Microbiological Evaluation of Opened Saline Bottles for Scleral Lens Use and Hygiene Habits of Scleral Lens Patients.

SIGNIFICANCE: Scleral lenses have become a widely used treatment option for patients with irregular corneas and ocular surface disease. Successful wear entails use of a nonpreserved saline solution to fill the lens before application on the eye. PURPOSE: The purposes of this study were to evaluate solution from opened bottles of multidose preservative-free saline for microbiological growth and to better understand study participant hygiene habits while handling these bottles for scleral lens wear. METHODS: Eligible study participants in this single-center prospective study were patients who routinely used multidose preservative-free saline solution for scleral lens rinsing and filling. Study participants completed a 12-question survey regarding their scleral lens hygiene habits and donated their opened multidose preservative-free saline bottle (PuriLens Plus; The Lifestyle Company, Inc., Freehold, NJ), which was processed for bacterial and fungal cultures. RESULTS: Thirty-five participants (19 males, 16 females) with ages ranging from 6 to 81 years (mean, 47.9 years) were included. Indications for scleral lens wear included those with irregular corneas and ocular surface disease. The overall rate of microbial contamination among saline samples was 62.9% (n = 22). Twenty-one different microorganisms were identified. The survey responses did not differ significantly (P > .05) for any of the questions with regard to likelihood of positive culture. There were no significant age or sex differences between participants with positive or negative culture results. No significant differences were found between isolation of specific microorganisms and any of the survey responses. CONCLUSIONS: This study suggests that off-label multidose preservative-free saline commonly used to rinse and fill scleral lenses before application on the eye may become contaminated with microorganisms once the bottle has been opened. Eye care practitioners and scleral lens patients should be aware of these potential contaminations and prioritize lens, hand, and environmental hygiene to minimize the risk of ocular complications.

Progestin-induced apoptosis in the Macaque ovarian epithelium: differential regulation of transforming growth factor-beta.

BACKGROUND: Oral contraceptive (OC) use is associated with a reduced risk of ovarian cancer. An OC component, progestin, induces apoptosis in the primate ovarian epithelium. One regulator of apoptosis is transforming growth factor-beta (TGF-beta). We determined the effect of progestin on TGF-beta expression in the primate ovarian epithelium and examined the relationship between TGF-beta expression and apoptosis. METHODS: Female cynomolgus macaques were randomly assigned to receive a diet for 35 months containing no hormones (n = 20); the OC Triphasil (n = 17); or each of its constituents, ethinyl estradiol (estrogen, n = 20) or levonorgestrel (progestin, n = 18 ), alone. Ovarian sections were immunostained with monoclonal antibodies against TGF-beta1 or TGF-beta2 plus TGF-beta3 (TGF-beta2/3) isoforms. The expression of TGF-beta isoforms in four ovarian compartments (epithelium, oocytes, granulosa cells, and hilar vascular endothelium) was compared among treatment groups. The association between TGF-beta expression and apoptosis, as determined by morphology and histochemistry, was examined in ovarian epithelium. All statistical tests were two-sided. RESULTS: Compared with ovaries from the control and estrogen-only-treated monkeys, the ovaries of progestin-treated monkeys showed 1) a marked decrease in the expression of TGF-beta1 and a concomitant increase in the expression of the TGF-beta2/3 isoforms in the ovarian epithelium (P<.001), 2) an increase in the expression of TGF-beta2/3 in the hilar vascular endothelium (P<.001), and 3) a marked decrease in TGF-beta2/3 expression in granulosa cells (P<.001). The apoptotic index of the ovarian epithelium was highly associated with the change in expression from TGF-beta1 (P<.001) to TGF-beta2/3 (P

The effect of posttranslational modifications on the in vitro activity of recombinant human thyroid-stimulating hormone.

Posttranslational modification can influence the biologic activity of recombinant proteins. The effects of beta-subunit C-terminal truncation, oligosaccharide heterogeneity, and chemical oxidation on the in vitro activity of recombinant human thyroid-stimulating hormone (rhTSH) were investigated. beta-Subunit C-terminal truncation up to residue 113 did not effect the in vitro activity of the hormone. The relationship between the heterogeneity of oligosaccharide structures on rhTSH and specific activity of the glycoprotein hormone was also examined. Oligosaccharide profiles were generated for preparations of rhTSH containing similar sialic acid levels. A weak correlation was observed between relative levels of monosialylated biantennary, bisialylated biantennary, and trisialylated triantennary oligosaccharide species and in vitro activity of the recombinant hormone (p < 0.05). To examine the effect of chemically induced methionine oxidation on the activity of rhTSH, the hormone was treated with tert-butyl hydroperoxide and then characterized. Using peptide mapping and mass spectrometry, the degree of oxidation of the five methionine residues within rhTSH was measured. Met-71 in the alpha-subunit was the most susceptible to oxidation whereas Met-9 in the beta-subunit was the most resistant. Also, after tert-butyl hydroperoxide treatment, levels of oxidation of Met-32 in the beta-subunit, and Met-29 and Met-47 in the alpha-subunit were less than half of that observed for Met-71. The in vitro activity of rhTSH initially declined with increasing oxidation; however, the loss in activity plateaued at approximately 50% of the control sample activity. In summary, despite the possible effects that posttranslational modifications may have on the bioactivity of a protein, a limited degree of variation in bioactivity was observed for the rhTSH preparations described in this study.

Modulation of ATP-induced calcium signaling by progesterone in T47D-Y breast cancer cells.

Extracellular ATP activates purinergic (P(2)) receptors with an increase in intracellular calcium and phosphorylation of MAPK. In this study we have investigated the effect of progesterone/progestin on ATP-induced calcium mobilization and phosphorylation of the kinase ERK in the T47D-Y breast cancer cell line that exhibits no detectable nuclear progesterone receptor expression. Brief pretreatment with progesterone/progestin results in a dose dependent inhibition of ATP-induced intracellular calcium mobilization, and inhibition of ERK phosphorylation. Response to a cell impermeable ligand and inhibition of the response by an inactivating antibody suggests a mechanism of action at the plasma membrane. These results in T47D-Y cells strongly suggest that progesterone can act in a rapid non-nuclear manner to inhibit extracellular ATP effects on intracellular calcium mobilization and ERK activation. This research provides an example of progesterone action in a breast cancer cell line lacking expression of the classical nuclear progesterone receptors.

Successful pregnancies in patients with estrogenic anovulation after low-dose human chorionic gonadotropin therapy alone following hMG for controlled ovarian hyperstimulation.

OBJECTIVE: To demonstrate that folliculogenesis can be sustained with 200 IU human chorionic gonadotropins (hCG) after FSH-priming and result in pregnancy in women with estrogenic ovulatory dysfunction and risk factors for severe ovarian hyperstimulation syndrome (OHSS). CASE REPORT: Three women with infertility associated with estrogenic ovulatory dysfunction and hyperinsulinemia who appeared to be at high risk for severe OHSS during gonadotropin therapy. INTERVENTIONS: After 10 days of receiving either 150 IU hMG or recombinant FSH, patients were switched to 200 IU hCG/day alone for 2-3 days. 5,000 IU of hCG was then administered followed by either home intercourse, intrauterine insemination or transvaginal oocyte retrieval-embryo transfer. MAIN OUTCOME MEASURES: Endovaginal ultrasound measurement of follicle number and size, serum estradiol levels, symptoms of ovarian hyperstimulation, pregnancy test, and evaluation of pregnancy by transvaginal ultrasound. RESULTS: After discontinuation of hMG or recombinant FSH, serum estradiol concentrations continued to rise, and follicles >14 mm continued to grow during low-dose hCG administration. All women conceived without developing symptoms of OHSS. Pregnancy outcomes achieved include a term singleton delivery, a term twin delivery, and triplets delivered at 31 weeks gestation. CONCLUSION: The use of low-dose hCG alone is sufficient for supporting the late stages of folliculogenesis in women with estrogenic ovulatory dysfunction. This ovulation induction regimen appears to support the follicular growth of larger follicles while decreasing the number of smaller preovulatory follicles, thereby reducing a known risk factor for OHSS. We report on the positive pregnancy outcomes in 3 women with estrogenic ovulatory dysfunction and clinically appeared to be at high risk for developing severe OHSS who safely underwent this protocol.

Residency programs and psychotherapy competencies: a survey of chief residents.

OBJECTIVE: To survey chief residents' opinion about various aspects of psychotherapy competency determination. METHODS: Chief residents of various psychiatry residency programs were surveyed. RESULTS: One hundred two chief residents were surveyed. Seventy two (70.58%) completed the survey. Eighty four percent of the respondents reported that they were aware of the competencies. The number of patients required for competency determination in five areas of psychotherapy varied widely among the programs. Global assessment by psychotherapy supervisors was the most commonly used method of competency determination (61%). Nineteen (26%) chief residents opined that not all the faculty members involved in teaching and assessing competencies are qualified to do so. Only 23 (31%) of respondents reported that competency criteria were well integrated into the residency curriculum. CONCLUSION: The little consistency in psychotherapy competency determination across various programs, the differential preparedness of programs for competencies and the lack of consistent integration of competencies into residency curricula call for development and implementation of more uniform assessment methods. This variability also calls into question the decision to establish a standard in five areas of psychotherapy competency.

Lysosomal acid alpha-glucosidase consists of four different peptides processed from a single chain precursor.

Pompe's disease is caused by a deficiency of the lysosomal enzyme acid alpha-glucosidase (GAA). GAA is synthesized as a 110-kDa precursor containing N-linked carbohydrates modified with mannose 6-phosphate groups. Following trafficking to the lysosome, presumably via the mannose 6-phosphate receptor, the 110-kDa precursor undergoes a series of complex proteolytic and N-glycan processing events, yielding major species of 76 and 70 kDa. During a detailed characterization of human placental and recombinant human GAA, we found that the peptides released during proteolytic processing remained tightly associated with the major species. The 76-kDa form (amino acids (aa) 122-782) of GAA is associated with peptides of 3.9 kDa (aa 78-113) and 19.4 kDa (aa 792-952). The 70-kDa form (aa 204-782) contains the 3.9- and 19.4-kDa peptide species as well as a 10.3-kDa species (aa 122-199). A similar set of proteolytic fragments has been identified in hamster GAA, suggesting that the multicomponent character is a general phenomenon. Rabbit anti-peptide antibodies have been generated against sequences in the proteolytic fragments and used to demonstrate the time course of uptake and processing of the recombinant GAA precursor in Pompe's disease fibroblasts. The results indicate that the observed fragments are produced intracellularly in the lysosome and not as a result of nonspecific proteolysis during purification. These data demonstrate that the mature forms of GAA characterized by polypeptides of 76 or 70 kDa are in fact larger molecular mass multicomponent enzyme complexes.