoichiometric 1:1 molar ration with cytochrome P450. Mitochondrial cytochrome P450 (0.62 nmol/mg mitochondrial protein) and aa3 (0.19 nmol/mg mitochondrial protein) levels were determined in a normal human adrenal A patient receiving ACTH (3d) demonstrated a doubling of P450 content, while cytochrome aa3 levels remained unchanged. ACTH plus hydrocortisone therapy (3d) increased both P450 and aa3 levels, however 24 h hydrocortisone therapy alone was without effect. An area of focal hyperplasia of the zona fasciculata also demonstrated P450 and aa3 levels elevated above the contralateral and normal human adrenal levels. The possibility of hormonal control of human adrenal mitochondrial cytochromes P450 and aa3 is suggested.


Biopsy study of hepatocarcinomas in Nigerian Igbos. Fifty-five hepatocarcinomas were found in a review of approximately 7,500 surgical biopsies done on Nigerian Igbos during a period of 6 years. The male: female ratio was 2.9:1 and the age peak was between 20 and 49 years. The main symptoms were abdominal swelling, pain, emaciation, jaundice, fever, anorexia and diarrhea. Physical examination revealed a palpable liver in nearly all patients. Two patients presented acutely with hemoperitoneum due to rupture of necrotic tumor nodule. Cirrhosis was found in 60% of the adequately sized specimens. In comparison with published data, this series from an ethnic group in Nigeria, West Africa, reveals both similarities and dissimilarities which are noteworthy.


Anti-HBc titer in relation to the etiological role of hepatitis B virus in primary hepatocellular carcinoma. The antibody to hepatitis B core antigen (anti-HBc) is belived to be a marker for natural infection with hepatitis B virus (HBV). In order to study the etiological role of HBV in relation to primary hepatocellular carcinoma (PHC), the anti-HBc in sera of 31 PHC patients was surveyed by the immune adherence hemagglutination method which was about 10 times more sensitive than the complement fixation method. Twenty two out of 31 PHC cases were positive for anti-HBc (71.0%). The is a higher rate of incidence than that of HBs-Ag (51.6%). However, high anti-HBc titer above 2(10) which might reflect current infection with HBV was observed in 15 of these 22 cases. The remaining 7 cases had a titer lower than 2(8); 5 of these patients had neither HBs-Ag nor anti-HBs in their sera. As control, 37 anti-HBs positive blood donors with no definite liver disease were surveyed for anti-HBc titer. Thirty-four of them were positive, but in the majority of cases, the serum titer was less than 2(9), which might only reflect previous infection with HBV. These findings indicate that not only the prevalence of anti-HBc, but also the determination of its real titer is quite important for studying the relationship of HBV to PHC.


Postmenopausal osteoporosis: the effect of parathormone and large dose vitamin D3 on the serum calcium level in sex hormone deficient rats. Rats deprived of adrenal and gonadel sex hormones are more sensitive than normal rats to the hypercalcaemic (osteolytic) effect of parathormone and toxic doses of vitamin D3. It is suggested that sex hormone deficiency and the consecutive decrease of calcitonin sensitivity in postmenopausal osteoporosis makes the patients unprotected against factors inducing increased bone resorption, and this leads over the years of osteoporosis.


Role of prostaglandin E2 in the stimulatory effect of ACTH on ovarian blood flow. It has been studied in dogs anaesthetized with chloralose-urethane whether Prostaglandin E2(PGE2) might act as a mediator of the stimulatory effect of ACTH on ovarian blood flow. Without any damage to the ovarian vessels, a heated thermocouple was inserted into the stroma of the ovaries and local blood flow was recorded continuously. Similarly to ACTH, PGE2 (100 ng/kg/min) infused into the ovarian bursa increased local blood flow. The ovaries were isolated in situ and perfused by a peristaltic pump from the femoral artery. Synthetic ACTH or PGE2 was infused into the ovarian artery for 15 min and perfusion pressure was measured. Both ACTH (12.5--25.0--50.0--100.0 ng/kg/min) and PGE2 (3912--6.25--12.50--25.00 ng/kg/min) reduced perfusion pressure, caused vasodilatation. There was a significant linear correlation between the degree of vasodilatation and the logarithm of the doses. If the doses of the two agents are expressed in terms of mols, the effect of both agents can be characterized by a common equation. Fifty--200 mug/kg indomethacin inhibited the effect of ACTH but increased the effect of PGE2. It is suggested that in dogs ACTH exerts its ovarian blood flow stimulatory effect by releasing PGE2 or some similar agent(s).


Cyclic AMP and segmental epidural analgesia during labour. The blood cyclic AMP level was determined just before, during and after induced labour in 27 healthy women. To achieve complete pain relief during the first stage of labour, 14 of them were given segmental epidural analgesia at the height of Th 10-12. The remaining parturients served as controls. Cyclic AMP was above the normal non-gravid level before induction in both groups. In the control group the cyclic AMP content decreased during the first stage of labour, then it increased and reached a peak at the moment of delivery. These changes were, however, not significant. In the epidural group the cyclic AMP level rose significantly during the first stage, and also reached its peak at the moment of delivery. There was a statistically significant difference between the groups at a cervical dilatation of 6-8 cm. After delivery the cyclic AMP rapidly declined to its initial value in both groups. The possible role of the decreased uterine contractions after the block in the increase of the cyclic AMP is discussed.


Effect of adrenocorticotrophic hormone on muscle acetylcholinesterase and nonspecific esterase. Acetylcholinesterases in the muscle homogenates of hamsters receiving ACTH intraperitoneally for six weeks were reduced as compared to the activities in the control samples. In polyacrylamide disc gel electrophoresis of the ACTH-treated muscles some acetylcholinesterase bands were absent. These observations may be significant in explaining the ACTH-induced improvement in myasthenia gravis patients and in the pathophysiology of steroid myopathy.


In vitro lymphocyte transformation of MS patients to paramyxovirus antigens. The immune response of 21 MS patients and 21 healthy volunteers was tested in vitro lymphocyte transformation assay. The stimulating antigens were measles, mumps, parainfluenza, E. coli, S. aureus and C. albicans. MS patients were found to respond similarly to the normal controls to all antigens tested with the exception of mumps antigen. The maximum responses of the MS patients to this antigen were significantly lower than those observed for the normals. However, since 95 per cent of the MS lower maximum responses were still within the positive range, the immunological significance of this phenomenon will require further clarification.


Some histological aspects of amyloid polyneuropathy. Two sporadic cases of amyloid polyneuropathy with clinical features corresponding to the Portuguese type of this disease were studied. Histological examination of sural nerve demonstrated a marked loss of myelinated and unmyelinated fibres in the case 1 due to axonal degeneration, high content of fibers with segmental demyelination and the occurrence of several enlarged axons filled with the 10 nm filaments (so-called giant axons). In the case 2 there was total loss of unmyelinated axons and myelinated fibers were nearly completely lacking. In the development of changes in the myelinated fibers their direct compression by amyloid deposits seems to play  an important role. It leads to the appearance of both axonal degeneration and segmental demyelination. The latter seems to be due to local compression and it may involved many fibers. In the  light of observations reported by other authors the mechanism of changes developing in unmuelinated fibers is explained by the presence of changes in the cells of posterior root ganglia, however the question whether some abnormalities seen in unmyelinated axons could not be related to the pressure exerted by amyloid deposits directly to these fibers, remains open.


The ultrastructure of early visna lesions. The ultrastructure of visna, a slowly progressive menigo-encephalomyelitis of sheep, was studied in animals sacrificed one month after intracerebral inoculation of visna virus. The major pathological changes, representative of those seen during the first year after infection, consist of inflammation and minor focal destructive lesions of grey and white matter. The inflammatory infiltrates, both subependymal and perivascular as well as of the choroid plexus, were composed mainly of lymphocytes and macrophages with varying numbers of plasma cells. The demyelination seen was of the secondary or Wallerian type. There was no evidence of primary demyelination. Visna virions were not seen in any of the CNS material studied. The ultrastructural findings are compatible with the view that lesions in visna may be induced by a cell-mediated immune response. However, changes characteristic of an autoimmune reaction to myelin antigens were not observed.


Intra-axonal corpora amylacea in the CNS. The distribution of intra-axonal corpora amylacea (IACA) in the CNS was studied in 2 cases of ALS, a hepatic encephalopathy and a Shy-Drager syndrome. A total of 149 IACAs were found. IACAs were observed far most frequently in the gracile nuclei, followed by the anterior horns of the spinal cord and in descending order; lateral geniculate bodies, reticular formation, sensory and motor nuclei in the brain stem, cerebellum and cerebral cortex. The basal ganglia, including the thalamus, contained only a few. IACA were not observed in the pyramidal tracts or in the spinal roots. About 3% of the IACA were sectioned longitudinally, together with the original small nerve fibres and abrupt enlargement of the axons due to the deposits was well demonstrated.


Light and electron microscopic studies of "nude" mice CNS after subcutaneous administration of the E variant of the encephalomyocarditis (EMC) virus. Sixteen 3 month old "nude" mice, 24 of their litter mates and 30 Swiss mice were injected subcutaneously with 0.1 ml suspension of the E variant of the encephalomyocarditis (EMC) virus. While the mortality rate of the litter mates and Swiss mice during 5-7 days after inoculation was more than 40%, none of the "nude" mice died during the experiment. The surviving animals were sacrificed at 24 h intervals from day one to seven days after injection. Brain suspensions assayed for the presence of the virus yielded significant titers at 24 h in all groups, which increased during 7 days. The litter mates and Swiss mice showed proliferation of lymphocytes and microglial cells in the perivascular areas of the brain during the fifth to the seventh day. The  "nude" mice, on the other hand, displayed no perivascular lymphocytic infilteration during the same periods. Ultrastructurally, all groups showed aggregates of ribosomes in the cytoplasmic matrix on the third day, which became enlarged in size on the 5th day. At 7 days, both litter mates and Swiss mice showed an increased number of necrotic cells, while these changes were not observed in the "nude" mice. These findings suggest that the high mortality rate in immunologically normal mice was related to the efforts of T cells to eliminate virus-infected cells and to produce extensive necrosis, while T cell-depleted animals showed good survival rates.


The diagnostic significance of Russell bodies in endoscopic gastric biopsies. The incidence of cells containing so-called Russell bodies in 315 benign single gastric biopsies taken from stomachs with proven carcinoma, but outside the carcinomatous area and 786 biopsies taken from stomachs without carcinoma were compared. The Russell body containing cells were counted in a fixed area and on the basis of the counting each biopsy was classified in one of four groups: Russell body containing cells absent or present in a number of 1-5, 6-10 or more than 10 in the area. The results showed that endoscopic biopsy examinations containing one single benign biopsy with more than 5 Russell body containing cells in the carcinoma. If half or more than half of the biopsies in the examination contained 1-5 Russell body containing cells significant differences were also found. The sensitivity of such examinations was low but their predictive values (PVpos) varied from 46 per cent to 95 per cent, meaning that such examinations may be of importance for the diagnosis of gastric cancer.


Relationship between lysosomal damage, fatty infiltration and hepatocellular necrosis in the course of acute liver injury induced by carbon tetrachloride in the rat. In the course of liver injury induced by CCl4 in rats the change of the endoplasmic reticulum takes 5 hours and that of the lysosomal membrane, 18 hours to develop. The latter change precedes hepatocellular necrosis. Elevation of plasma free fatty acids and fatty infiltration of the liver can be observed at 3 hours after CCl4 administration. The maximum of fatty infiltration, hepatocellular necrosis and the highest degree of lysosomal damage develop at the same time. Since CCl4 is eliminated in a few hours, it must initiate a cellular process which then leads to lysosomal membrane damage and hepatocellular necrosis.


Effect of cyclic adenosine monophosphate on renal function and renin secretion. The effect of cAMP, infused into the renal artery, on renal function and on the release of renin was studied in dogs displaying a normal serum Na level. Intrarenal infusion of 1 mg per kg per minute of cAMP caused a significant increase in RBF, dilatation of the efferent arterioles, the redistribution of renal blood flow and a significant increase in tubular Na-reabsorption. The antidiuretic action of cAMP is due partly to a hemodynamic and partly to a direct tubular effect. Decreased renin secretion was explained by the Na-load on the macula densa segment, being decreased by the intrarenal shifting of renal blood flow and with an eventual decomposition of cAMP.


Changes in ATP and cyclic nucleotide levels during sympathetic nerve stimulation in canine subcutaneous adipose tissue in situ. Subcutaneous adipose tissue in fed, female dogs was isolated. Biopsies of the tissue (30-150 mg) were taken and rapidly frozen in liquid nitrogen before, during and after nerve stimulation (3-4 Hz). In unstimulated adipose tissue the levels of ATP1 were 74+/-7 nmol/g, of cyclic AMP 90 +/- 12 pmol/g and of cyclic PGMP 18 +/- 3 pmol/g (mean+/-S.E.). During sympathetic nerve stimulation the levels of ATP and cyclic GMP fell by 30 and 50% respectively (p less than 0.01), while the cyclic AMP content increased by 50% (p less than 0.05). After nerve stimulation there was a marked increase in glycerol release, and the levels of all three nucleotides returned to control. The fall in ATP during nerve stimulation was essentially eliminated by prior adrenergic alpha-receptor blockade. It is concluded that 1) sympathetic nerve stimulaton induces a rapid, reversible fall in tissue ATP content, which may be related to hypoxia secondary to the vasoconstriction, and 2) lipolytic responses to sympathetic nerve stimulation in vivo are preceeded by small increases in the tissue cyclic AMP level, and a 3-fold increase in the cyclic AMP/cyclic GMP ratio.


Family study of high density lipoprotein cholesterol and the relation to age and sex. The Tromso Heart Study. A family study of serum high density lipoprotein (HDL) cholesterol and total serum cholesterol concentration has been undertaken, and the relation to age, sex, cigarette smoking, physical activity and familial occurrence of myocardial infarction (MI) was examined. HDL cholesterol was determined in 251 females and 194 males and total serum cholesterol in 677 females and 657 males, all aged 0-49 years. With respect to HDL cholesterol, significant sex differences were observed both in absolute level and in age-related change. A negative correlation between HDL cholesterol and total serum cholesterol was observed in all age groups except females aged 0-19 years, supporting the hypothesis of HDL as a ""clearing'' lipoprotein. HDL cholesterol showed a positive correlation only in pairs of first-degree relatives involving the mother and in sib-sib paris of the same sex. On the other hand, for serum cholesterol a positive correlation was found among all family members, although significantly higher between first-degree relatives than between spouses. No relation was found between cigarette smoking, physical activity of familial occurrence of MI and the HDL cholesterol or total serum cholesterol concentrations. In accordance with the ""HDL hypothesis'', the present finding could partly explain the higher incidence of ischaemic heart disease (IHD) in males than in females, and partly also the high risk which is transmitted from women with IHD to their first-degree relatives.


Substance P and primary afferent transmission. Substance P has a powerful excitant action on spinal motoneurons of the newborn rat, and the action is specifically antagonized by Lioresal, which also readily blocks spinal reflexes involving primary afferent transmission. Substance P is highly concentrated in bovine and feline dorsal roots as well as in the dorsal part of the dorsal horn of cat spinal cord. There is evidence that substance P is synthesized in spinal ganglia, transported through the dorsal roots and stored in the axon terminals of primary afferent neurons. These and other results suggest that the undecapeptide substance P, or one of its shorter C-terminal analogues, functions as an excitatory transmitter of the primary afferent fibers.


Changes of rat cerebellar guanosine 3',5'-cyclic phosphate by dopaminergic mechanisms in vivo. Apomorphine produced a dose-dependent increase of the cGMP level in the cerebellum. Haloperidol and scopolamine completely prevented this increase, and reserpine reduced the increase to 50%. LSD elevated cerebellar cGMP, and this effect was also abolished by haloperidol. It is suggested that the primary site of action of these drugs is the caudate nucleus, from which two neuronal pathways could trigger the increase of cGMP in the cerebellum.


Opiates, cyclic nucleotides, and xanthines. In rat brain homogenate, opiates exert a dose-related inhibition of PGE-stimulated cyclic (AMP) formation. This effect is stereospecific and is correlated with agonist potency. Naloxone antagonizes heroid in a dose-related way, without, at effective concentrations, itself inhibiting PGE-stimulated cyclic AMP formation. In morphine-dependent rats, intracerebroventricular injection of cyclic AMP intensifies and of dibutyryl cyclic GMP diminishes precipitated abstinence effects. In naive rats, the xanthines, theophylline and 3-isobutyl-1-methylxanthine, produce a quasimorphine-abstinence syndrome that is readily suppressed by heroin and intensified by nalosone. In rat brain homogenate, these xanthines inhibit cyclic AMP phosphodiesterase. These findings are concistent with these views: (a) The opiates specifically inhibit an adenylate cyclase of morphine-sensitive neurons that is sensitive to stimulation by PGEs. (b) Opiate agonist action is associated with the lowering of a neuronal cyclic AMP. (c) Both the morphine-abstinence syndrome in dependent rats and the quasi-abstinence syndrome in naive rats are associated with a rise in this neuronal cyclic AMP. (d) There are two types of endogenous humoral mediator acting on morphine-sensitive neurons, one of which is morphine-like and the other antimorphine-like in action.


Repartition and drug sensitivity of dopamine and L-isoproterenol-sensitive adenylate cyclases in rat brain homogenates. The characteristics of dopamine, 1-isoproterenol, and d-LSD stimulated adenylate cyclases were studied in homogenates of fresh or frozen tissues. In rat striatum, when the assay was done in the presence of 1 mM MgSO4, dopamine (10(-4) M) stimulated the enzyme activity by 3.5-fold. This effect was completely blocked by fluphenazine (10(-5) M; Ki = 9 X 10(-9) M) and by phentolamine (Ki = 3 X 10(-7) M). d-LSD stimulated the adenylate cyclase activity (Km = 1.4 X 10(-7) M) by interacting with the dopaminergic receptors. Maximal adenylate cyclase stimulation by d-LSD was 1.4-fold; as a matter of fact, this compound acted as a partial agonist on the dopaminergic receptors. l-Isoproterenol (Km = 10(-6) M) activated an adenylate cyclase present in rat striatum homogenates through a receptor distinct from the dopaminergic receptor; this stimulation was not affected by addition of fluphenazine or phentolamine but suppressed by dl-propranolol (10(-4) M). The topographical distributions of dopamine adenylate cyclase activity and endogeneous dopamine content were examined in homogenates prepared from discs punched out from serial frozen (-7 degrees C) slices of the striatum. A 4.8-fold progressive decrease in the amount of cyclic AMP produced in the presence of dopamine (10(-4) M) was observed from the rostral to the caudal part of the structure. The d-LSD-sensitive adenylate cyclase followed a similar distribution. It should be noted that the topographic distribution of endogeneous dopamine is quite comparable to the distribution of the dopamine-sensitive adenylate cyclase, suggesting that this enzyme is an integral part of the dopamine synapses. We also reported that the frontal cortex contains a dopamine-sensitive adenylate cyclase. In conclusion, we trust that the micromethod described for adenylate cyclase assay will be of some use in the study of the precise topographic distribution of catecholamine sensitive adenylate cyclases in different structures of brain.


Translational regulatory mechanisms in neoplasia in vivo. It is clear that regulation of genetic expression of cells need not involve alterations of the basic DNA structure. Between the production of messenger RNA and enzyme degradation there are a number of steps, any of which can be modulated by hormones and other factors to yield cellular abnormalities associated with malignancy. It is important to identify the exact step at which a carcinogen operates and to discover how this alteration is transmitted to the progeny of malignant cells.


Serum lipids and proteins in lactose malabsorption. It has been suggested that dietary lactose may reduce the intestinal absorption of fat and protein in individuals with lactase deficiency. On the other hand, it is known that a high carbohydrate diet increases serum lipids. The purpose of this study was to examine whether there are differences in the fasting serum lipid and protein concentrations between people with lactose malabsorption and people with normal lactose absorption. Therefore in the connection of a family study serum lipids and proteins were measured in 409 subjects belonging to 11 families. Of these 288 were relatives of the 11 index persons and 121 were spouses or relatives of the spouses. The weight, height, and milk consumption of each person were recorded. When the age, sex, relative weight and milk consumption effects were taken into account there was a statistical difference between the lactose malabsorption and lactose absorption groups for the concentration of serum triglycerides, but not for the other variables. Besides, serum triglyceride values of over 200 mg/100 ml were significantly fewer in people with lactose malabsorption. It was hypothesized that increased intestinal motility may disturb the absorption of fats and cause the observed difference at least in the Finnish population.


Chromium deficiency, glucose intolerance, and neuropathy reversed by chromium supplementation, in a patient receiving long-term total parenteral nutrition. A white female, now age 40 and receiving total parenteral nutrition for more than 5 years, developed unexpected 15% weight loss after 3 1/2 years of regimen, together with peripheral neuropathy confirmed by nerve conduction measurements. An intravenous glucose tolerance test showed that the fractional rate (K) had decreased to 0.89%/min (normal greater than 1.2). There was observed during this glucose infusion a borderline normal insulin response with a fall in plasma free fatty acids and in plasma leucine. During daily infusion of well over 400 g of glucose, the respiratory quotient was 0.66. Chromium balance was negative. Chromium levels were, in blood 0.55 ng/ml (normal 4.9 to 9.5) and in hair 154 to 175 ng/g (normal greater than 500). Regular insulin daily (45 micron) in the infusate nearly maintained euglycemia but despite this, and even with further glucose intake to restore weight loss, intravenous glucose tolerance test (K) and respiratory quotient were unchanged. Administration of insulin was then stopped and 250 microng of Cr added to the daily total parenteral nutrition infusate for 2 weeks. After this the intravenous glucose tolerance test (K) and respiratory quotient became normal (1.35 and 0.78, respectively). Over the next 5 months insulin was not needed and glucose intake had to be reduced substantially to avoid overweight. In this period nerve conduction and well-being returned to normal. With a maintenance addition of chromium to the total parenteral nutrition infusate (tentatively this addition is 20 microng/day) the patient has remained well for 18 months (to July 1976). These results suggest that relatively isolated chromium deficiency in man, hitherto poorly documented, causes 1) glucose intolerance, 2) inability to utilize glucose for energy, 3) neuropathy with normal insulin levels, 4) high free fatty acid levels and low respiratory quotient and, 5) abnormalities of nitrogen metabolism.


Effects of dibutyryl cyclic adenosinemonophosphate and prostaglandin E1 on platelet aggregation and shape changes. Prostaglandin E1 (PGE1) exerted an immediate inhibitory effect upon the induction of platelet aggregation by adenosinediphosphate, epinephrine, thrombin, bovine fibrinogen, or sodium fluoride, whereas dibutyryl cyclic adenosinemonophosphate (DBcAMP) produced a delayed type of inhibition of aggregation induced by these same agents. In addition, however, both PGE1 and DBcAMP, as well as adenosinetriphosphate (ATP) and Mg++, restored, with incubation, the washed platelets to a more discoid shape. Platelet aggregability also correlated well with platelet shape. All of these findings suggest the possibility of mediation of the delayed type of inhibition through restoration of the platelet membrane by increasing the level of metabolically active ATP or the availability of cations. They tend, however, to exclude the possibility that PGE1 exerts its immediate inhibitory effect through the increase of platelet cAMP.


Screening of mammary carcinoma for hormone dependency in vitro. Enzymatic activity in short-term organotypic cultures of breast biopsies from 62 patients. Enzymatic activity in short-term organotypic cultures of breast biopsies from 62 patients. Am J Clin Pathol 67: 393-396, 1977. Mammary carcinomas from 62 patients were assessed for pentose shunt dehydrogenase activity initially and after 24-72 hours in organotypic cultures with or without exogenous hormones. Hormones tested were (1) estradiol, (2) testosterone, and (3) prolactin. Thirty-seven (60%) were judged hormone-independent, in vitro; 14 (23%) were judged hormone-dependent, in vitro; 11 (17%) were classed as "indeterminant." Clinical results of endocrine management of 13 cases and an appraisal of the usefulness of the method are presented.


Fructose-1,6-diphosphatase deficiency.

Fructose-1,6-diphosphatase deficiency. A girl aged 3 years and 11 months, with recurrent episodes of unexplained metabolic acidosis, hepatomegaly, and fasting hypoglycemia unresponsive to glucagon, showed profound falls in blood glucose levels in response to oral fructose and glycerol challenge. In vitro analysis of her hepatic glycolytic and gluconeogenic enzymes demonstrated absent fructose-1,6-diphosphatase activity. A therapeutic trial of orally given folic acid, 30 mg daily, did not improve her tolerance for fructose and glycerol. Over the next two years she showed improvement in tolerance to fasting, and to fructose and glycerol loading on dietary management.


Cholangiocarcinoma associated with biliary cirrhosis due to congenital biliary atresia. An 11-year-old girl developed cholangiocellular carcinoma in association with biliary cirrhosis due to congenital biliary atresia. An exploratory laparotomy and an operative cholangiogram at 3 months of age had confirmed the diagnosis of extrahepatic biliary atresia. A liver biopsy specimen taken at 6 months of age showed biliary cirrhosis. The subsequent clinical course was characterized by persistent moderate jaundice, anemia, malnutrition, rickets, pathologic fractures, and recurrent gastrointestinal bleeding. The presence of cholangiocellular carcinoma of the liver with advanced biliary cirrhosis was established at an exploratory laparotomy a week before her death. We discuss here the pathogenesis of biliary cirrhosis and carcinoma of the liver; there may be a relation between the two in the childhood population.


Obstructive jaundice secondary to hepatoma. Case report and literature review. A 67-year old male, who presented a clinical picture of obstructive jaundice with cholangitis, was found to have hepatoma tumor embolus blocking the common duct. The site of invasion was the right hepatic duct. There are 15 such cases reported in the literature. Hepatoma rarely presents with obstructive jaundice. A clinical picture of biliary colic or cholangitis is frequently misleading. Invasion of a hepatic duct large enough to allow formation of a tumor embolus implies tumor close to the bifurcation of right and left hepatic ducts and has precluded effective surgical treatment in all but one case.


Long-term follow-up studies on herpes simplex antibodies in the course of cervical cancer: patterns of neutralizing antibodies. The kinetics of neutralization was used to study antibodies to herpes simplex virus type 2 in 90 patients with invasive carcinoma of the cervix uteri. The first sera were drawn before treatment and consective sera were drawn at intervals thereafter. Control groups consisted of 90 age-matched healthy women and 70 cancer patients suffering from malignancies other than cervix carcinoma. Patients surviving during the observation period of 12-60 months had a significantly higher frequency of neutralizing antibodies than control groups. Twenty-six of the women with cervix carcinoma died during the observation period and 13 others had a recurrence of the cancer. Survivors with advanced cervical cancer had higher antibody titers than survivors with less severe cervical cancer while patients whose cancer caused death had low titer or to a great extent lacked neutralizing antibodies. Women with recurrences and progressing cervical lesions had no detectable antibodies while women with recurrences and regressing lesions all had antibodies. There was a rise in K2-values 6-18 months after treatment. In serum samples collected later than 18 months, there was a return to original levels.


The Seattle Virus Watch. VII. Observations of adenovirus infections. The following findings were made from observations of adenovirus (AV) infections in Seattle VW families, 1965-1969, which extended the 1961-1965 New York VW studies: That infections are predominantly enteric, may be abortive or invasive and followed by persistent intermittent excretion was confirmed. That such excretion is most characteristic of types 1, 2, 3 and 5 viruses may explain why these types were usually endemic. However, since observed duration of excretion was not increased despite a longer average observation period, persistent excretion appears not to continue indefinitely and generation-to-generation transmission now seems improbable. Unilike New York, alternate cycling of types 1 and 2 viruses was not seen. Among homotypic susceptibles, infection rates for the endemic types were highest in infants (greater than 90% for types 1 and 2), decreased with age in older children but increased in parents, perhaps because of closer contact with infants. Development of serum neutralizing antibody was most frequent (about 90%) after types 1 and 2 infection; in all cases, titers decayed over time. While delayed virus spread related to persistent intermittent excretion did occur, spread closely following new or renewed (after larger than or equal to 3 months) excretion was more important. Sibling introducers were more effective spreaders than infants, and duration of excretion was more important than mode. These data indicate that homotypic immunity is 85% protective against infection. A protective effect of heterotypic immunity could not be shown. Illness (chiefly respiratory and often febrile) was associated with 49% of infections in susceptibles and with 65% when respiratory shedding occurred. The contribution of AV to all infectious illness, based on virus-positive infections only, was 5% in infants and 3% in the 2-4-year age group; for febrile illness, the corresponding contributions were about 10% and 5%. Inclusion of infections discovered only be serology (49% of all infections) would greatly increase the contribution of AV to illness.


Partial and total iodide organification defect in different sibships in a kindred. Identical twins with goiter but without intellectual retardation and with slightly delayed bone age were found to have defective iodide organification as demonstrated by incomplete perchlorate discharge tests. They are grandnieces of a normal member of a sibship which included four children with severe retardation and complete thyroid iodide organification defect. The parents and grandparents are not consanguine. Possible explanations are considered for the problem of why the disorder is manifest completely in one sibship and only partially in the other.


Chronic lymphoproliferative disorder with unusual clinical, morphologic, ultrastructural and membrane surface marker characteristics. Four of 105 patients with chronic lymphocytic leukemia (CLL) manifested clinical, morphologic, ultrastructural and membrane surface marker characteristics that differed from those found in patients with typical CLL of demonstrated B-lymphocyte origin. These four patients presented with moderate increases in absolute lymphocyte counts, absolute neutropenia, polyclonal hypergammaglobulinemia and hepatosplenomegaly without lymphadenopathy. Two of them were unusually young, 19 and 25 years old, at the time of diagnosis. The proliferating lymphocytes carried receptors for sheep erythrocytes, a T-lymphocyte marker. In the three patients tested, the lymphocytes also carried Fc receptors. Ultrastructurally the lymphocytes contained cytoplasmic inclusion bodies consisting of parallel tubular arrays. The parallel tubular arrays corresponded to prominent cytoplasmic azurophilic granules on light microscopy. Parallel tubular arrays were found in less than 1 per cent of the lymphocytes in eight patients with typical B-lymphocyte CLL. The process in these four patients may be a distinctive chronic lymphoproliferative disorder originating in T lymphocytes with Fc receptors found in small numbers in the blood of normal persons.


The epidemiology of viral hepatitis: an overview. Type A hepatitis maintains itself in human populations without either an extrahuman or human reservoir. Intestinal carriers do not appear to be epidemiologically important; viremic carriers have not been demonstrated. Person-to-person transmission by the fecal-oral route is the usual mechanism. Epidemic and endemic occurrence is usually recognizable by well-defined characteristics; a "hyperendemic" patern has been documented. In some countries, but not all, the long-term trend of type A disease has been downward in recent years. Type B hepatitis is worldwide in distribution, and capable of maintaining itself by the carrier reservoir. Transmission is by multiple mechanisms, with the percutaneous route still of major importance when adequately defined in terms of subtle exposures. Contact-associated transmission probably occurs by both the oral-oral and venereal routes. The long-term trend of type B hepatitis is unknown, but some evidence suggests an increase in the United States apart from the epidemic associated with drug abuse. The existence of additional viruses of human hepatitis is suggested by data concerning transfusion-associated disease and multiple episodes in the same individual.


Cholangiocarcinomas induced by feeding 3'-methyl-4-dimethylaminoazobenzene to rats. Histopathology and ultrastructure. Thirty-three male Sprague-Dawley rats were fed a carcinogenic (0.064% 3'-methyl-4-dimethylaminoazobenzene, 3'-Me-DAB) ground meal normal diet. After 12 weeks the ground meal diet was replaced with a normal pellet diet, and the 30 surviving animals were divided into three equal groups. One group was sacrificed at the twelfth week and the other groups 4 and 8 weeks later. Control animals were also run. Based on previous studies which used "tumor-promoting" diets and 3'-Me-DAB, we expected a less than 100% incidence of predominantly hepatocellular carcinomas. However, we found mucin-producing cholangiocarcinomas in all 30 animals and, in addition, a small hepatocellular component in 3 of the animals. By electron microscopy the intestinal mucosal features of microvillous border cells, goblet cells, and endocrine-like cells were found. We suggest that the tumors produced as described here provide a good animal model of mucin-producing cholangiocarcinomas.


Canine hyperlipoproteinemia and atherosclerosis. Accumulation of lipid by aortic medial cells in vivo and in vitro. Dogs maintained for 1 year on a semisynthetic diet containing hydrogenated coconut oil and cholesterol developed hypercholesterolemia. In those cases where plasma cholesterol levels exceeded 750 mg/100 ml, the animals also developed severe atherosclerosis. This atherogenic hyperlipoproteinemia was characterized by the presence of beta very low density lipoproteins (B-VLDL), increased levels of low density lipoproteins (LDL), and the occurrence of the HDLc lipoproteins. In all of these cholesterol-rich lipoproteins the arginine-rich apoprotein (ARP) was prominent. Moreover, the HDLc (d = 1.006-1.02) contained the ARP as the only detectable apoprotein. The atherosclerosis involved the abdominal aorta, coronary and cerebrovascular arteries, and many of the peripheral arteries. Histologically, the aortic lesions were characterized by a variable intimal proliferative response and extensive medial lipid deposition. In the peripheral, coronary, and cerebral arteries, the lesions were more extensive and involved primarily the media of the vessel wall, with little intimal reaction in many cases. The correlation between the in vivo disease process and the response of aortic smooth muscle cells (SMC) grown in tissue culture to the various cholesterol-induced lipoproteins was examined. B-VLDL, LDL, and HDLc (but not HDL2) caused a marked accumulation of free and esterified cholesterol in the SMC. The cholesterol accumulation was found to be more extensive in canine SMC than in swine smooth muscle cells or smooth muscle cells of other species in response to a similar lipoprotein cholesterol concentration. The enhanced sterol uptake appeared to be a property of canine smooth muscle cells rather than a property of the canine lipoproteins. These in vitro results may be related to the observed propensity for the development of medical disease that was demonstrated in the in vivo studies.


Gastrointestinal and metabolic function in patients with the carcinoid syndrome. We evaluated gastrointestinal absorption in six consecutive patients with metastatic serotonin-secreting carcinoid tumors. One patient had a consistent defect in fat absorption and two other patients malabsorbed fat during spontaneous or dopamine-induced exacerbation of the carcinoid syndrome. The steatorrhea of the patient with the persistent defect in fat absorption was reduced when tumor serotonin production was reduced by the tryptophan hydroxylase inhibitor parachlorophenylalanine. The six patients had normal hemoglobin levels and the serum concentration of the following urinary constituents was normal in most of the patients: albumin, carotene, 25-hydroxycalciferol, parathyroid hormone, calcitonin, calcium, phosphorous, osteogenous alkaline phosphatase, cholesterol, triglycerides, and serum lipoproteins. The excretion of the following urinary constituents was also normal in most of the patients: creatinine clearance, tubular reabsorption of phosphorous, calcium, D-xylose, cyclic 3'5' monophosphate and hydroxyproline. We conclude that patients with the carcinoid syndrome may have steatorrhea, and that their hyperserotoninemia plays a role in this process.


Ultrastructural localization of membrane phosphatases in teratocarcinoma and early embryos. Ectodermal cells of the two- and three-germ layer-thick mouse egg-cylinders are considered to be the progenitors of embryonal carcinoma cells in embryo-derived teratocarcinomas. In an attempt to find differences between the tumor cells and equivalent embryonic cells, we have studied the electron microscopic cytochemical localization of alkaline phosphatase, 5'-nucleotidase, and Mg2+-activated adenosine triphosphatase (ATPase) in embryo-derived teratocarcinomas and mouse egg-cylinders. Alkaline phosphatase was detected in both embryonic and tumor cells, but its activity appeared much more intense in the tumor cells. No ATPase was demonstrated in embryonic ectodermal cells of 6-day-old embryos and only in occasional  cells of 7- and 8-day-old embryos. No 5'-nucleotidase activity could be demonstrated in 6- to 8-day-old cylinders. There was marked ATPase and 5'-nucleotidase activity in the membranes of embryonal carcinoma cells. These data point out some differences on the plasma membrane between the embryonal carcinoma cells and equivalent embryonic cells. The potential significance of these differences is discussed with regards to the transformation of embryonic cells in tumor cells. (Am J Pathol 87:297-310, 1977).


Effects of hormones on 3', 5' -cyclic adenosine monophosphate in choroid plexus. Choroid plexus of rabbit and rat was incubated for 2-30 min at 37 degrees C under 95% O2-5% CO2 in Tyrode solution containing 10 mM glucose and 1 mM theophylline with these agents: epinephrine, norepinephrine, isoproterenol, dopamine, histamine, serotonin, arginine, and lysine vasopressins, oxytocin, angiotensin, adrenocorticotropin (ACTH), beta-melanocyte-stimulating hormone, and choroid plexus peptide IIF. After incubation, tissue and medium were analyzed for 3', 5' -cyclic adenosine monophosphate (cAMP) content. Each amine or peptide was tested initially at 1,000 microng/ml. Only ACTH and serotonin affected cAMP content of rabbit choroid plexus. At 1,000 microng/ml, these agents caused a 10 and 4 times (respectively) increase in cAMP content of tissue + medium at 2-10 min with decline in content at 10-30 min. More than 90% of the increment was located in  tissue, less than 10% in medium. Minimal effective dose (MED) to cause a significant (P less than .05) accumulation of cAMP was 0.1 microng/ml (2.2 x 10(-8) M) for ACTH and 10 microng/ml (5.7 x10(-3) M) for serotonin. Only isoproterenol, epinephrine, and norepinephrine influenced cAMP content of rat choroid plexus. MED's for this effect by isoproterenol, epinephrine, and norepinephrine were .001, .01, and 10 microng/ml (4.7 x 10(-9), 5.5 x 10(-8), and 5.9 x 10(-5) M), respectively.


Oxidative response to aldosterone of pyridine nucleotide in rat kidney in situ. In an attempt to elucidate early biochemical events in the action of aldosterone on rat kidney in situ, the response of the pyridine nucleotide oxidation-reduction state of surface cells was directly and continuously recorded with an organ-fluorometer. Intravenous administration of aldosterone induced a rapid (maximum response in 10 min) and dose-related (2.5-25 microng/100 g rat) oxidative response that was specific to aldosterone and to the kidney. The oxidative response was 1) detectable with minute dise (approximately 0.2 microng/100 g rat) of hormone: 2) reproduced by other mineralocorticoids; 3) enhanced by a maneuver for expanding the extracellular fluid compartment or by adrenalectomy; and 4) prevented by spironolactone, progesterone, actinomycin D, and cycloheximide. All of these data argue that the redox response is related to the subsequent changes in aldosteronemediated ion transport. Experiments with an uncoupler and with redox substrates showed that mitochondrial NADH was the major nucleotide pool responding to hormone. The oxidation was not accompanied by changes in the adenylate energy charge level of the whole organ. These observations support the view that aldosterone acts on energy metabolism of tubular cells before developing apparent cation transport effects.


Effects of insulin, ADH, and cyclic AMP on sodium transport in the toad bladder. These studies further define the mechanisms by which insulin stimulates Na transport in the toad bladder. Serosal but not mucosal addition of insulin, 100-1,000 mU/ml, stimulated short-circuit current (SCC) by 25-50%. The initial rise in SCC occurred at 5 min and the peak response at 15-25 min. Doses of insulin greater than 250 mU/ml increased SCC values for up to 3 h. Actinomycin D did not block the early rise in SCC produced by insulin, but it blocked the delayed effects. Insulin increased SCC in substrate-depleted bladders, although the increase in SCC was less (P less than 0.01) than in nonsubstrate-depleted bladders. Pyruvate addition to substrate-depleted bladders restored to normal the rise in SCC observed after insulin. Simultaneous addition of ADH and insulin led to an increase in SCC that was greater than the sum of the responses observed when each hormone was added independently. Synergistic effects on SCC were also obtained with cyclic AMP and insulin. Insulin did not increase cyclic AMP levels in toad bladder epithelial cells. It is suggested that insulin stimulates active Na transport by two mechanisms: 1) a rapid phase, which may involve unmasking of pump sites within the membrane, and 2) a delayed effect which seems to require protein synthesis. The synergism of which seems to require protein synthesis. The synergism of insulin with ADH or cyclic AMP may reflect a facilitative effect of insulin on ADH or cyclic AMP-sensitive pump sites or, alternatively, the uncovering of latent pump sites that then may be available to stimulation by ADH or cyclic AMP.


Effect of sodium fluoride on concentrating and diluting ability in the rat. Mechanisms for the concentrating defect produced by fluoride were examined in the rat. Free-water clearance at all levels of delivery was normal after 5 days of chronic fluoride administration in the hereditary hypothalamic diabetes insipidus rat. In the Sprague-Dawley rats, during moderate fluoride administration (120 micronmol/kg per day), urine osmolality and cyclic AMP excretion decreased and urine volume increased, but after exogenous vasopressin, volume decreased and osmolality and cyclic AMP increased appropriately. During larger daily doses of fluoride (240 micronmol/kg per day) urinary osmolality and cyclic AMP decreased and volume increased, which was similar to the changes seen during lower fluoride dosages, but these parameters did not change after exogenous vasopressin. These data suggest that ascending limb chloride reabsorption is unaltered by fluoride administration; in the presence of sufficient fluoride, collecting tubular cells apparently do not generate cyclic AMP or increase permeability appropriately in response to vasopressin. The postulated defect is felt to be due to either a decrease in ATP availability or to a direct inhibitory effect of fluoride on the vasopressin-dependent cyclic AMP generating system.


Cyclic AMP in action of antidiuretic hormone: effects of exogenous cyclic AMP and its new analogue. Exogenous cyclic 3',5'-adenosine monophosphate (cAMP) stimulates the effect of the antidiuretic hormone, vasopressin (VP), only in pharmacologic quantities and results have often been inconsistent. The present study examined the ability of a new analogue, 8-[p-Cl-phenylthio]cyclic 3',5'-adenosine monophosphate (C1-PheS-cAMP) to mimic the effect of VP, both biochemically (protein kinase activation) and functionally (hydrosomatic response of perfused collecting tubules) in mammalian kidney tissue. C1PheS-cAMP was found to be about 100 times as effective as cAMP both biochemically and functionally. The increased effectiveness of C1PheS-cAMP is probably is probably due to a greater permeability across the cell membrane and to the resistance of C1PheS-cAMP to enzymatic degradation, Cyclic AMP phosphodiesterase inhibition was observed with C1PheS-cAMP, but its contribution to overall effect was minor. C1PheS-cAMP was found to be more effective than exogenous vasopressin, an effect probably due primarily to its resistance to catabolism. The results provide further new evidence that cAMP and protein kinase are involved in the cellular action of vasopressin. C1PheS-cAMP proved to be a useful tool in the study of hormone action, especially in steps subsequent to cAMP generation.


Computer simulation of ischemic rat heart purine metabolism. I. Model construction. A model is proposed for the partial depletion of the adenine nucleotide pool in the ischemic perfused rat heart which involves seven enzymes: adenylate cyclase, 3',5'-cyclic AMP phosphodiesterase, 5'-nucleotidase, adenosine kinase, adenosine deaminase, purine nucleoside phosphorylase, and inorganic pyrophosphatase. The computer implementation of this model is in terms of rate laws, several of which were obtained by a systematic least-squares fitting procedure. Depletion of the adenine nucleotide pool is initiated by the release of endogenous noradrenaline into the interstitial fluid, which results from a fall in tissue PO2, and the subsequent activation of adenylate cyclase. In this model the substrate for 5'-nucleotidase is a membrane-bound AMP pool formed by hydrolysis of extracellular fluid and functions as a vasodilator; excess adenosine is incorporated into the tissue by a "permease" with Michaelis-Menten kinetics and converted to AMP, inosine, and hypoxanthine. Alternative mechanisms, such as the deamination of AMP by adenylate deaminase and conversion of AMP to adenine by AMP pyrophosphorylase, were rejected primarily on qualitative biochemical grounds.


Human poxvirus disease after smallpox eradication. A 5-year-old boy living in a small camp in the rural Ivory Coast had a disease resembling smallpox. This occurred 4 years after smallpox had been eradicated from the Ivory Coast and 1.5 years after the last case of smallpox was detected in West and Central Africa. Clinical, serological, and epidemiological evidence indicated this disease was probably monkeypox, a poxvirus of the variola/vaccina subgroup. A serologic survey of poxvirus antibodies in the wild animal population detected neutralizing antibodies in rodents, larger mammals, primates, and birds. The laboratory and ecological characteristics of poxviruses require further elucidation, especially those which have been found in animals near human monkeypox cases.


Studies on the epidemiology of sandfly fever in Iran. I. Virus isolates obtained from Phlebotomus. A total of 62 virus isolates was obtained from 12,485 male and female sandflies (predominately Phlebotomus papatasi) collected in a village in central Iran during the summer of 1975. The overall virus isolation rate from female sandflies was 1 per 177 insects processed. Of the 62 virus strains obtained, 49 were identified as Sicilian virus, 11 as Karimabad, and 2 as a new member of the vesicular stomatitis serogroup. One isolate each of Sicilian and Karimabad virus was made from pools of male sandflies. The  three virus types were active in the sandfly population simultaneously. Sicilian virus isolation rates showed little variation during the study period, suggesting continuous virus activity during the sandfly season. The implications of these findings for the epidemiology of sandfly fever are discussed.


Studies on the epidemiology of sandfly fever in Iran. II. The prevalence of human and animal infection with five phlebotomus fever virus serotypes in Isfahan province. Human and animal sera from an endemic area of sandfly fever in Iran were tested by plaque reduction neutralization method against five different Phlebotomus fever virus serotypes (Naples, Sicilian, Karimabad, Salehabad, and I-47). The overall prevalence of Naples, Sicilian, and Karimabad virus antibodies among the human population was 17%, 25%, and 66%, respectively. All sera were negative against Salehabad and I-47 viruses. Age-specific antibody rates suggested that Sicilian and Karimabad viruses were endemic in the study area but that Naples virus activity was sporadic. These observations were confirmed by isolations of Sicilian and Karimabad viruses from sandflies collected in the study area. Among the animal sera tested, evidence of Phlebotomus fever virus infection was detected only in gerbils. Of 38 Rhombomys opimus tested, 34% had neutralizing antibodies against Sicilian virus and 32% against Karimabad. These results indicate that gerbils are infected with these two viruses and possibly might serve as reservoirs or amplifying hosts. The serologic studies also suggest that the ecology of Sicilian and Karimabad viruses involves chiefly sandflies, gerbils, and man, an epidemiologic pattern previously demonstrated for cutaneous leishmaniasis in the same region of Iran.


Isfahan virus, a new vesiculovirus infecting humans, gerbils, and sandflies in Iran. The characteristics and ecology of Isfahan virus, a new vesicular stomatitis virus (VSV) serotype, are described. Two strains of the agent were isolated from pools of Phlebotomus papatasi collected in Iran in 1975. Its animal pathogenicity, growth rate, cytopathic effect, and plaque morphology are similar to those of the other VSV serotypes. Electron microscopic examination of the virus demonstrated a bullet shape, the presence of truncated particles and maturation at plasma membranes. Antigenic relationships between Isfahan virus and three other VSV serotypes (Cocal, Piry, and Chandipura) were demonstrated by complement fixation or neutralization tests. A high prevalence of Isfahan neutralizing antibodies was found in human sera from several regions of Iran, suggesting that the virus may be of some public health importance. All of the residents over 5 years of age in the village where the virus was isolated had been infected. Neutralizing antibodies to Isfahan virus were also found in sera of Iranian gerbils but not in domestic animals. Results of this study suggest that the ecology of Isfahan virus is distinct from the other VSV serotypes and involves chiefly humans, gerbils, and sandflies, a pattern also observed with cutaneous leishmaniasis and sandfly fever in Iran.


Overwintering mechanism for bluetongue virus: biological recovery of latent virus from a bovine by bites of Culicoides variipennis. Bluetongue virus (BTV) was biologically transmitted by the bites of colonized Culicoides variipennis gnats to recipient sheep from a Hereford bull with latent infection. Four biological recoveries of BTV were mediated over 2 years by multiple feedings of the vector during a 4- to 72-hour interval. Initial stimulation by gnat bites at 0 hour permitted biological recovery of BTV by gnats that fed at later intervals. The 4th biological recovery of the virus from the bull clearly indicated a vector-mediated viral recovery mechanism in which initial vector bites at 0 hours stimulated a "showering" of BTV into the blood stream of the bull. The BTV carrier bull developed no overt signs of illness during these studies. The pertinent BTV assay and serologic results for the 4 1/3 years of the bull's life are summarized in this report.


Hazards and safeguards during carotid endarterectomy. Technical considerations. The hazards and safeguards of carotid endarterectomy are presented in a consecutive series of 130 patients over a fourteen month period. The mortality rate of 1.5 per cent was due to myocardial infarction. Two patients (1.5 per cent) suffered minor but permanent neurologic deficit. There was an incidence of 4.6 per cent transient neurologic deficits, all of which cleared within 24 hours. Special emphasis is placed on peripheral nerve injuries (12.3 per cent). Local neurologic anatomy is reviewed to facilitate safe dissection and an avoidance of nerve injury.


Nonautonomy of parathyroid hormone and urinary cyclic AMP in primary hyperparathyroidism. This study demonstrates that appreciable changes in serum parathyroid hormone and urinary cyclic AMP occur during experimentally induced hyper- and hypocalcemia in almost all patients with primary hyperparathyroidism regardless of histology. A single patient with tertiary hyperparathyroidism also demonstrated a significant elevation of serum parathyroid hormone and urinary cyclic AMP in response to EDTA induced reduction in ionized calcium. Thus, total autonomy of hormone secretion was not present in the great majority of the patients with a parathyroid adenoma, parathyroid hyperplasia, or the single patient with tertiary hyperparathyroidism. Therefore, preoperative evaluation of the rsponse of urinary cyclic AMP and serum parapthyroid hormone to EDTA or calcium infusion will not distinguish parathyroid adenomas from hyperplasia on the basis of total autonomy of hormone secretion. If a difference in secretory control is present between parathyroid adenomas and parathyroid hyperplasia, it is more subtle than total autonomy for adenomas and nonautonomy for hyperplasia.


Primary benign solid tumors of the liver. The benign solid primary tumors of the liver separate into three major groups: (1) the most common lesions--small, nodular tumors, often found incidentally with histologic features suggesting a reactive etiology, that never rupture or metastasize, that seem to grow most rapidly in children and pregnant women, and that are probably unrelated to birth control medication; (2) the less common adenomas that are purely epithelial, that most often occur in menstruating females, that often show necrosis and rupture, and that are closely associated with oral contraceptive agents; and (3) very rare tumors, usually mesenchymal, occurring solely in children. The therapeutic and prognostic implications of these tumors are defined based upon a study of 111 patients, all but one of whom have undergone resection.


Anaesthetic management of insulinoma.

Anaesthetic management of insulinoma. The anaesthetic management of two cases of insulinoma treated by distal pancreatectomy is presented. Stress is laid on pre-operative and intra-operative glucose administration in sufficiently high quantity. Pre-operative corticosteroid therapy is better avoided. Dextrose infusion should be stopped soon after removal of the tumour to prevent excessive hyperglycemic response in the postoperative period. Methyoxyflurane was used because it tends to raise the blood sugar level. Pancuronium bromide was preferred because of the stable cardiovascular conditions it produces.


Effects of halothane, thiopental, and lidocaine on fluidity of synaptic plasma membranes and artificial phospholipid membranes. The effects of halothane, thiopental, and lidocaine were studied with spin-labeling methods in synaptic plasma membranes (order parameter) and artificial phospholipid membranes (lateral diffusion). Halothane had a biphasic action, low concentrations (0.64 mM) ordering and high concentrations (2.9 mM) fluidizing both types of membranes. A biphasic effect in phospholipid membranes was also seen with thiopental, 0.1 mM ordering and 10 mM fluidizing, whereas in synaptic plasma membranes both low and high concentrations caused an increased order in the lipid bilayer region. At high thiopental concentrations, a considerable number of molecules may have reacted with membrane proteins or accumulated in the highly fluidic hydrophobic interior region of the membrane without affecting the rotational movement of the labeled fatty acid. Lidocaine alone, or together with calcium chloride, at various concentrations to 10 mM had no significant effect, and a fluidizing effect of 1 mM calcium chloride was possibly a result of interaction of calcium chloride with the label. The results indicate that the three lipid-soluble anesthetics interact differently with the lipid part of membranes. Lidocaine did not seem to affect bilyer lipids, while thiopental and halothane in phospholipid vesicles and halothane alone in synaptic membranes caused a dose-dependent biphasic effect.


Quantitative transmissible gastroenteritis virus shedding patterns in lactating sows. To test the role of sows in spreading transmissible gastroenteritis (TGE), 11 sows were intravenously, intranasally, or intramammarily inoculated with virulent virus within 5 days of farrowing. Six of the sows were separated from their offspring, and 5 were allowed to nurse their litters. All sows became clinically ill with sign of anorexia, depression, and fever that persisted until postinoculation day 4 or 5. They shed virus through milk, nasal secretions, and feces, with individual variations occurring in degree and duration of shedding in the 1st week after inoculation. Of 40 pigs separately fed milk samples from the 6 inoculated sows, 19 pigs (47.5%) became sick in 24 to 40 hours, and virus was isolated from them at necropsy. Of 43 pigs in the 5 litters that nursed exposed dams, all became sick with typical signs of TGE, and 29 (67.4%) died in 2 to 9 days. Sows given the single intramammary inoculation of virus developed statistically significant higher levels of TGE virus-neutralizing antibodies than did sows inoculated intravenously or intranasally.


Serotyping avian adenoviruses by a microneutralization procedure. A microneutralization procedure, using chicken kidney cell monolayers as an indicator system, was developed and applied to the serotyping of isolates characterized as avian adenoviruses. The method was determined to be reproducible, since coefficients of variation were low for 12 replicate titrations of homologous reagents of 9 prototype avian adenoviruses. Prototype reagents were specific according to results of reciprocal end point-neutralization tests and comparison of antigenic relatedness, using results obtained by previous researchers. Forty-two avian adenovirus isolates were classified into 6 serotypes by one-side end point-neutralization tests against antiserums made to 9 prototype avian adenoviruses. An additional 20 isolates were antigenically related to prototype viruses, but they could not be specifically types with the typing criteria. Different serotypes were isolated from birds having similar clinical diagnostic signs and lesions of disease.


Isolation and characterization of epizootic hemorrhagic disease virus from white-tailed deer (Odocoileus virginianus) in eastern Washington. A virus was isolated from the spleen of a white-tailed deer (Odocoileus virginianus) that had died during an epizootic in Washington state in 1967. Inoculation of a 10% spleen suspension from the deer caused hemorrhagic disease in normal white-tailed deer. Studies were conducted on the biological, physicochemical, and serologic properties of the Washington isolate. An in vitro assay system, utilizing a cultured primary of white-tailed deer fetal cells from an entire fetus, was employed for isolation and propagation of the virus. Cytopathic effect was characterized by focal development of rounded and clumped cells. Propagation was unsuccessful in suckling mice, BHK-21, and Vero cell cultures. The virus was resistant to treatment with ether, sodium deoxycholate, trypsin, oxytetracycline hydrochloride, and was sensitive to chloroform. Virus yield was not affected when infected cultures were treated with 5-iodo-2'-deoxyuridine, but dactinomycin (actinomycin D) treatment of infected cultures reduced virus yield. The virus was inactivated when heated at 70 C for 5 minutes or when exposed to pH 5 for 18 hours at 4 C. The virus was completely excluded from the filtrate by a 0.10- micronm (APD) membrane filter. Staining of infected cells with acridine orange indicated the presence of double-standard nucleic acid in the cytoplasm. Serum-neutralization tests with antiserums against the homologous virus and the New Jersey and Alberta strains of epizootic hemorrhagic disease virus resulted in neutralization of the Washington isolate. The Washington virus was not neutralized by bluetongue virus antiserum. Cells infected with the Washington isolate exhibited intracytoplasmic fluorescence by the indirect fluorescent antibody method with New Jersey and Alberta epizootic hemorrhagic disease antiserums but not with bluetongue antiserum.


Equine antibody to bovine serum induced by several equine vaccines as a source of extraneous precipitin lines in the agar gel immunodiffusion test for equine infectious anemia. Precipitin lines not associated with equine infectious anemia (EIA) were observed in routine agar gel immunodiffusion (AGID) testing for the infection. The serums which produced these lines were obtained from horses which had been given multiple vaccinations with commercially available cell culture-origin equine virus vaccines as part of a comprehensive herd health program. The lines formed against cell culture-derived, but not spleen-derived EIA viral antigens. Investigation revealed that bovine serum proteins in the vaccines induced precipitating antibodies which reacted with bovine serum proteins in cell culture-derived antigens. A vaccination trial, utilizing 4 commercially available vaccines in various combinations, indicated that as few as 2 vaccinations could induce AGID-detectable antibodies to bovine serum proteins in individual ponies. These antibodies were very transitory, usually lasting no longer than a week. Some horses, however, which had been given 4 vaccinations developed similar antibodies which persisted 3 months beyond the last vaccination. The extraneous precipitin lines produced by these antibodies in the AGID test for EIA were readily distinguished from true EIA-associated reactions and did not result in false-positive interpretations of the test. However, heavy percipitin lines due to strong antibovine serum activity did mask weakly positive EIA reactions.


Nonlethal experimental inoculation of Columbia black-tailed deer (Odocoileus hemionus columbianus) with virus of epizootic hemorrhagic deer disease. Intramuscular or intravenous inoculation of 5 Columbia black-tailed deer (Odocoileus hemionus columbianus) with virus of epizootic hemorrhagic deer disease (EHD) did not produce overt clinical disease. Two white-tailed deer (Odocoileus virginianus) exposed identically died in 5 to 6 days. There were no significant lesions in 1 black-tailed deer euthanatized on postinoculation day 5. The EHd virus was not isolated from the spleen of that deer. Seroconversion occurred in black-tailed deer, from zero EHD virus antibody titer before inoculation to titers of 1:128 to 1:256 after inoculation.


Demonstration and quantitation of immunoglobulins in bovine serum, follicular fluid, and uterine and vaginal secretions with reference to bovine viral diarrhea and infectious bovine rhinotracheitis. Immunoglobulin concentrations (IgG, IgM, and IgA) in bovine serum, follicular fluid, and uterine and vaginal secretions were determined. The specificities of IgG, IgM, and IgA for virus-neutralizing antibody against bovine viral diarrhea (BVD) and infectious bovine rhinotracheitis (IBR) viruses were also examined. High concentrations of IgG were present in both serum and follicular fluid. The IgG, IgM, and IgA concentrations were low in uterine and vaginal secretions. There was more IgG in the uterus during estrus than at any other time. Virus-neutralizing antibodies against BVD and IBR in serum of cows were mainly the IgG class. There was positive correlation between serum and follicular fluid virus-neutralizing antibody titers fro BVD and IBR. These antibodies may provide some protection for recently ovulated ova.


The relationship between nicotinamide adenine dinucleotide concentration and antibacterial activity of isoniazid in Mycobacterium tuberculosis. The relationship between the antibacterial effect of isoniazid and the intracellular concentration of nicotinamide adenine dinucleotide (NAD) was investigated in Mycobacterium tuberculosis strain H37Rv given continuous and pulsed exposures to the drug. Depletion of NAD to a plateau value occurred rapidly during exposure, and recovery after a pulse of isoniazid was also rapid. It seemed unlikely that NAD depletion was the direct cause of the antibacterial activity because (1) insufficient depletion occurred at low isoniazid concentrations; (2) antibacterial activity, but not NAD depletion, was proportional to the product of isoniazid concentration and the exposure period; and (3) NAD depletion was not related to antibacterial activity in cultures of differing physiologic state.


Generalized amyloid in a family of Swedish origin. A study of 426 family members in seven generations of a new kinship with neuropathy, nephropathy, and central nervous system involvement. We report a new kinship with systemic amyloid presenting as peripheral neuropathy in the fourth and fifth decades of life. A progressive sensory and motor loss starting in the lower extremities occurs from this disease, and there is subsequent renal, cardiac, gastrointestinal, ocular, and cutaneous involvement. Histologic studies show that amyloid deposition is mainly in connective tissue structures; there is an unusual infiltration of the meninges and central nervous system. Review of records of 426 family members in seven generations showed that this disease is inherited as an autosomal dominant. The absence of immunoglobulin disorders in two affected family members studied in depth suggests that this is not the primary type of amyloid in which the deposits are composed of fragments of immunoglobulin light chains. Similarly the absence of elevated levels of protein SAA (the serum precursor of secondary amyloid) suggests that this is not a secondary form of amyloid.


[Purification and some properties of "Clostridium perfringens" delta toxin (author's transl)]. Delta toxin, a hemolytic exocellular protein excreted by C. perfringens type C has been purified to homogeneity, assessed by polyacrylamide disc gel electrophoresis. Purification steps involved successively calcium phosphate gel formation in culture supernatant fluid, salting-out of unadsorbed material by ammonium sulfate to 50 % saturation, isoelectric focusing and gel filtration on Sephadex G75. Purified toxin appears as a basic protein occuring in two forms with isoelectric points of 8.8 and 9.4 as disclosed by isoelectric focusing. Molecular weight estimated by SDS-polyacrylamide disc gel electrophoresis was found to be close to 42,000 for the two forms. The lytic activity of delta toxin is inhibited by Ca++ and EDTA. The toxin is activated by short-term treatment with low concentration of trypsin.


Fibrous histiocytoma of the subglottic larynx. Histiocytic tumors are being reported with increasing frequency in the medical literature. In spite of confusion regarding nomenclature, appropriate diagnostic and treatment modalities exist. A malignant fibrous histiocytoma of the subglottic larynx is reported. Discussion of the present criteria for a diagnosis of malignancy and a rationale of treatment is presented.


The role of metals in enzyme activity.

The role of metals in enzyme activity. Metal ions play important roles in the biological function of many enzymes. The various modes of metal-protein interaction include metal-, ligand-, and enzyme-bridge complexes. Metals can serve as electron donors or acceptors, Lewis acids or structural regulators. Those that participate directly in the catalytic mechanism usually exhibit anomalous physicochemical characteristics reflecting their entatic state. Carboxypeptidase A, liver alcohol dehydrogenase, aspartate transcarbamoylase and alkaline phosphatase exemplify the different roles of metals in metalloenzymes while the nucleotide polymerases point to the essential role of zinc in maintaining normal growth and development.


Crystalline muscle inclusions in atypical postinfectious polyradiculoneuropathy. Crystalline inclusions were found in the muscle of a previously healthy 63 year old male with atypical postinfectious polyradiculoneuropathy. Approximately 12 days after the onset of the illness, biopsy of biceps femoris and vastus lateralis muscles revealed, in addition to changes consistent with early neurogenic atrophy, inclusions measuring 22 micronm in greatest dimensions. In plastic-embedded toluidine blue stained sections the translucent ovoid to angular inclusions were seen in a subsarcolemmal position, adjacent to nuclei. Ultrastructural examination revealed a crystalline structure consisting of parallel dense lines measuring approximately 70 A in diameter with an axial periodicity of 130 A. Unlike rods, the crystalline inclusions were not continuous with Z-discs; rather they displaced contractile and other cellular elements. Their appearance was similar to intramuscular crystals in diabetic amyotrophy and to vincristine-induced and naturally occurring intracytoplasmic crystalline structures. The nature of the crystalline inclusions and their relationship to the basic disease process was not established.


Determination of liver microsomal glucose-6-phosphatase. A procedure for the determination of liver microsomal glucose-6-phosphatase is described. Homogenization and ultracentrifrigation were used to prepare a precipitate whose character was defined by monitoring the desire enzyme activity which serves as a marker. Activity of the enzyme was determined by means of a sensitive colorimetric reaction for the product, inorganic phosphate. Non-enzymatic hydrolysis problems with the substrate are minimized in this procedure by the masking action of citrate. The final heteropoly blue color appears to be considerably sensitized by interaction of phosphomolybdous ion with arsenite. The stability of the relatively labile enzyme was ensured by chelating any metals present with ethylene diamine tetraacetic acid. The overall results obtained by the procedure appear to be useful as an aid in the diagnosis of Type I glycogenosis, a glycogen storage disease called Von Gierke's disease.


[Action of distension upon uterine growth (author's transl)]. The uterine horn growth has been studied in castrated female rats under the influence of distension. One of the two horns was used as a control horn, to insure us that no endogenous hormone was responsible for the uterine growth. Morphological studies, as well at low as at high resolution, demonstrated the ability of distension to initiate an epithelial growth. The use of 3H thymidine, or H3 uridine also provided some evidence for RNA synthesis 30 mm after distension initiation, and DNA synthesis 24 to 48 hours after distension onset. In these cases all the uterine tissues were concerned.


[Immunoglobulins and sheep red cell receptors in human lymphoblastoid cell lines simultaneously carrying Epstein-Barr virus and cytomegalovirus genomes]. In this communication we attempt to characterize the principal immunological properties of 3 human lymphoblastoid cell lines (LCL) established in our laboratory which carry both the EBV and CMV genomes. The data obtained suggests that two LCL are representative of B lymphocytes while the other LCL has a mixed cell population with the majority being of the B type. The reasons for this heterogeneity are discussed. It is suggested that CMV may also transform thymus-independent (B) lymphocytes.


[Interferon inhibition of lymphocyte mitogenesis]. Interferon inhibits the early phase of mitogenic action of concanavalin A on lymphocytes. There was no evidence for a role of cyclic AMP in the process of inhibition. Dialysis of interferon against high salt and urea did not separate cell inhibitory activity from antiviral activity.


[Monoclonal immunoglobulins, cytomegalovirus infection and malignant observations blood diseases]. Monoclonal immunoglobulins (Eg) are detected in the serum of 7/29 patients treated for leukemia (6 cases) or lymphoma (1 case). All of them developped cytomegalovirus (CMV) infection. The 7 patients were children or young adults; 6/7 were in complete remission of the hematological disease. They completed chemotherapy before the onset of the viral infection and had received blood components. The monoclonal Ig are IgG (6 cases) and IgM (1 case), In 2 sera there are two M components. The level of the peak was under 1 g/100 ml (5 cases) and reached 1.5 and 1.7 g/100 ml in the 2 other sera. Polyclonal Ig are normal except in one case where there is moderate hypo-Ig; Plasmocytic infiltration was observed in none of the 7 patients. Evidence of CMV infection was confirmed by viremia and/or significant rise of complement fixing antibodies. In four cases, the monoclonal Ig disappeared in less than 6 months. The significance of the monoclonal Ig associated with CMV infection is discussed.


Mechanisms of hepatic bile formation.

Mechanisms of hepatic bile formation. It should be evident from this review of recent investigations that we are still very far from a consistent description of bile formation, much less a satisfactory understanding. Nevertheless certain broad conclusions emerge. Four distinct kinds of active solute transport can be identified, and because bile always has nearly the same osmotic pressure as plasma, each of them is a determinant of bile flow. 1. Concentrative transport of water-soluble organic constituents, of which bile acids are quantitatively most important, occurs in the canaliculi accompanied by the passive flow of water and inorganic electrolytes. Owing to micelle formation the osmotic force for this flow is largely attributable to Na+ ions that accompany the bile acids anions. 2. The canalicular flow obligated by the excretion of bile acids is supplemented by the entry of additional fluid, the so-called bile acid-independent canalicular fraction. Because no organic component has been identified to account for this phenomenon, the active transport of one or more inorganic ions is probably responsible. The limited evidence available at present suggests that Na+ ions is the most likely candidate. 3. The extralobular biliary epithelium can modify the flow and composition of bile by the reabsorption of inorganic ions--a process which resembles reabsorption from the gallbladder in the sense that bile in the lumen remains iso-osmotic with plasma while bile acids and the other organic constituents are concentrated. 4. Under the influence of secretin, and to a lesser degree other intestinal hormones, the ducts or ductules can secrete additional fluid in which HCO3- is concentrated with respect to plasma. A fifth component of bile is generated by the canalicular excretion of phospholipid and cholesterol, but these are insoluble in water and are incorporated into micelles, and, therefore exert no osmotic force. The existence of these processes is inferred from studies of many different species, and it should be emphasized that the picture is a composite one. For example, distal fluid reabsorption has been convincingly demonstrated only in dogs and monkeys, and secretin is not a choleretic in rats or rabbits. It should also be clear that the actual mechanisms of solute transport remain poorly defined. Thus the term active transport in the present context should be thought of in its general thermodynamic sense rather than as denoting any particular transport mechanism. For the future, the most pressing problems are methodologic. To mention only three that seem especially important: ways must be found to sample bile closer to its origin; the proper interpretation of studies with isolated liver cell membranes will require unambiguous methods to certify their source; and descriptions of transport kinetics must somehow be refined to reflect the effective intracellular concentration of solutes as well as their distribution within the liver lobule.