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Biomedical subjects

J M Ward

Publications and source records attributed to J M Ward.

At least 253 records · Page 14Linked to original sources
PubMed1988

Mouse papillary lung tumors transplacentally induced by N-nitrosoethylurea: evidence for alveolar type II cell origin by comparative light microscopic, ultrastructural, and immunohistochemical studies.

A histogenetic study was designed to evaluate controversial findings on the cell of origin of tubular/papillary lung tumors in mice, i.e., bronchiolar Clara cell versus alveolar type II cell. N-Nitrosoethylurea (0.5 mmol or 0.74 mmol/kg) was given to pregnant C3H (C3H/HeNCr MTV-) and Swiss Webster [Tac:(SW)fBR] mice as a single i.p. injection on Day 14, 15, 16, or 18 of gestation. The offspring were studied at various ages ranging from 7 days to 52 wk. Serial sections of the whole lung (100 to 200 sections per mouse) showed that solid/alveolar and papillary tumors arose from the pulmonary acinus, invading the bronchioles only as the tumors grew. Furthermore, a mixture of solid and papillary patterns within a single module did not represent a merging of two tumors but a progression from the solid to the papillary form. By use of two rabbit antisera against mouse lung surfactant apoproteins found in normal alveolar type II cells, it was shown by the avidin-biotin peroxidase complex procedure, by the peroxidase-antiperoxidase technique, and by indirect immunofluorescence that both solid and papillary tumors contained these proteins that are specific markers for alveolar type II cells. With a rabbit anti-rat Clara cell antiserum, none of the tumors studied was immunoreactive while normal Clara cells were reactive. The nitroblue tetrazolium formazan stain for dehydrogenase enzymes, found particularly in Clara cells, did not reveal these enzymes in any lung tumors from either strain. Ultrastructurally, no typical features of the mature Clara cell were detected in papillary or other pulmonary neoplasms. However, all tumors showed characteristic alveolar type II cell structures such as various stages of lamellar body formation, although these features were less well differentiated in the papillary tumors. Argentaffin dense bodies, representing lysosomes and immature forms of lamellar bodies, were commonly observed in papillary tumors. Some features of the papillary tumors such as cell shape, high glycogen content, and primary cilia were equivalent to those seen in pulmonary epithelial precursor cells during fetal development. With age, the papillary tumors became invasive, accumulated neutral lipids, and developed bizarre cleaved nuclei and lamellated nuclear pseudoinclusions. In conclusion, the papillary lung tumors of the mouse, at least those induced transplacentally by N-nitrosoethylurea, constitute less well-differentiated or poorly differentiated alveolar type II cell adenomas or carcinomas with fetal morphological and biochemical properties.

Age Factors↗
PubMed1988

The chronic hepatic or renal toxicity of di(2-ethylhexyl) phthalate, acetaminophen, sodium barbital, and phenobarbital in male B6C3F1 mice: autoradiographic, immunohistochemical, and biochemical evidence for levels of DNA synthesis not associated with carcinogenesis or tumor promotion.

Male B6C3F1 mice, 6 weeks of age, were fed diets or water containing di(2-ethylhexyl) phthalate (DEHP) at 12,000 or 6000 ppm, acetaminophen (ACT) at 10,000 or 5000 ppm, sodium barbital (BBS) at 1000 ppm, or phenobarbital (PB) at 500 ppm for 40 weeks. Groups of six mice were terminated at 2, 8, 24, and 40 weeks for evaluation of liver and kidney weights, histopathology, and thymidine kinase (TK) activity in liver and kidney and levels of DNA synthesis, measured by tritiated thymidine [( 3H]T) autoradiography or bromodeoxyuridine (BrdU) immunohistochemistry. Liver weights, as percentage of body weight, were significantly elevated at most time intervals for mice exposed to all chemicals at each dose. The hepatocyte labeling indices (LI) with [3H]T autoradiography or BrdU immunocytochemistry were significantly elevated in mice fed DEHP at 12,000 ppm at 24 and 40 weeks or BBS and ACT at 2 weeks. LI were not elevated in mice fed PB. Hepatic TK activity was significantly elevated in mice fed DEHP, BBS, or ACT at Weeks 2 and 8. Histopathologic hepatic lesions were associated with these elevations, while hepatic lesions were not associated with changes in TK activity in PB-treated mice. In contrast, only DEHP and BBS induced toxic renal lesions. Persistent or transient elevation of the renal LI and TK activity accompanied renal toxicity. Thus, the hepatic toxin DEHP induced chronic renal hyperplasia without evidence of renal carcinogenicity or tumor promotion in previous studies at the doses used. ACT, a hepatotoxin, produced transient chronic hepatic hyperplasia without evidence of carcinogenicity in B6C3F1 mice in earlier studies at the same doses used. Thus, persistent or transient hepatic or renal hyperplasia was associated with carcinogenic or tumor promoting activity of these chemicals in some cases but not in others.

Acetaminophen↗
PubMed1988

Rapid development of hepatocellular neoplasms in aging male C3H/HeNCr mice given phenobarbital.

One hundred and nineteen male C3H/HeNCr mice, 12 months of age, with spontaneous preneoplastic and neoplastic hepatocellular lesions were given phenobarbital (PB) at 500 ppm in drinking water. Groups of 9-10 mice were sacrificed after 12, 24 and 36 weeks of PB exposure. Identical numbers of untreated controls were used. A group of 6-week-old C3H/HeNCr mice were also given PB and sacrificed at 12, 24 or 36 weeks. In aging mice, PB exposure significantly increased the number of gross tumors or microscopic foci, adenomas or carcinomas per liver at all time periods, especially unique eosinophilic proliferative lesions, while young mice did not develop any focal proliferative lesions by 36 weeks. These findings suggest that in aging mice a fraction of the hepatocyte population (normal, spontaneously-initiated or preneoplastic) is more highly susceptible to phenobarbital 'carcinogenesis' than are hepatocytes of younger mice.

Age Factors↗
PubMed1988

Detection of activated proto-oncogenes in N-nitrosodiethylamine-induced liver tumors: a comparison between B6C3F1 mice and Fischer 344 rats.

DNA from B6C3F1 mouse and Fischer 344 rat liver tumors induced by N-nitrosodiethylamine (DEN) were examined for the ability to induce morphological transformation of NIH3T3 cells. DNAs from 14 of 33 of the mouse liver tumors induced by a single injection of DEN at 12 or 15 days of age were positive in this assay while DNA from only one of 28 DEN-induced rat liver tumors was active. Southern blot analysis of the NIH3T3 transformants derived from the mouse liver tumors revealed amplified and/or rearranged restriction fragments homologous to the H-ras proto-oncogene. DNA from two independent foci induced by the rat tumor DNA did not hybridize to probes for members of the ras gene family or c-raf. Activating mutations in the H-ras genes from the DEN-induced mouse liver tumors were characterized by selective oligonucleotide hybridization and the detection of a new XbaI restriction site by Southern blot analysis. In activated H-ras genes from the DEN-induced mouse liver tumor DNA, seven of 14 had a CG----AT transversion at the first base of the 61st codon, three of 14 had an AT----GC transition and four of 14 had the AT----TA transversion at the second base of codon 61. This spectrum of mutations is very similar to that recently observed in activated H-ras genes found in spontaneously occurring B6C3F1 mouse liver tumors. Taken together, the data suggest that the DEN-induced rat and mouse liver carcinogenesis may involve genetic targets other than or in addition to the H-ras gene.

Animals↗
PubMed1988

H-ras p21 and peanut lectin immunoreactivity of hyperplastic, preneoplastic and neoplastic urinary bladder lesions in rats.

Hyperplastic, preneoplastic and neoplastic urinary bladder lesions induced by bladder carcinogens and toxins in the rat were evaluated for immunoreactivity with polyclonal or monoclonal antibodies to H-ras p21 or binding to peanut lectin with avidin-biotin immunocytochemistry. A low proportion (less than 20%) of hyperplastic and neoplastic bladder lesions induced by N-butyl-N-(4-hydroxybutyl)-nitrosamine and fixed in Bouin's fixative only were immunoreactive on the cell membrane with the antibodies to H-ras p21. Lectin binding was found for these lesions, as well, even in formalin-fixed tissue and for lesions induced by other carcinogens, but not in regenerative bladder hyperplasias after cyclophosphamide exposure or in bladder exposed to bladder tumor promoters. The latter lesions were also not immunoreactive with antibodies to p21. Our results suggest that this relatively simple technique might be used for identification and screening of tumors for involvement of ras oncogenes and carcinogen initiation.

Animals↗
PubMed1988

Inoculation of baboons and macaques with simian immunodeficiency virus/Mne, a primate lentivirus closely related to human immunodeficiency virus type 2.

A primate lymphotropic lentivirus was isolated on the human T-cell line HuT 78 after cocultivation of a lymph node from a pig-tailed macaque (Macaca nemestrina) that had died with malignant lymphoma. This isolate, originally designated M. nemestrina immunodeficiency virus (MnIV) and now classified as simian immunodeficiency virus (SIV/Mne), was inoculated intravenously into three juvenile rhesus monkeys (Macaca mulatta), three juvenile pig-tailed macaques (M. nemestrina), and two juvenile baboons (Papio cynocephalus). All six macaques became viremic by 3 weeks after inoculation, whereas neither of the baboons developed viremia. One pig-tailed macaque died at 15 weeks with suppurative peritonitis secondary to ulcerative, necrotizing colitis. Immunologic abnormalities included a marked decrease in CD4+ peripheral blood lymphocytes. Although five macaques mounted an antibody response to SIV/Mne, the animal that died at 15 weeks remained antibody negative. Three other macaques (two rhesus and one pig-tailed) died 66 to 87 weeks after inoculation after exhibiting progressive weight loss, anemia, and diarrhea. Histopathologic findings at necropsy included various manifestations of immune deficiency, nephropathy, subacute encephalitis, pancreatitis, adenocarcinoma, and lymphoid atrophy. SIV/Mne could be readily isolated from the spleens and lymph nodes of all necropsied macaques, and from the cerebrospinal fluid, brains, bone marrow, livers, and pancreas of some of the animals. SIV antigens were localized by avidin-biotin immunohistochemistry to pancreatic islet cells and to bone marrow endothelial cells. The data suggest that African baboons may be resistant to infection by SIV/Mne, whereas Asian macaques are susceptible to infection with this pathogenic primate lentivirus.

Acquired Immunodeficiency Syndrome↗
PubMed1988

Mouse hepatoblastomas: a histologic, ultrastructural, and immunohistochemical study.

Hepatoblastomas from B6C3F1 and BALB/c mice were examined by light and electron microscopy and by immunohistochemical reactions for alpha-fetoprotein, keratin, and vimentin. Tumors occurred in one group of a chronic bioassay for the interaction of diet, genetic strain, and the carcinogen, 2-acetylaminofluorene. Tumors had several populations (including epithelial and mesenchymal cells) in various stages of differentiation. Neoplastic epithelial cells had features of embryonal hepatocytes, such as sparse cytoplasmic organelles, absence of glycogen, abundant free ribosomes, occasional bile canaliculi, and peroxisome-like dense bodies. Embryonal fibroblast-like cells had pleomorphic and folded nuclei with prominent perinuclear chromatin and dispersed cytoplasmic organelles. Fibroblast-like cells were surrounded by bundles of collagen fibrils. Intermediate or transitional types of cells were seen. No tumor cells were immunoreactive for mouse alpha-fetoprotein (AFP) antibody, unlike those in hepatocellular adenomas or carcinomas. Epithelial and mesenchymal tumor cells contained intermediate filaments throughout the cytoplasm; some of these cells stained for keratin but not for vimentin. These findings suggest that mouse hepatoblastomas are derived from bipotential liver blastema cells and are composed of a mixture of several cell populations.

Animals↗
PubMed1988

Progressive multifocal leukoencephalopathy and retroviral encephalitis in acquired immunodeficiency syndrome.

Antigens of human polyomaviruses, the etiologic agents of progressive multifocal leukoencephalopathy (PML), and of human immunodeficiency virus were localized in paraffin sections from brains of six patients who died with the acquired immunodeficiency syndrome. Immunostaining revealed polyomaviral antigens in oligodendrocytes and in some astrocytes. Human immunodeficiency (retro) virus antigens were immunostained in mononuclear macrophages, glial cells, and vascular endothelial cells. Both viral types were found ultrastructurally. The lesions of PML were more destructive than is usually seen in cases without the acquired immunodeficiency syndrome. The retroviral encephalitis could have occurred before the onset of PML. However, a secondary retroviral encephalitis could have resulted if the monocytes responding to an initial polyomaviral lesion were already infected with human immunodeficiency virus before they differentiated into macrophages.

Acquired Immunodeficiency Syndrome↗
PubMed1988

Aspergillus rhinitis in Wistar (Crl:(WI)BR) rats.

In two separate 24 month studies on the carcinogenic effect of single cadmium chloride injections in male Wistar (CRl:(WI)BR) rats, a total of 22% (129/597) of animals studied histologically were found to have chronic suppurative rhinitis caused by Aspergillus fumigatus. The diagnosis was based on characteristic conidial heads present in the sections, and positive methenamine-Grocott (GMS) staining of septate hyphae with dichotomous branching at angles of 45 degrees. Fungal hyphae balls, surrounded by a wall of neutrophilic granulocytes, were found in areas of the naso- and maxilloturbinates and occasionally caused complete blockage of the nasal passages. The underlying tissue showed an inflammatory response. In sections from 32 of the 129 cases (25% of the cases), epithelial necrosis and hemorrhage were indicative of fungal tissue invasion, but without dissemination to other organs. The infection rate was unaffected by the cadmium treatment or the location of rats in different cages. Positive antibody titers to Sendai and sialodacryoadenitis viruses suggested that transient inflammation of the upper respiratory tract rendered the mucosa susceptible to the fungal infection. The infection appeared to be sustained by growth around foreign bodies (hairs and plant material). Although focal squamous cell metaplasia of the respiratory epithelium with hyperplasia and hyperkeratosis occurred more frequently in rats with Aspergillus rhinitis, the incidence of tumors of the nasal cavities was not affected.

Animals↗
PubMed1987

Immunohistochemical determination of inducibility phenotype with a monoclonal antibody to a methylcholanthrene-inducible isozyme of cytochrome P-450.

A monoclonal antibody (MAb) to a methylcholanthrene (MC)-induced cytochrome P-450, designated MAb 1-7-1, was used for immunohistochemical staining of formalin-fixed tissues from oil- and MC-treated C57BL/6, DBA/2, and [(C57BL/6 X DBA/2) F1 X DBA/2] F2 mice. An avidin-biotin-peroxidase complex immunohistochemical technique was used. For controls, the tissues were also exposed to MAbs 1-48-5 and HyHel-9 (to egg white lysozyme). In liver, MAb 1-7-1 specifically stained the cytoplasm of centrilobular hepatocytes of C57BL/6 mice treated with MC (80 mg/kg) 48 h before kill; staining was not observed with vehicle-treated C57BL/6 mice, with oil- or MC-treated DBA/2 mice, or with comparable antibody concentrations of control MAbs 1-48-5 or HyHel-9. In the F2 mice, about 50% were expected to be MC inducible (AhbAhd). Inducibility phenotype was determined by measuring the conversion of [14C]MC to oxidized and conjugated products by liver homogenates. In freshly fixed material from MC-treated mice, those livers shown by the determination of phenotype to be inducible also stained with MAb 1-7-1, whereas those not induced were immunohistochemically negative. Furthermore, there was a significant positive correlation between degree of staining and the level of MC-metabolizing activity measured biochemically. The immunohistochemical procedure was also accurate in determination of inducibility phenotype of livers that had been in paraffin blocks for up to 2 yr if more concentrated antibody was used. In lung, MAb 1-7-1 stained specifically the alveolar walls and endothelium of blood vessels in MC-induced C57BL/6 mice only; the control MAbs and other mice gave negative results. Similarly, in kidney MAb 1-7-1 stained only glomeruli and interstitial tissue of MC-induced C57BL/6 mice and only endothelium of blood vessels in the colons of these mice. These observations are consistent with induction of the cytochrome P-450 recognized by MAb 1-7-1 in the endothelial cells of extrahepatic tissue. Immunohistochemical staining with MAb thus shows great promise for highly specific localization of particular species of cytochromes P-450 in tissues, for in situ quantification of these enzymes, and for determination of inducibility phenotype with fixed material.

Animals↗
PubMed1987

Nickel--magnesium interactions in carcinogenesis: dose effects and involvement of natural killer cells.

The effects of magnesium carbonate (MgCarb) on carcinogenesis and natural killer (NK) cell modulation by nickel subsulfide (Ni3S2) were studied. Male Fischer F344/NCr rats, 50-90 g body wt, 20 rats per group, received single i.m. injections into both thigh muscles of 2.5 mg Ni3S2 alone or combined with different proportions of MgCarb; the Mg/Ni molar ratio ranged from 0.25 to 4.0. Control rats received i.m. injections of normal saline or magnesium acetate (MgAcet), or s.c. MgCarb at a site distant from Ni3S2. The animals were observed over 79 weeks for the development of tumors. The NK cell activity was determined over the first 3 weeks of the experiment in separate groups of rats treated as above, with the use of the 51Cr/YAC-1 release assay for blood and spleen cells and the peroxidase localization of Ox-8-immunoreactive lymphocytes at the injection site. I.m. administration of MgCarb mixed with Ni3S2 up to the Mg/Ni molar ratio of 1.0 inhibited the carcinogenicity of Ni3S2 in a dose-related manner; final incidence of sarcomas decreased from 100 to 55% and the appearance of first tumors was delayed from 25 to 39 weeks. Higher doses of MgCarb did not exert further effect. Distant s.c. injection of MgCarb or local i.m. application of MgAcet did not change the carcinogenic potency of i.m. Ni3S2. MgCarb or saline alone did not produce any tumors. I.m. Ni3S2 had no significant influence on the activity of NK cells in blood and spleen, while i.m. MgCarb alone did not affect the NK activity in blood but doubled it transiently in the spleen 24 h after injection. In the injected muscle, Ox-8-positive cells became abundant around MgCarb but could not be found close to Ni3S2. This inhibitory effect of Ni3S2 was partially reversed by MgCarb. Also, numerous multinucleated giant cells infiltrated the sites of injection of MgCarb alone and MgCarb + Ni3S2 but not Ni3S2 alone. The results indicate a dose-dependent and strictly local character of the inhibition by MgCarb of Ni3S2 carcinogenesis, as well as a possible involvement of NK and phagocytic cells in this inhibition.

Animals↗
PubMed1987

The agarase gene (dag A) of Streptomyces coelicolor A3(2): affinity purification and characterization of the cloned gene product.

The coding and regulatory sequences of the agarase gene of Streptomyces coelicolor A3(2) were cloned in Streptomyces lividans 66 on the plasmid vector pIJ61, resulting in a several hundred-fold increase in the production of the secreted protein. Subcloning experiments localized the sequences required for agarase production and for the mediation of carbon catabolite repression to a segment of about 1.2 kb. A simple protein purification procedure that uses affinity binding of agarase to agarose beads was developed. Preliminary characterization of the enzyme, together with the results of in vitro transcription-translation studies, suggest that the intracellular form of agarase (about 34 kDa) possesses a signal sequence that is cleaved upon secretion across the cell membrane to produce an extracellular protein of about 29 kDa.

Chromatography, Affinity↗
PubMed1987

Immunohistochemical localization of human and simian immunodeficiency viral antigens in fixed tissue sections.

Antigens of human (HIV) or simian immunodeficiency viruses (SIV) were identified with polyclonal or monoclonal antibodies and avidin-biotin complex (ABC) immunohistochemistry in fixed surgical pathology and autopsy specimens of humans or monkeys with the acquired immunodeficiency syndrome. With B-5 fixative, viral antigens were readily detected in lymph nodes of 8 of 13 patients with follicular hyperplasia, but in only 1 of 12 patients with follicular atrophy. Antigen was detected in follicular dendritic reticular cells and rare blastlike cells, extracellularly, and in postcapillary venules, medullary lymphocytes, sinus histiocytes, and macrophages in some lymph nodes. In the brain at autopsy, antigen could be found in gliomesenchymal-cell nodules, astrocytes, vascular endothelial cells, multinucleated cells, and astrocytes and macrophages associated with demyelination. In contrast, 4 rhesus monkeys with experimental SIV infection had abundant antigen in sinus histiocytes, macrophages, and multinucleated giant cells of lymph nodes and spleen and in thymic epithelial cells. Brain lesions of monkeys resembled those of humans, with antigen found in macrophages and multinucleated giant cells. Antibodies to HIV also were immunoreactive in formalin-fixed tissue sections of monkeys containing SIV antigens. The ABC technique provided a fast and efficient method for localizing HIV and SIV antigens in fixed surgical and autopsy specimens. These findings are consistent with those found with in situ hybridization, ultrastructural studies, frozen sections of lymph nodes, and permanent sections of brain.

Acquired Immunodeficiency Syndrome↗
PubMed1986

Effects of a single dose of polychlorinated biphenyls to infant mice on N-nitrosodimethylamine-initiated lung and liver tumors.

Polychlorinated biphenyls (PCBs) are widespread, chemically-stable environmental contaminants; some congeners are commonly found in human adipose tissue and breast milk. We investigated the effects of a single dose of one PCB mixture (Aroclor 1254) on tumors initiated by N-nitrosodimethylamine (NDMA), also a common environmental agent. Infant outbred Swiss male mice were treated with NDMA (5 mg/kg) i.p. on the 4th day of life, to initiate lung and liver tumors. Four days later each received a single intragastric dose of PCBs (50, 250, or 500 mg/kg of Aroclor 1254) or oil. Groups were killed 16 and 28 weeks later. At both endpoints the mice given 500 mg/kg PCBs after NDMA developed twice as many lung tumors (alveologenic adenomas) as those treated with NDMA only, a significant difference. The PCBs alone did not cause lung tumors. This is the first demonstration of tumor promotion by PCBs in an extrahepatic organ, and it occurred after a single exposure. There were also complex, multiple effects on NDMA-caused liver tumors (adenomas and carcinomas) and on focal hepatocellular proliferative lesions: PCB treatment after the NDMA was associated with decreased number but increased size of these tumors and foci. All of these changes were accompanied by retention in the bodies of 0.1-6 ppm PCBs, as indicated by gas chromatography with electron capture detection. Of this, 80% or more consisted of 2,4,5,2',4',5'-and 2,3,4,2',4',5'-hexachlorobiphenyls in about equal amounts for periods up to 28 weeks. These results point to a need for both experimental and epidemiological studies of the effect of PCB body burden on tumor development.

Animals↗
PubMed1986

The role of iodine in carcinogenesis.

Iodine is an essential nutrient for the normal growth and development of humans and animals and is necessary for normal metabolism and regulation of thyroid hormones. Iodine excess can produce thyrotoxicosis but not cancer. However, radioiodine is carcinogenic for the thyroid gland. Dietary iodine deficiency is associated with goiter in humans and animals. The goiter develops because of a feedback system between thyroid hormones, the pituitary gland, and the hypothalamus, and it regulates the synthesis and release of thyroid-stimulating hormone. Chronic hypersecretion of thyroid-stimulating hormone causes profound goiter (diffuse thyroid hyperplasia), which appears to be related to carcinogenesis. Chronic dietary iodine deficiency in rats leads to thyroid follicular adenomas by 12 months and follicular carcinomas by 18 months. An increased risk of thyroid cancer has been reported in humans with goiter and those living in some iodine-deficient areas of the world. In very recent animal studies, iodine deficiency, chemical goitrogens, and thyroid toxins have been shown to have potent tumor-promoting effects. In rats, iodine deficiency is a much more effective tumor promoter than it is a carcinogen, suggesting that a similar relationship may exist in human populations. These studies suggest that a major role of iodine is to prevent the formation of thyroid tumors in humans and animals.

Animals↗
PubMed1986

Construction and characterisation of a series of multi-copy promoter-probe plasmid vectors for Streptomyces using the aminoglycoside phosphotransferase gene from Tn5 as indicator.

Several versatile, multi-copy, promoter-probe plasmid vectors have been constructed that replicate in a wide range of Streptomyces species. Transcriptional activity is detected by the expression of a promoter-less aminoglycoside phosphotransferase gene (neo) derived from the transposon Tn5; expression of this gene confers kanamycin and neomycin resistance on Streptomyces lividans. An efficient transcriptional terminator from E. coli phage fd has been inserted upstream of the neo coding region to prevent significant transcriptional read-through from vector promoters. A translational stop codon situated downstream from the site(s) used for cloning and preceding and in frame with the ATG start codon of the neo gene ensures the detection of transcriptional, rather than translational, fusions. Relative promoter strengths can be determined by gradient plate assays of kanamycin resistance, by measuring the amount of aminoglycoside phosphotransferase produced or by estimating neo mRNA synthesised. The high copy number of the vectors facilitates the rapid isolation and characterisation of promoter-active fragments and convenient restriction sites are available for DNA sequencing and S1 mapping of cloned inserts. Some derivatives contain a polylinker that facilitates the insertion, excision and analysis of cloned fragments and which enhances the use of these plasmids as general cloning vectors.

Cloning, Molecular↗
PubMed1986

Lack of effect of phenobarbital on hepatocellular carcinogenesis initiated by N-nitrosodiethylamine or methylazoxymethanol acetate in male Syrian golden hamsters.

Subchronic toxicity and long-term tumor-promoting effects of phenobarbital (PB) were investigated in male Syrian golden hamsters. In subchronic studies, PB was administered in drinking water to 5-week-old male hamsters for periods of 8 or 16 weeks at dosage levels of 250, 500, or 1000 ppm. No significant change in the ratio of liver weight to body weight was observed at 8 weeks; however, at 16 weeks there was a dose-dependent increase in the ratio of liver weight to body weight and a significant decrease in body weight gain among animals that received PB at 1000 ppm. The effect of PB on hepatic cytochrome P-450 and P-450-dependent aminopyrine N-demethylase activity was compared in male Syrian golden hamsters, F-344/NCr rats, and B6C3F1 mice. PB enhanced cytochrome P-450 activity in all three species; however, a significant increase (p less than 0.05) in aminopyrine N-demethylase activity was observed only in rats and mice. Potentially preneoplastic hepatocellular hyperplastic foci and hepatocellular neoplasms were studied in weanling male Syrian golden hamsters that received a single ip injection of either 100 mg N-nitrosodiethylamine (DEN)/kg body wt or 20 mg methylazoxymethanol acetate (MAM)/kg body wt at 5 weeks of age, followed by administration of 500 ppm PB in drinking water that began 2 weeks after the carcinogen injection and continued to 69 weeks of age. Groups of hamsters were killed at 25, 52, and 69 weeks of age; portions of liver and other organs with gross lesions were fixed in Formalin and examined histologically. MAM was a more potent hepatocarcinogen than DEN in male Syrian golden hamsters. PB failed to promote the development of either preneoplastic hepatocellular foci or hepatocellular neoplasms (adenomas or carcinomas) in either DEN- or MAM-initiated hamsters. Also, PB had no effect on the development of nonhepatic lesions occurring either spontaneously or induced by DEN or MAM in these animals.

Aminopyrine N-Demethylase↗