The expression of HMGA genes is regulated by their 3'UTR.

Many benign mesenchymal tumors are characterized by chromosomal abnormalities of the regions 12q15 or 6p21.3 leading to aberrant expression of either HMGA2 (formerly HMGIC) or HMGA1 (formerly HMGIY). The proteins of both genes belong to the HMGA (formerly HMGI(Y)) family of architectural transcription factors. As a rule, aberrant HMGA transcripts found in a variety of benign tumors have intact coding regions at least for the DNA binding domains with a truncation of their 3' untranslated regions. Adding this to the finding that an altered HMGA protein level is not always correlated with an increased amount of corresponding mRNA indicates a posttranscriptional expression control mediated by regulatory elements within the 3'UTR. To check if HMGA expression is under control of such elements we performed luciferase assays with several HMGA2 and HMGA1 3'UTRs of different length. Experiments showed that an up to 12-fold increase in luciferase activity is obtained by the truncation of the 3'UTRs suggesting that the expression of HMGA2 and HMGA1 is controlled by negatively acting regulatory elements within their 3'UTR. Chromosomal aberrations affecting the HMGA genes may therefore influence their expression by an altered stability of the truncated transcripts as a result of the cytogenetic aberrations.

Regulation of the expression of HMG1, a co-activator of the estrogen receptor.

HMG1 is a protein of high clinical significance. Besides shielding of DNA adducts against repair enzymes making cells more sensitive to cisplatin therapy, it is also a co-modulator of the activity of steroid hormone-regulated genes. Although HMG1 is regulated by various factors, including steroid hormone estrogen, nothing was known about regulatory sequences. Also the sequence of parts of HMG1 including its promoter remains still unknown. We have completed the genomic organization of human HMG1 and characterized its regulatory region by luciferase assay for promoter activity. Insights into the regulation of HMG1 expression are given by the promoter analysis showing a strong functional promoter, enhancing and negative regulatory regions together with a CpG island in intron 1 and several transcription factor binding sites. Furthermore, the finding of two estrogen responsive elements within intron 1 is relevant as they indicate a direct mechanism of HMG1 up-regulation by estrogen making the presence of an estrogen receptor a significant marker for a combined treatment of special tumor types with estrogen and the anticancer drugs cisplatin or carboplatin.

Variation of HMGB1 expression in breast cancer.

The amount of steroid hormone receptor proteins does not always correlate with the response of breast cancers to endocrine therapy. This may partly be due to the fact that binding of the estrogen receptor (ER) to estrogen responsive elements (ERE) of its target genes is mediated by additional cellular proteins. One of these is the high mobility group protein HMGB1, known to interact with ER thus dramatically increasing its binding to ERE. This is the first report analysing the expression patterns of HMGB1 in breast cancer cells. Northern blot analyses of the 1.4 kb and the 2.4 kb transcripts of HMGB1 in 13 breast cancer samples revealed a strong intertumoural variation by a factor of 8.5 and 14.5, respectively. This variation may contribute to the different response, of estrogen receptor-positive breast tumours to endocrine therapy, making HMGB1 a marker of considerable clinical interest.

HMGIY is the target of 6p21.3 rearrangements in various benign mesenchymal tumors.

Specific chromosomal abnormalities of chromosomal region 6p21.3 have been described in subsets of many benign mesenchymal tumors. In the presented study, we investigated a series of 36 such cases by FISH, and Southern blot analyses for HMGIY rearrangements. FISH results revealed that the chromosomal breakpoints of 11 pulmonary chondroid hamartomas (PCHs), 12 endometrial polyps (EPs), one lipoma, and two uterine leiomyomas (ULs) were located within a 80 kb region surrounding the HMGIY gene. In 11 PCHs and one UL the breakpoints were located 3' of HMGIY, and one PCH showed a breakpoint 5' of HMGIY. Southern blot analyses with intra- and extragenic probes were performed of primary tumor material or cell lines from one UL, three PCHs, and five EPs. In none of these cases was an intragenic rearrangement found. Finally, we were able to detect expression of truncated HMGIY transcripts by 3'-RACE PCR. Our data clearly show the role of a further member of the HMGI family in the development of benign mesenchymal tumors. Although most of the breakpoints of the chromosomal translocations involving HMGIY are located outside the gene, aberrant transcripts resembling the structure of those observed in the case of HMGIC have been found. Our molecular investigations thus led to the identification of the molecular mechanism by which rearrangements of either of two closely related genes lead to the development of frequent benign mesenchymal tumors in humans.

Ofloxacin in human serum, urine, and tear film after topical application.

We evaluated the systemic absorption of ofloxacin eyedrops in humans and their availability in the tear film. Serum, urine, and tear film concentrations of ofloxacin were measured in 30 healthy women topically treated with 0.3% ofloxacin, in both eyes, four times daily for 10 1/2 days. Serum was collected before the first daily dose on days 1 and 11 and at 18 time points before the second dose. Maximum serum ofloxacin concentrations (1.89 +/- 1.13 ng/ml) after 10 1/2 days of topical dosing were more than 1,000 times lower than those reported after standard oral doses of 300 mg ofloxacin. Urine was collected for the 24-h period after the first daily dose on days 1 and 10. Topical ofloxacin was excreted in the urine primarily in unmodified form and recovery rates were significantly higher on day 10 (76.1 +/- 41.5%) than on day 1 (56.6 +/- 31.6%) (p less than 0.05). Both serum and urine data give evidence to accumulations of ofloxacin over a 10 1/2-day period. The low serum concentration at steady state suggests an extremely low potential for producing systemic effects. No systemic side effects attributable to topical ofloxacin were observed. Mild ocular irritation was reported by two patients while under treatment. Tears were collected 4 h after the first treatment on day 11 and at 5, 10, 20, 30, and 40 min after the second treatment.(ABSTRACT TRUNCATED AT 250 WORDS)

The comparative in vitro activity of ofloxacin and selected ophthalmic antimicrobial agents against ocular bacterial isolates.

The in vitro activity of ofloxacin, a new fluoroquinolone anti-infective agent, was evaluated against 419 ocular bacterial isolates of 55 species to determine its potential as a topical agent for the treatment of ocular infections. Other agents tested in this study, in which a modified tube-dilution procedure was used, include norfloxacin, gentamicin, tobramycin, chloramphenicol, and polymyxin B. Ofloxacin demonstrated good to excellent activity against a variety of gram-positive and gram-negative pathogens. The minimum inhibitory concentration against 90% of all bacterial strains tested (MIC90) of ofloxacin was 0.5 microgram/ml for Staphylococcus aureus and S. epidermidis, 2 micrograms/ml for Streptococcus pneumoniae, and 4 micrograms/ml for Pseudomonas aeruginosa. These species were more susceptible to ofloxacin than to any of the nonquinolones tested. The MIC90 of ofloxacin was lower than that of norfloxacin, another quinolone, against S. aureus, S. epidermidis, and St. pneumoniae and equal to that of norfloxacin against P. aeruginosa. Because of its broad spectrum of activity and excellent in vitro activity, we concluded that ofloxacin has the potential for development into a superior topical treatment for ocular infection.

[Dipivefrin: its mechanism of action, clinical efficacy and good tolerance]. / Dipivéfrine, son mécanisme d'action, son efficacité clinique, sa bonne tolérance.

Dipivefrin (DPE) is the first prodrug in general use in ophthalmology. We will discuss the metabolism of DPE to epinephrine within the eye, and the efficacy and safety of DPE in the treatment of elevated intraocular pressure. In addition, we will address the incidence of adverse systemic effects, ocular tolerance and allergy, and the ocular hypotensive efficacy when DPE is used alone or in combination with other antiglaucoma medications.

Ophthalmic rods. New ocular drug delivery devices.

Ophthalmic Rods, new drug delivery devices for ophthalmic medications, are 2-inch-long plastic rods coated with an ocular diagnostic or therapeutic agent. When the drug-coated tip of the rod is brought into contact with the conjunctiva, the medication dissolves into the tear film. We evaluated the safety, comfort, and ease of use of Ophthalmic Rods coated with fluorescein (30 micrograms) in 28 volunteers. Seventy-nine percent (22 of 28) of the patients rated the device as superior to eyedrops, citing cleanliness, comfort, and ease of application as the primary advantages. Ophthalmic Rods effectively delivered fluorescein to the eye and were found to be safe in patients with various refractive conditions, including those with compromised near vision and accommodation.