Ultrastructure of synapses in the central nervous system of lamellibranch molluscs. Fine-structural characteristics of synaptic contacts were investigated in the central nervous system of different species of lamellibranch molluscs. Neuropile of the ganglia is characterized by regular occurrence of ultrastructurally well-defined polarized chemical synapses resembling those described in other invertebrate species and vertebrates. In addition to the generally observed membrane thickenings, enhanced density of synaptic membranes, cleft material and vesicle clustering on the presynaptic membrane, synapses are occasionally characterized by other and pinocytotic invaginations. Synaptic connections were distinguished on the basis of the vesicle content of the presynaptic terminal. Different forms of synaptic configurations (divergence, convergence, presynaptic modification) were observed in the ganglia.


Effects of an intravenous injection of tetracosactid on plasma corticosteroid and testosterone levels in unstressed male rabbits. Administration of tetracosactid into male rabbits, fitted with permanently indwelling jugular catheters, resulted in a rapid rise of plasma corticosteroids and plasma testosterone. Corticosteroid concentrations were significantly elevated at 40 and 60 min and testosterone concentrations 20 min after the iv injection of tetracosactid (2.5, 5.0, and 10.0 micrograms/kg body weight), in comparison to pre-treatment levels. Corticosteroid values in plasma were elevated as long as 120 min after tetracosactid injection. In contrast, testosterone levels were lower at 60-120 min after tetracosactid injection than corresponding pre-treatment values. However, these differences were not significant. At the doses used no tetracosactid-dose-dependent corticosteroid or testosterone release could be found; apparently, testosterone release is only dependent upon basal plasma levels but not upon the dose of tetracosactid applied. From these studies it is concluded that tetracosactid may bring about an increase or decrease of testosterone concentration in plasma in the buck depending upon the length of time elapsing between injection of tetracosactid and blood withdrawal.


Replacement oral ethinyloestradiol therapy for gonadal dysgenesis: growth and adrenal androgen studies. We have studied growth and adrenal dehydroepiandrosterone (DHA) responses to iv synthetic adrenocorticotrophic hormone (ACTH, Cortrosyn) in 6 girls with gonadal dysgenesis before and during treatment with low-dose ethinyloestradiol (EOe2). In all patients there was a statisfactory induction of secondary sexual characteristics including increase in breasts and public hair and onset of withdrawal bleeding within 6 months of therapy. Height velocity increased from 2.8 +/- 0.9 cm/year pre-treatment to 5.3 +/- 1.5 cm/year (P less than 0.02) in the first year. There was deceleration to 1.9 +/- 1.1 cm/year in the second year. There was no disproportionate advancement in bone age and thus, presumably, no loss of ultimate height. We could demonstrate no change in basal or ACTH-stimulated levels of DHA, a specific adrenal androgen, to account for the increased public hair and growth in these patients.


Receptors for gonadotrophin and prostaglandin F2 alpha in bovine corpora lutea of early, mid and late luteal phase. A total of 15 corpora lutea representing early (day 3), mid (day 13) and late luteal phase (day 20 and 21-24) were obtained by ovariectomy on cycling cows. The luteal weights and peripheral plasma progesterone levels just prior to ovariectomy, were consistent with the above luteal phases. The specific binding of [125I]human chorionic gonadotrophin to membranes prepared from corpora lutea was significantly higher (P less than 0.01) for days 13 and 20 than for days 3 and 21-24. The binding in day 21-24 corpora lutea was higher (P less than 0.01) than day 3. Although there was no different either in number or affinity (apparent dissociation constant (Kd) = 0.04 nM) of gonadotrophin receptors in days 13 and 20 corpora lutea, only in the former did the binding correlate well with plasma progesterone levels. The specific binding of [3H]prostaglanding (PG)F2 alpha to the membranes of the same corpora lutea showed a progressive increase (P less than 0.01) from day 3, reached the highest value at a time when corpora lutea were actively regressing (day 20) and the decline (P less than 0.01) by day 21-24. Although a considerable number of PGF2 alpha receptors existed at day 13, the affinity of these same receptors was 203 times lower (Kd = 3458 nM) than the affinity of receptors in day 20 corpora lutea (Kd = 17 nM). In summary, the above results show that gonadotrophin receptors correlate with luteotrophic, whereas PGF2 alpha receptors correlate with luteolytic phases in bovine corpora lutea.


Thermal stability of F-actin as studied by spin labelling. An analysis of the EPR spectra of maleimide spin-labelled actin was undertaken. We estimated a rotational correlation time of (13 +/- 2) nsec for the five-membered maleimide spin label bound to G-actin. The polarity of the environment of the bound labels indicated a strong polar character. The temperature dependence of the EPR spectral parameters of the attached label for F-actin exhibited rapid changes between 60-70 degrees C, which might be due to changes of protein structure. The conformational changes were reversible below 65 degrees C. The spin label spectra showed that the polymerization and depolymerization could be accomplished on actin thermally treated in F-form for 10 minutes at a temperature not higher than 60 degrees C. The findings suggest thermal stability of the spin-labelled sites in F-actin below 65 degrees C.


[Surgical treatment of tumors of the parotid gland (author's transl)]. One hundred and thirty-one cases of patients operated for a parotidal tumor were reviewed. We report on 85 cases that had no previous treatment, including 68 benign and 17 malignant tumors. The surgical treatment applied is analysed according to the benign or malignant nature of the tumor and to the results. For benign mixed tumors (43 cases) there were 29 total resections, 12 superficial lobectomies and 2 simple enucleations (intraparotidal location unforseen). For tumors of Warthin (19 cases) the treatment was a superficial lobectomy in all cases. For malignant tumors (17 cases) 5 superficial lobectomies and 12 total resections of the gland were performed. In the latter, the facial nerve was severed in 6 patients and left intact in the other 6. The minimal operation realizing the resection of a parotidal tumor, is the superficial lobectomy realized in once, the deep lobe only being resected if invaded. Preservation of the facial nerve should always be tried, sacrificing it only in cases of infiltrating malignant tumor.


Fingerprint inclusions and circular structures in the muscle. Report of a case. Fingerprint inclusions, circular structures of unknown origin, mitochondrial changes, dilatation of the sarcoplasmic reticulum, rods of Z-line material, elongated T-systems, honeycombs, and myelin figures were present in some damaged muscle fibres. Many damaged fibres showed loss of myofilaments and groups of atrophied fibres occurred. The muscle biopsy was from the left biceps brachii and presented also the changes of polymyositis. The patient had muscle weakness, malignant hypertension, alcoholism, polyarthropathy, and evidence of mild peripheral neuropathy.


Studies on the early changes in acute isoniazid neuropathy in the rat. Large single doses if isoniazid by mouth (1--2g/kg) have been shown to produce in rats Wallerian degeneration visible with the light microscope from the third day onwards. By contrast, changes in axons are seen from 24 h onwards by electron microscopy. The earliest ultrastructural changes are associated with vacuoles appearing between axon and Schwann cells. These are large and focal, and often compress the axon. The adjacent axon may show changes in smooth ER, and in microtubular arrangement. Alterations in smooth membranes and in mitochondria are visible in Schwann cell cytoplasm, not necessarily related to the vacuole formation and axonal features. It is suggested that INH neuropathy is essentially a multifocal axonal lesion.


Histopathology of the lens capsule in fibrillopathia epitheliocapsularis (FEC) or so-called senile exfoliation or pseudoexfoliation. An electron microscopic study. Twenty lenses from cataractous eyes have been subjected to enzymatic breakdown by collagenase and alpha-chymotrypsin in order to remove part of the lens capsule and the zonular apparatus. Twelve of the lenses had clinical fibrillopathia epitheliocapsularis (FEC syndrome). In addition four FEC lenses and four ordinary cataractous lenses served as controls. A characteristic fibrillar substance was found in the FEC lenses located to circular epithelium-near areas varying in size from 10 micrometers to 150 micrometers. The circular areas were found in zona germinativa in front of the bow region and corresponded to the discoid plaques of the deep layer. The ultrastructure of the finely fibrillar material of the deep layer were indistinguishable from that of the surface material of the peripheral band and the central disc. A connection between the discoid plaques of deep layer and the peripheral granular band were demonstrated in the form of radial "cobble-stone" sectors seen in partially digested FEC capsules. An increased amount of age dependent spindleshaped formed bodies were also found in FEC lenses. The evidence presented points towards a lenticular origin of the Busacca-bushes in the peripheral ganular band of the lens capsule.


The prognosis in congenital lower limb hypertrophy. Twenty-eight patients with congenital total hypertrophy and ten patients with lower limb hypertrophy with congenital vascular abnormalities are reviewed. The pattern of increase in leg length discrepancy during growth and its influence on surgical management is discussed and the clinical features of the affected limbs are described. In congenital total hypertrophy the maximal increase in leg length discrepancy of more than 2.5 cm at age 4 years are likely to develop significant limb overgrowth that will require eventual surgical correction. In patients with congenital vascular abnormalities the change of leg length discrepancy was variable in degree and unpredictable in pattern, even in those with similar venous anomalies. The outcome for the limb was detergiography.


Primary carcinoma of the liver. A histological study of 52 cases from Denmark. Among 7763 autopsies performed in Greater Copenhagen in 1973, there were 309 cases of cirrhosis of the liver and 52 cases of primary carcinoma of the liver (PCL). Of the latter, 45 were hepatocellular carcinoma (HCC), 4 combined HCC and cholangiocarcinoma (CCC) and 3 CCC. HCC was found in 7.8 per cent of the cirrhotic livers and was in 57.1 per cent accompagnied by cirrhosis. The criteria of WHO, Peters (modified) and Anthony were used for classification. The degree of differentiation of the tumours was estimated using the criteria of WHO and Edmondson. The apparently small number of CCC may be due to the fact that this tumour is often overdiagnosed at the expense of HCC. The incidence of combined tumours is probably higher than generally assumed. The reticulin stain was found very valuable in HCC, both for descriptive and diagnostic purposes. In contrast to the situation in sub-Saharan Africa where hepatitis B virus is incriminated as the most important etiologic factor of HCC, it was found in the present study that alcoholism was a very essential cause of cirrhosis and thereby of HCC.


Increased brain uptake of copper and zinc in mice caused by diethyldithiocarbamate. Mice given 64CuCl2 and 65ZnCl2 (10 mumol/kg) were treated with sodium diethyldithiocarbamate (0.5 mmol/kg). The treatment increased the brain level of radioactive copper five-fold and that of radioactive zinc three-fold. Such redistribution of metal ions may be explained from the formation of lipophilic metal chelates. The increased brain levels may involve neurotoxic effects.


Effect of toxogonin and P2S on the toxicity of carbamates and organophosphorus compounds. Toxogonin (80 mg/kg intraperitoneally) given 15 min. prior to the administration of organophosphorus insecticides dimethoate, malathion, parathion and azinphos-methyl, organophosphorus warfare agents soman and tabun, or carbamates physostigmine, pyridostigmine and aldicarb, reduced the toxicity in mice of these agents by increasing their LD50 dose 1.5-3 fold. The toxicity of the carbamate insecticide carbaryl, however, was significantly increased by toxogonin. Similar results were obtained for P2S (150 mg/kg intraperitoneally) with respect to the toxicity of dimethoate, soman and pyridostigmine, whereas no effect could be detected on the toxicity of tabun. Only a slight reduction in the toxicity of physostigmine was observed. The acetylcholinesterase activity in erythrocytes, cerebrum and diaphragm of surviving mice 20 hours after organophosphate intoxication was similar both in toxogonin and P2S treated animals and untreated animals.


The role of alpha-adrenergic receptors in the vasoconstrictor response induced by indomethacin in the kidney. The effect of indomethacin, an inhibitor of prostaglandin (PG) synthesis, was studied on the renal circulation, Na+ and water excretion in anaesthesized dogs during alpha-receptor inhibition. Indomethacin decreased cortical blood flow (CBFcontr, 454 +/- 142; CBFindo, 332 +/- 51 ml per min per 100 g; p less than 0.02) as well as medullary blood flow (OMBFcontr, 339 +/- 95; OMBFindo, 183 +/- 46 ml per min per 100 g; p less than 0.001), salt and water excretion, further it caused a shift in the intrarenal blood flow distribution toward the cortex. Alpha-blockade prevented the indomethacin-induced vasoconstriction in the cortex (CBF alpha inhibition + indo, 455 +/- 76 ml per min per 100 g) but not in the medullar (OMBF alpha inhibition + indo, 259 +/- 102 ml per min per 100 g, p less than 0.05). Alpha-blockade failed to prevent the indomethacin-induced antidiuresis, antinatriuresis and the intrarenal blood flow redistribution. GFR remained unaffected in all three series of studies. Our experimental findings are in line with the presumption that alpha-receptors are involved in the renal circulatory changes caused by indomethacin, probably as a result of an enhanced NE release during the inhibition of PG production. A NE--PG feed back mechanism is suggested in the regulation of renal circulation. The reduction of salt and water output induced by indomethacin appears to be independent of the alterations in renal haemodynamics, and seems rather to be the result of enhanced Na+ reabsorption, predominantly at the distal segment of the nephron, in the absence of PG, and/or a direct action of indomethacin.


Cardiac output and peripheral resistance of swim-trained rats under urethan anesthesia. Several circulatory variables were determined in control and swim-trained white rats under urethan anesthesia. Trained animals exhibited significantly lower heart rate and cardiac index and significantly higher peripheral resistance than control animals. Blood pressure, systolic index and left ventricular work output were not statistically different in the two groups. These results suggest an alteration of resting cardiovascular regulation in trained animals. Beta adrenergic mechanisms are suppressed by increased parasympathetic tone, and the resting circulatory balance is maintained mainly by increased alfa adrenergic mechanisms.


Effects of macrophages in resistance to murine cytomegalovirus infection. In a preliminary experiment, the protective effects of peritoneal macrophages was shown by transferring macrophages from adult mice to newborn and to 7 and 14 days old mice. It was demonstrated that such transplantation protect suckling mice from intraperitoneal infection with MCMV by reducing the mortality rate from 100% to 27%.


Syncytia formation of human transformed cell lines by simian sarcoma virus type I (SSV-I/SSAV-I). Human cells derived from malignant tumors (HeLa, HEp-2 and KB) and human cells transformed by tumor viruses (KCand RSb) formed syncytia by simian sarcoma virus type I (SSV-I/SSAV-I), but human diploid or non-transformed cells (WI-38, HEL and HEC) did not.


Demonstration of EBNA (Epstein-Barr virus nuclear antigen) antibodies of different immunoglobulin classes. Anit-EBNA IgM, a previously unknown antibody, was detected by the antihuman globulin anticomplement immunofluorescence (ACIF) method in serum samples from acute infectious mononucleosis (IM) of Epstein-Barr virus (EBV) origin. The antibody disappears from the serum in some weeks during convalescence. It was absent in anti-EBV=positive sera of healthy donors and in serum samples taken from patients with IM caused by cytomegalo-virus. The antibody appears simultaneously with anti-EBV IgmM and, reaching a lower titre than the latter, its titre curve runs parallel with the anti-EBV IgM curve. Since in acute EBV infections, anti-EBNA IgM always appeared, its presence may serve as an additional evidence of the acuteness of EBV infection. In EBV-seropositive healthy subjects, the bulk of antibodies belongs to the IgG class, non-complement-fixing IgA antibodies occur only sporadically.


Transformation of hamster embryonic fibroblast cells by UV-irradiated human cytomegalovirus. Infection of hamster embryonic fibroblast (HEF) cells with UV-irradiated human CMV resulted in appearance of transformed clones. The established cell line (87-TRH-5) when inoculated subcutaneously induced tumour in newborn hamsters. The 87-TRH-5 cell line and the cell lines (87-TRH-5-Tsc1, 87-TRH-5-Tsc2, 87TRH-5-Tsc3) developed from the induced tumour, proved to bear CMV-spcific cytoplasmic and surface antigens. Demonstration of infectious CMV or MCV-specific nuclear antigen in 87-THR-5-cells has failed.


Neoplastic response of turkey liver to MC29 leukosis virus. Twelve to fourteen days after intravenous inoculation of MC29 virus (1 x 10(5)LD50) into 1-day-old turkeys, liver tumour developed in 100% of the infected animals and led to death of birds. The tumour proved to be hepatomas histologically and electron microscopically. Numerous C-type particles were detectable among the tumour cells, as well as budding from the cell membrane. C-type particles were also observed in the tumour-free liver tissue in the spaces between the cells, budding was not detectable. Large number of virus particles were found in spleen extracellularly and intracellular vacuoles. Kidney tumours did not develop, but a few extracellular virus particles were located in the tissue. The MC29 virus-induced primary liver tumour in the turkey seems to be a suitable model for the morphological study of the relationship between oncogenic viruses and eukaryotic cells.


Experimental cerebral concussion. A histochemical study. The activity of mitochondrial enzymes (succinic dehydrogenase and cytochrome oxidase) and enzymes associated with blood-brain barrier function (butyrylcholinesterase and alkaline phosphatase) in the CNS of rats was studied from 5 minutes to 62 hours after cerebral concussion. There was a transient increase in succinic dehydrogenase activity during the first hour after concussion in the neurons of the structures close to the impact. The alkaline phosphatase activity, strongly positive in the walls of normal blood vessels, decreased within five minutes after concussion; it virtually disappeared in 15 minutes but returned to normal level after 62 hours. These findings are in good correlation with previous electron microscopic observations. Their significance is discussed.


Nerve cell calcium as a messenger for opiate and endorphin actions. The studies outlined in this chapter provide convincing evidence for the role of calcium in pharmacological and biochemical actions of opiates on the central nervous system. The ability of the synaptosomes to accumulate excess calcium during narcotic exposure represents a singular major example of cellular adaptation at the level of the membrane. These findings together with the possibility that CDR and calcium may mediate a number of processes involving neurotransmitter activity provide many avenues for the future study of opiates and other central nervous system depressants.


Histochemical localization of opiate receptors and the enkephalins. It is possible to localize opiate receptors and opioid peptides by light microscopic histochemical methods. The receptors have been mapped by autoradiography with specifically bound 3H-diprenorphine. They are found in anatomical areas associated with physiologic functions known to be altered by opiate administration, as well as in other areas. There appear to be opiate receptors on axons and nerve terminals; some of these are probably for axo-axonic synapses in glomeruli. Some electrophysiologic studies are supportive of the autoradiographic results. Enkephalin-containing neurons can be revealed by immunohistochemistry. Often, although not always, the distribution of enkephalin-like immunoreactivity is similar to that of the opiate receptors. These results are compatible with the notion that the enkephalins are the endogenous substrates of the opiate receptors.


The obesity of middle age: a common variety of Cushing's syndrome due to a chronic increase in adrenocorticotrophin (ACTH) and beta-endorphin activity. The common obesity of middle age presents a set of features that strongly resembles the cardinal symptoms of Cushing's syndrome: obesity of the face (moon face), upper back (buffalo hump) and trunk (pot belly) accompanied by signs of protein-wasting. In non-obese individuals who remain at a constant weight throughout life, the proportion of adipose tissue increases with age at the expense of lean tissue loss. Thus, a mild version of Cushing's syndrome may be part of the normal aging process. A more intense version of this process may occur in overweight adults. Excess and chronic activity of two pituitary hormones may contribute to this adiposity. Both hormones are produced in the same pituitary cell by cleavage from a common large precursor known as pro-opiocortin. One hormone is adrenocorticotrophin (ACTH), which stimulates the release of the glucocorticoid hormones. These hormones promote the conversion of bodily proteins to glucose (gluconeogenesis). The other pituitary hormone is beta-endorphin, a stimulant of appetite that causes the release of insulin. This pancreatic hormone promotes the conversion of glucose and fatty acids to triglycerides (lipogenesis). Three different etiologies are suggested for the excessive and chronic action of these two pituitary hormones: tumors that increase the number of cells that synthesize pro-opiocortin; mutant strains that produce excessive amounts of ACTH and beta-endorphin such as the genetically obese mouse (ob/ob) and rat (fa/fa); and an age-determined shift in the type of cleavage enzymes present in the pro-opiocortin cell that favors ACTH and beta-endorphin production.


Glucocorticoid receptor inactivation and activation by phosphorylation mechanisms. The specific glucocorticoid binding capacity of cytosol preparations is rapidly lost on incubation at 25 degrees in the absence of ligand. We have examined this process in cell-free preparations from rat thymus, rat liver and mouse fibroblasts (L 929 cells), and we have found that the unoccupied receptor is inactivated by endogenous enzymes to a form that does not bind steroids. The inactivation can be prevented by inhibitors of phosphatase action such as molybdate, fluoride and glucose-1-phosphate. On the basis of this type of evidence we propose that the receptor activity of cytosol can be rendered inactive by a dephosphorylation process. We have now been able to partially reactivate the receptor in both L cell and rat thymus cytosols in an ATP dependent manner. If fibroblast (L cell) cytosol is preincubated to permit receptor inactivation by endogenous enzyme, further inactivation can be prevented by the addition of 10 mM molybdate and reactivation of the binding capacity can be obtained by adding 5 to 10 mM ATP in addition to molybdate. ATP dependent activation is prevented with EDTA and this block is overcome by added magnesium. ADP, CTP, GTP, and UTP are inactive. After inactivating the glucocorticoid binding capacity of rat thymocyte cytosol by incubation for 45 minutes at 25 degrees, considerable reactivation is obtained by addition of dithiothreitol and ATP. This system does not absolutely require the presence of a phosphatase inhibitor in order to show activation. Thymocyte cytosol can also be activated to a steroid binding state by addition of DTT and heat-treated (90 degrees for 15 min.) cytosol from a variety of cell types. The heat-treated cytosol contains ATP, reducing equivalents, and a relatively small molecular weight heat-stable activator(s) that potentiates the reactivation process. Maximum receptor activation is obtained by adding dithiothreitol, heat-stable factor, ATP, and molybdate to the inactivated thymocyte cytosol.


Relationship between histamine-induced changes of cyclic AMP and mechanical activity on smooth muscle preparations of the guinea-pig ileum and the rabbit mesenteric artery. On guinea-pig ileum and rabbit mesenteric artery contracted by high potassium (100 mM) histamine produced relaxations which were inhibited by the H2-receptor antagonist metiamide. These results are thus indicative for the role of H2-receptors in mediating relaxation and for H1-receptors in mediating contraction on smooth muscle. Time course studies for the relaxing and cyclic AMP responses to histamine showed that the cyclic AMP increase preceded the H2-receptor mediated relaxation. The cyclic AMP increase in response to histamine was prevented by metiamide, but remained unaffected by mepyramine on both the guinea-pig ileum and the rabbit mesenteric artery. In addition, dose-response curves obtained on the mesenteric artery demonstrated that the H2-receptor mediated depressor responses coincided with cyclic AMP increases. Thus, these results gave clear-cut evidence that cyclic AMP is an intracellular metabolic event only implicit in the H2-receptor mediated relaxation, but not in the H1-receptor mediated contraction on smooth muscle preparations.


Effects of D-penicillamine on lymphocyte modulation of synovial collagenase production. While D-Penicillamine is effective in the treatment of rheumatoid arthritis, its mechanism of action is unknown. In this study, effects of D-Penicillamine on collagenase production by adherent rheumatoid synovial cells were investigated. D-Penicillamine did not directly affect the synovial collagenase production. However, lymphocyte-free-supernatant (LFS) recovered from lymphocytes exposed to D-Penicillamine in vivo and in vitro significantly reduced collagenase production by adherent synovial cells. LFS from lymphocytes of normal subjects and from non-D-Penicillamine treated rheumatoid patients stimulated collagenase production. These investigations indicate that D-Pencillamine indirectly affects collagenase production by cultured synovial cells and suggests beneficial effects on controlling the primary disease process.


Differential susceptibility of peripheral nerves of the hen to triorthocresyl phosphate and to trauma. The nerve fibres of largest diameter and of greatest length are considered to be the most vulnerable to triorthocresyl phosphate (TOCP). In this study, the differential vulnerability of the particular sciatic nerve branches was determined in the course of TOCP neuropathy and of Wallerian degeneration. The branch innervating the lateral gastrocnemius muscle, made up predominantly of large-diameter fibres, proved most susceptible to TOCP. By contrast, after proximal sciatic-nerve transection, degeneration commenced in the lateral nerve of the third digit, containing long nerve fibres of small diameter.


Effects of the vestibular system on human development, part 2: Effects of vestibular stimulation on mentally retarded, emotionally disturbed, and learning-disabled individuals. In Part 1 of this two-part series, studies concerned with the effects of vestibular stimulation on human development and function were reviewed and some implications for therapy were suggested. In Part 2, three categories of dysfunction with possible links to the vestibular system are discussed. Studies in the category of mental retardation evaluate motor development and sensory preference. Possible vestibular associations with emotional disturbance are examined by review of studies concerned with etiology, motor activity, speech, and clinical observations. A brief review of studies concerned with early identification and speech and language factors of learning-disabled children constitutes the third category. Interpretations are drawn and some implications for therapy are made.


Formation and fate of ethionine-induced cytoplasmic crystalloids in rat parotid acinar cells. The formation and fate of cytoplasmic crystalloids in rat parotid acinar cells were investigated during ethionine intoxication and recovery. By day 3 of ethionine treatment, acinar cells had numerous autophagic vacuoles containing recognizable secretory granules and fragments of rough endoplasmic reticulum. By day 5, immature crystalloids were present in many of the autophagic vacuoles, and as the crystalloids matured, a 7-nm periodicity became apparent. Crystalloids were never observed in the Golgi saccules or in any other organelle associated with secretory granule formation. When ethionine treatment was stopped, the acinar cells rapidly returned to their normal morphology. The majority of the crystalloids and autophagic vacuoles were lost from the cells during the first two to three days of recovery. At this time annulate lamellae were present intracellularly, and macrophages, many containing crystalloids, were associated with the basal surface of the acinar cells. These results indicate that the cytoplasmic crystalloids are formed in autophagic vacuoles, and do not represent an abnormal secretory product. Additiontionally, during recovery crystalloids may be removed from the acinar cells by interaction with macrophages. The sequence of autophagic vacuole formation, development of crystalloids, macrophage infiltration and phagocytosis of acinar cell debris appears to be a non-specific response of the rat parotid gland to cellular injury occurring in a variety of experimental and pathological conditions.


The isolation of single cells from the avian salt gland. Collagenase, hyaluronidase, and trypsin were used to isolate single cells from the avian salt gland. Three cell types were distinquishable in the resultant cell suspension: peripheral, intermediate, and principal cells. The fine structure of these cells is described and related to the morphology of the intact gland.


Structure of the endodermal epithelium of the chick yolk sac during early stages of development. The structure of the areas pellucida and vasculosa of the early chick embryo (stages 11-29) was examined by light, transmission and scanning electron microscopy. The most striking feature of the endodermal cells of these areas is the presence of large intracellular yolk drops which are characteristic of the regions in which they are found; lipid-like homogeneous drops in the area pellucida, heterogeneously composed pleomorphic drops in the mid-region of the area vasculosa and granular drops at the periphery of the area vasculosa in the region of the sinus terminalis. On morphological criteria it is postulated that granular drops may arise by direct engulfment of extracellular yolk, but this does not appear to be true for pleomorphic or homogeneous drops. Since the apical junctions between endodermal cells across the yolk sac are tight, they seal off the extraembryonic compartment from the vitelline circulation and presumably prevent intercellular passage of the yolk constituents. Thus the endodermal epithelium must mediate the transport of nutrients from the yolk mass to the developing embryo. Endodermal cells exhibit a variation across the yolk sac in the presence and number of structures associated with uptake of extracellular materials. The mid-region of the area vasculosa appears to be the most endocytotically active region with an abundance of microvilli, bristle-coated pits and vesicles and apical canaliculi and vacuoles. There is a close association between the endoderm and vitelline blood vessels and this association is maintained, as the yolk sac develops, by the formation of small vessels juxtaposed between the vascular surface of the endoderm and the walls of the large vitelline vessels.


The effect of a natural high-fiber diet on serum lipids, fecal lipids, and colonic function. In a cross-over experiment,  46 young healthy volunteers consumed in succession a high-fiber and a low-fiber diet for 3 weeks at two levels of dietary cholesterol. Half of the dietary fiber came from fruits and vegetables, and the rest from bread and other cereal products. On the high-fiber diet, concentrations of serum cholesterol decreased on average by 0.44 mmole/liter with high-cholesterol and 0.31 mmole/liter with low-cholesterol regimes; high density lipoprotein-cholesterole decreased by 0.1 mmole/liter; on average fecal weight increased by 115 g/day and mean transit time through the gut was decreased by 18 hr. Only part of the decrease in serum cholesterole may be due directly to the high intake of dietary fiber components. The remainder is due to differences in fat intake: during the high-fiber period subjects consumed less fat and cholesterol than had been planned.


Infectious mononucleosis test performance on CAP survey specimens. The participant performances on 1977 and 1978 infectious mononucleosis survey specimens are tabulated. The various methods utilized gave similar results on negative specimens. Commercial reagent kits incorporating differential absorption performed somewhat better on positive specimens than kits without differential absorption. Many of the commercial reagent kits performed very poorly on the false postitive specimen.


Somatomedin and growth hormone studies in pediatric renal allograft recipients who receive daily prednisone. Plasma somatomedin (SM) activity and growth hormone (GH) concentration were measured in ten growth-retarded, well-nourished pediatric renal allograft recipients who received daily prednisone therapy. The SM activity ranged from 0.21 to 1.22 micron/mL, and it was subnormal in three patients. A significant correlation was found between SM activity and creatinine clearance. Serial SM levels were determined during a 24-hour period in five patients; SM activity decreased at six and 12 hours and returned to normal values by 24 hours. The 24-hour plasma GH concentrations ranged from 1.5 to 7.6 mg/mL. Peak GH concentrations ranged from 2.1 to 14.2 ng/mL after insulin-induced hypoglycemia and from 1.8 to 24.6 ng/mL after oral glucose loading. Sleeping GH peaks were absent in two patients. These results suggest that growth failure in renal allograft recipients who receive daily prednisone may result from (1) partial GH deficiency, (2) reduced SM levels owing to diminished allograft function, and (3) daily transient decrease in plasma SM levels after prednisone administration.


Surgical management of insulinoma.

Surgical management of insulinoma. Three patients with insulinoma have been presented. Accurate diagnosis, localization and multiplicity are possible employing angiography, sonography, CAT and insulin concentration determinations of specimens obtained by percutaneous, transhepatic simultaneous sampling of the portal, splenic and superior mesenteric veins. Meticulous exploration of the gland will permit enucleation rather than major resection to be a feasible solution and the operation of choice.


Prevalence of antibody to hepatitis A and hepatitis B viruses in selected populations of the South Pacific. Five island populations representing the three major cultural groups of the South Pacific--Polynesia, Micronesia, Melanesia--were studied for prevalence of antibody to hepatitis A virus (anti-HAV) and of antibody to the core antigen of hepatitis B virus (anti-HBc). Sera were collected in the late 1950s and early 1960s, selected where possible for appropriate age and sex distributions, and were tested by radioimmunoassay. Rather marked differences in prevalence were observed. Anti-HBc patterns confirmed that HBV is endemic in the Pacific populations. Furthermore, the patterns differed somewhat from each other and did not correlate with ethnogeographic area. Prevalence of anti-HAV was high in all populations studied. A Unique pattern was found for the island of Ponape (Micronesia): In a Ponape population bled in 1963, anti-HAV was not detected in any individual under 20 years of age, but almost all individuals over that age were found to be seropositive. On testing a second group of sera collected in 1975, all individuals aged 14--21 years were found to be antibody positive, indicating that HAV had returned to Ponape sometime prior to 1975. There was no significant difference in the prevalence of HAV or HBV infection between males and females in any of the populations studied.


A seroepidemiologic study of infection with HAV and HBV in five Pacific Islands. A batch of 1025 serum samples, obtained from healthy subjects and hospital patients from five Pacific islands (Viti Levu, Funafuti, Niue, Rarotonga and Upolu) were tested for antibody to hepatitis A virus (anti-HAV), hepatitis B surface antigen (HBsAG) and antibody to hepatitis B surface antigen (anti-HBs) by solid phase radioimmunoassay (SPRIA). High frequencies of anti-HAV were observed in each population (Funafuti, 79.9%; Upolu, 81.6%; Viti Levu, 84.3%; Rarotonga, 95.0%; and Niue, 95.2%), and the age-specific prevalence suggested that the majority of infections with this virus had occurred in the first decade of life. Hepatitis B was endemic in each population, although the total infection rates (as measured by the sum of HBsAG and anti-Hbs frequencies) were lower than for hepatitis A. While infections with each virus tended to run in parallel, peak prevalence of anti-HAV was usually reached in the second decade of life and peak prevalence of HBsAg and anti-HBs was not attained until a decade later. The pattern of infection with HBV was similar in each of the groups studied with the exception of Indians living in Viti Levu, among whom unusually low levels of antigen and antibody were demonstrated.


Comparative epidemiology of two rotavirus serotypes and other viral agents associated with pediatric gastroenteritis. Human rotavirus (HRV) type 1 or 2, adenovirus, or non-cultivatable 27 nm virus-like particles were demonstrated by electron microscopy and/or rotavirus ELISA in fecal samples from 45.5% of 604 gastroenteritis inpatients, 25.0% of 200 gastroenteritis outpatients and 6.0% of 812 control subjects, all sampled at Children's Hospital National Medical Center. Washington, DC. Rotaviruses were the most common pathogens detected as 39% and 22% of gastroenteritis inpatients and outpatients, respectively, shed HRV. About three-fourths of the rotaviruses were type 2, which was prevalent during five successive epidemic years from January, 1974, through June, 1978. HRV type 1 was detected in the last four successive epidemic years and represented nearly half of the HRV infections observed among gastroenteritis inpatients during the year 1977--1978. Both rotavirus serotypes were detected most often in the month of January, when 71% of 123 gastroenteritis inpatients and 62% of 34 gastroenteritis outpatients shed one of these viruses. Uncultivatable adenoviruses were detected significantly more frequently in stools from patients with gastroenteritis (3.9%) than from control subjects (0.6%), suggesting that these viruses played a role in acute enteric disease. The frequency of detection of 27 nm particles was not significantly different in gastroenteritis and control patients. Numerically, HRV infection was detected most often in gastroenteritis inpatients who were 10 through 12 months of age. The group of gastroenteritis inpatients with the highest percentage of HRV infection was 13 through 15 months of age. The excess of type 2 HRV infection relative to type 1 infection was especially large in those aged 7 through 24 months. Lower socioeconomic status or greater crowding appeared to be associated with the occurrence of rotavirus infection earlier in life and earlier in the epidemic year.


The association of total cholesterol, triglycerides and plasma lipoprotein cholesterol levels in first degree relatives and spouse pairs. HDL, VLDL, and LDL cholesterol as well as total cholesterol were measured in young adult offspring of Framingham Heart Study participants. Parental total cholesterol levels measured in the early 1950s were found to be significant predictors of present total cholesterol in the offspring. While both maternal and paternal cholesterol levels made an independent contribution to the prediction of the offspring's cholesterol, the mother's contribution was significantly greater than the father's for male offspring with a similar but less striking relationship for female offspring. The correlation for LDL cholesterol levels for siblings was higher than for HDL cholesterol or log VLDL choles terol. When adjustments were made for age, body weight, alcohol intake and cigarette smoking, slight reductions in the correlations were noted, but all adjusted correlations remained significantly different from zero. The spouse correlations for lipoproteins, however, did not differ from zero after adujustment. Since the significant association for lipoproteins in spouse pairs disappears on adjustment for correlates but the association for siblings does not, it is likely that the sibling lipoprotein associations result from either genetic or environmental factors shared early in life.


Familial absorptive hypercalciuria and renal tubular acidosis. Hypercalciuria was considered as a secondary condition when associated with familial renal tubular acidosis. Later studies suggested that hypercalciuria could lead to renal tubular acidosis and nephrocalcinosis. Selected members of a family spanning five generations were studied. Renal tubular acidosis was present in eight subjects in three consecutive generations. Increased 24-hour urinary calcium excretion was present in nine subjects in three consecutive generations, alone in the younger generation, and in combination with renal tubular acidosis and nephrocalcinosis in the older generation. Calcium loading tests showed the absorptive nature of hypercalciuria in nine of 18 subjects studied. This report suggests that in this family the absorptive hypercalciuria is an autosomal dominant genetic defect with complete penetrance and variable expressivity which leads to renal tubular acidosis and nephrocalcinosis.


Genital bacteriology: a comparative study of premenopausal women with postmenopausal women. Premenopausal women are reportedly at a higher risk than postmenopausal women of postoperative infection following vaginal hysterectomy. A study was designed to determine whether a relationship exists between bacterial flora and patient age that may explain the difference in the risk of postoperative infection. No statistically significant difference was found in the number or type of bacterial species isolated from the cervix or vagina of premenopausal and postmenopausal women. Postmenopausal women on a regimen of conjugated estrogens had genital flora similar to that of the other women studied.


Polypeptide hormones in argentaffin and argyrophil gastroduodenal endocrine tumors. The morphology and histochemistry of gastroduodenal endocrine tumors from 16 patients were studied. All patients underwent operation, in most cases with a preoperative diagnosis of nonendocrine tumor, ulcer, o polyp(s). The argentaffin reaction was positive in three tumors, and the Hellerström--Hellman argyrophil reaction was positive in four tumors. All tumors reacted positively to the Grimelius argyrophil stain, and 13 were positive with the Sevier--Munger argyrophil stain. Gastrin immunoreactivity was found in eight tumors, and substance-P immunoreactivity in seven tumors. No enteroglucagon, adrenal cortex hormone, or pancreatic polypeptide was observed in any of the tumors. Three patients with Sevier--Munger-positive gastric tumors had concurrent pernicious anemia, and 2 patients with gastrin-immunoreactive tumors had acute or chronic gastroduodenal ulceration. The results indicate that the gastroduodenal endocrine tumor as a rule gives no endocrine symptoms and that the tumor type is an unexpected finding at operation. The tumors may contain gastrin, substance P, somatostatin, and serotonin and tend to be multihormonal.


Selective mitochondrial degeneration in renal tubules following hyperbaric oxygen exposure. To study the ultrastructural effects of hyperoxia on the kidney, young adult Sprague Dawley rats were exposed to 3 atmospheres absolute (ATAs) of pure oxygen for 5 hours and were killed in a time sequence varying from immediately to 30 days after exposure. Their renal cortices were processed for electron microscopy. Selective mitochondrial changes were observed within sublethally and transiently altered proximal tubular epithelial cells. The most consistent finding was the accumulation of 0.08 mu to 0.5 mu round to ovoid homogeneous matrical inclusions which frequently formed larger confluent amorphous masses. The inclusions stained intensely with lead and uranium but appeared homogeneously electron-lucent in unstained sections. Energy-dispersive x-ray analysis revealed that they did not contain calcium or phosphorus. The inclusions were different from the innately electron-opaque flocculent densities commonly found in pathologically altered mitochondria. Since the mitochondria containing them were removed by autophagocytosis, it is suggested that the inclusions were associated with selective mitochondrial degeneration induced by hyperoxia. No glomerular lesions were found.


Na+,K+-ATPase enzyme units in lean and obese (ob/ob) thyroxine-injected mice. The possible involvement of Na+,K+-ATPase in the etiology of obesity in the obese (ob/ob) mouse was explored. The number of Na+,K+-ATPase enzyme units in skeletal muscle, liver, and kidneys from 4- and 8-wk-old obese and lean mice was estimated from saturable [3H]ouabain binding to particulate fractions. Neither phenotype nor age altered the Kd value for ouabain binding in these three tissue preparations. The total number of [3H]ouabain binding sites in hindlimb muscles was 35--55% lower in 4- and 8-wk-old obese mice than in their lean counterparts. However, the total number of [3H]ouabain binding sites in liver and kidneys of obese mice was similar to values observed in their lean counterparts. Because it has been suggested that ob/ob mice are hypothyroid, we investigated the response of Na+,K+-ATPase in these mice to thyroid hormone treatment (approximately 5 microgram thyroxine/day for 2 wk). The number of [3H]ouabain binding sites in the three tissues increased in both obese and lean mice injected with this relatively large dose of thyroxine, but the obese mice were 2--3 times more responsive than lean mice.


Mucosal gastrin receptor. I. Assay standardization and fulfillment of receptor criteria. Using iodinated gastrin with demonstrable biological activity, this study has shown that optimal specific gastrin binding occurs in rat gastric mucosal 270--30,000 g membrane preparations after an incubation period of 30 min at 30 degrees C (pH 7.4) with a protein concentration of 150--200 micrograms per assay tube. The gastrin binding was shown to be saturable with an equilibrium Ka of approximately 0.25 X 10(10) M-1 and an equilibrium Kd of approximately 4 X 10(10) M. The binding capacity was approximately 4 fmol/mg protein. Specific gastrin binding was shown to be present in the oxyntic gland and duodenal mucosa and to be absent from the antral mucosa, liver, spleen, and kidney. In order to decrease the specific binding of gastrin by 50% the competitors in order of potency are 15-Leu G-17 greater than cholecystokinin greater than caerulein greater than pentagastrin; secretin did not display a response similar to the other four competitors tested, indicating that its inhibition may be non-competitive. Fasting decreased the binding capacity of the gastrin receptor and refeeding brought the receptor levels back to control range; this result parallels the decrease seen in serum gastrin after fasting and the return to normal levels with refeeding. This suggests that rat gastric mucosal gastrin receptors may exhibit autoregulation. This study is the first to meet all the criteria for establishing the existence of a mucosal gastrin receptor.


Mucosal gastrin receptor. II. Physical characteristics of binding. Gastrin binding to rat gastric mucosal membrane preparations revealed a linear Scatchard plot, suggesting the existence of a single class of binding sites. Further studies revealed the absence of a positive cooperative effect. Because the system appeared to satisfy the conditions for Michaelis-Menten type kinetics, the determination of a Kd of approximately 3.6 X 10(-10) M was justified. These studies also demonstrated the efficacy of the specific activity of our iodinated gastrin. Trypsin treatment abolished specific binding of gastrin to the membrane preparation, indicating the protein nature of the receptor. The importance of maintaining extremely low temperatures during the incubation and wash period is demonstrated by the decrease in both the rate and the amount of dissociation at 0 degrees C compared with 30 degrees C.


Dissociation between plasma, urine, and renal papillary cyclic AMP content following vasopressin and DDAVP. The effects of in vivo physiologic doses of vasopressin and 1-deamino-8-D-arginine vasopressin (DDAVP) on the cyclic AMP content of plasma, urine, and renal papillary tissue were determined in the ADH-deficient Brattleboro rat. During clearance studies, plasma cyclic AMP concentrations and both total and nephrogenous urinary cyclic AMP excretion in vasopressin- and DDAVP-treated rats were similar to the values in time-matched controls. In contrast, in situ renal papillary cyclic AMP content was higher (P less than 0.001) in both vasopressin- (35.7 +/- 3.6 pmol/mg protein) and DDAVP- (29.7 +/- 2.2 pmol/mg protein) treated rats compared to controls (15.1 +/- 1.3 pmol/mg protein). Endogenous stimulation of vasopressin by dehydration in normal rats increased both papillary cyclic AMP content (27.1 +/- 2.7 pmol/mg protein) and urine osmolality, whereas no change in papillary cyclic AMP was observed following dehydration in Brattleboro rats (13.6 +/- 0.8 pmol/mg protein) despite an increase in urine osmolality. The results demonstrate that changes in cyclic AMP following in vivo vasopressin are best demonstrated by measurement of in situ cyclic AMP content of the renal papilla, whereas total urinary cyclic AMP and nephrogenous cyclic AMP are not useful indices of tubular sensitivity to this hormone.


Effect of fasting on tubular phosphorus reabsorption. In the thyroparathyroidectomized (TPTX) rat, fasting increased urinary phosphorus excretion by decreasing the tubular reabsorption of P1 (TRP) and resulted in hypophosphatemia. The administration of either sucrose or NaHCO3 prevented the metabolic acidosis associated with fasting and decreased the phosphaturia, indicating that the phosphaturia in fasting is in part due to metabolic acidosis. In rats on partial reduction of P1 intake selectively, the phosphaturic response to parathyroid hormone (PTH) was completely suppressed. On the other hand, the fasting rat partially retained the phosphaturic response to PTH, although dietary P1 intake was totally absent. These findings suggest that the renal P1 wasting in fasting may take place by dual mechanisms: a) the PTH-independent decrease in TRP, and b) an inability to totally suppress the response to PTH. Cyclic AMP generation in response to PTH, determined both in vivo and in vitro, was not measurably altered in fasting. However, the phosphaturic response to cyclic AMP was decreased in fasting, suggesting that the mechanism of partial resistance to PTH is probably not at but after cyclic AMP generation.


Effects of repetitive bursts of vagal activity on atrioventricular junctional rate in dogs. A stable atrioventricular (AV) junctional rhythm was produced in open-chest dogs by injecting pentobarbital into the sinus node artery. When the cervical vagus nerves were stimulated repetitively, the junctional pacemaker cells tended to become synchronized with the vagal activity. During such synchronization, the junctional rate varied directly rather than inversely with the frequency of vagal stimulation. The magnitude of the chronotropic response depended on the timing of the vagal stimuli within the cardiac cycle. In 9 dogs, when the mean heart periods were plotted as a function of the R-st intervals (i.e., the time from the beginning of ventricular depolarization to the beginning of the stimulus burst), the mean heart periods varied from a maximum of 1,815 ms to a minimum of 1,160 ms, depending on the R-st interval. A small change in the R-st interval was capable of evoking a relatively large change in cycle length. Therefore, the impulses from various efferent vagal fibers to the AV junction must arrive almost synchronously, the released acetylcholine must be removed rapidly, and the sensitivity of the pacemaker cells to acetylcholine must change rapidly at some critical time during the cardiac cycle.


Presynaptic inhibition of canine renal adrenergic nerves by acetylcholine in vivo. Experiments were performed in anesthetized dogs to determine whether previously reported in vitro inhibition of sympathetic neurotransmitter release by acetylcholine could be demonstrated in the renal vasculature of the intact animal. Vasoconstrictor responses to renal sympathetic nerve stimulation at varying frequencies were compared to intra-arterial injections of norepinephrine before and during intra-arterial infusions of acetylcholine, 2.5--80 micrograms/min. The vasoconstrictor responses to nerve stimulation were inhibited to a greater extent than were responses to norepinephrine during infusions of acetylcholine. The inhibitory effects of acetylcholine on nerve stimulation were dose and frequency dependent. The inhibition was blocked by atropine but not altered by physostigmine. Changes in renal blood flow per se did not contribute to the inhibitory effect of acetylcholine, since another vasodilator agent, sodium acetate, did not affect the nerve stimulation-norepinephrine vasocontriction relationship. Thus, acetylcholine produced inhibition of in vivo renal sympathetic vasoconstrictor responses, and the receptor involved appears to be muscarinic.


Ascending spinal pathways for the somatosympathetic A and C reflexes. Sympathetic unit nerve activity was recorded from the cervical sympathetic trunk of anesthetized, vagotomized, and carotid sinus-denervated cats. Single pulses or short trains of pulses with suprathreshold intensity for afferent C fibers were applied to the common peroneal nerve to elicit somatosympathetic A or simultaneous A and C reflexes, respectively. Bilateral lesions of an ascending spinal pathway in the dorsolateral sulcus area (DLS) of the T12 spinal cord eliminated the sympathetic C reflex. The remaining somatosympathetic A reflex was abolished by the subsequent lesion of an ascending spinal pathway in the dorsolateral funiculus (DLF) bilaterally. The duration of the silent period was found to be mainly influenced by an ascending spinal pathway in the DLF. It was concluded that excitation of afferent A fibers activates ascending pathways mainly in the DLF and a smaller contribution in the DLS of the spinal cord. This produces a small reflex excitation (A reflex) followed by a long silent period in the sympathetic nerves resulting in a net decrease of total sympathetic nerve activity. On the other hand, excitation of afferent C fibers activates the ascending spinal pathway in the DLS. This produces a reflex excitation (C reflex) which exceeds the decreased activity due to the silent period resulting in a net increase of total sympathetic nerve activity.


Liver glycogenolysis during exercise without a significant increase in cAMP. Liver glycogenolysis may be controlled by glucagon or catecholamine-induced changes in cAMP or by cAMP-independent mechanisms. The purpose of these experiments was to determine whether an increase in liver cAMP occurs during exercise at a time when the rate of liver glycogenolysis is greatly accelerated. Rats were taught to run on a treadmill 10 min/day for 6 wk. They were then run continuously for periods of time ranging from 0 to 120 min at 0.8 mph up a 15% grade. Liver glycogen was depleted by the end of 90 min in fed animals and by 20 min in overnight-fasted animals. Liver cAMP was not significantly increased in fed animals during the first 60 min of exercise. The major increase in liver cAMP occurred after liver glycogen was depleted, at which time the rat must rely entirely on gluconeogenesis for maintenance of blood glucose. This increase in cAMP corresponded to large increases in plasma glucagon and catecholamines. We conclude that liver glycogenolysis in the rat can occur during exercise independently from significant detectable increases in cAMP concentrations.


Computer simulation of metabolism in pyruvate-perfused rat heart. II. Krebs cycle. A realistic metabolic model of the tricarboxylic acid cycle in the perfused rat heart was constructed to help explain the sequence of biochemical events regulating the metabolism of exogenous pyruvate following a large increase in work load. The unchelated Mg2+ level was the most important controlling factor. The resulting mixture of chelated and unchelated nucleotides and tribasic acids effected coordinated control of citrate synthase, aconitase, isocitrate dehydrogenase, succinyl CoA synthetase, fumarase, and nucleoside diphosphokinase, because Mg2+-chelates are generally substrates whereas unchelated species are inhibitors. Succinate dehydrogenase is largely controlled by the ubiquinone redox potential. The fluxes through alpha-ketoglutarate and malate dehydrogenases are largely dependent on thepyridine nucleotide redox potential, but the succinyl CoA-to-CoASH ratio strongly affects the former enzyme as well. The model predicts an accumulation of succinate during the transition to higher work output.


Computer simulation of metabolism in pyruvate-perfused rat heart. V. Physiological implications. The results of a simulation of metabolism in the pyruvate-perfused rat heart subjected to a sudden increase in work load are interpreted to provide a coherent explanation for the observed physiology. Respiration is most closely correlated with the mitochondrial phosphate potential, calculated from the MgATP and MgADP levels. No correlation between respiration and the pH gradient across the mitochondrial membrane was found. The transient falls in pH in the cytosol and perhaps the mitochondria are due largely to carbonic and lactic acidosis and appear to be only weakly coupled. The heart maintains a high ATP level during the transition to increased work by utilizing its energy reserves in order of decreasing availability in response to physiological signals mediated by Mg2+, Ca2+, and cAMP.


Results of hepatic lobectomy for primary epithelial cancer in 31 adults. Hepatic lobectomy for primary epithelial cancer was performed in 31 adults from 1964 through 1977 in the surgical departments of six Scandinavian hospitals. Twenty-three patients were discharged and had a 2 year survival rate of 62 per cent and a 5 year survival rate of 16 per cent. Alternatives to surgery have not yet emerged. Further progress requires centralization.


[The fine structure of the exocrine pancreas of the mink (author's transl)]. The pancreas of female mink has been investigated by transmission electron microscopic means. The following results can be summarized: The pancreas of the mink is built up by the well known gland lobules as found in many other species; each lobule contains branched ducts with acini. The acinar cells are characteristically packed with granular endoplasmic reticulum, large Golgi apparatuses, and zymogen granules. Particularly interesting are large vacuoles, which seem to emerge directly from the endoplasmic reticular cysternae. The centroacinar cells form relatively extended protrusions or pseudopodia, which frequently penetrate into the intercellular spaces between neighbouring acinar cells. The peripheral isthmic parts of the ducts are covered by an isoprismatic epithelium. The adventitial tissue of the intralobular ducts contain mucous glands. Within the loose connective tissue between the exocrine cells, blood vessels as well as numerous nerves can be found. These results are compared to earlier reports on the same subject, and are also discussed together with available data from the literature.


Morphology of osteoclasts in resorbing fetal rat bone explants: effects of PTH and AIF in vitro. Osteoclastic bone resorption was studied using 45Ca-labeled fetal rat bones cultured in the presence of parathyroid hormone (PTH) and an anti-invasion factor (AIF) derived from bovine hyaline cartilage which is enriched in a collagenase inhibitor. The specific morphological expressions of osteoclasts cultured in PTH and AIF were observed in both light and electron microscopy and analyzed cytometrically. Stimulation of bone resorption with PTH revealed significant increases in the numbers and activity of osteoclasts, whereas bones cultured in the presence of AIF showed significant decreases in numbers of osteoclasts and altered cell features including the loss of osteoclast contact with bone surfaces. These structural modifications were evaluated with 45Ca release data derived from matched-pair explants of fetal rat bones, revealing the existence of a relationship between resorptive states of the cultured bones and morphological expressions of osteoclastic activity.


Lipoprotein particles in the jejunal mucosa of postnatal developing rats. The jejunal mucosa of neonatal rats contains lipid particles of the same size, electron density and intracellular and extracellular distribution as particles identified by others in adult jejunum as lipoprotein particles. As in fetal jejunum obtained during the last three days of gestation, the jejunal mucosa of unsuckled newborn rats contains exclusively lipoprotein particles the size of very low density lipoproteins (VLDL). Within one day after initiation of suckling, there is in the mucosa a spectrum of lipoprotein particles ranging widely in size from those of VLDL particles to those of chylomicrons. These particles are seen in the endoplasmic reticulum and Golgi material of absorptive cells and within interepithelial cell spaces, the extracellular spaces of the lamina propria and lymphatic lacteals. VLDL-sized and chylomicron-sized particles are also seen, although in decreasing number, in the jejunal mucosa of 18-day-old suckling rats. However, in rats of comparable age, fasted for 48 or 72 hours, only VLDL-sized particles are seen in the jejunal mucosa. Ligation and transection of bile duct followed by fasting in rats of this age results in a marked decrease in the number of lipoprotein particles in absorptive cells. The results indicate that endogenous lipid contributes to the formation of VLDL particles whereas dietary triglycerides are needed for formation of chylomicrons.


Bovine trypanosomiasis: effect on the immune response of the infected host. Yearling cattle were inoculated with a recently isolated field strain of Trypanosoma congolense. Dinitrophenylated ovalbumin, a bacteriophage, or bovine parainfluenza-3 virus injected into the cattle during the first 5 weeks of infection resulted in peak serum antibody titers lower but not much lower than those produced by noninfected cattle. Primary and secondary antibody responses of inoculated cattle required more time to reach peak titers. Peripheral blood lymphocyte concentrations decreased to 42% of preinfection base-line value 1 1/2 weeks after the onset of T congolense infection and thereafter plateaued at less than 75% of base-line value. Lymphocyte cultures prepared from infected and noninfected cattle gave no marked differences in [(3)H]thymidine uptake after stimulation with phytohemagglutinin or pokeweed mitogen. Differences in lymphpocyte responsiveness were not noticed in one-way mixed lymphocyte culture reactions, using mitomycin C-treated lymphocytes from noninfected cattle as stimulator cells.


Concurrent porcine rotaviral and transmissible gastroenteritis viral infections in a three-day-old conventional pig. A 3-day-old suckling pig with diarrhea was necropsied, and immunofluorescent microscopic examination of the small intestinal mucosa, together with immune electron microscopic examination of the large intestinal contents, provided a presumptive diagnosis of a concurrent infection with transmissible gastroenteritis (TGE) virus and porcine rotavirus. Immunofluorescent microscopic, immune electron microscopic, and serologic data obtained from gnotobiotic pigs experimentally inoculated with the large intestinal contents of the suckling pig confirmed this diagnosis. Two gnotobiotic pigs, convalescent from previous TGE viral infections, became infected with porcine rotavirus only. However, another gnotobiotic pig, convalescent from a previous porcine rotaviral infection, became infected with TGE virus only, following inoculation with the large intestinal contents of the suckling pig.


Adrenal gland function in the horse: effects of cosyntropin (synthetic) and corticotropin (natural) stimulation. The plasma concentration of hydrocortisone was determined in mares given either cosyntropin (100 IU, given IV) or corticotropin (200 IU, given IM). Plasma hydrocortisone concentrations of the mares treated with cosyntropin increased by 46%, 57% and 80% at 30, 60, and 120 minutes, respectively, when compared with base-line values; these values returned to base line at 240 minutes. In mares treated with corticotropin, mean plasma hydrocortisone concentrations increased by 42%, 143%, 101% and 155% at 30, 60, 120, and 240 minutes, respectively, when compared with base-line values. Differences in total leukocyte count, total eosinophil count, and plasma concentrations of electrolytes (calcium, sodium, magnesium, potassium) of cosyntropin- and corticotropin-treated mares, and these values in control animals were not significant. Results of the present study indicated that the horse responds to small dosages of cosyntropin (IV) in a prompt and reproducible manner as determined by plasma hydrocortisone values. Response to corticotropin was slow and less consistent. Thus, administration of cosyntropin to the horse, according to test results with paired samples collected (before administration and again at 2 hours after injection), was found to be a prompt and meaningful test of adrenal gland function.


Comparative thermometric coagulation studies of plasmas from normal outbred Swiss Webster mice and persons. The functional capabilities of a thermometric clot-timer have been demonstrated in a comparative study of human and mouse plasma coagulation. The influence of some variables on coagulation times of mouse and human plasmas were examined in activated partial thromboplastin time, one-stage prothrombin time, and Russell's viper venom time assays. Mouse plasma coagulation times were generally shorter and more reproducible than those of human plasma. Optimal assay conditions are also described.


Occurrence of anaerobic bacteria in diseases of the dog and cat. A survey for anaerobic bacteria was conducted in 314 clinical specimens from dogs and cats. A total of 187 anaerobic isolates in pure and mixed culture were isolated from 111 of the specimens that contained anaerobic bacteria. Common isolated included Actinomyces (9.1%), Clostridium perfringens (19.3%), other Clostridium spp (11.2%), Peptostreptococcus anaerobius (7.5%), Bacteroides melaninogenicus (13.4%), other Bacteroides spp (17.6%), and Fusobacterium necrophorum (5.3%). Anaerobic bacteria were involved in serious lesions that often were life threatening to the animals. Antibiotic susceptibility data indicated that the lincomycin family, the penicillin family, chloramphenicol, and cephaloridine are preferred drugs for treatment of anaerobic infections. Data from the survey were used in formulation of a table to aid practitioners in clinical diagnosis of disease caused by anaerobes. Clostridium perfringens was isolated in large numbers from five of six dogs with a clinical diagnosis of canine hemorrhagic gastroenteritis and from one cat with hemorrhagic diarrhea. Experimental infections were induced in rats, using caine feces as inoculum. Induced lesions contained aerobic and anaerobic bacteria similar to those bacteria isolated in the clinical survey, indicating that feces may serve as a major source of these bacteria in clinical infections of the dog.


Etiology of liver disease in renal-transplant patients. The etiology of 72 episodes of liver disease that developed in 62 of 162 renal-transplant recipients was evaluated. Infection with hepatitis B virus was a minor problem, and none of our patients had evidence of infection with hepatitis A. Cytomegalovirus infection was ubiquitous in the population and probably accounted for many episodes of acute liver disease. This agent's role in causing chronic hepatitis is less secure. Infections with other viruses including Epstein-Barr virus, adenovirus, and the herpes viruses were only rarely associated with hepatic disease. Azathioprine was responsible for some episodes of acute cholestasis but could not be incriminated as a direct cause of chronic disease. A cause could be identified for the majority of episodes of acute hepatic dysfunction, but the cause of most of the chronic hepatitis remains undetermined. It is likely that infection with non-A, non-B hepatitis virus accounts for much of this serious, often fatal, complication of renal transplantation.


[Expression of ts-3 mutant of polyoma virus in mouse cells (author's transl)]. A thermosensitive mutant (ts-3) of polyoma virus produces T antigen, viral DNA and V antigen in BalbC/3T3 cells only at the permissive temperature (31 degrees C). At the non-permissive temperature (39 degrees C) these cells are unable to complement the viral defect. The pulse technique reveals only transitory cellular DNA induction between 15 and 25 h after infection at 39 degrees C. However, at both temperatures, cytoplasmic penetration is observed, and the viral DNA reaches the nuclei. Similarly, the myoblast cell line PCD2 does not complement the viral defect. In other idfferent types of cells tested (3T6, mouse kidney or secondary embryo cultures), the ts-3 mutant is expressed equally at 31 degrees C and 39 degrees C.


Phosphodiesters in muscular dystrophies. Phosphorus nuclear magnetic resonance studies of various normal and dystrophic muscles have demonstrated differences in phosphodiester contents. Dystrophic chicken pectoralis muscle contains elevated levels of the diester SEP, and Duchenne dystrophic human leg muscle lacks the diester GPC. It is shown that SEP may be characteristic of slow fiber types, indicating imcomplete maturation of the dystrophic pectoralis muscle. The activities of SEP-metabolizing enzymes have been determined in microsomes of pectoralis muscle, kidney, and intestinal mucosa of normal and dystrophic chickens. There was little difference between normal and dystrophic values of SEP phosphodiesterase and SEP synthase in kidney and mucosa. Dystrophic pectoralis muscle microsomes possessed significantly elevated levels of both enzymes. The increase in SEP synthase activity of dystrophic muscle may explain the increase in the SEP level if the phosphodiesterase is regulated in vivo. It is shown that human muscle disease processes may raise, leave unchanged, or lower GPC levels. In certain diseases when GPC is markedly elevated, such as Werdnig-Hoffmann, or depressed, such as Duchenne muscular dystrophy, it may serve as a marker.


Experimental hyperlipidemia in rats.

Experimental hyperlipidemia in rats. Implantation of MtT-F4 tumor, a mammotropic tumor that secretes large quantities of ACTH, GH and prolactin, into male Fisher rats induced the development of hyperlipidemia. Free fatty acid, triglyceride and cholesterol levels in the plasma were significantly increased at 31 days after tumor implantation. Blood glucose and glycerol levels remained normal, while uric acid concentration in the blood was significantly decreased. The concentrations of the serum lipoproteins were significantly increased, while, only small changes in the distribution of the serum lipids and the composition of the lipoproteins were observed. Following stimulation of isolated adipose tissue cells with ACTH, the lipolytic response and the accumulation of cyclic AMP was higher in cells derived from the rats with the tumor, although the accumulation of cyclic GMP was not different from control adipocytes. Further, when the isolated adipose tissue cells were stimulated with dibutyryl cyclic AMP no difference was observed between the control and tumor bearing groups. Clofibrate administered in the diet resulted in a complete elimination of the tumor effect on serum triglycerides and to a great extent prevented the rise in serum cholesterol. The tumor-induced increase in the concentration of the high density lipoproteins was not affected, but the elevation of the d less than 1.063 lipoproteins was not affected, but the elevation of the d less than 1.063 lipoproteins was partially reversed. The increased lipolytic response and accumulation of cyclic AMP following stimulation by ACTH was not altered in adipocytes derived from tumor bearing rats. However, clofibrate treatment resulted in a significantly greater accumulation of cyclic GMP in fat cells stimulated with ACTH from both control and tumor bearing rats. Clofibrate in the diet did not alter the levels of GH or prolactin or serum lipids in the control rats nor were the elevated hormone levels of the tumor bearing rats changed.


[Pathology of the polymorphonuclears in dermatology. Methods of investigation. Clinical pictures (author's transl)]. A remarkable increase in the number of recognized clinical abnormalities of neutrophil function has occurred within the 8 years past. Of major importance in the delineation of these disorders is the establishment of appropriate methodology of their characterization. This review discusses phagocytosis and its disorders by dividing it into stages that encompass the way in which phagocytes ferret out injection, how they recognize pathogens and the intracellular events leading to engulfment and killing of micro-organisms. The pathology of chemotaxis and random mobility is described: the lazy leukocyte syndrome, the Chediak-Higashi syndrome, the familial chemotaxis pecular dermatosis, and a variety of serum abnormalities of the serum complement system associated with deficient generation of chemotactic activity. The second part of these disorders is dailing with the inherited abnormalities of the oxygen dependent killing mechanisms of phagocytes, the chronic granulomatous disease, the glucose-6-phosphate deshydrogenase deficiency, the myeloperoxydase deficiency and the glutathione peroxydase deficiency.


Studies on specific antigen content in different fractions of preparations from BLV-producing cell cultures. Different protein fractions of BLV-antigens were examined with serological and chemical methods. The fractions of ether resistant and dual antigens were obtained by means of QAE-50A Sephadex chromatography and the fractions of gp-antigen were collected in the course of the affinity chromatography on Concanavalin A-Sepharose. All protein fractions examined by means of polyacrylamide gels electrophoresis occurred to be heterogenous. Some of them showed positive RID or GID reactions with sera of cattle or sheep affected with leukosis. By means of thinlayer chromatography on Sephadex, there were estimated following molecular weights of proteins in the serologically positive fractions: 12.500, 67.000, 70.000 and 150.000.


Subunit and fine structure of the glycoprotein of bovine leukemia virus. The structure of the glycoprotein of bovine leukemia virus (BLV) was investigated. Analysis of BLV under nonreducing conditions revealed two viral glycoproteins with apparent molecular weights of 162,000 (VGP I) and 85,000 (VGP II). After reduction and alkylation, two glycosylated polypeptide chains of apparent molecular weights of 60,000 (gp 60) and 30,000 (gp 30) could be demonstrated in VGP I and VGP II; VGP I appears to contain two molecules of gp 60 and two molecules of gp 30, and VGP II consists of only one molecule of gp 60 and one molecule of gp 30 linked by disulfide bonds. Comparison of the tryptic peptides of gp 60 and gp 30 provided evidence for the existence of partial sequence relationship between gp 60 and gp 30. The structural similarity of the BLV glycoprotein molecule to immunoglobulins is discussed in regard to the possible function of this protein in the pathogenesis of bovine leukemia.


Detection of a precursor of bovine leukemia virus structural proteins in purified virions. Gel filtration chromatography of disrupted bovine leukemia virus (BLV) resulted in the isolation of the 25,000 dalton major virion internal protein and two previously uncharacterized virion proteins of molecular weight 65,000 and 12,000 respectively. The 65,000 dalton protein does not bind to concanavalin A and its antigenicity is ether-resistant. Therefore, this polypeptide is different from the previously described glycoprotein associated with BLV. Radiommunoprecipitation and competitive radioimmunoassays indicated that the 65,000 molecular weight polypeptide shares antigenic determinants with the 25,000, 15,000, and 12,000 dalton BLV proteins, respectively. Thus, the 65,000 dalton polypeptide may represent the precursor of these three smaller virion proteins.


Bilateral hypoglossal nerve paralysis after irradiation therapy. Cancericidal doses of irradiation therapy to be the head and neck may be followed by paralysis of cranial nerves due to fibrosis of tissue about the nerves and infiltration of the nerves with fibrous tissue. The hypoglossal nerve is particularly prone to this delayed effect of irradiation therapy and a case is presented here in which there was bilateral hypoglossal paresis appearing three years after therapy.


Heterogeneous properties of BLV-precipitating immunoglobulins in bovine leukemic sera. Purified precipitating serum antibodies against bovine leukemia virus present in cattle suffering from various stages of leukosis were identified as immunoglobulins of the IgG 1 subclass and IgA class. Each of these precipitating immunoglobulins exhibited several electrical charges as well as molecular sizes the origin of which will be discussed. Differences in precipitating activities against antigen gp69 and p24 could be observed comparing serum from a suspect animal and a highly leukemic one. In disrupted BLV preparations four antigens were detected immunoelectrophoretically, and two of them could be analyzed. Antigen gp69 less basic than p24 showed a considerably faster mobility.


Bovine leukemia virus (BLV) specific RNA in infected cells. BLV specific RNA sequences were detected by hybridization to the single stranded DNA copy of the viral RNA in fetal lamb kidney cells infected with bovine leukemia virus (FLK-BLV). The DNA synthesis was carried out in vitro by an endogenous reaction. It was found that uninfected lamb cells do not contain RNA sequences related to the BLV genome, whereas BLV-DNA completely annealed to FLK-BLV RNAs. RNA from FLK-BLV cells contains three species of virus specific RNA which sediment respectively at 35 S, 20--24 S and 12--4 S. In view of their EDTA-sensitive association with ribosomes, viral specific RNA sequences are viral messenger RNAs and their polarity is identical to that of the 70 S viral RNA.


Further investigations on the porcine lymphoma C-type particle (PLCP) and the possible biological significance of the virus in pigs. The possible biological significance of the procine lymphoma C-type particle (PLCP) was investigated serologically. The screening of 1 200 field sera of apparently normal breeder pigs, performed with an immunodiffusion test, revealed negative results. The same results were obtained with sera from a herd with hereditary lymphosarcoma and with sera from experimentally infected piglets. In cases of doubtful precipitation lines additional tests with a more sensitive indirect immunofluorescence assay were performed. In no case a specific cytoplasmic fluorescence could be observed. The same sera of the 1 200 pigs were tested with BLV antigen as well as 350 bovine sera from BLV-positive cattle were tested with PLCP antigen. A possible antigenic relationship between BLV and PLCP could not be detected by the assays mentioned above. Purified PLCP was compared in SDS-PAGE with MuLV, FeLV, and BLV. The electrophoretical pattern presents suggestive evidence for a typical oncornavirus polypeptide composition. Apart from the presumable major internal protein with a molecular weight of 28 000 d, three low molecular weight proteins appeared at 12 000 d, 11 000 d, and 10 000 d. High molecular weight proteins could not be detected yet. The antigenic specificity of these bands has still to be confirmed.


Studies on bovine leukosis. VI. Enzyme linked immunosorbent assay for the detection of antibodies to bovine leukosis virus. The results of examination of 2 784 sera in the AGIDT-gp were compared with those of the ELISA. A discrepancy was seen in 0.5% of the cases. It was concluded that the ELISA was another useful serological method added to the list of serological techniques presently available for the detection of antibodies to BLV. The advantages of the technique are discussed.


Detection of complement-dependent lytic antibodies in sera from bovine leukemia virus-infected animals. Cattle and sheep infected by Bovine Leukemia Virus (BLV) harbor complement-dependent cytotoxic IgG1 antibodies. These antibodies lyse BLV producing cells but are inactive against noncultured tumorous lymphocytes of animals with persistent lymphocytosis. The target structure in the cell membrane contains antigenic determinants of BLV gp 60. The level of cytotoxic activity increases with progression of the disease towards the tumor phase. Sheep vaccinated with inactivated BLV have no cytotoxic activity in the serum but seem to resist infectious BLV challenge. Such cytotoxic activities do not protect the host against tumor growth but strongly control viremia and thus limit BLV spread within bovine and ovine populations.


Indirect hemagglutination test for detection of antibody against bovine leukemia virus. An indirect hemagglutination test was developed for detecting antibody against Bovine Leukemia Virus. Medium from Bovine Leukemia Virus infected monolayer fetal lamb kidney cell line cultures was concentrated by a combination of ammonium sulfate precipitation and freezedrying. Tanned sheep red blood cells were exposed to p24 and GP antigens, separately and together. The GP antigen did not attach to the sheep red blood cells, while the p24 antigen did. On testing 150 samples from suspect animals with the crude antigen-coated sheep red blood cells, the indirect hemagglutination test detected almost the same number (74) of positive animals as the agar gel immunodiffusion test with GP antigen (79), and by each of these test a far greater number of cases of leukosis were detected than by hematologic examination (58) or agar gel immunodiffusion test with p24 antigen (44). These results suggest that the indirect hemagglutination test may detect almost the same number of positive as the agar gel immunodiffusion test with GP antigen. Moreover, its results can be read two days earlier, and an impression of the antibody level in the sera is obtained. In routine or large-scale work the indirect hemagglutination test is too troublesome and time-consuming compared to the agar gel immunodiffusion test. The indirect hemagglutination test may be improved if it becomes possible to sensitize sheep red blood cells with GP antigen.


Characterization of antibodies responsible for the inhibition of BLV induced early polycaryoxytosis. A quantitative in vitro assay (EPI) was developed for the serological detection of bovine leukemia virus (BLV) infection in cattle. Positive sera react in inhibiting early polycaryocytosis (EP) induced by BLV. We report here experiments which demonstrate that BLV induced antibodies are responsible for this inhibition. Serum fractionation by gel filtration and affinity chromatography indicates that EP inhibiting activity is mediated by immunoglobulins of the IgG class but not by those of the IgM or other classes. Furthermore, in view of their ion-exchange chromatographic behaviour, these antibodies were attributed to the IgG1 subclass. We also ruled out the participation in the EP inhibition rection of the other compounds like polypeptides, including serum complement and interferon or DNA and RNA. These results demonstrate that EP inhibition by bovine sera is specific of BLV induced antibodies.


Early polykaryocytosis inhibition test: evaluation of its performance in a seroepidemiological survey of bovine leukemia virus induced antibodies in cattle. We report the results of a seroepidemiological survey, using the recently described early polykaryocytosis inhibition (EPI) assay to detect bovine leukemia virus (BLV) infection in cattle. At the herd level, there is a good correlation between epidemiological, hematological, anatomopathological data and of both the presence and titer of EP inhibiting antibodies. The EPI procedure revealed to be more sensitive than two other routine serological techniques, complement fixation and immunodiffusion. In addition, at the individual level the average EPI titers of the sera increase progressively from normal to hematologically-suspect, -- positive and finally to lymphosarcomatous animals; when the age is considered, there is a very significant increase of the amount of EP inhibiting antibodies among lymphocytotic animals, while there is only a very moderate increase of the EPI titer among hematologically normal contact animals. This suggests a loss of control of BLV expression in most lymphocytotic animals and may explain the more frequent occurrence of lymphosarcomas in this category of animals. The horizontal transmission of BLV throughout the entire life span is supported by the observation of an increase with age of the percentage of animals with EP inhibiting antibodies.


An infectivity assay for the bovine leukemia virus based on the induction of the major internal virion antigen in susceptible cell cultures. This report described the development of a 7-day infectivity assay for the bovine leukemia virus (BLV) which is based on the induction of the major internal virion antigen (p25) in susceptible indicator monolayer cell cultures. In this assay the antigen is detected in the indicator cells by the immunoperoxidase antibody technique using a monospecific anti-BLV serum. The immunoperoxidase infectivity assay (IPIA) is specific, quantitative, reproducible and more sensitive than the previously developed syncytia induction assay. The IPIA can be applied for the detection of BLV-infected cells and provides a reliable method for the direct diagnosis of BLV infection in cattle.


Bovine leucosis in Ireland.

Bovine leucosis in Ireland. An abbatoir tumour survey showed that lymphosarcoma cases arose at a rate of approximately 2/100,000 cattle slaughtered. No multiple cases occurred in herds. A limited haematological survey showed no evidence of E.B.L. E.B.L. was made a notifiable disease in October 1977. Since then all tumour cases were back-traced and the herd of origin tested by A.G.I.D. test. All herds proved negative. A survey of herds containing imported cattle was started. To date seven herds containing Canadian Holsteins have proved positive serologically. Little spread of infection within the herds has taken place.


Epidemiology of bovine leukemia in Venezuela. Approximately 20 thousand bovines of autochtonious and imported breeds, located in different regions of the Country were serologically tested by Immuno-diffusion (I.D.) in agar, using BLV GP60 antigens, for the purpose of determining the prevalence of infection by Bovine Leukemia Virus. Hematologic and histopathologic assays were made with the object of determining variations in hematologic values and the determination of stages of persistent lymphocytosis, and of studying the morphocytological changes that accompany malignant transformations of lymphoid elements, in order to make studies on the incidence of clear tumoral lesions in herds highly affected by Leukemia. From 16 thousand samples of serum, tested by the I.D., 6 thousand were positives (37,5%) pertaining to 150 herds distributed in all 20 states of the Country. The sera correspond to different breeds: dairy cattle, beef cattle, brahman, zebú hybrids, and buffalo. Hematologically, persistent lymphocytosis induced by the BLV was verified, by previously discarding other pathologic stages with transitory lymphocytosis. Factors which explain this high incidence and the spread of the Bovine Leukemia in Venezuela were detected.


Enzootic bovine leukosis: an epidemiological survey among cattle in the west of France by precipitating antibody detection. In order to inform the sanitary veterinaty authorities, an epidemiological survey was carried out in the West of France to appreciate the level of infection by BLV in cattle. More than 10,000 sera from 763 herds were tested by agar gel immuno-diffusion. The animals were more than one-year-old dairy cattle. 23 sera (0.22%) from 11 herds (1.44%) were found positive. In the 11 herds the levels of serologically positive animals ranged from 1.6 to 40% with a 9% average. These levels are comparatively low and are almost in correspondence with the number of leukotic tumours recorded in the slaughter houses and knackeries.


Epidemiologic study on enzootic bovine leukemia in Italy. The results obtained from serologic and hematologic investigations conducted on 1.124 cattle belonging to 14 herds located in Central Italy are reported. The results demonstrate a considerable number of animals with antibodies to Bovine Leukemia Virus (BLV) and/or lymphocytosis, coming from Friesian herds with or without history of leucosis. Furthermore it is demonstrated the absence of animals with circulating antibodies to BLV in herds made up of the Chianina breed even with a herd history of leucosis.


Results of an epidemiological survey of enzootic bovine leukosis in the northern part of Lower Saxony and a preliminary communication of an examination into relationship between BLV-antibody development and calving. The Agar-Gel-Immunodiffusion-Test as proposed by EEC has been carried out during the last 2 1/2 years within a defined test area of Lower Saxony. About 500 leukosis-infected herds with a total of about 20.000 head of cattle have been tested five times now within three to six months sequences. The AGIDT has proved to be an easy and very practical test for routine diagnosis in large scale surveys. Our results described again show the superiority of the glykoprotein-antigen over the ether-treated p24-antigen in the AGIDT. Slaughter of AGIDT-reactors within this population led to a fast decrease of leukosis-infection as detected by the test. Epidemiological herd-data show, that spreading of infection within a herd depends largely on direct contact between infected and noninfected animals. Moreover our data give rise to the suspicion that the manipulations during blood-sampling seem to be implicated in the spread of the disease within a herd. Possible reasons for inconsistencies of antibody-titer in infected animals are discussed.


Sequential studies of bovine leukemia virus antibody development in dairy cattle over a four-year period. Serological examinations using the agar gel immunodiffusion and immunofluorescence tests were undertaken to study: a) the development of bovine leukemia virus infections in a given herd under field conditions monitored by antibody development, b) the reproducibility of the results of serological examination during 22 sequential tests, c) the antibody titre variations amongst individual animals. A total of 286 Friesian cattle representing 2,352 serum samples was tested at regular intervals over a period from 1974/75 to 1978. The percentage of animals with bovine leukemia virus antibodies continuously increased during the observation period from an initial 4.8% (1974/75), 9.7% (1976), 34.2% (1977) to 52.6% (1978). On the basis of 22 sequential tests representing 1,672 serum samples obtained from 76 animals no false positive reactions were seen in the immunodiffusion test. Thirty-six (14.2%) out of 254 animals, however, showed variations of the antibody titre leading to a shift from positive to negative or negative to positive results, respectively. AnothFr three cattle (1.2%) shifted from a positive to negative immunodiffusion result and maintained their negative reactivity for at least two following tests. But they still had specific antibodies if tested by the immunofluorescence test. A detailed analysis of the group of variable reactors supports the view that a more sensitive and quantitative test system may be helpful to complement the presently used immunodiffusion test for the eradication of enzootic bovine leukosis.


BLV-antibodies in serial sampling over five years in a bovine leukosis herd. The sequential monthly bleedings of bovine leukosis herd animals showed that a majority of the animals was positive for BLV-antibodies at the age of 1 1/2 years or over and remained reactors throughout the observation period. Two phases of BLV-antibodies were observed in the calves born in the herd. In the first phase the calves remained reactors for two to seven months, indicating residual maternal antibodies acquired through colostrum. In the second phase, the animals became permanent reactors reflecting an active concomitant infection with BLV. The degree of spread of BLV infection among the leukosis herd animals appeared to depend upon the ratio between reactors and negative animals.


An evaluation of the role of milk in the natural transmission of BLV. In order to evaluate the role of milk in the transmission of BLV, the presence of the virus and viral antibodies was investigated in cattle that were born to, and nursed on, infected dams and then raised in isolation or in contact with infected animals. Only 3 of the 17 isolated cattle became infected during 25-29 months of observation. During the same period of time, all 17 cattle raised in contact with BLV-positive animals developed BLV infection. From these results, it is apparent that milk-borne transmission of BLV, if it occurs, is much less frequent than contact transmission.


Horizontal transmission studies on enzootic bovine leukosis. Four groups of sero-negative cattle were brought into contact for various periods with either sero-positive and antigen-negative or sero- as well as antigen-positive animals. A seroconversion as evidence of transmission was only detected in animals that had been in contact with sero- and antigen-positive cattle. Shortest period of contact for those was 2 months after which a seroconversion was found. A distinct elevation of the number of lymphocytes was only detected in two animals which had been transported shortly after evidence of the seroconversion had occured.


Tumor-associated antigen and cell surface marker in cells of bovine lymphosarcoma. The detection of tumor-associated antigen and B cell surface marker was attempted in lymphoid cells of peripheral blood (PBL) and lymph nodes obtained from cattle with lymphosarcoma, cattle infected with bovine leukosis virus (BLV), and clinically normal cattle. As a result, specific tumor-associated antigen was found in PBL and lymphoid tumor cells from cattle with the adult form of lymphosarcoma (ALS) using anti-ALS serum. The antigen was also detected in PBL of BLV-infected cattle. A weak reaction for the antigen was found in 2 of 5 cases of calf form. No significant reaction for the tumor-associated antigen of neoplastic lymphoid cells from ALS was found in the thymic and skin forms. The percentage of B cells in PBL and lymph nodes from ALS was higher than that in normal cattle. However, a loss of B cell surface marker was suggested in some ALS cases. A good relation was obtained between an increased B cell percentage and an appearance of the tumor-associated antigen in PBL of BLV-infected cattle. In lymphoid cells of PBL and lymph nodes from sporadic cases, the percentage of B cells was lower than that observed in normal cattle.


Distribution of bovine leukemia virus proviral sequences in tissues of bovine, ovine and human origin. Bovine leukemia virus (BLV) single-stranded cDNA was used to study the distribution of DNA sequences in tissues (normal or malignant) from bovine, ovine and human origin. After recycling against normal bovine DNA, BLV (3H) cDNA hybridized with bovine enzootic tumor DNA but did not hybridize with normal bovine DNA. These results indicate that BLV is an exogenous RNA oncogenic virus and confirm that enzootic bovine leukosis (EBL) is an infectious disease. Proviral BLV sequences were also detected in buffy coat cells of animals in persistent lymphocytosis (PL) and carrying antibodies to BLV but no tumors. In animals at the tumor stage of EBL, the proviral sequences were found in buffy coat cells, in solid tumors (lymphosarcomas) and in organs infiltrated with tumoral lymphoid cells but not in apparently normal organs. No hybridization above background was observed between BLV (3H) cDNA and DNAs extracted from buffy coat cells and tumors corresponding to sporadic forms of bovine leukosis and some human leukemias and sarcomas.


Development of bovine leukosis virus infection in cattle. A preliminary study. A serological study in three herds for 5 to 7 years indicated that BLV infection usually developed when cows where 2 to 4 years old, thereafter, the rate of new infection was somewhat less. The reactor status of the dam had no influence on whether or not and when progeny became reactors. In only one herd 3 calves from 57 reactor cows become persistent reactors, thus there was little evidence of vertical transmission. Such can occur where there is a break in the placental barrier.