Effect of hyperprolactinemia induced by pituitary grafts or castration on porphyrin content of the mouse Harderian gland.

The histology and porphyrin concentrations of Harderian glands and plasma prolactin levels were examined in B6C3F1 mice castrated or isografted with pituitaries in combination with the administration of bromocriptine (2-bromo-á-ergocryptine), a potent suppressor of prolactin. Four pituitaries were transplanted underneath the bilateral kidney capsules of each mouse. Furthermore, we investigated Harderian porphyrins and plasma testosterone levels in mice that were castrated or treated with neuroleptic butyrophenone (timiperone), a potent accelerator of prolactin, and treated concurrently with bromocriptine. Light microscopically, pituitary grafts increased the concretion of porphyrin pigments within the Harderian gland lumina. Pituitary grafts or castration increased both Harderian porphyrin concentrations and plasma prolactin levels compared with the intact control mice. In pituitary-grafted or castrated mice, bromocriptine distinctly prevented the rise in both porphyrins and prolactin levels. Administration of butyrophenone did not result in any marked change in testosterone levels, although Harderian gland porphyrins were significantly increased. The present results indicate that in mice prolactin stimulates the porphyrin production of the Harderian gland and has an important role in the regulation of Harderian gland porphyrins.

Morphological classification of dental lesions induced by various antitumor drugs in mice.

To characterize and compare maxillary incisor lesions caused by various antitumor drugs, male BALB/c mice were given a single intravenous injection of an estimated 10% lethal dose (LD10)) of 5-fluorouracil (5-FU), adriamycin (ADR), mitomycin C (MMC), vinblastine sulfate (VBL). taxotere (TXR), irinotecan hydrochloride (CPT-11), DX-8951f, or cisplatin (CDDP). After 3, 5, 10, 15, and 60 days, the animals were sacrificed, and the maxillary incisors were examined microscopically. The dental lesions observed were classified into 4 different types on the basis of their morphological features. The lesion due to 5-FU was characterized by focal defects in the dentin, and this injury was reversible (transient dentin injury). ADR- or MMC-induced lesions were defined by abnormal structure of the apical aspect of the tooth and irregular odontogenesis, lasting for a long period (persistent apical injury). Treatment with VBL or TXR showed irregular enamel formation and abnormal dentinogenesis. Their targets were considered to be both immature and mature odontogenic cells (diffuse dental injury). Exposure to CPT-11, DX-8951f, or CDDP elicited minor reductions in a few precursor cells in the epithelial sheath on day 3, but no prominent dental abnormalities were seen thereafter (nontoxic injury). In conclusion, antitumor drugs can cause a variety of dental lesions that vary temporally and spatially, making histopathological examination of the maxillary incisor an important component of the safety assessment process for novel antitumor drugs.

Thyroid hypertrophic effect of semotiadil fumarate, a new calcium antagonist, in rats.

We studied the effects of semotiadil fumarate (SF), a new calcium antagonist with a unique benzothiazine structure, on the thyroid gland and liver in rats and compared them with those of another calcium antagonist, nicardipine (NCD), a well-known thyroidal hypertrophic agent and microsomal enzyme inducer, phenobarbital (PB), and goitrogen propylthiouracil (PTU). In oral 2-week treatment, SF caused increases in hepatic microsomal protein levels, uridine diphosphate glucuronosyltransferase (UDPGT) activity and an increase in serum thyroid stimulating hormone (TSH) level together with decreases in serum thyroid hormone levels. These results suggest that SF accelerates peripheral disposition of thyroid hormones and subsequently stimulates secretion of TSH from the pituitary gland as a compensatory response. PB and NCD had similar effects on the thyroid gland and the liver. PTU showed obvious thyroid hyperplasia and an increase in relative liver weight. Drastic increase in TSH level was observed in the PTU-treated group together with significant decreases in serum thyroid hormone levels and an increase in hepatic UDPGT activity. Histopathologically, PTU depleted the colloids in follicles, suggesting the inhibition of thyroid hormone synthesis. SF, PB and NCD showed thyroidal hyperplasia, but the extent of the change was far more moderate than that induced by PTU. These results indicate that the effect of SF is similar to those of PB and thyroid hypertrophy seen in the oral 2-week treatment with SF, and may be caused by indirectly elevated TSH levels which resulted from the induction of hepatic UDPGT activity.

Invasive pituitary tumors in female F344 rats induced by estradiol dipropionate.

To clarify the histopathological progression of invasive tumors in the pituitary pars distalis due to estrogen, female Fischer 344 (F344) rats were treated subcutaneously with 5 mg/animal of estradiol dipropionate (ED) once every 2 wk for 13 wk. The animals were killed serially at 2-wk intervals during the investigation. The pituitaries with surrounding tissues were examined light microscopically. At week 7, pituitary cells showed proliferation and atypia with formation of blood-filled spaces. Lesions with these characteristics were diagnosed as adenomas. At week 9 or later, neoplastic cells exhibited extensive proliferation and infiltration into the surrounding tissues, suggesting development of carcinoma. Both proliferating cell nuclear antigen (PCNA) and 5-bromo-2'-deoxyuridine (BrdU) labeling index, markers of cell proliferation, were significantly increased in animals with adenoma or carcinoma. To detect sequential changes in pituitary weight, its signal intensity was periodically monitored in identical rats by using magnetic resonance (MR) imaging. The estimated pituitary weights revealed by MR imaging were comparable to the tumor weights obtained from rats at scheduled sacrifices. These results indicate that ED possesses the potential to cause carcinoma in rat pituitary and MR imaging is an effective tool for estimating the pituitary weight.

Epigenetic variation illustrated by DNA methylation patterns of the fragile-X gene FMR1.

Genomic methylation patterns of mammals can vary among individuals and are subject to dynamic changes during development. In order to gain a better understanding of this variation, we have analyzed patterns of cytosine methylation within a 200 bp region at the CpG island of the human FMR1 gene from leukocyte DNA. FMR1 is normally methylated during inactivation of the X chromosome in females and it is also methylated and inactivated upon expansion of CGG repeats in fragile-X syndrome. Patterns of methylation (epigenotypes) were determined by the sequencing of bisulfite-treated alleles from normal males and females and alleles from a family of five brothers who are methylation mosaics and are affected to various degrees by the fragile-X syndrome. Our data indicate that: (i) methylation of individual CpG cytosines is strikingly variable in hypermethylated epigenotypes obtained from a single individual, suggesting that maintenance of cytosine methylation is a dynamic process; (ii) methylation of non-CpG cytosines in the region studied may occur but is rare; (iii) mosaicism of methylation in the analyzed fragile-X males is remarkably similar to that found for the active X and inactive X alleles in normal females, suggesting that the methylation mosaicism of some fragile-X males reflects similar on and off states of FMR1 expression that exist in normal females; (iv) hypermethylation is slightly more pronounced on fragile-X alleles than on normal inactive X alleles of females; (v) the general dichotomy of hypo- and hypermethylated alleles persisted over the 5 year period that separated samplings of the fragile-X males; (vi) methylation variability was most pronounced at a consensus binding sequence for the alpha-PAL transcription factor, a sequence that may play a role in regulating expression of FMR1.

Effect of hyperprolactinemia induced by neuroleptic agent, timiperone, on porphyrin content of mouse harderian gland.

The histology and porphyrin content of the Harderian gland and the serum prolactin levels were examined in male B6C3F1 mice treated with neuroleptic butyrophenones (timiperone and haloperidol) and treated concurrently with timiperone and 2-bromo-alpha-ergocryptine (bromocriptine), a potent suppressor of prolactin. Light-microscopically, both timiperone and haloperidol increased the number of accretions of porphyrin pigment within the Harderian gland lumina. Timiperone treatment of mice increased both the porphyrin content of the Harderian gland and the serum prolactin levels. Administration of bromocriptine to timiperone-treated mice distinctly prevented the rise in both tissue porphyrin and serum prolactin levels. In intact mice, bromocriptine also exerted inhibitory effects on both the Harderian gland porphyrin content and the serum prolactin level. Electron microscopic investigation revealed that the cytoplasm of type A cells in the Harderian glands of mice treated with timiperone contained trilaminar profiles similar to those seen in the intraluminar pigment masses. These results indicate that timiperone accelerates porphyrin secretion from the type A cells of the mouse Harderian gland by increasing the serum prolactin levels.

[Effects of intravitreal levofloxacin on the rabbit retina].

Effects of intravitreal injection of levofloxacin (LVFX) on the electroretinogram (ERG), visual evoked potential (VEP), and retinal histology were studied in 23 albino and 23 pigmented rabbits to establish the non-toxic intravitreal dosage of LVFX. Doses of 200, 500, 1,000 or 2,000 micrograms of LVFX were injected intravitreally. The ERG and VEP were recorded before injection, and 3 hours, 2 days, 1 week, 2 weeks and 4 weeks after injection. The oscillatory potential transiently deteriorated with 1,000 and 2,000 micrograms doses of LVFX in albino and pigmented rabbits. No ERG changes were observed with 200 and 500 micrograms doses. No abnormal changes were observed in the VEP or retinal histology with any doses of LVFX. These results indicate that intravitreal injections of 200 and 500 micrograms of LVFX are nontoxic to the rabbit retina.

[Increased serum alkaline phosphatase induced by DT-5061, an oral contraceptive, in rats].

DT-5061, a steroid oral contraceptive which contains norethisterone (NET) and ethinyl estradiol (EE) in a ratio of 100:7, was administered orally to rats for 14 days in order to investigate a possible origin of the increased serum alkaline phosphatase (ALP). Increased serum total ALP was noted in rats receiving DT-5061 at 5.35 mg/kg/day or EE at 0.35 mg/kg/day. Electrophoresis of serum ALP revealed that both liver and bone-type ALP isozymes were increased in the DT-5061 5.35 mg/kg and EE 0.35 mg/kg groups, and the ratio of increase in the liver-type was greater than that in the bone-type although the increase in concentration of the bone-type was greater as compared to the liver-type. ALP level in the liver was elevated together with an increase in liver weight, consistently with the increased serum liver-type isozyme. However, neither histological changes indicative of cholestasis nor increase in serum leucine aminopeptidase, bilirubin, GOT or GPT were seen. No changes were observed in bone ALP activity; hence inconsistent with the increased serum bone-type isozyme. From these results, it is considered that the increased serum ALP induced by this drug was due to the increased liver-type isozyme induced in the liver and to the increased bone-type isozyme, and among the ingredients of this oral contraceptive, EE was mainly involved in the increased serum ALP induced by this drug.

Oncogenicity studies of the cognition-enhancing agent nefiracetam in mice and rats.

Oncogenicity studies of nefiracetam (N-(2,6-dimethylphenyl)-2-(2-oxo-1-pyrrolidinyl)acetamide, DM-9384, CAS 77191-36-7), a new cognition-enhancing agent, were carried out in male and female mice and rats. The compound was administered in diet for 104 weeks at dosage levels of 30, 90 and 270 mg/kg/d for mice and of 200, 600 and 1800 ppm for rats. The administration of nefiracetam produced no effects on survival, appearance or behavior. Body weights of the high dose male mice and rats were occasionally significantly decreased when compared to the controls. When calculated on a g/animal/d basis, food consumption was sometimes decreased for these male groups. At necropsy, there was no evidence of treatment related changes, nor were these seen on histopathological examination. All microscopic changes seen in mice and rats were of the usual type commonly occurring in untreated aged B6C3F1 mice and F344 rats. In conclusion, the administration of nefiracetam for 24 months to B6C3F1 mice and F344/DuCrj rats produced only slight effects on body weight in the high dose males with a no-effect level of 90 mg/kg/d for mice or 600 ppm for rats. There was no evidence of an oncogenic effect of nefiracetam.

Effect of the new quinolone antibacterial agent levofloxacin on multiple organ carcinogenesis initiated with wide-spectrum carcinogens in rats.

A multiple organ carcinogenesis model was used in male F344 rats to test the carcinogenic potential of (-)-(S)-9-fluoro-2,3-dihydro-3-methyl-10-(4-methyl-1-piperazinyl)-7-oxo- 7H-pyrido[1,2,3-de][1,4] benzoxazine-6-carboxylic acid hemihydrate (levofloxacin, DR-3355, CAS 10086-85-4). After sequential treatment with diethylnitrosamine (DEN: carcinogen for the liver), N-methylnitrosourea (MNU: carcinogen for the esophagus, forestomach and thyroid) and dihydroxy-di-N-propylnitrosamine (DHPN: carcinogen for the lungs, kidney and urinary bladder), rats were treated with DR-3355, N-butyl-N-(4-hydroxybutyl)-nitrosamine (BBN), N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), catechol (CC) or phenobarbital (PB) to examine whether these compounds modified the carcinogenesis in multiple organs. As a result of histopathological examinations at study week 20, DR-3355 did not induce neoplastic lesions, nor did it enhance the occurrences of proliferative preneoplastic lesions. In contrast, BBN increased the incidences of hyperplasias and papillomas of the urinary bladder. CC enhanced the occurrences of hyperplasias and papillomas of the forestomach as well as submucosal glandular growth for the glandular stomach. PB increased the number of altered cell foci in the liver and the incidence of follicular cysts and hyperplasias of the thyroid. These results indicate that DR-3355 is not capable of promoting the development of tumors in rat multiple organs.

Ophthalmotoxicity and ototoxicity of the new quinolone antibacterial agent levofloxacin in Long Evans rats.

An ophthalomo- and ototoxicity study of a new quinolone antibacterial agent, (-)-(S)-9-fluoro-2,3-dihydro-3-methyl-10-(4-methyl-1- piperazinyl)-7-oxo-7H-pyrido[1,2,3-de] [1,4]benzoxazine-6-carboxylic acid hemihydrate (levofloxacin, DR-3355, CAS 100986-85-4) was investigated in Long Evans rats. The rats were orally administered 100 mg/kg of DR-3355, ciprofloxacin (CPFX), norfloxacin (NFLX) or nalidixic acid (NA) for 2 weeks, and the effects on visual and auditory functions were examined. Examination of electroretinograms (ERGs) revealed a decrease in the amplitudes of the a- and b-waves, a prolongation of the latency and diminution or disappearance of oscillatory potential waves in NA treated rats. Similar but milder changes were also noted in the NFLX treated rats. ERGs from DR-3355 or CPFX treated rats were normal. Histopathological examination revealed no changes suggestive of ophthalmotoxicity or ototoxicity in the rats treated with DR-3355, CPFX or NFLX. On the other hand, NA treated rats showed partial loss of the outer hair cells of the organ of Corti in the cochlea, suggesting that NA had slight ototoxicity. DR-3355 did not show any deleterious visual or auditory effects at the dose used in this study.

Transverse-mode stabilized Ga(1-x)Al(x)As visible diode lasers.

Transverse-mode stabilized visible diode lasers in the 0.7-microm wavelength region are fabricated by growing a Ga(1-x)Al(x)As double heterostructure on a grooved GaAs substrate (channeled-substrate-planar structure). The diodes operate stably in the fundamental transverse mode and provide nonstigmatic laser beams. They can be collimated with a 0.5-1-mrad beam divergence using a simple graded-index fiber lens. Corresponding to single-longitudinal-mode operation at current levels above 1.1 times the threshold, a coherence length as long as 14 m is obtained in Michelson interference experiments.