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TAN-999 and TAN-1030A, new indolocarbazole alkaloids with macrophage-activating properties.

Two new indolocarbazole alkaloids, TAN-999 and TAN-1030A, were isolated from culture broths of Nocardiopsis dassonvillei C-71425 and Streptomyces sp. C-71799, respectively. Their structures were elucidated on the basis of their reactions, spectroscopic analyses and in particular, comparison of spectral data with that of staurosporine. These metabolites induced spreading of a murine macrophage cell line, Mm 1. They also augmented the phagocytic activity, Fc gamma receptor expression and beta-glucuronidase activity of murine macrophage cell lines, Mm 1 and J774A.1. When proteose-peptone elicited peritoneal macrophages from mice were incubated with these metabolites for 2 days, the phagocytosis-dependent respiratory burst of these cells was enhanced. Similar enhancement was also observed when the peritoneal macrophages in mice were modulated by intraperitoneal administration of these metabolites. These results reveal that TAN-999 and TAN-1030A can activate macrophage functions in mice.

Alkaloids

North Carolina tanning operators: hazard on the horizon?

BACKGROUND: Previous small studies have demonstrated that operators of commercial tanning equipment may not be adequately trained. Most states have no training requirements for tanning operators. This study was designed to assess the medically relevant knowledge of cosmetic tanning salon operators and relate these to demographic variables. METHODS: A written, anonymous questionnaire was distributed to 984 attendees of the first required operator training seminars conducted in North Carolina. Between July 1990 and June 1991, proprietors and their employees of the commercial tanning industry who attended these seminars were surveyed. No conference attendee refused to complete the questionnaire. Demographic information and responses to the medical information section of the questionnaire were scored by computer. RESULTS: Important findings included that 71% of surveyed operators had never received any training for their current position. Operators of cosmetic tanning equipment were often unaware of contraindications to tanning such as photosensitizing drugs and specific medical conditions. Few operators recognized methoxsalen (26%) or trisoralen (34%) or their respective proprietary names as contraindications to tanning. Operator knowledge was not related to their educational achievement, previous tanning salon experience, or prior tanning industry-sponsored training. Children younger than 10 years old were tanned by 2.6% of the operators. CONCLUSION: These data suggest that the public may be at risk for UV light-induced diseases and burns because of operator lack of knowledge.

Adult

Immunogenicity of glutaraldehyde-tanned bovine pericardium.

Glutaraldehyde-tanned bovine pericardium was tested for its ability to induce immunologic responses in vivo. Sections of glutaraldehyde-tanned bovine pericardium were implanted between the abdominal muscles of rats and guinea pigs. Control animals received Dacron implants. Lymphocytes and sera from animals were isolated at 2 and 4 weeks after implantation (four animals per group per time). Tritiated thymidine incorporation and an enzyme-linked immunosorbent assay were used to measure T- and B-lymphocyte responses to glutaraldehyde-tanned bovine pericardium antigens. At the same time points, implants and surrounding tissue from all animals were processed for histologic data. Results show that T-lymphocytes from animals with glutaraldehyde-tanned bovine pericardium implants responded significantly (p less than 0.001) to glutaraldehyde-tanned bovine pericardium antigens in vitro but not to Dacron. In contrast, lymphocytes from animals with Dacron implants failed to respond to glutaraldehyde-tanned bovine pericardium or Dacron preparations. Results of enzyme-linked immunosorbent assay show that animals with glutaraldehyde-tanned bovine pericardium implants produced antibody directed against glutaraldehyde-tanned bovine pericardium antigens. Histologic study revealed a dense mononuclear and multinuclear giant cell infiltrate at the interface between glutaraldehyde-tanned bovine pericardium and surrounding host tissues, with focal degradation of implant collagen. Dacron elicited a nonspecific lymphocytic and foreign body-type reaction. These results indicate that glutaraldehyde-tanned bovine pericardium can induce immunologic responses in vivo consistent with a host-versus-graft reaction.

Abdominal Muscles

Synthesis and biological activity of (S)-2-amino-3-(2,5-dihydro-5-oxo-4-isoxazolyl)propanoic acid (TAN-950 A) derivatives.

(S)-2-Amino-3-(2,5-dihydro-5-oxo-4-isoxazolyl)propanoic acid (TAN-950 A (1)) is a novel amino acid antibiotic which shows a high affinity for glutamate receptors of the central nervous system. To improve the affinity for glutamate receptors, the structure-activity relationships of TAN-950 A derivatives 6a--o, 15a--o were investigated. Optically active TAN-950 A analogs 15a--h were synthesized starting with methyl (S)- and (R)-N-Boc-pyroglutamate (8) via acylation at the C-4 position followed by isoxazolone formation with hydroxylamine and subsequent deprotection reactions. The lactam 16, prepared from (RS)-aminoadipic acid, and dimethyl esters 19 of (R)- and (S)-aspartic acid were converted to (RS)-3-methyl-homo-TAN-950 A (15i) and optically active nor-TAN-950 A derivatives 15j--o, respectively, utilizing a similar sequence of reactions. Most of TAN-950 A derivatives 6a--o, 15a--o showed an affinity for glutamate receptors. The 3-alkyl derivatives 15b, d--g, especially, showed a high affinity for the quisqualate subtype-receptor and had a strong activating effect on the hippocampal neurons (glutamate agonistic activity). The (R)-enantiomer 15a of TAN-950 A had increased selectivity for the N-methyl-D-aspartate (NMDA) subtype-receptor. This selectivity was further enhanced by removal of the methylene group in the amino acid moiety of 15a. The most potent and selective NMDA agonistic activity was observed with (R)-3-methyl-nor-TAN-950 A (15m).

Animals

Longwave ultraviolet radiation (UVA, 320-400 nm)-induced tan protects human skin against further UVA injury.

The protective effect of a UVA (320-400 nm) induced tan against cutaneous injury by further UVA-irradiation was studied by evaluating the histopathologic changes in tanned and untanned normal human buttock skin 24 h after exposure to 2 and 4 minimal erythema doses of UVA. In each subject there were fewer polymorphonuclear leukocytes and less endothelial cell prominence and vessel wall necrosis in the UVA tanned skin than in the untanned UVA-irradiated skin. In the tanned control and tanned UVA-irradiated skin there was a prominent mononuclear cell inflammatory infiltrate that was much greater than in untanned skin. In immunoperoxidase stained tissue sections, the mononuclear cells were predominantly T cells, and in all of the specimens the number of phenotypic helper/inducer cells exceeded the phenotypic cytotoxic/suppressor cells. This demonstrates that a UVA tan provides photoprotection against acute UVA exposure. In addition, tanning, with or without further UVA-irradiation, was associated with a mononuclear cell inflammatory infiltrate.

Adult

TAN-931, a novel nonsteroidal aromatase inhibitor produced by Penicillium funiculosum No. 8974. I. Taxonomy, fermentation, isolation,characterization and biological activities.

A novel nonsteroidal aromatase inhibitor, TAN-931, was isolated from the culture filtrate of a soil isolate fungus, No. 8974. The strain was identified as Penicillium funiculosum No. 8974. TAN-931 inhibited human placental and rat ovarian aromatase activity, and the IC50 value was 17.2 and 162 microM, respectively. The inhibition of human placental aromatase was uncompetitive with respect to androstenedione conversion with a Ki value of 40 microM. When TAN-931 was subcutaneously administered at doses of 25, 50 and 100 mg/kg (once/day, x4) to 20-day-old female Sprague-Dawley rats treated with gonadotropin, the plasma estradiol-17 beta level and the weight of ovaries and uterus were markedly reduced in a dose-dependent manner. The in vivo inhibitory activity of TAN-931 was more potent than that of 4-hydroxyandrostenedione. Consecutive administration of TAN-931 (100 mg/kg, sc, twice/day, x 7) to 9-week-old male Sprague-Dawley rats did not induce any adrenal hypertrophy even though administration of aminoglutethimide caused 2-fold enlargement of the adrenal under the same conditions. Specific binding of TAN-931 to the estrogen receptor from a human breast cancer cell line, MCF-7, was not detected.

Adrenal Glands

Photocarcinogenesis in hairless mice induced by ultraviolet A tanning devices with or without subsequent solar-simulated ultraviolet irradiation.

The carcinogenic effect of 3 commercially available ultraviolet A (UVA) tanning sources was studied in lightly pigmented hairless mice. The tanning sources (Bellarium-S SA-1-12 and Philips TL 09R and TL 10R) have different emission spectra and emit different quantities of UVB. The tanning sources were administered either alone, or before irradiation with solar-simulated UV (solar UV). All 3 UVA tanning sources were able to induce skin tumors when administered in daily doses resembling those used in tanning salons (20 min/d, 5 d/week). Irradiation with Bellarium-S during 32 weeks induced skin tumors in all mice; a similar response was seen after 66 weeks of irradiation with Philips TL 09R. Irradiation with Philips TL 10R during 98 weeks induced tumors in 6 of 20 mice. Nine groups of 20 mice were pretreated 20 min/d, 5 d/week during 13 weeks with one of the UVA tanning sources. Three groups were irradiated with Bellarium-S, 3 groups with Philips TL 09R and 3 groups with Philips TL 10R in daily doses ranging from 0.2 to 1.8 minimum erythema doses (MED). The highest daily doses were equivalent to the doses received during one session in a commercial solarium. Subsequently all 9 groups were irradiated with 3.1 MED/d solar UV 10 min/d, 4 d/week until all mice had died. Time to first tumor was compared. All groups pretreated with Bellarium-S and Philips TL 09R showed an enhanced tumor development compared with a group irradiated with solar UV only. Pretreatment with Philips TL 10R did not enhance the carcinogenic effect of solar UV.

Animals

UVA tanning devices interact with solar-simulated UV radiation in skin tumor development in hairless mice.

The carcinogenic effect of three UVA tanning sources was studied in lightly pigmented hairless mice. The three tanning sources (Bellarium-S SA-1-12, Philips TL 09R and Philips TL 10R) have different emission spectra, and emit different amounts of UVB. Radiation from the tanning sources was administered for 20 min/day, 5 day/week in daily doses equivalent to those used in suntan salons. The radiation was given alone or after 12 weeks of exposure to solar-simulated UV radiation (SOLAR UV) (10 min/day, 4 day/week; daily dose, 19.5 kJ/m2 UVA and 3.9 kJ/m2 UVB). Irradiation with Bellarium-S SA-1-12 for 47 weeks and Philips TL 09R for 74 weeks induced skin tumours in 20/20 and 13/20 of the animals, respectively. When irradiation with Bellarium-S SA-1-12 and PHilips TL 09R was administered after 12 weeks of SOLAR-UV exposure, a strong enhancement of SOLAR-UV-induced photocarcinogenesis was observed (p < 0.001). Irradiation with Philips TL 10R was only slightly carcinogenic, and during 85 weeks of irradiation only one skin tumor appeared in a group of 20 mice. However, when irradiation with Philips TL 10R was administered after 12 weeks of exposure to SOLAR UV, an enhancement of SOLAR-UV-induced carcinogenesis was observed (p < 0.001). Our results suggest that the hazards of exposure to commercial tanning devices are increased when they are used after a period of natural sun exposure. Even tanning sources with a low carcinogenic potential are able to increase SOLAR-UV-induced carcinogenesis significantly.

Animals

Photoprotective effect of a psoralen-UVA-induced tan.

To determine whether a tan produced by 8-MOP and UVA protects from subsequent solar light irradiation, volunteers were irradiated with unfiltered Xenon arc light before and 10 days after a 1 week's course of four 8-MOP-UVA treatments. Evaluation of the minimal erythema doses and of histological changes before and after 8-MOP-UVA treatment revealed that the 8-MOP-UVA induced tan protected against the erythemogenic and cell damaging effects of Xenon arc light. Unscheduled repair DNA synthesis, used as a measure for UVB-induced DNA damage and repair, was also investigated in skin irradiated with the Xenon arc before and after 8-MOP-UVA induced tanning. Both the number of grains per sparse labeled cell and the number of sparse labeled cells per 1000 cells, were found to be significantly lower in tanned skin; taking decreased unschedules repair DNA synthesis as a measure for decreased DNA-damage, these findings also demonstrate a photoprotective effect of the 8--MOP-UVA induced tan.

DNA Repair

The effect of the anoxic radiosensitizing agent TAN on induction of revertants by gamma-rays and helium ions in Salmonella tester strains.

The modification effect of the anoxic radiosensitizer TAN on the mutagenesis in various Salmonella tester strains after gamma-ray and helium ion irradiation was studied. The oxygen enhancement ratios (OER) for all 3 strains on the lethal assay after gamma-irradiation are approximately equal to 2. The induction of reversions in TA98 and TA100 does not modify under anoxia. The value of OER on the mutagenic assay in TA102 equals 1.6. The OER after helium ion irradiation on the lethal and mutagenic assays was less than after gamma-irradiation. The mutagenesis in 3 strains after irradiation under anoxia is enhanced by TAN. The value of the TAN modification effect after gamma-irradiation increases from 2.1 +/- 0.2 for TA102 to 5.2 +/- 0.4 for TA100. However, the TAN influence on mutagenesis in TA100 after helium ion irradiation decreases to 3.1 +/- 0.3. We conclude that peculiarities of mutagenesis in various tester strains under anoxia with TAN can be explained by considering the nature of premutational DNA damages.

Anaerobiosis

TAN-1, the human homolog of the Drosophila notch gene, is broken by chromosomal translocations in T lymphoblastic neoplasms.

Previously we described joining of DNA in the beta T cell receptor gene to DNA of an uncharacterized locus in a t(7;9)(q34;q34.3) chromosomal translocation from a case of human T lymphoblastic leukemia (T-ALL). We now show that the locus on chromosome 9 contains a gene highly homologous to the Drosophila gene Notch. Transcripts of the human gene, for which we propose the name TAN-1, and its murine counterpart are present in many normal human fetal and adult mouse tissues, but are most abundant in lymphoid tissues. In t(7;9)(q34;q34.3) translocations from three cases of T-ALL, the breakpoints occur within 100 bp of an intron in TAN-1, resulting in truncation of TAN-1 transcripts. These observations suggest that TAN-1 may be important for normal lymphocyte function and that alteration of TAN-1 may play a role in the pathogenesis of some T cell neoplasms.

Amino Acid Sequence

Tanning salons: an area survey of proprietors' knowledge of risks and precautions.

An area survey of tanning salon proprietors was conducted in a medium-sized midwestern city. Proprietors reported they were in compliance with federal safety regulations, but not all had age, frequency, or duration restrictions. Similarly, proprietors were not uniformly informing patrons of potential tanning hazards, including the possibility of skin cancer, and were not knowledgeable about the risk and benefits of tanning. Some establishments reported selling psoralens to patrons to enhance tanning. More explicit guidelines regarding the use of tanning equipment and more accurate consumer information are needed.

Adolescent

A novel glutamate agonist, TAN-950 A, isolated from streptomycetes.

A novel antifungal amino acid antibiotic, TAN-950 A ([S]-2-amino-3-(2,5-dihydro-5-oxo-4-isoxazolyl)propanoic acid), was found to have affinity for three excitatory amino acid (EAA) receptors and to inhibit [3H]alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid ([3H]AMPA), [3H]kainate and [3H]3-(2-carboxypiperazin-4-yl)propyl-1-phosphonic acid ([3H]CPP) binding competitively. It caused excitation of rat hippocampal CA1 neurons in vitro, an effect that was antagonized by an AMPA/kainate antagonist, 6,7-dinitroquinoxaline-2,3-dione (DNQX). Chemical modification of TAN-950 A brought about a large change in its pharmacological activity. Alkylation at the C-3 position of the isoxazolone ring markedly increased the ability to elicite neuronal firing. This agonistic effect was also antagonized by DNQX. The (R) enantiomer of TAN-950 A had increased selectivity for the N-methyl-D-aspartate (NMDA) receptor subtype. This selectivity was further enhanced by removal of the methylene group from the amino acid moiety. The most potent NMDA agonistic activity was observed with [R]-2-amino-2-(2,5-dihydro-3-methyl-5-oxo-4-isoxazolyl)acetic acid. These derivatives of TAN-950 A might be useful agents for investigating the pharmacological and physiological roles of EAA receptors.

Animals

Effect of traditional Chinese herbal medicines on the pharmacokinetics of western drugs in Sprague-Dawley rats of different ages (II): Aminophylline-huan shao tan and aminophylline-pu chung yi chi tang.

The effect of Chinese herbal medicines (Huan Shao Tan and Pu Chung Yi Chi Tang) and western drugs (sodium phenobarbital and cimetidine) on the serum concentration and pharmacokinetic parameters of theophylline and cytochrome P-450 of Sprague-Dawley (SD) rats of three different ages were examined. The older rats without pretreatment with Chinese herbal medicines and western drugs exhibited higher serum theophylline concentration and lower pharmacokinetic parameters of theophylline than middle-aged and younger rats (P < 0.05), but there was no difference in cytochrome P-450 activity among the three different ages of rats. All rats when pretreated with sodium phenobarbital showed lower serum theophylline concentration and higher pharmacokinetics parameters of theophylline. Also, the activity of cytochrome P-450 was higher (P < 0.05). When cimetidine was pre-administered in SD rats of three age groups, all rats exhibited lower serum theophylline concentration and higher pharmacokinetics parameters (P < 0.05), but the activity of cytochrome P-450 remained unchanged (P > 0.05). The results were opposite to other studies, probably because the dose and dosing intervals were different. No single effect occurred on the younger and middle-aged rats after pretreatment with Huan Shao Tan and Pu Chung Yi Chi Tang: their serum theophylline concentration, pharmacokinetics parameters and cytochrome P-450 activity were the same as the control group. However, the older rats after pretreatment with Huan Shao Tan or Pu Chung Yi Chi Tang showed lower serum theophylline concentration and higher pharmacokinetics parameters than the younger and middle-aged rats pretreated with similar Chinese herbal medicines. This indicates that Huan Shao Tan and Pu Chung Yi Chi Tang may perhaps improve the elimination of theophylline in older rats. This might be attributed to the increase in hepatic blood flow or in liver volume, since the activity of cytochrome P-450 was not affected by the administration of Chinese herbal medicines.

Aging

Production and biological activities of a new antifungal antibiotic, TAN-950 A.

A novel antifungal antibiotic, TAN-950 complex, was isolated from the culture filtrate of Streptomyces platensis A-136 (IFO 14603, FERM BP-1786). The water-soluble amphoteric substances in this complex were purified by chromatography using ion-exchange resins, QAE-Sephadex and adsorptive resins and were designated TAN-950 A and TAN-950 A-E mixture. The molecular formula of TAN-950 A was determined to be C6H7N2O4Na for the sodium salt. This new amino acid antibiotic showed antifungal activity against Candida albicans in vitro and in vivo, and had low toxicity in mice.

Animals