Role of pituitary hormones in the growth of human breast cancer. Hypophysectomy was performed in 28 women with Stage IV breast cancer who were treated initially with antiestrogens. Six of 13 patients who responded to tamoxifen and 2 of 12 who failed to benefit from tamoxifen obtained remissions from hypophysectomy. The remissions average 11+ months. Three of 8 patients treated initially with antiestrogens have responded to androgen therapy. The results suggest that hormones other than estrogen, which appears to play a major role, may be involved in stimulating the growth of some human breast cancers. Prolactin receptors were detectable in 51% of human breast cancers and were detected in both estrogen receptor-positive and -negative tumors. Preliminary clinical correlations suggest that prolactin receptors will not be useful in predicting response to antiestrogen therapy.


Human breast carcinoma cells in continuous culture: a review. A comprehensive listing of putative human breast carcinoma cell lines and the extent to which each has been characterized is presented. Criteria used to certify the human, mammary, and malignant origin of a cell line include: (a) a reliable histopathological diagnosis; (b) interspecies specificity established by human karyotype, isoenzyme profiles, and/or cell surface antigenicity; (c) intraspecies specificity, demonstrated by genetic evidence of a unique, human donor distinct from other cells including HeLa cells; and (d) organ specficity, supported by morphological evidence of epithelial structure and secretory activity, and especially by the expression of differentiated functions; these include presence of receptors for sex steroid hormones, hormone responsiveness, and production of milk proteins, fatty acids, or milk-specific antigens. Of the 47 cell lines for which data are here reported, 22 have been shown to be derived from human non-HeLa donors and to have epithelial morphology as revealed by light or electron microscopy. Differentiated function has been recorded for 19 cell lines. Additional human breast cancer cell lines have been reported, but characterization of some of these has been insufficient to judge the legitimacy of their predigrees. For others mammary origin is questionable. Six purported breast cell lines are in reality HeLa cells, and one is of nonhuman origin.


Inhibition of spermiation in the Syrian hamster using dibutyryl cyclic-AMP. Young adult male Syrian hamsters were given intraperitoneal injections of 50 mg dibutyryl cyclic AMP twice daily for a period of three days. On the fourth day the animals were sacrificed and their testes were processed for light and electron microscopy. The results indicate that the mature spermatozoa were retained within the seminiferous epithelium after the stage in the seminiferous cycle in which spermiation normally occurs. The unreleased spermatozoa were ultrastructurally normal. Typical Sertoli-spermatid junctional specializations remained associated with the retained spermatozoa. These findings indicate that normally spermiation is initiated by the disappearance of the junctional specializations. In addition, the present results demonstrate that spermiation can be controlled.


Structure of the Adenohypophysis in parturient female and neonate harp seals, Pagophilus groenlandicus. The pars distalis of parturient harp seals (Pagophilus groenlandicus) is divisible into three distinct zones based on the major cellular components of the different regions. A clear zonation is lacking in neonate seals, the medical "basophilic wedge" and the anterior gonadotrophic were small and difficult to identify. Five cell types were identified based on cytological characteristics: somatotrophs, mammotrophs, thyrotrophs, gonadotrophs and stellate cells; corticotrophs could not be identified, with any certainty, in any of the preparations, nor could the gonadotrophs be classified into LH, FSH, and ICSH cells. The pars intermedia enlarged during the early post-partum period and was large in the parturient females.


The complete sequence of a unique RNA species synthesized by a DI particle of VSV. The 2S RNA synthesized in vitro by the RNA polymerase of a defective interfering (DI) particle of vesicular stomatitis virus was labeled at its 3' terminus with 32P-cytidine 3', 5' bisphosphate and RNA ligase. Analysis of the labeled RNA showed that it was a family of RNAs of different length but all sharing the same 5' terminal sequence. The largest labeled RNA was purified by gel electrophoresis, and the sequence of 41 of its 46 nucleotides was determined by rapid RNA sequencing methods. The assignment of the remaining 5 nucleotides was made on the basis of an analysis of one of the smaller RNAs and published data. A new approach in RNA sequencing based on the identification of 3' terminal nucleotides of rna fragments originally present in the DI product or generated during the ligation reaction confirmed most of the sequence. The complete sequence of this 46 nucleotide long plus-sense RNA is: ppACGAAGACCACAAAACCAGAUAAAAAA UAAAAACCACAAGAGGGUC-OH. This RNA anneals to the RNA of the DI particle from which it was synthesized, indicating that its synthesis is template-specified. At least the first 17 and possibly all of the nucleotides are also complementary to sequences at the 3' end of two other VSV DI particles which were derived independently and whose genomes differ significantly in length. These data suggest a common 3' terminal sequence among all VSV DI particles which contain part of the Lgene region of the parental genome.


Characterization of T antigens in polyoma-infected and transformed cells. Polyoma-infected 3T6 cells contain a number of proteins precipitable by serum from rats carrying polyoma-induced tumors. The virus codes for three species having apparent molecular weights of 90,000, 60,000 and 22,000 daltons, as determined by polyacrylamide gel electrophoresis (90K, 60K and 22k). The 90K and 22K species produced by a large plaque and a small plaque wild-type polyoma have similar mobilities, but the 60K species produced by the large plaque wild-type. In cells infected by each of seven polyoma tsA mutants, the 90K species is unstable at the nonpermissive temperature, while the 60K and 22K species are stable. In cells infected by a mutant carrying a deletion between roughly 98 and 3 map units in the early region of the viral genome, the 22K species is present, but the 90K and 60K species are absent. Tryptic peptide analysis of the isolated 90K, 60K and 22K species shows that the three species have common N terminal regions. The 60K and 22K species contain amino acid sequences not found in the 90K species , and the 60K species has several unique, methionine-containing peptides not found in either the 22K or 90K species. Two polyoma-transformed BHK cell lines do not have detectable amounts of the 90K protein.


At least 104 nucleotides are transposed from the 5' terminus of the avian sarcoma virus genome to the 5' termini of smaller viral mRNAs. Cells producing avian sarcoma virus (ASV) contain at least three virus-specific mRNAs, two of which are encoded within the 3' half of the viral genome. Each of these viral RNAs can hybridize with single-stranded DNA(cDNA5') that is complementary to a sequence of 101 nucleotides found at the 5' terminus of the ASV genome, but not within the 3' half of the genome. We proposed previously (Weiss, Varmus and Bishop, 1977) that this nucleotide sequence may be transposed to the 5' termini of viral mRNAs during the genesis of these RNAs. We now substantiate this proposal by reporting the isolation and chemical characterization of the nucleotide sequences complementary to cDNA5' in the genome and mRNAs of the Prague B strain of ASV. We isolated the three identified classes of ASVmRNA (38, 28 and 21S) by molecular hybridization; each class of RNA contained a "capped" oligonucleotide identical to that found at the 5' terminus of the ASV genome. When hybridized with cDNA5', each class of RNA gave rise to RNAase-resistant duplex hybrids that probably encompassed the full extent of cDNA5'. The molar yields of duplex conformed approximately to the number of virus-specific RNA molecules in the initial samples; hence most if not all of the molecules of virus-specific RNA could give rise to the duplexes. The duplexes prepared from the various RNAs all contained the capped oligonucleotide found at the 5' terminus of the viral genome and had identical "fingerprints" when analyzed by two-dimensional fractionation following hydrolysis with RNAase T1. In contrast, RNA representing the 3' half of the ASV genome did not form hybrids with cDNA5'. We conclude that a sequence of more than 100 nucleotides is transposed from the 5' end of the ASV genome to the 5' termini of smaller viral RNAs during the genesis of these RNAs. Transposition of nucleotide sequences during the production of mRNA has now been described for three families of animal viruses and may be a common feature of mRNA biogenesis in eucaryotic cells. The mechanism of transposition, however, and the function of the transposed sequences are not known.


Complete nucleotide sequence of the leader RNA synthesized in vitro by vesicular stomatitis virus. The complete nucleotide sequence of the leader RNA synthesized in vitro by the Indiana serotype of vesicular stomatitis virus is presented. The sequence was determined by the technique described by Donis-Keller, Maxam and Gilbert (1977) in combination with the standard two-dimensional fingerprint techniques described by Barrell (1971). The leader RNA contains 48 nucleotides variably terminating at the 3' terminus with cytosine (68%) and adenosine at position 47 (32%). Since the leader RNA is complementary to the 3' terminal portion of the viral genome RNA, the first 48 nucleotides from the 3' end of the genome RNA can be decuded. The leader RNA contains repetitive and palindromic sequences with a polypurine sequence at its 3' terminus. The possible role of some of the sequences is discussed.


Dexamethasone-mediated induction of mouse mammary tumor virus RNA: a system for studying glucocorticoid action. We have investigated the mechanisms by which dexamethasone (a synthetic glucocorticoid) stimulates the production of mouse mammary tumor virus (MMTV) by cell cultures derived from mammary carcinomas of GR mice. Treatment of these cells with dexamethasone stimulates a rapid accumulation of intracellular virus-specific RNA which is dependent upon RNA synthesis but not upon DNA or protein synthesis. The effect of dexamethasone is probably mediated by a specific and saturable glucocorticoid receptor. We conclude that the accumulation of MMTV RNA is a primary response to dexamethasone and that the rate of synthesis of MMTV RNA is probably accelerated by treatment with dexamethasone.


Regulation of the activity of the low density lipoprotein receptor in human fibroblasts. A specific receptor on the surface of cultured human fibroblasts binds plasma low density lipoprotein (LDL) with high affinity, and thereby initiates a cellular process by which the LDL is internalized and degraded within lysosomes and its cholesterol component is made available for cellular membrane synthesis. Current studies demonstrate that the activity of this LDL receptor is under feedback regulation. Prior incubation of fibroblast monolayers with cholesterol, 25-hydroxycholesterol, or LDL progressively reduced the ability of the cells to bind 125I-labeled LDL at the high affinity site. A series of kinetic studies indicated that this reduction in binding was due to a decrease in the number of LDL receptors. From measurements of the rate of decline in 125I-LDL binding activity after administration of cycloheximide, the LDL receptor was calculated to have a half-life of about 25 hr. LDL appeared to reduce 125I-LDL-binding activity by suppressing the synthesis of receptor molecules. Thus cultured human fibroblasts regulate their intracellular cholesterol content by regulating the activity of the LDL receptor, which in turn controls the rate of cellular entry of cholesterol derived from plasma LDL contained within the culture medium.


Genetic determination of the alpha-galactosidase developmental program in mice. The expression of alpha-galactosidase in liver, heart, and brain during postembryonic development has been examined in several inbred mouse strains. In most strains, the developmental patterns of alpha-galactosidase are coordinate with those of two other acid hydrolases, beta-glucuronidase and beta-galactosidase. Certain inbred mouse strains, including members of the C57-C58 family, have a tissue-specific alteration in the temporal expression of alpha-galactosidase activity. This alteration shows additive inheritance and appears to be controlled by a single genetic locus. The altered developmental expression of the enzyme is not accompanied by any discernible change in its physical properties.


Ovarian plasminogen activator: relationship to ovulation and hormonal regulation. A technique is described for detecting fibrinolytic activity of single cells in culture. This method was applied to the analysis of rat ovarian granulosa cells. Cells obtained from follicules shortly before ovulation show high levels of fibrinolytic activity. This activity is plasminogen-dependent, indicating that it is due to plasminogen activator. The appearance of this activity is correlated with ovulation by temporal and functional criteria, and can be demonstrated both in immature animals primed with hormones and in mature cycling animals. Granulosa cell cultures can be stimulated to release plasminogen activator by exposure in vitro either to luteinizing hormone or to dibutyryl cyclic AMP.


Specific binding proteins for cyclic AMP and cyclic GMP in Dictyostelium discoideum. In Dictyostelium discoideum both cyclic AMP and cyclic GMP are regulated by chemotactic stimuli. Binding proteins specific for cAMP and cGMP have been found in aggregation competent cells as well as in cells harvested during growth. The activity of binding proteins was, on the average, lower in the growth phase cells. cAMP binding proteins were separated into 3 fractions, whereas the cGMP binding activity appeared in 1 major peak both on DEAE-cellulose and Sephadex G-200. Protein kinase activity was present in most but not all cyclic necleotide binding fractions; evidence for a relationship is however missing.


[An unusual case of polydactylia].

[An unusual case of polydactylia]. A woman of 85 had a symetrical malformation of the hands manifested as a combination of syndactyly of the soft parts of the 3rd and 4th fingers and an incomplete bifurcation of the bones of the 4th finger.


Chemical carcinogen alteration of SV40 virus induced transformation of normal human cell populations in vitro. The frequency of simian papovirus 40 (SV40) induced transformation of human cells was enhanced after pretreatment with either napthylamine-2,N-methyl-N'-nitrosoguanidine (MNNG), N-acetyl-2-fluorenylacetamide (N-Ac-AAF), benzo[a]pyrene (BP), aflatoxin B1, propane sultone (PS), beta-propiolactone, 4-nitroquinoline oxide (4-NQO), methylmethane sulfonate (MMS) or diethyl nitrosamine (DEN). Posttreatment with 4-NQO, MMS, MNNG or DEN inhibited transformation; while posttreatment with either aflatoxin B1, beta-propiolactone or napthylamine-2 did not alter transformation similar to the action of N-Ac-AAF and BP. All carcinogens that altered transformation after pretreatment damaged cellular DNA. Pretreatment or posttreatment with carcinogens 3-methylcholanthrene (3-MCA) or 7,12-dimethylbenzanthrene (7,12-DMBA), that did not damage cellular DNA also did not enhance transformation. Moreover, pre- or posttreatment with other weak or non-carcinogens that did not damage cellular DNA did not alter virus induced transformation. All foci formed in the co-carcinogen treated cultures whether the carcinogen inhibited or enhanced transformation were virus directed. While a similar pattern of response existed for carcinogens that either enhance or inhibit transformation, each of the carcinogens that enhanced or inhibited foci formation damaged cellular DNA. Moreover, those carcinogens that enhanced focus formation, compared to the carcinogens that inhibited focus formation, exhibited similar DNA damage profiles.


Application of alkaline sucrose gradient sedimentation to the study of DNA damage and its repair in mammalian cells treated with methylmethanesulfonate and 4-nitroquinoline-1-oxide. KB cells and L cells were treated with methylmethanesulfonate (MMS) or 4-nitroquinoline-1-oxide (4 NQO) and the resulting damage to DNA and its repair were examined by sedimentation in an alkaline sucrose gradient. The sedimentation profiles obtained were found to be the resultant of a complex interrelationship between drug dosage, duration of the lysis period and the repair capacity of the cells. A systematic study of these variables was made which led to a plausible and useful interpretation of the sedimentation profiles. Both drugs produce two kinds of DNA modifications which show up as a single-strand breaks but affect the sedimentation profile in characteristic ways. One of these modifications which is quite alkali-labile can be studied using a 30-min lysis period. The other modification is less alkali-labile and can be studied using a long lysis period. Both KB cells and L cells can repair the former type of damage but only KB cells can repair the latter type of damage.


[Umbilical angiofibroma (apropos of a case)]. A woman aged 15 without any notable pathological history, was submitted to an operation because of a gradual growth of her umbilical region for a month. A tumor as large as an hazel-nut, was an angiofibroma of the round ligament of the liver. The round ligament may give cysts and tumors as the allantois does. If the tumors are generally malignant when situated in allantois, they would not always be mild when in the round ligament, as attested by a late observation in litterature.


A study of filling defects in the liver and spleen with multiple radionuclides. Studies were made to determine if examination with multiple radiopharmaceuticals would improve the sensitivity and specificity of colloid liver spleen scans. Increased uptake of Ga-67 citrate and In-111 bleomycin was found in most Tc-99m sulfur colloid scan defects caused by hepatocellular hepatoma or lymphoma. Increased uptake of these agents was found in some defects caused by malignant melanoma, breast carcinoma and carcinoma of the lung, and was rarely seen in defects caused by cholangiocarcinoma or gastrointestinal neoplasms. Gallium was useful in the followup of patients with hepatoma. Procedures designed to evaluate the gall bladder fossa, renal impression, or blood pool activity of an apparent tumor were found to be helpful and simple to perform. Iodine-131 as NaI was useful in studying functioning liver metastases from thyroid carcinoma as were bone scanning agents in evaluating hepatic metastases from osteogenic sarcoma. Multiple radiopharmaceutical evaluation of the physiologic and biochemical characteristics of liver lesions supplements current radiologic examinations and increases diagnostic specificity.


Mechanism for lipid abnormalities of erythrocyte membranes in biliary obstruction: lecithin content and its fatty acyl composition. The lipid compositions of erythrocyte membranes, plasma and bile of 16 patients with biliary obstruction were analysed to obtain information regarding the origin of excess lecithin which is usually found in the erythrocyte membranes in obstructive jaundice. Phospholipids and free cholesterol were found to be increased proportionally to the degree of biliary obstruction with an elevation in the free cholesterol/phospholipid ratio in the red cell membranes. The increase in phospholipid content is primarily due to lecithin. There was a highly significant alteration in the fatty acid composition of lecithin of erythrocyte membranes, plasma and bile from patients with severe jaundice. Red cell membrane lecithin amounted to more than 40% of the total phospholipid in these patients. Interestingly, the fatty acid composition of lecithin showed a similar pattern in erythrocyte membranes, plasma and bile. In addition, the fatty acyl chain composition of lecithin in lipoprotein-X was very similar to that of the red cell membrane. Freeze-fracture electron microscopy showed an alteration in membrane morphology and a reduced number of membrane-associated particles in the fractured faces. From these findings, we suggest that the lecithin of lipoprotein-X is derived from abnormal bile lecithin, which is incorporated into erythrocyte membranes by fusion with lipoprotein-X. On the other hand, the fatty acid composition of bile lecithin from patients with mild jaundice, whose erythrocyte membrane lecithin amounted to less than 31% of total phospholipid, was not different from that of normal individuals. However, in sharp contrast to the bile content, the fatty acid composition of erythrocyte membranes and plasma in these same patients showed a similar but small change compared to that of patients with severe biliary obstruction. The red cells of patients with mild jaundice were almost normal, biconcave disc-shaped, as observed by scanning electron microscopy and no abnormalities in the distribution or number of membrane particles were detected by freeze-fracturing. We propose that the abnormal lecithin content of erythrocyte membranes in patients with mild jaundice can be explained by the gradual exchange of lecithin between red blood cells and plasma lipoprotein.


Low urinary oestrogen excretion during pregnancy, due to an impairment of fetal adrenocorticotrophic hormone secretion. Four cases of newborn infants whose mothers had a low urinary oestrogen excretion during pregnancy are reported. Placental sulphatase deficiency in placental insufficiency were excluded by a dehydroepiandrosterone sulphate loading test. Postnatally, they developed a clinical picture characterized by an inappropriate secretion of cortisol which, by the results of an adrenocorticotrophic hormone stimulation test, appeared to be due to an impairment of adrenocorticotrophic hormone secretion. The prenatal and postnatal steroid metabolism is discussed.


Methods for the estimation of high density cholesterol, comparison between two laboratories. The determination of HDL by quantification of HDL-cholesterol was investigated using different methods and employing the Abell method as reference. The routinely applied direct method of Huang (calibrated on Abell standardized sera) and the enzymatic method of Röschlau were tested. Results show that with the Huang method, as compared to Abell values, a total laboratory error of less than 4.5 mg/100 can be achieved in 95% of the cases. The enzymatic method gave rise to erroneous results. Comparisons between two laboratories in experiments using random and pool sera are presented.


The vitamin D3 metabolite-type activity of Solanum malacoxylon. 1. Administration of an aqueous extract of the dried leaves of Solanum malacoxylon (DLSM) to rats causes a rapid hyperphosphataemia and a decrease in plasma alkaline phosphatase activity; the two effects are typical of 1,25(OH)2D3, the hormonally active metabolite of vitamin D3. 2. DLSM, like both vitamin D3 and parathyroid hormone, increases plasma calcium and citrate levels in rats. The effect of DLSM in influencing plasma citrate, and the role of this important metabolite in mineral metabolism is discussed. 3. A decrease of plasma magnesium levels occurs in rats following treatment with DLSM. This decrease, which is associated with a renal loss of this cation, is remarkably similar to that produced by hypervitaminosis D3. 4. Prolonged administration of DLSM to vitamin D deficient rats causes a polyuria, hypercalciuria, hyperphosphaturia, hypermagnesuria, an increase in urinary total hydroxyproline, an increase in plasma total hexosamines, and a corresponding decrease in the bone total hexosamines. These effects, some of which can also be produced by hyperparathyroidism, or following the administration of parathyroid extract (PTE), large doses of vitamin D3, or 1,25(OH)2D3, suggest that DLSM, like the latter compounds, is capable of causing bone mineral mobilization, and the dissolution of bone organic matrix.


The chemistry and conformational and biological analysis of vitamin D3, its metabolites and analogues. The chemical properties, stereochemical relationships and solution conformation, as assessed in part by proton NMR spectroscopy, for vitamin D3, its major metabolites [including 1alpha,25-(OH)2D3, its hormonally active form] and a number of A-ring and side chain analogues are evaluated and discussed in relation to their biological activity. In particular the relative ability of many of these seco-steroids to compete both with 25-OHD3 for its chick serum binding protein and 1alpha,25-(OH)2-D3 for its chick intestinal cytosol-chromatin receptor system was quantitated, in vitro. Further, the relative effectiveness of all these metabolites and analogues to mediate in vivo intestinal calcium absorption and bone calcium mobilization was determined. Collectively these chemical and biological studies constitute a "systems analysis" of the various steroid structural parameters both required and tolerated by the multi-stepped endocrine system associated with the biological actions of vitamin D.


The assay of 1alpha,25-dihydroxyvitamin D3: physiologic and pathologic modulation of circulating hormone levels. A sensitive radioreceptor assay for 1alpha,25-dihydroxyvitamin D3 (1alpha,25-(OH)2D3) is utilized to quantitate the circulating concentration of this sterol in experimental animals and humans. When weanling rats are grown for 2 weeks on low calcium or low phosphate diets, limited availability of either ion elicits a five-fold increase in the plasma level of 1alpha,25-(OH)2D3. The enhancement of 1alpha,25-(OH)2D3 in calcium deficiency is dependent upon the presence of the parathyroid and/or thyroid glands, which is consistent with parathyroid hormone (PTH) mediation of this effect. In contrast, the response to phosphate deficiency is independent of these glands and may result from a direct action of low phosphate on the renal synthesis of 1alpha,25-(OH)2D3. Studies in humans indicate that the normal level of 1alpha,25-(OH)2D is 2.1--4.5 ng/100 ml plasma. Patients with chronic renal failure have markedly lower circulating 1alpha,25-(OH)2D and this kidney hormone is undetectable in anephric subjects, but returns to normal within 1 day after successful renal transplantation. Hypoparathyroidism and pseudohypoparathyroidism are associated with reduced plasma 1alpha,25-(OH)2D while patients with primary hyperparathyroidism have significantly elevated sterol hormone levels. Thus, from measurements in rats and humans, it appears that circulating 1alpha,25-(OH)2D3 is regulated by PTH and/or phosphate and that abnormal plasma 1alpha,25-(OH)2D3 is a part of the pathophysiology of renal osteodystrophy and parathyroid disorders.


VIP measurement in distinguishing Verner-Morrison syndrome and pseudo Verner-Morrison syndrome. 1. Plasma VIP immunoreactivity is always diagnostically raised in patients with pancreatic tumour causing the Verner-Morrison syndrome. 2. Human tumour VIP is physico-chemically similar to porcine VIP. 3. The only other situation in which plasma VIP is very elevated is in patients with ganglioneuroblastomas associated with diarrhoea. 4. VIP is not elevated in patients with diarrhoea associated with pancreatic islet hyperplasia, designated the pseudo Verner-Morrison syndrome.


Mode of action of peptide hormones.

Mode of action of peptide hormones. The mode of action is discussed of the peptide hormones which trigger neosynthesis of a specific product in their target cells without being involved in any release step. Particular attention is paid to the early events elicited by ACTH in isolated adrenocortical cells. It is shown that extracellular calcium ions at physiological concentrations can serve as first messenger activating steroidogenesis if the isolated cells are pretreated in an appropriate ionic environment. Among other factors the extracellular calcium/phosphate ratio seems to be of importance. A model is proposed where calcium serves as direct messenger in the physiological activation by ACTH, cyclic AMP being a subserving factor maintaining full steroidogenesis.


Concepts in ectopic hormone production.

Concepts in ectopic hormone production. Non-endocrine tumours are capable of production of ectopic hormones, precursor molecules subunits and hormone-related fragments. The measurement of these may provide biochemical markers of malignancy, allowing presymptomatic diagnosis which may lead ultimately to an improved prognosis for the patient. The study of ectopic humoral syndromes may yield new insights into the nature of cellular differentation and the fundamental process of malignant change.


Hormone receptors and cyclic nucleotide metabolism in cancer cells. The possibility of a relationship between cyclic AMP formation and metabolic processes in tumours has been investigated. Changes in basal activity and hormone-responsiveness of adenylate cyclase were demonstrated in plasma membranes and intact cells from pre-cancerous liver of rats fed a diet containing the carcinogen 3'-methyl-4-dimethylaminoazobenzene. Basal adenylate cyclase activity in hyperplastic parathyroid gland membranes was 200% higher than that in parathyroid adenoma membranes, corresponding with their relative rates of parathyroid hormone secretion in vitro. Membrane adenylate cyclase activity in hypernephromas was consistently 100--300% higher than in adjacent human renal cortex. Furthermore the adenylate cyclase activity of the tumour membranes was not influenced by a wide range of hormones which were effective stimulants in 'normal' renal cortex membranes. Conversion of 25-hydroxycholecalciferol to 1,25-dihydrocholecalciferol could not be demonstrated in either hypernephroma or adjacent renal cortical tissue. However, three of the four hypernephromas tested secreted a bone-resorbing factor. Cyclic AMP formation was increased by salmon, human and porcine calcitonins in both plasma membranes and intact cells from a poorly differentiated epidermoid cell carcinoma which was itself secreting calcitonin in culture. This phenomenon might be related to a feedback regulation of calcitonin production in this cell line. The observations are consistent with the concept of a relationship between cyclic AMP formation and certain metabolic functions (e.g. hormone production) in tumour cells.


Ectopic hormone production by bronchial carcinomas in culture. Monolayer tissue culture has been used as a system in which to study aspects of ectopic hormone secretion. Of a series of twenty-four human bronchial carcinomas, nineteen were successfully established in culture and the supernatant medium from each tested for peptide hormones by radioimmunoassay. Six tumours were found to produce adrenocorticotrophin (ACTH), four to release calcitonin (CT) and one to release both of these hormones. No growth hormone or insulin was detected throughout the series. Net in vitro synthesis of both ACTH and CT was demonstrated by recovery of more hormone during culture than was originally contained in the explanted tumour tissue. The production of hormone by four out of six proliferative cultures established, and its persistence through many subculture passages, confirms ectopic hormone production as a stable heritable characteristic of some lung tumours. The ability of hormone-producing bronchial tumour cells to respond to factors known to influence hormone output from normal endocrine cells was tested. ACTH release was stimulated in one tumour by Pitressin and CT in another by gastrin. In addition, the release of CT from the same tumour cell line was shown to be inhibited by the accumulation of high external concentrations of CT as has been reported for normal C-cells.


Hormone-receptor interactions. The message sequence of alpha-melanotropin: demonstration of two active sites. The purpose of this investigation was to elucidate the biological significance of lysine11 and of the tripeptide sequence =Lys-Pro-Val-NH2 for the biological activity of alpha-melanocyte-stimulating hormone. To this end the in vitro melanotropic activities of twenty-four synthetic peptides related to the hormone were determined. Extension or reduction of the length of the lysine11 side chain results in a marked decrease of the melanotropic potency of the respective analogue. The C-terminal tripeptide (11--13), the tetrapeptide (10--13), and the pentapeptide (9--13) were found to be hormonally active in the same order of magnitude as the central hexapeptide (5--10). The following conclusion was drawn: alpha-MSH possesses (in contrast to ACTH) two message sequences (active sites), (i)-Glu-His-Phe-Arg-Trp-, and (ii)-Gly-Lys-Pro-Val-NH2 which are capable of independently triggering the hormone receptor responsible for melanin dispersion. Thus, despite the close structural similarity of the two hormones, alpha-MSH and ACTH appear to react with their respective target cell receptors by quite different chemical mechanisms, implying different receptor structures.


On the metabolism of two adrenocorticotrophin analogues. The metabolism of Synacthen (corticotrophin-(1--24)-tetracosapeptide) and C-41795-Ba ([D-Ser1,Lys17,Lys18]-corticotrophin-(1--18)-octadecapeptide amide) have been compared in the rat following intravenous injection. Using Synacthen and C-41795-Ba labelled with tritium it was possible to follow the tissue distribution of the two peptides. The kidney was shown to concentrate intact peptide and fragments of both peptides. Autoradiography of perfused kidney sections demonstrated the uptake of radioactivity into the lysosomes of the kidney proximal tubule cells. Comparison of the distribution of radioactive products formed from the tetracosapeptide labelled in different positions indicates that, prior to renal uptake, metabolism is taking place at other sites in the body. It is suggested that rapid cleavage occurs extracellularly at or near the N- and C-termini. Then large fragments remaining in the circulation, together with any intact material, are filtered through the glomerulus and are taken up by endocytosis into the proximal tubule cells of the kidney. The synthetic N- and C-terminal protection of the octadecapeptide appears to inhibit the extracellular attack and so the concentration of intact peptide in the kidneys is initially higher than the tetracosapeptide. Although concentrations of radioactivity in other tissues are low in comparison with the kidneys, the quantities are quite high when the weight of the tissues are considered. Chromatographic analysis of this radioactivity reveals that the octadecapeptide gives rise to much higher tissue levels of intact peptide and we believe that this acts as a depot and gives rise to the sustained blood concentrations and prolonged biological effects observed with this peptide.


Regulation of vitamin D: an evolutionary view. The regulation of vitamin D metabolism by the kidney is now known to be multifactorial. Three regulatory factors are discussed: 1,25(OH)2D3 itself; parathyroid hormone, and phospate. The influence of 1,25(OH)2D3 probably depends on new protein synthesis, while the mode of action of phosphate is unknown. Parathyroid hormone is not essential for regulation but may have an important biological role. The direction of its effect varies, perhaps due to a complex interrelation of changes in calcium and phosphorus metabolism and the level of kidney adenyl cyclase activity.


A possible ACTH secreting tumour of the pituitary developing in a conventionally treated case of Addison's disease. A patient with Addison's disease, treated with conventional corticosteroid therapy, developed endocrine and radiological features suggestive of an ACTH secreting pituitary tumour. The negative feedback control of ACTH secretion by the inhibitory effect of hydrocortisone was shown to be preserved although attenuated. Retiming and alteration of corticosteroid therapy reinforced this feedback control, without the need for supraphysiological amounts of steroid, and resulted in the regression of the endocrine disorder.


Stimulation of amniotic fluid cells by fibroblast growth factor. Amniotic fluid cells were grown in modified Dulbecco's medium with and without the following additives in different combinations: fibroblast growth factor (FGF), dexamethasone (DME), insulin and cyclic GMP under controlled conditions. Experiments were also conducted with varying amounts of foetal calf serum and FGF to establish optimum levels. Comparative growth, determined by measuring 3H-thymidine uptake, showed that the combination of FGF, DME and insulin in medium with 20% foetal calf serum produced the highest growth rate. These experiments indicate that the culture time of amniotic fluid cells for chromosome studies could be reduced by 25%. FGF did not have any deleterious effect on the chromosomes.


Alteration of body distribution of 99mTc-pyrophosphate by radiographic contrast material. Abnormal distribution of 99mTc-PYP was found in an elderly patient who had received 100 ml of radiographic contrast material between the time of injection of the radiopharmaceutical and the performance of the bone scan. Reasons for these alterations are postulated.


Effect of unconjugated and conjugated phenol and uraemia on the synthesis of adenosine 3' :5' -cyclic monophosphate in rat brain homogenates. 1. The effects of phenol and phenyl glucuronide on the responses of normal rat brain adenyl cyclase to noradrenaline and dopamine have been investigated. Neurotransmitter responses have also been examined in brains from uraemic and normal rats. 2. A depressive effect of phenol on the adenosine 3' :5' -cyclic monophosphate response of the neostriatum to dopamine was shown to be completely abolished if the toxin was present in the conjugated form; the response of the cortex to noradrenaline was stimulated by the presence of phenyl glucuronide, even though the unconjugated form had no effect. 3. The uraemic state in the rat also resulted in a depression of the neostriatum response to dopamine, yet an enhancement of the cortical response to noradrenaline. 4. The action of phenols of the brain is relevant to hepatic and uraemic coma.


Use of 99Tcm-DMSA as a static renal imaging agent. The use of 99Tcm-labelled DMSA as a static renal imaging agent has been analyzed semi-quantitatively in 366 patients. Study with this agent proved to be of most value in patients with equivocal space-occupying lesions of the kidney, provided useful information in various destructive diseases of the kidney when used for determining divided renal function but was of little value in chronic renal failure. In 33 patients, the uptake of 99Tcm-DMSA at 3 h as a measure of divided renal function was compared with the uptake of 99Tcm-DTPA from 30 to 150 sec following injection and was found to correlate well. In a series of 7 dogs with induced unilateral renal impairment, divided function determined with 99Tcm-DMSA was found to correlate well with results obtained using 51Cr-EDTA.


Cellular interactions between vasopressin and prostaglandins in the mammalian kidney. Current experimental evidence indicates that endogenous renal medullary prostaglandins modulate the antidiuretic response to vasopressin in the mammalian kidney. The predominant effect of prostaglandins is to attenuate the antidiuretic response to vasopressin; inhibition of prostaglandin synthesis potentiates the renal effect of vasopressin. Prostaglandins likely antagonize the renal effects of vasopressin at the cellular level of hormone-dependent cyclic adenosine 3,5-monophosphate metabolism, but the exact molecular mechanism is not known. Likewise, it is not known whether such modulatory effect is due to primary prostaglandins, prostaglandin precursors or to other metabolites of arachidonic acid. Vasopressin itself could stimulate intrarenal prostaglandin synthesis; this effect may represent a negative-feedback regulatory pathway for the antidiuretic response to the hormone. Recent experimental evidence suggests that modulatory effect of prostaglandin may be a factor in pathogenesis of some types of urinary concentrating defects.


The kallikrein-kinin system in the kidney. To understand the role of the kallikrein-kinin system in the kidney all components of the system and their localization need to be considered. About half the kallikrein in urine occurs as the proenzyme which arises in the distal tubule. Kinins are formed in the distal tubule and collecting duct from urokinnogen which is found throughout the tubule. Urine contains about twice as much lysyl-brandykinin as bradykinin. A third kinin, methionyl-lysyl-bradykinin, also can occur in urine. It is probably produced by uropepsin as the kinin is largely formed in acidified urine and its formation is inhibited by pepstatin. The significance of the three kinins is unknown. Kinins are normally slowly (few hours) destroyed in urine. The importance of kallikrein, urokinogen and kininases in regulating the level of kinins needs to be determined.


Contribution to the problem of the relationship of infection and rejection in patients with kidney transplants. The paper deals with the incidence of infectious complications and rejection crises in patients with renal transplants. It was revealed by statistical methods that there exists a certain correlation between infections and rejections, which depends on the type of the infectious agent. In patients where infectious complications were caused only by gram-negative flora an indirect relationship was revealed, while in patients with infectious complications where gram-positive flora, viruses or fungi participated, a marked direct correlation was found. Statistical methods proved also a higher incidence of rejection crises in patients with active cytomegalic infection.


Granular-cell tumors (myoblastomas) of the anal region. Three cases of granular-cell tumor of the perianal region are reported. The clinical and pathologic features of these tumors are reviewed. Their precise histogenesis remains uncertain. The association with pseudoepitheliomatous hyperplasia of the overlying epithelium, which was well illustrated in two of the three cases, is discussed. The lesions are nearly always benign, and the treatment of choice is local excision.


Submucosal tumors of the colon: experience with twenty-five cases. Submucosal tumors of the colon are rare and require alertness on the part of the physician for early diagnosis. These tumors may not cause symptoms before attaining large size. In most cases the clinical histories are atypical. Because of the possibility of malignancy, adequate diagnosis and treatment are necessary. This report deals with 25 cases of submucosal tumors of the colon. The tumor series included seven leiomyomas, one granular-cell myoblastoma, two leiomyosarcomas, seven carcinoid tumors, six lipomas, one lymphangioma, and one neurofibroma. Five of the tumors were asymptomatic. The endoscopic appearances of the tumors, although not diagnostic, may give some indications as to their natures. Endoscopic removal is possible, although it is indicated for high-risk patients only. Because submucosal tumors may be malignant, and because differentiation from secondary tumors (metastases) or tumor-like lesions is difficult without histologic examination, complete removal of the tumor is the treatment of choice.


Multiple granular-cell tumors of the ascending colon: microscopic study. Light and electron microscopic study of a case of multiple granular-cell tumor of the ascending colon is presented, and the relevant medical literature is reviewed. It seems that the biologic behavior of this tumor in the colon does not differ from that in other locations. Histologically, a few granular cells are seen in neural tissue in the vicinity of the tumor, suggesting a close relation of the tumor to the neural tissue. Electron-microscopically, the granular cells resemble Schwann cells more closely than neural cells and axons. Histochemically, the granules of the tumor cells show autofluorescence and high activity of acid phosphatase, which was located electron-microscopically in the limiting membranes and matrices of the granules. It is suggested that the granular-cell tumor is a neoplasm of Schwann-cell origin with a unique metabolism that causes acceleration of autophagocytosis and accumulation of ceroidlipofuscin.


[Changes in 3H-noradrenaline activity in the adrenals and serum of rats under the influence of histamine and adrenocorticotropic hormone]. The authors examined the changes in the distribution of 3H-noradrenaline in the adrenal glands and serum of white rats-control and experimental-after administration of histamine and ACTH. The results from the experiments revealed characteristic curves of accumulation of 3H-noradrenaline in the adrenal glands of the examined animals, with maximum at the tenth minute and characteristic changes in the activity of the labeled noradrenaline in the adrenals and serum after administration of histamine and ACTH, followed up to the second hour. On the basis of the obtained results from the radiometric examination and from the follow-up of the level of the blood sugar, the authors make inferences for the sequence and character of the processes of incorporation and release of catecholamine from the adrenal glands under the action of ACTH and histamine.


Cyanide-insensitive NADH oxidation by subcellular fractions isolated from human polymorphonuclear blood cells. The biochemical triad, NADH oxidation, oxygen (O2) uptake and hydrogen peroxide (H2O2) formation, by subcellular fractions of human blood polymorphonuclears (PMNs) was investigated. It was found that this biochemical triad (1) was under the control of the granule-rich fraction (GRF) only; (2) was not inhibited by cyanide; (3) occurred stoichiometrically for its three components, and (4) accounted quantitatively for the respiratory burst of the stimulated PMN. It was also shown that the above biochemical triad (1) involved an enzymatic step; (2) was enhanced by acidic pH (0.5) and Mg++; (3) was inhibited by Cu++ or low concentration of Mn++; (4) was dependent on H2O2, perhydroxyl radical (HO2) and hydroxyl radical (HO) since either catalase or superoxide dismutase or scavengers of HO2 or HO were inhibitor, and (5) involved multistep reactions. Evidence is provided that the sequence of the reactions is first a generation of H2O2, (spontaneously from NADH in our incubation medium), secondly the production of HO from H2O2, thirdly the oxidation of NADH with further production of HO2,O2 uptake and H2O2 formation, probably through a chain reaction. The identification of the enzyme(s) involved in these multistep reactions needs further studies.


Hysteretic nature of NADH substrate inhibition of bovine liver glutamate dehydrogenase. NADH substrate inhibition of bovine liver glutamate dehydrogenase appears to be eliminated at enzyme concentrations above 0.5mg/ml. Since the inhibition cannot be restored by preincubation of the enzyme with any substrate or product combination, the release of inhibition had previously been considered the result of enzyme polymerization. Benzene-saturated solutions, however, increase the extent of enzyme polymerization without affecting the NADH inhibition. These and related control measurements demonstrate that the release of substrate inhibition is the result of a hysteretic transition of an enzyme central and transitory complex.


Molecular biology of metabolic disease: defects in the regulation of enzymic activity. The Jacob-Monod model for the regulation of enzymic activity has been used in the analysis of some types of metabolic disease. Three lesions have been considered: (1) loss of allosteric inhibition of phosphofructokinase by citrate in the condition of lipomatosis; (2) failure of covalent modification of triglyceride lipase from inactive to active forms in the condition of triglyceride storage disease, and (3) failure of repression of HMG-CoA reductase by a mutant low-density liprotein in a new variant-of familial hypercholesterolaemia. Defects in enzymic regulation are contrasted with catalytic defects (the inborn errors); the major difference being an accumulation of a normal metabolic end-product of an unregulated pathway, rather than accumulation of an unusual intermediary metabolite as in the inborn errors.


Transient-phase studies on the arginine kinase reaction. 1. The initial formation of arginine phosphate by arginine kinase was studied in the time range 2.8--50 ms by the quenched-flow method. 2. A transient burst phase of product formation was obtained, the amplitude of which was temperature-dependent. At 35 degrees C it was 0.64 mol arginine phosphate/mol arginine kinase and at 12 degrees C, 0.25 mol/mol. 3. These results show that for the reaction pathway of arginine kinase the rate-limiting step follows the formation of arginine phosphate on the enzyme. This is in contrast to the creatine kinase reaction where no transient phase was observed [Engelborghs, Y., Marsh, A. & Gutfreund, H. (1975) Biochem. J. 151, 47--50]. 4. The rate-limiting step on the arginine kinase reaction pathway is only slightly affected by temperature: the change in Kcat with temperature is due to a change of an equilibrium constant pertaining to at least two previous steps.


Mitochondrial transport processes and oxidation of NADH by hypotonically-treated boar spermatozoa. 1. After hypotonic treatment spermatozoa have metabolic characteristics of mitochondria isolated from other cells. Ejaculated boar spermatozoa treated in this way can oxidise external NADH via both a lactate-pyruvate shuttle and a malate-aspartate cycle; this oxidation is coupled to the phosphorylation of ADP. 2. The dicarboxylate transport inhibitors butylmalonate, phenylsuccinate and bathophenanthroline sulphonate inhibit NADH oxidation dependent on added malate, glutamate and aspartate. alpha-Cyanocinnamate, a strong inhibitor of pyruvate transport, inhibits lactate-dependent NADH oxidation. 3. NADH oxidation dependent on malate, glutamate and aspartate is inhibited by uncoupling agents, but lactate-dependent NADH oxidation is stimulated. 4. Lactate-dependent NADH oxidation is inhibited by oxamate, an inhibitor of lactate dehydrogenase. Aminooxyacetate, an aminotransferase inhibitor, inhibits glutamate, malate and aspartate-dependent NADH oxidation. 5. Hypotonically-treated spermatozoa retain radioactivity after incubation with L-[U-14C]malate, [1,5-14C]citrate or [2-14C]malonate. Exchanges of retained radioactivity with various substrates indicate that dicarboxylate and tricarboxylate exchange carriers exist in the mitochondrial membrane.


Oxygen activation in isolated chloroplasts. Mechanism of ferredoxin-dependent ethylene formation from methionine. Low-potential electron acceptors of photosystem I of chloroplast lamellae produce superoxide anions (0-2) and hydrogen peroxide by autoxidation, but have no effect on ethylene formation from methionine; equimolar amounts of ferredoxin are less active in photosynthetic O-2 and H2O2 production but strongly stimulate ethylene production from methionine. 2. Ten to fifty units of superoxide dismutase inhibit fifty to two hundred units of superoxide dismutase stimulate ethylene formation from methionine by chloroplast lamellae in the presence of ferredoxin. This stimulation is stronger at pH 7.0 than at pH 7.8. Catalase inhibits ethylene formation from methionine. 3. Pulse-radiolytic production of nitrite (NO-2) from hydroxylamine, initiated by hydroxyl radicals (.OH) or O-2, shows no difference in the presence or absence of ferredoxin, nor do the decay kinetics of O2. 4. From the above observations and from model reactions (xanthine/xanthine oxidase; iron salts in the presence of H2O2), it is concluded that reduced ferredoxin in the presence of H2O2 forms a Fenton-type oxidizing species for methionine, generating ethylene in the presence of pyridoxal phosphate. 5. Inhibitory effects of both superoxide dismutase and catalase in oxygen-dependent reactions need not necessarily indicate the participation of the 'Haber-Weiss' reaction.


Collagenase sensitivity and aggregation properties of Electrophorus acetylcholinesterase. Tailed forms of Electrophorus acetylcholinesterase, mainly A (9 S) and C (14.2 S) forms, have been subjected to collagenase treatment. Several steps have been identified, yielding molecules which have lost different portions of the tail, and eventually resulting in separation of the isolated tetramers. These modifications result in the disappearance of the low-ionic strength aggregating properties. The molecules which have retained relatively large fragments of the tail do not aggregate in the same conditions as the intact forms, but still form small aggregates in the presence of high levels of polyanions. A model of the tailed molecules, illustrating the existence of discrete collagenase-sensitive regions in the tail, is discussed.


Parathyroid hormone and calcitonin secretion in uraemic infants. In six infants aged between 4 and 8 months with chronic renal failure, we have studied blood levels of calcium (Ca), phosphorus (P), alkaline phosphatase, immunoreactive parathyroid hormone (PTH), and calcitonin (CT), as well as urinary excretion of Ca, P, hydroxyproline and cyclic AMP under basal conditions and during an infusion of 20 mg/kg of 10% Ca gluconate in normal saline over 4 h. Under basal conditions four infants had normal serum Ca and P values, alkaline phosphatase levels at the upper limit of normal, and very high PTH (range: 1450--2550 pg Eq/ml) and CT (range: 700--1900 pg/ml) levels. The urinary Ca excretion was low, whereas the urinary excretion of P, hydroxyproline and cyclic AMP was high. During Ca infusion, the total serum Ca and CT levels increased, PTH fell without however reaching the normal upper limit, and urinary P and cyclic AMP excretion decreased. In two infants with osteodystrophy and the highest levels of PTH (2900 and 3500 pg Eq/ml respectively) there was no suppression of PTH during Ca infusion.


Influence of thyroid hormone on the positive inotropic effects mediated by alpha- and beta-adrenoceptors in isolated guinea pig atria and rabbit papillary muscles. Effects of thyroxine treatment for 7--11 days on the positive inotropic effects mediated by alpha- and beta-adrenoceptors were studied in isolated guinea pig atria and rabbit papillary muscles. In guinea pig atria, the thyroxine treatment inhibited the positive inotropic effect of lower concentrations of phenylephrine (PHE), and attenuated the inhibitory effect of phentolamine on the PHE response. The effect of isoproterenol (ISO) was potentiated by the thyroxine treatment. In rabbit papillary muscles, the thyroxine treatment shifted the dose--response curve for PHE to the right and attenuated the inhibitory effect of phentolamine on the PHE response. Propranolol, in both guinea pig atria and rabbit papillary muscles, inhibited the PHE response more effectively in preparations from thyroxine-treated animals than in controls. In guinea peg atria, the attenuation of the PHE response mediated by alpha-adrenoceptors was observed after the thyroxine treatment for only 2 days, whereas the potentiation of the ISO response required the thyroxine treatment for a longer period. It was concluded that the thyroxine treatment attenuated the positive inotropic effect mediated by alpha-adrenoceptors and potentiated that mediated by beta-adrenoceptor-mediated positive inotropic effects due to the thyroxine treatment may be independent of each other.


Prednisolone-3,20-bisguanylhydrazone: the mode of interaction with rat brain sodium and potassium-activated adenosine triphosphatase. The mechanism of interaction between prednisolone-3,20-bisguanylhydrazone (PBGH) and Na+,K+-ATPase (ATP phosphohydrolase, EC 3.6.1.3) was studied using partially purified rat brain enzyme preparations. PBGH inhibited Na+,K+-ATPase rapidly and reversibly. The enzyme-inhibiting action of PBGH was competitively antagonized by potassium. PBGH inhibited Na+,Mg2+ and ATP-supported binding of (3H)-ouabain to the enzyme. When PBGH was added to the incubation mixture at the time when the (3H)-ouabain binding was close to its equilibrium state, the concentration of (3H)-ouabain complex was rapidly reduced and shifted to a lower equilibrium state. A double reciprocal plot analysis of the (3H)-ouabain binding data indicates that the inhibition of ouabain binding by PBGH is apparently competitive. Binding of (3H)-ouabain in the presence of Tris-phosphate and Mg2+ was also inhibited by unlabeled PBGH. Thus, it appears that the binding of PBGH precludes the binding of ouabain to Na+,K+-ATPase.


Hypotension induced by inhibition of angiotensin-converting enzyme in pentobarbital-anesthetized dogs. The mechanism of the hypotensive response produced by inhibition of the angiotensin converting enzyme was studied in pentobarbital anesthetized dogs. A recently developed potent inhibitor of the converting enzyme, SQ 14,225 (D-3-mercapto-2-methyl propanoyl-L-proline), administered i.v. to intact dogs resulted in a rapid marked decrease in blood pressure. In nephrectomized dogs, SQ 14,225 retained significant hypotensive activity, although the absolute magnitude of the decreases in blood pressure were less than had been observed in dogs with intact kidneys. SQ 14,225 also lowered blood pressure when administered to intact dogs in which angiotensin II receptors had been blocked with the receptor antagonist Sar1,Ala8-angiotensin II. This apparent ability of SQ 14,225 to decrease blood pressure in the absence of a functional renin angiotensin system was shared by a structurally dissimilar, nonapeptide, angiotensin converting enzyme inhibitor, SQ 20,881 (Glu-Trp-Pro-Arg-Pro-Gln-Ile-Pro-Pro). SQ 20,881 also produced significant decreases in blood pressure in nephrectomized dogs. These findings indicate that the angiotensin converting enzyme inhibitors, SQ 14,225 and SQ 20,881 may lower blood pressure in anesthetized normotensive dogs via a mechanism unrelated to either the renin angiotensin system or the renal kinin system.


Reserpine-induced supersensitivity to the inotropic and phosphorylase activating effects of glucagon in the perfused rat heart. Glucagon (0.125--2.0 microgram) produced a dose-dependent increase in cyclic AMP, % phosphorylase a and force of contraction in the isolated perfused rat heart. Pretreatment of animals with reserpine (2.5 mg/kg, 24 h) resulted in an enhancement of the inotropic and phosphorylase activating effects of glucagon but the effect on cyclic AMP was not altered. It is suggested that reserpine-induced supersensitivity in the rat heart occurs at a point beyond the cyclic AMP step.


3H-Catecholamine binding to alpha-receptors in rat brain: enhancement by reserpine. (+/-)-3H-Epinephrine and (-)-3H-norepinephrine bind to rat cortex membranes in a saturable manner with dissociation constants of 16.7 and 27 nM respectively. The maximum number of 3H-catecholamine binding sites, 10--12 pmoles/g tissue, and the pharmacological characteristics of (+/-)-3H-epinephrine binding, indicate that the catecholamines label the same alpha-noradrenergic receptor in the rat as does 3H-clonidine. At 25 degrees, (+/-)-3H-epinephrine binding associates rapidly to equilibrium, and dissociates in a biphasic manner. The affinities of alpha-agonists at the 3H-catecholamine binding site are 2--4 fold weaker in the rat than in the calf cortex under the same experimental conditions. Ergot alkaloids and phenoxybenzamine have similar affinities in the two tissues, whereas phentolamine and WB-4101 are 8--10 times weaker in the rat. Reserpine (0.25 mg/kg s.c. per day for 3 weeks) causes 25 and 46% increases in the numbers of (+/-)-3H-epinephrine and 3H-WB-4101 alpha-receptor binding sites respectively, and a 51% increase in the number of 3H-dihydroalprenolol beta-receptor sites, in rat forebrain. Reserpine pretreatment does not alter the affinities of either alpha- or beta-receptor 3H-ligands.


Membrane-associated adenosine triphosphatase in normal and injured hypoglossal nuclei. The hypoglossal nuclei of adult male albino rats, either normal, or from 1 to 70 days after left hypoglossal nerve transection, were studied cytochemically with a lead method for sodium- and potassium-activated membrane ATP-ase, using light and electron microscopy. Reaction product was measured at the light microscopical level by microdensitometry, and significance determined statistically. Light microscopy revealed a brown reaction product in the neuropil, but none in cell bodies. Blood vessel walls were more strongly coloured. Reaction product was undiminished by ouabain pre-incubation, or by incubation without sodium or potassium, or by incubation with calcium instead of magnesium. Reaction product was diminished by absence of magnesium if calcium was also absent, and abolished if sections were boiled before incubation, or if substrate was absent. Axotomy caused a statistically significant increase in neuropil reaction product in injured nuclei, maximal at 35 days postoperatively, and subsequently decreasing to normal at 70 days. Electron microscopy showed a predominantly surface membrane reaction product, with occasional positive intracellular cisternae. Basal lamina and intra-endothelial vacuoles were also positive. Axotomy resulted in the arrival and disappearance of microglis (2 to 35 days), followed by astrocyte hypertrophy (35 days), and increase in thickness and homogeneity of surface membrane reaction product. The results suggest the presence of one or more calcium- or magnesium-dependent membrane ATP-ases. The peak of the increase after axotomy is probably partly attributable to hypertrophic astrocytes, and partly to the surfaces of neuronal processes. Increase of membrane movements might explain such enzyme activity.


The ventral spino-olivocerebellar system in the cat. IV. Spinal transmission after administration of clonidine and L-dopa. 1. The transmission from the flexor reflex afferents (FRA) to the spino-olivocerebellar paths ascending through the ventral funiculus (VF-SOCPs; Oscarsson and Sjölund, 1977) was compared with the transmission from the FRA to segmental reflex arcs in cats with the spinal cord transected in the third cervical segment sparing only the ventral funiculus on one side. The climbing fiber responses evoked in Purkinje cells by electrical stimulation of limb nerves were monitored by recording the mass activity at the cerebellar surface simultaneously from several termination zones, while the flexion reflex was recorded from a flexor nerve and the primary afferent depolarization (PAD) from a dorsal root filament. 2. Changes in the segmental reflex responses were produced by i.v. injections of l-Dopa and Clonidine. L-Dopa produced the expected depression of the flexion reflex and the FRA-induced PAD in the FRA, whereas Clonidine depressed only the flexion reflex but did not affect or even enhanced the FRA-induced PAD in the FRA. 3. The changes in transmission to one of the ascending paths, the a-VF-SOCP, parallelled that in the flexion reflex arc. The transmission to another path, the b2-VF-SOCP, parallelled that in the pathway responsible for the FRA-induced PAD in the FRA. 4. The remaining hindlimb paths, the c1- and c3-VF-SOCPs, monosynaptically activated from primary afferents, were little influenced by the changes induced in the segmental reflex arcs. 5. It is suggested that the a- and b2-VF-SOCPs carry information related to interneuronal activity in segmental centers, whereas the c1- and c3-VF-SOCPs forward information mainly related to peripheral events.


Conditioned changes of synaptic transmission in the motor cortex of the cat. Intracellular recordings were made from 117 neurons in the motor cortex of anesthetized cats. The pyramidal tract (PT) and VL nucleus of thalamus were stimulated in order to activate the neurons from two directions. 1. PT cells were conditioned by antidromic trains (10--50 cps for 4--15 s) and by paired PT and VL stimuli with different intervals and sequences. The VL-EPSPs were examined before and after conditioning, to find differences in efficacy in giving rise to spikes. The conditioning procedures resulted in a remarkable facilitation of VL-EPSPs, manifesting itself as a significant rise of efficacy in generating spikes, a shortening of peak latency and in some cases, an enhancement of background firing. 2. In non-PT neurons the same conditioning procedures elicited heterosynaptic facilitation and a rise in firing activity. 3. Intracellularly injected square wave pulses also resulted in facilitation of VL-EPSPs. 4. Pairings of PT and VL stimuli were more effective than trains in evoking conditioned changes. 5. Plastic modifications were observed in the 13.7% of the neurons subjected to conditioning procedures. 6. The authors assume that synchronous activity of the pre- and postsynaptic neurons is a highly important condition for plastic changes in the efficacy of synaptic transmission.


The relationship of phosphodiesterase and cyclic AMP to the process of fluid secretion in the salivary glands of the ixodid tick Amblyomma americanum. Phosphodiesterase (PDE) activity in the salivary glands of the female. Amblyomma americanum decreased as the tick progressed from a slow to a rapid phase of feeding, while the rate of fluid secretion increased when glands were stimulated with cyclic AMP and theophylline. Dopamine stimulated PDE activity and an 'inhibitory' factor was found in glands obtained from rapidly engorging ticks which decreased PDE activity. These findings are discussed as they relate to the process of fluid secretion by salivary glands of feeding ixodid ticks.


Lipoprotein lipase activator deficiency in very low density lipoproteins in rat nephrotic syndrome. Marked urinary loss of lipoprotein lipase activator in experimental rat nephrotic syndrome may be partly responsible for its deficiency in plasma very low density lipoproteins.


Flavonoid compounds are potent inhibitors of cyclic AMP phosphodiesterase. The inhibitory activity of 19 flavonoid molecules on cyclic AMP breakdown by a commercial beef heart phosphodiesterase preparation is reported. 7 compounds are active in the micromolar range, 2 of which have a potency equivalent to that of papaverine. Some structure activity relationships are drawn.


Relationship between A-type and C-type particles in Ehrlich ascites tumor cells. Intracisternal and intracytoplasmic A-type particles were discovered in Ehrlich ascites tumor cells. In addition, 'mature' and 'immature' C-type particles were also seen in the intercellular space. It is believed that A particles may represent a precursor or a formative stage of the C particles.


The EM immunocytochemical demonstration of lysozyme in macrophage giant cells in sarcoidosis. The giant cells (multinucleate macrophages) of human sarcoidosis have been shown by the unlabelled antibody immunoperoxidase technique at electron microscope level to contain lysozyme within cytoplasmic granules.


Biochemical studies of pigments from a pathogenic fungus Microsporum cookei. V. Evidence for the transmembrane permeability of xanthomegnin across phospholipid bilayer membranes. Direct evidence is provided for the transmembrane permeation of xanthomegnin across phospholipid bilayer membranes using ascorbate-loaded liposomes. This process may be associated with an uncoupling effect on the oxidative phosphorylation of mitochondria.


Inhibition of phosphodiesterase activity of human spermatozoa by spermine. Sperm motility and metabolism are dependent upon the levels of cyclic AMP. Our studies have demonstrated that spermine by inhibiting sperm phosphodiesterase activity could regulate sperm physiology.


[Effect of nicotinamide, adenosine and nicotinamide-adenine dinucleotide on the NAD and NAD.H2 content in the liver and brain of intact rats and in hypoxia]. Nicotine-amide (25 mg/kg) in the liver and adenosine (50 mg/kg) in the liver and brain of intact rats enhance the NAD+NAD'H2 content. These drugs when used in the same doses raise the NAD+NAD'H2 level in the organs reduced in hypoxia and their proportion only with the combined application of these drugs. NAD (30 mg/kg) increases the NAD+NAD'H2 concentration in the liver and brain of intact and in the liver of "hypoxic" rats. It appears that in the mechanism of the antihypoxic action of nicotine-amide, adenozine and NAD a definite part plays their action on the level of nicotine-amide co-enzymes.


Chemical grafting of functional NAD in the active site of a dehydrogenase: regeneration in situ. A functional NAD molecule was immobilized at the active site of Alcohol dehydrogenase within a proteic membrane. The presence and the functionality of the cofactor was checked by fluorescence analysis. The dehydrogenase NAD membrane does not require addition of soluble cofactor for its activity. The system represents a new worthwhile approach because both problems of retention and regeneration of cofactor are solved. The method can be used not only for industrial and analytical applications but also to try to get a better understanding of the kinetics and mechanisms of the catalytic action of dehydrogenase.


The photobiogenesis and metabolism of vitamin D. Provitamin D3 (7-dehydrocholesterol) is converted to previtamin D3 by the action of ultraviolet radiation on the skin. Previtamin D3 thermally isomerizes to vitamin D3 in the skin and the vitamin is then transported to the liver on the vitamin D-binding protein. Although there are extrahepatic vitamin D-25-hydroxylases, the liver is the major site for the 25-hydroxylation of vitamin D. In response to hypocalcemia and hypophosphatemia, 25-OH-D is metabolized by a renal-cytochrome. P450-dependent mixed function oxidase system is 1alpha,25(OH)2D. When calcium and phosphate homeostasis prevails the renal 25-OH-D-1alpha-hydroxylase activity is limited and instead a non-cytochrome P450 mixed function oxidase metabolizes 25-OH-D to 24R,25(OH)2D. Parathyroid hormone has clearly been shown to be a trophin for the renal synthesis of 1,25(OH)2D; however, the role and significance of the adrenal steroids, or gonadal and pituitary hormones, on the renal 25-OH-D-1alpha-hydroxylase is not well defined. The regulation of the photometabolism of provitamin D3 to vitamin D3, the role and significance of the side-chain metabolism of 1,25(OH)2D by the small intestine, and the metabolism of 25-OH-D to 24R,25(OH)2D by chondrocytes and its stimulation of protein synthesis in these cells are just a few issues that will require further investigation.


Foodborne hazards of microbial origin.

Foodborne hazards of microbial origin. Foods can serve as vehicles of many pathogenic and toxigenic agents of disease. Bacterial agents comprise three groups: 1) those that grow in the food and produce an active toxin before consumption (e.g., clostridium botulinium); 2) those that merely exist as contaminants in the food but are able to initiate infection when swallowed (e.g., Salmonella spp.); and 3) those that multiply and produce large numbers of vegetative cells in the food, then release an active enterotoxin when they sporulate in the gut. A few parasitic (e.g., Trichinella spiralis) and viral agents (e.g., hepatitis A) also can be transmitted by food. Botulinum poisoning is the deadliest foodborne disease. The potential danger of botulism from cured meats is a major factor in the argument over use of nitrites as meat curing agents. A new disease called infant botulism has been recognized since 1976. Apparently it is not foodborne but results from intraintestinal growth of C. botulinum in very young infants. Salmonellosis is the most important of the foodborne diseases from the standpoint of overall human health. The primary vehicles are contaminated raw meat, poultry, and eggs. Faulty food handling practices are responsible for most food poisoning in the United States.


Acetylcholine-induced ionic channels in rat skeletal muscle. This paper briefly reviews the evidence for ionic channels mediating the conductance increase caused by acetylcholine application to the end-plate of skeletal muscle fibers. "Membrane noise" observed during application of constant low concentrations of acetylcholine to an end-plate is thought to arise from the random superposition of many elementary events corresponding to the opening and closing of discrete ion channels. Statistical analysis of acetylcholine-induced noise reveals an elementary conductance event of of 34 pS (1 S = 1 omega-1) amplitude and 1 msec duration at room temperature in rat muscle fibers. Both size and duration of the elementary event are temperature dependent. Analysis of currents induced by application of acetylcholine to the extrasynaptic membrane of chronically denervated fibers shows that the elementary conductance has a similar size but is of much longer duration. Direct recording of square pulse-like currents by a patch clamp method confirms some of the conclusions drawn from fluctuation analysis.


Large-scale production, concentration and purification of Rous Sarcoma Virus in tissue culture. A method is described for long-term standard production of purified virus from cultures of PR-RSV-C-transformed chicken cells. The mean yields were 4.91 mg of purified virus per liter of culture medium. Cells were grown in long-term cultures for 26 weeks. A combination of polyethylene glycol 6 000 precipitation and zonal centrifugation has been found satisfactory for virus concentration and purification.


Fractionation of proteins from Rous sarcoma virus and avian myeloblastosis virus by isoelectric focusing. The isoelectric focusing technique in the pH gradients was used for a preparative isolation of proteins from Rous sarcoma virus and avian myeloblastosis virus. The purified major gs protein, p27 (pI = 9.1) and the gP86 (pI = 5.3) were obtained after disruption of virus with 1% non-ionogenic detergent in the presence of 6M urea. The p10, p15, and p19 were present in the same range of pH (pI = 6.8). A strongly basic protein, immunologically active, presumably the p12, was found in the alkaline region of the pH gradient 10.8. These proteins fully retained their immunological activity. On the other hand, in the acidic region of the pH gradient between pH 4 and 5, strong precipitates were regularly found. These precipitates were complexes which were formed by interaction of the acid components of ampholines with the viral proteins during isoelectric focusing. Almost all viral proteins were present, differing only in quantity. The complexes were stabile in 1% non-inogenic detergent and 6M urea. They were dissociated with 1% SDS and 5M urea, and had no immunological activity. The methods of virus disruption and possibilities of formation of the ampholine-viral protein complexes are discussed.


Long-term karyological study of RVP3 tumour cells grown in vitro. The RVP3 cell line established from an in vivo passaged RVP3 tumour, which originally arose in RSV-injected mice, was analyzed at different passage levels and characterized virologically and cytogenetically. The RVP3 cell line is characterized by the presence of 2 to 5 medium-sized to large metacentric chromosomes, the majority of the mitoses contain about 1 subtelocentric chromosome, and in addition, every mitosis contains several chromosomal structures corresponding to the group called m-ms that comprises both minute chromatin bodies and microchromosomes. In three monocellular clones derived from the RVP3 cell line with a similar karyological picture it was established that the number of m-ms increased proportionally to the total number of chromosomes in individual metaphases. It was verified that: the RVP3 cell population does not contain any transmissible agent producing chromosomal changes in mouse cells; RVP3 cells are highly malignant; no RSV could be rescued from the clone tested. The significance of m-ms is discussed.


Effects of phosphoenolpyruvate, other glycolytic intermediates and methylxanthines on calcium uptake by a mitochondrial fraction from rat pancreatic islets. 45Ca2+-accumulation by a mitochondrial fraction from isolated rat pancreatic islets was stronly stimulated by ATP. The ATP-dependent uptake was inhibited by phosphoenolpyruvate in a dose-dependent manner over a wide variety of conditions. Inhibition by phosphoenolpyruvate was non-completitive with respect to calcium, competitive with respect to magnesium, and antagonised by high Mg-ATP2- concentrations; fructose 1,6-diphosphate also decreased 45Ca2+-uptake. Other glucose metabolites were either less effective or ineffective in diminishing mitochondrial 45Ca2+-accumulation. The ATP-dependent uptake was also inhibited by xanthine derivatives (caffeine and 3-isobutyl-1-methylaxanthine) which potentiate the effects of glucose on insulin secretion. Cyclic AMP had no effect. It is thought that the rate of insulin secretion is a function of the cytosolic calcium concentration in the B-cell. These data show that phosphoenolpyruvate, fructose 1,6-diphosphate and methylxanthines might influence exocytosis by direct effects on mitochondrial calcium accumulation, and thus the intracellular distribution of calcium.


86Rb+ fluxes and K+-stimulated nitrophenyl phosphatase activity in the pancreatic islets of genetically diabetic mice (C57BL/KsJ-db/db). Fluxes of 86Rb+ and hydrolysis of p-nitrophenyl phosphate were measured in collagenase-isolated islets of diabetic C57BL/KsJ-db/db-mice and normal controls (C57BL/KsJ-+/+). Both types of islets accumulated Rb+ avidly, as originally reported for hand-dissected islets of non-inbred ob/ob-mice. KsJ-db/db-mouse islets showed enhanced accumulation of Rb+ and normal activity of K+-activated nitrophenyl phosphatase. D-glucose, 20 mmol/l, inhibited Rb+ efflux in normal islets but not in those from KsJ-db/db-mice. The glucose insensitivity of Rb+ efflux was observed in young animals, which exhibit glucose-induced insulin release, as well as in old animals, which do not secrete insulin in response to glucose. The anomalous regulation of Rb+ efflux already present in young animals may bear on the liability of KsJ-db/db-mouse B-cells to develop defective control of membrane potential, an abnormal metabolism of cyclic AMP, and a marked failure of insulin secretory capacity.


The ultrastructure of functional mouse adrenal cortical tumor cells in vitro. Cells derived from a transplantable mouse adrenal cortical tumor maintain their differentiated function in vitro and secrete steroids in response to ACTH and other stimulatory agents. The cell line has been widely employed for various biochemical investigations but there have been few attempts to correlate this work with morphologic data. This communication describes the electron microscopic appearance of the tumor transplant in vivo and primary cultures derived from it at various intervals after the cells are placed in culture. Tumor cells in vivo bear considerable resemblance to normal adult mouse adrenal cortical cells. Organelles generally considered to be directly involved in steroid biosynthesis (mitochondria, smooth endoplasmic reticulum and lipid droplets) are not drastically altered. Certain modifications of the vasculature and cell membrane, seemingly related to steroidogenesis, are present in both the tumor and normal adrenal cortex. Within 2 days after the tumor cells are introduced to culture, their cytoplasm assumes a more simplified appearance. Smooth endoplasmic reticulum is less conspicuous and free ribosomes and polysomes are very abundant. Mitchondrial inner membranes are reorganized from a saccular arrangement in the cells in vivo into distinct lamellar cristae. The tumor cells now resemble undifferentiated embryonic adrenal cells, or cultured adrenal cells from various mammalian sources which have dedifferentiated in the absence of ACTH. In their morphologically unspecialized state, the normal cells are incapable of functional responses to ACTH. In contrast, the cultured, dedifferentiated tumor cells respond within minutes to this hormone and can demonstrate 5-20 fold increases in their basal steroid output. These data suggest that substantial steroidogenic activity can occur although the characteristic appearance of adrenal mitochondria is absent.


Ultrasound guided fine-needle aspiration biopsy of abdominal masses. Twenty-six cases of ultrasonically guided percutaneous fine-needle aspiration biopsies of abdominal masses are reported. Included are masses in the pancreas, retroperitoneum, liver, kidney, and pelvis. Aspiration biopsies accurately diagnosed or excluded malignancy in 21 patients (81%). The procedure, performed under local anesthesia, is rapid, simple and almost painless. No complications occurred; specifically, there was no hemorrhage, peritonitis, or spread of tumor along the needle tract.


Multiple concentric ring sign in the ultrasonographic diagnosis of intussusception. Gray scale ultrasound examination of two patients with abdominal space-occupying lesions demonstrated characteristic ring shadows on longitudinal scans. In one case a multiple concentric ring shadow resulted from an ileocolic intussusception, and in the other a "bull's eye" image was due to an infiltrating carcinoma of the pyloric antrum. A study of these patients confirms the suspicion [1] that there is a characteristic ultrasonic appearance of intussusception in bowel loops.


ERCP update: diagnostic and therapeutic applications. Endoscopic retograde cholangiopancreatography (ERCP) is currently utilized to evaluate jaundice, both intra- and extrahepatic, and can differentiate between these by demonstrating specific anatomic lesions. ERCP is definitive in the evaluation of gallbladder disease when other modalities are indefinite. For the first time outside the operating room, the pancreatic duct can be opacified by ERCP. The newest application of ERCP, endoscopic papillotomy (EP), has revolutionized the treatment of common bile duct gallstones and stenosis of the duct, offering a safe and effective alternative to repeated surgery, especially in those patients at risk. Because of these diverse and effective applications, ERCP has become firmly established in our diagnostic armamentarium.


Cyclic nucleotide-dependent protein kinase activity in acinar cells from guinea pig pancreas. Acinar cells from guinea pig pancreas possess two distinct protein kinase activities. Cyclic GMP-dependent kinase elutes as a single peak on diethylaminoethyl (DEAE)- cellulose chromatography, is not inhibited by protein kinase inhibitor, and has a greater affinity for cyclic GMP (half-maximal activation at 20 nM) than for cyclic AMP (half-maximal activation at 100 nM). Cyclic AMP-dependent kinase elutes as two peaks on DEAE-cellulose chromatography, is inhibited by protein kinase inhibitor, and has a greater affinity for cyclic AMP (half-maximal activation at 20 nM) than for cyclic GMP (half-maximal activation at 7 micrometer). Binding of cyclic 3H-nucleotides to the enzyme preparation was rapid, specific, temperature-dependent, and reversible, and there was a close correlation between the ability of a particular cyclic nucleotide to inhibit binding of cyclic 3H-nucleotide and its ability of a particular cyclic nucleotide to inhibit binding of cyclic H-nucleotide and its ability to activate protein kinase. Binding of cyclic 3H--nucleotide could not be described as a simple bimolecular reaction and native cyclic nucleotides accelerated the dissociation of bound, labeled cyclic nucleotide. Vasoactive intestinal peptide or secretin, each of which increases cellular cyclic AMP, caused endogenous activation of protein kinase and inhibition of cyclic [3H]AMP binding but did not alter bindings of cyclic [3H]GMP or cyclic [3H]AMP.


A familial neuronal disease presenting as intestinal pseudoobstruction. The purpose of this paper is to describe 2 siblings who had a generalized neurological disease which presented as intestinal pseudoobstruction. The siblings had 40-year histories of abdominal pain, distention, and vomiting as well as gait ataxia, small, irregular, poorly reactive pupils, dysarthria, absent deep tendon reflexes, and impaired vibratory and position senses. Compared with age-matched controls, they had inappropriate blood pressure responses to phenylephrine, the Valsalva maneuver, and upright posture, lack of sweating on warming, and pupillary denervation hypersensitivity. Radiographs revealed hyperactive, nonpropulsive contractions of a dilated esophagus and small intestine and extensive colonic diverticulosis. Esophageal manometry recorded repetitive, spontaneous, nonperistaltic waves and positive Mechyolyl tests. Postmortem examinations showed degeneration of the myenteric plexuses of the esophagus, small intestine, and colon of both patients. Myenteric plexus neurons were significantly reduced in number compared with 7 controls. About one-third of the siblings' neurons contained round, eosinophilic intranuclear inclusions, which, by histochemistry, were composed of protein by lacked RNA, DNA, carbohydrate, and fat. By electron microscopy the inclusions consisted of an irregular array of nonviral, nonmembrane-bounded filaments. Neurons and glial cells of the brain, spinal cord, dorsal root, and celiac plexus ganglia contained identical intranuclear inclusions. Intestinal smooth muscle was normal. These 2 siblings represent a unique disease in which degeneration of the myenteric plexus resulted in hyperactive but uncoordinated smooth muscle activity and the clinical syndrome of intestinal pseudoobstruction, the presenting manifestation of their neurological disease.


[Estrogen treatment of excessively tall girls (author's transl)]. 20 girls, their age being between 8 to 13 years, with an actual height above the 97th percentile received 10,0 mg conjugated estrogens per day cyclicly or 0,3 mg ethinylestradiol continuously over a period of 0,3 to 3,8 years, in order to inhibit excessive growth. The duration of therapy ranged from 0,6 to 3,8 years regularly. Before and during treatment the patients were examined, and each time the expected final height was calculated based on the actual height and the bone age. The stage of puberty was defined according to Blizzard, and the determination of the bone age was performed as described by Tanner and Whitehouse. The extragenital side effects were also recorded. The effectiveness of the estrogen treatment, i. e. the reduction of the expected final height, depends on the chronological age, the skeletal age and the stage of puberty before therapy. It ranged between 0,5 and 5,1 cm. The earlier treatment was started the better were the results. The effectiveness was more obvious in cases with continuous estrogen treatment. No serious side effects could be observed.


Changes in the activity of hydrolytic enzymes in the brain of rats intoxicated by ethyl-mercury-p-toluene-sulfanilide. A histochemical study concerning the activity of phosphatases and esterases of the brain has been undertaken in rats experimentally intoxicated by the fungicide ethyl-mercury-p-toluenesulfanilide (EMTS). The results have shown that compared with other mercury compounds, both organic and inorganic ones, such as corrosive sublimate and calomel, EMTS proved to be a less induced of alterations in the activity of cerebral hydrolases. The brains of animals intoxicated by EMTS revealed a notable decrease of ATP-ase and acid phosphatase activity as well as a moderate drop of AChE activity. Instead, the neuronal TPPase activity was distinctly elevated. Degenerative changes of neurons were observed in various regions of the experimental brains, the pyramidal cells of the Ammon's horn being affected most severely.


[Electromyographic and electroneurographic findings in bronchial carcinoma]. Twenty one patients with confirmed bronchial carcinoma were checked by clinical, electromyographical and electroneurographical methods. None of these patients underwent cytostatic therapy. Clinical findings were: 4 patients suffered from polyneuropathy (PNP) and myelopathy (MP). 3 patients had PNP alone, 3 had MP. We could support these findings by EMG in 7 patients with PNP. 4 of these cases had a lowered motor nerve conduction velocity (MNCV). In 4 cases without clinical symptoms we could show a lowered MNCV at the distal legs and affection of the motor neuron by EMG. 3 further cases without clinical symptoms showed only a lowered MNCV. 5 patients had proximal pareses. Signs of MP were not found in these cases. In conclusion we found a significantly lowered MNCV in 5 cases: only two of these showed clinical signs of PNP. Pathological EMG changes were mostly found in patients with squamous cell carcinoma, sometimes also in patients with oat cell carcinoma. Thus, clinical, electromyographic and electroneurographic findings did not always correlate in our cases.


Homozygosity of adenylate kinase allele 3: two cases. The phenotype AK 3.3 in the isoenzyme system of human adenylate kinase has been found in two members of the Wayampi population of French Guiana.


Mitogen-induced activation of stable E-rosettes by human lymphocytes. Following on an earlier study which demonstrated that PHA can activate human lymphocytes to form temperature-stable (37 degrees C.) E-rosettes with sheep red blood cells, a number of other mitogens were tested. These included Concanavalin A, pokeweed mitogen, soybean agglutinin, sodium periodate, and galactose oxidase. All except soybean agglutinin demonstrated the capability to activate lymphocytes to form stable E-rosettes.


Antibody-dependent cellular cytotoxicity against Epstein-Barr virus-associated antigens in African patients with nasopharyngeal carcinoma. Antibody-dependent cellular cytotoxicity (ADCC) was performed on serum samples from African patients with nasopharyngeal carcinoma (NPC) to determine whether antibody titers determined by this assay might be of prognostic value in this disease. The serum donors were divided into two groups: (1) those individuals who died within 2 years following diagnosis of NPC; and (2) individuals who responded well to therapy and surivived for more than 2 years following diagnosis. The ADCC GMT for the survivor group was significantly higher than the GMT for non-survivors (%5,410 versus 615). Interestingly, there were a number of discordant sera in the non-survivor group that had very low ADCC titers (less than 240) at diagnosis in the presence of high VCA titers. When ADCC titers were compared with anti-EA or IgA antibody titers to VCA, a statistically significant inverse correlation was noted. These data suggest that ADCC titers might be of prognostic importance in African NPC.


Some photoresponses of isolated tissue preparations to ultraviolet light in the presence of photosensitizers. Responses of isolated tissue preparations to ultraviolet (UV) light were studied with and without the presence of photosensitizers like eosin, fluorescein and sodium nitrite. Exposure to UV light in the presence of sodium nitrite induced consistent relaxation of rat duodenum. The photorelaxation was found to be related to the concentration of sodium nitrite. Adrenergic or cholinergic mechanisms do not seem to be involved. The isolated rat duodenum preparation exhibited quantitatively consistent photoresponse for 3 to 4 hr at its normal tone obviating the need for additional spasmogens as needed with other preparations. The preparation is a suitable test model for the study of photobiologic response evoked by UV light.


Antibody patterns to herpesviruses in Kaposi's sarcoma. II. Serological association of American Kaposi's sarcoma with cytomegalovirus. The prominent finding of this extended serologic analysis on American and African Kaposi's sarcoma (KS) patients and appropriately matched control groups is the detection of a specific serologic association of cytomegalovirus (CMV) with American KS patients. All American KS sera contained CMV antibodies and their geometric mean titers (GMT) were significantly higher than those in sera of melanoma patients (GMT ratio k = 5.3 to 7.7 by complement fixation [CF], k = 8.9 by indirect hemagglutination [IHA]) or in sera of age- and sex-matched healthy controls (k = 12.6 to 16.0 by CF, k = 12.6 by IHA). The result is strongly reminiscent of the data obtained previously for European KS. Although the GMT to CMV of African KS patients were similar to the GMT of the American KS groups, their significance cannot be demonstrated due to the high background of CMV infections in the control groups. Complex mechanisms are hypothesized, by analogy with the Epstein-Barr virus (EBV) involvement in Burkitt's lymphoma (BL), for a CMV involvement in the development of KS.


Genetics of SV40 T-antigen expression: studies of twins, heritable syndromes and cancer families. Susceptibility of human skin fibroblasts to SV40 virus infection has been suggested as a marker of cancer risk. To evaluate the role of heritable factors in the regulation of SV40 T-antigen, fibroblasts from 9 pairs of identical twins and 129 members of cancer-prone families, including 16 with cancer, were tested in a 3-day immunofluorescence assay. In the twin study, the variance of T-antigen values was significantly less in identical than in fraternal or non-twin sibs, suggesting a heritable component in the regulation of SV40 infection. In the families, T-antigen values of parents and children were compared to models of Mendelian inheritance. At least three modes of inheritance--autosomal dominant, recessive, and X-linker--were observed. The distribution of offspring values compared to those of their parents suggested that interaction of multiple genetic factors influences the T-antigen value in individual patients. With the exception of Fanconi's anemia, the values for patients with cancer or predisposing syndromes were not uniformly elevated. The utility of this assay as a marker of cancer risk appears limited because of the complexity of factors that influence T-antigen expression in individual cases.


Interaction of the B95-8 and P3HR-1 substrains of Epstein-Barr virus (EBV) with peripheral human lymphocytes. Two substrains of the Epstein-Barr virus derived from the B95-8 and P3HR-1 cell lines were studied for their interaction with human peripheral lymphocytes. It has been previously shown that B95-8 virus has and P3HR-1 virus lacks lymphocyte-transforming ("immortalizing") properties. DNA stimulation induced by B95-8 virus showed a good correlation with the number of surface Ig-positive cells. P3HR-1 virus added before B95-8 virus completely abolished the stimulation of DNA synthesis. It also prevented EBNA induction by B95-8 virus. P3HR-1 virus added after B95-8 virus diminished DNA stimulation by the latter in a time-dependent fashion. P3HR-1 virus did not inhibit DNA stimulation by phytohaemagglutinin but was inhibitory if added before a B cell mitogen (Staphylococcus Aureus). The origin of P3HR-1 virus and its relationship to the transformation process are discussed.


Differences in the capacity of simian virus 40 (SV40) tumor antigens on cells, membranes and in soluble form to induce transplantation immunity in hamsters and mice. The immunogenicity of the SV40 tumor-specific transplantation antigen (TSTA) on cells, cell particulates and solubilized membranes was studied in mice and in Syrian hamsters. Immunizations were done with various concentrations of tissue-culture-passaged, non-virus-releasing transformed cells, purified cell membranes and in some cases purified nuclei and papain-solubilized membranes obtained from several species, including the mouse, hamster, man, and sheep. All transformed cell lines were T-antigen-positive. The immunosensitive mKSA line of BALB/c mice and the immunosensitive SV34 cell line of the hamster were used for tumor challenge. All materials, regardless of source and of type of preparation, were strikingly immunogenic in the mouse but only SV40 virus and SV34 (hamster) cells provided protection against tumor cell challenge in the hamster. Also, in a limited study, BKV-transformed hamster cells and purified cell membranes and JCV-transformed hamster cells were found to be immunogenic by the tumor rejection assay in the mouse but not in the hamster. SV40 immunization did not protect the hamster against BKV- and JCV-transformed hamster cells. These results are discussed in terms of possible different specificities resident on the TSTA molecule.


Virus-induced papillomas of the alimentary tract of cattle. An abattoir survey was carried out to determine the incidence and aetiology of squamous papillomas of the alimentary tract of cattle in Scotland and North England as they were suspected of being involved in the genesis of alimentary carcinoma in certain localized geographical areas. A total of 7,746 cattle of a wide age range was examined. Various subsets of this number were subjected to analyses of certain specific factors. The calculated overall incidence was 19% and the detailed site incidence and tumour multiplicity are given. The sites at which papillomas were found were identical with those at which carcinoma had been noted in animals from a high-cancer area. The number of sites affected by papilloma and the tumour multiplicity were much lower in the general population than in the high-cancer area. Inclusion bodies, identified by electronmicroscopy as virus, were found in tumour cell nuclei and a typical papilloma virus was purified from the tumours. The structure of the tumours is described and the possible plurality of bovine papilloma-viruses is discussed in the light of recent findings in the human viruses. The general interest of a naturally occurring and geographically localized oncogenic system, in which an environmental carcinogen and a virus might be involved, is extended.