Detection of direct mutagenicity of cigarette smoke condensate in mammalian cells

Mutagenicity of cigarette smoke condensate (CSC) and the acidic, basic and neutral fractions of CSC was examined in the AL hybrid cell, a Chinese hamster ovary cell containing one human chromosome 11. Since the human chromosome 11 is not necessary for survival of the AL cells, mutations involving large deletions and chromosomal loss by non-dysjunction are non-lethal events that are detectable by loss of human cell surface antigens (a1, a2 and a3) encoded by genes on chromosome 11p (a1 and a3) and 11q (a2) through an antibody-complement lysis assay. Exposure of AL cells to CSC without exogenous metabolic activation caused a dose-dependent cytotoxicity and mutagenicity. Mutagenicity also increased with time of incubation up to 3 h with a maximum of 300 a1- mutants/10(5) survivors (250% above background; P less than 0.0005) after incubation with 100 micrograms/ml CSC. Cytotoxicity and mutagenicity of CSC were inversely proportional to cell density. Fifty percent lethal doses for the acidic, basic and neutral fractions of CSC after 3 h of incubation were 30, 100 and 240 micrograms/ml respectively, and the acidic fraction at a concentration of 25 micrograms/ml induced 350 a1- mutants/10(5) survivors (230% above background; P less than 0.0005); the basic and neutral fractions were less mutagenic. These results indicate that CSC and fractions of CSC can directly produce a spectrum of mutations, through both deletional and non-dysjunctional mechanisms of a kind known to lead to inactivation of tumor suppressor genes.     

Activity of Eight Fluoroquinolones against Both Methicillin-Susceptible and -Resistant Staphylococcus aureus Isolated from Skin Infections

The in vitro susceptibility of Staphylococcus aureus to eight fluoroquinolones, norfloxacin, ofloxacin, enoxacin, ciprofloxacin, lomefloxacin, tosufloxacin, sparfloxacin, and nadifloxacin was established by agar dilution tests, 71 isolates of methicillin-susceptible (MSSA) and 74 isolates of -resistant S. aureus (MRSA) isolated from skin infections. Among all of the fluoroquinolones, nadifloxacin exhibited the lowest MIC for both MSSA and MRSA. In addition, there were no resistant S. aureus, neither MSSA and MRSA, to nadifloxacin. With the exception of nadifloxacin, the incidence of MRSA resistant to fluoroquinolones has gradually increased in recent years. Over half of the MRSA strains were resistant to norfloxacin, ofloxacin, enoxacin, ciprofloxacin, and lomefloxacin.     

Phosphorothioate analogs of oligodeoxynucleotides: inhibitors of replication and cytopathic effects of human immunodeficiency virus

Nuclease-resistant phosphorothioate analogs of certain oligodeoxynucleotides have been tested in vitro as antiviral agents against human immunodeficiency virus (HIV) in human T cells. Phosphorothioate analogs complementary to HIV sequences, as well as noncomplementary analogs including homooligomers, exhibited potent antiviral activity. The antiviral activity was related to the base composition of the analogs, and longer phosphorothioates were more effective than shorter ones. A 28-mer phosphorothioate oligodeoxycytidine (S-dC28) at a concentration of 1 microM exhibited potent antiviral activity and inhibited de novo viral DNA synthesis as shown by Southern blot analysis. However, S-dC28 failed to inhibit gag expression in chronically infected T cells assessed by immunofluorescent assay at concentrations up to 25 microM. An N3-methylthymidine-containing phosphorothioate analog, which does not hybridize efficiently in vitro to complementary normal DNA, showed no antiviral activity. A 14-mer phosphorothioate oligodeoxycytidine (S-dC14) synergistically enhanced the antiviral activity of 2',3'-dideoxyadenosine, an anti-HIV nucleoside. Therefore, phosphorothioate analogs of oligodeoxynucleotides could represent a unique class of experimental therapeutic agents against the acquired immunodeficiency syndrome and related diseases. However, their mechanism of action is likely to be complex.     

Induction of gastric carcinomas in nonhuman primates by N-ethyl-N'-nitro-N-nitrosoguanidine

N-Ethyl-N'-nitro-N-nitrosoguanidine [(ENNG) CAS: 63885-23-4] was administered to 5 Macaca monkeys (Macaca mulatta and M. irus) at a concentration of 200 or 300 micrograms/ml for 11-26 months in their drinking water. Gastric carcinomas in the pyloric region were observed in all 5 monkeys between experimental months 11 and 38. Histologically, these carcinomas were mainly poorly differentiated adenocarcinomas and signet-ring cell carcinomas, and a few moderately and well-differentiated adenocarcinomas were also found. The macroscopic and histologic appearances of these carcinomas were similar to those in humans.     

Effect of a high-protein diet on insulin and glucagon secretion in ventromedial hypothalamic (VMH) lesioned rats

The effects of a high-protein diet on insulin and glucagon secretion in ventromedial hypothalamic (VMH) lesioned and sham-operated (sham) rats were studied in vivo as well as in perfusate from isolated pancreas. Two weeks after VMH destruction or sham operation, the rats were given either a balanced diet (protein 27%, carbohydrate 61%, fat 12%) or a high-protein diet (protein 55%, carbohydrate 30%, fat 15%) for the following 2 weeks. The calorie intake and body weight changes after the commencement of the diets were almost the same in the groups of VMH lesioned rats, but these were much greater than those in the two sham-operated groups. Fasting blood glucose, plasma insulin, and plasma glucagon concentrations were also similar between the two VMH groups, but in the sham-operated rats fasting blood glucose and plasma insulin concentrations of those rats on high-protein diet were significantly increased when compared to those on balanced diet. In the isolated, perfused pancreas, an arginine-induced excess insulin and glucagon secretion was not significantly different between the VMH lesioned rats. An arginine-induced rise in insulin concentration in the sham-operated rats on high-protein diet was significantly higher than for rats on balanced diet. We therefore suggest that hyperinsulinemia already produced in the VMH lesioned rats may not be influenced by the change in the composition of the dietary protein and carbohydrate.     

Novel enzyme immunoassay (Immune Complex Transfer Enzyme Immunoassay) for anti-thyroglobulin IgG in human serum

A novel enzyme immunoassay (immune complex transfer enzyme immunoassay) for anti-thyroglobulin IgG in human serum is described. Anti-thyroglobulin IgG in human serum was reacted with dinitrophenyl thyroglobulin, and the complex formed between anti-thyroglobulin IgG and dinitrophenyl thyroglobulin was trapped onto an affinity-purified rabbit (anti-dinitrophenyl bovine serum albumin) IgG-coated polystyrene ball. After eliminating nonspecific IgG in test serum by washing, the complex was eluted from the polystyrene ball with dinitrophenyl-L-lysine and transferred to a clean polystyrene ball coated with rabbit anti-thyroglobulin IgG. The amount of human anti-thyroglobulin IgG in the complex on the rabbit anti-thyroglobulin IgG-coated polystyrene ball was estimated using rabbit (anti-human IgG 7-chain) Fab'-horse-radish peroxidase conjugate. The present enzyme immunoassay was 1,000–3,000-fold more sensitive than the conventional enzyme immunoassay, in which a thyroglobulin-coated polystyrene ball was incubated with serum containing anti-thyroglobulin IgG and subsequently with rabbit (anti-human IgG 7–chain) Fab'-horseradish peroxidase conjugate. By the present enzyme immunoassay, anti-thyroglobulin IgG was demonstrated in all patients with Graves' disease and chronic thyroiditis.     

Structural analysis of human beta-defensin-1 and its significance in urinary tract infection

Human beta-defensin-1 (hBD-1) is a 36-amino-acid antimicrobial peptide that functions in the host innate defense. We developed a highly sensitive radioimmunoassay for hBD-1 and identified several hBD-1 peptides in human kidney, urine, and plasma by amino acid sequencing and mass spectrometry. Large quantities of hBD-1 peptides are produced in the kidney, are released into the tubular lumen as 47-amino-acid pro-hBD1, and then undergo proteolytic processing and generate multiple truncated forms. The respective urine and plasma concentrations of hBD-1 in patients with pyelonephritis are 48.1 +/- (SEM) 15.7 pmol/mg creatinine and 2.66 +/- 0.41 pmol/ml, 3.1-fold and 1.8-fold those of normal individuals. hBD-1 is thought to contribute to mucosal defense in the urinary tract. Our findings provide a better understanding of the biosynthesis of this peptide and its pathophysiological significance in infectious diseases.     

Allelic loss in ovarian cancer

Loss of heterozygosity (LOH) was examined at 86 loci distributed on every chromosomal arm in 50 human ovarian tumors. Frequent allele losses were observed on chromosomes 13q (42%), 17p (42%), 17q (45%), and Xp (41%). Deletion mapping on chromosome 17 revealed a candidate gene on the long arm distal to D17S41/S74 for ovarian cancer which is distant from the locus for early onset breast cancer. LOH on chromosome 17q was found to be concordant with LOH on chromosomes 3p, 13q, 17p and Xp suggesting that it may be an early event in neoplastic development. These findings indicate that multiple tumor-suppressor genes for ovarian cancer possibly exist on chromosomes 13q, 17, and/or Xp and provide the basis for the identification of candidate gene(s) associated with ovarian cancer. The chromosomal mechanisms resulting in allele losses in ovarian cancer include deletion, deletion/duplication, mitotic recombination and monosomy, in concordance with the developed genetic model.     

Antinociceptive effect of the combination of pentazocine with morphine in the tail-immersion and scald-pain tests in rats

We investigated the antinociceptive effect of pentazocine hydrochloride (pentazocine) in combination with morphine hydrochloride (morphine) using two antinociceptive tests; i.e., the tail-immersion and scald-pain tests, in rats. In the tail-immersion test, the rat's tail was immersed in warm water at 47 degrees C, and the latency to a nociceptive response was measured. In the scald-pain test, the right hind foot was scalded by immersion into hot water at 57 degrees C. Two hours later, additional thermal stimulus was applied to the same foot, and the latency to a nociceptive response was measured. Subcutaneous treatment with either pentazocine (6, 12, 24 mg/kg) or morphine (1.5, 3, 6 mg/kg) alone dose-dependently showed antinociceptive effects in both tests. The ED50 values (95% confidence limit) of pentazocine and morphine were 13.0 (5.4-31.5) and 2.4 (1.6-3.7) mg/kg in the tail-immersion test and 11.0 (4.5-26.6) and 3.8 (1.8-7.2) mg/kg in the scald-pain test, respectively. Simultaneous treatment with pentazocine at the similar dose augmented the morphine (1.5 mg/kg)-induced antinociception, but did not diminish the morphine (6 mg/kg)-induced antinociception in both tests. These results suggest that the simultaneous administration of pentazocine at the antinociceptive dose and morphine exerts additional antinociceptive activity against thermal and scald-induced inflammatory pain.