Efficiency of two enucleation methods connected to handmade cloning to produce transgenic porcine embryos.

The purpose of our work was to establish an efficient-oriented enucleation method to produce transgenic embryos with handmade cloning (HMC). After 41-42 h oocytes maturation, the oocytes were further cultured with or without 0.4 microg/ml demecolcine for 45 min [chemically assisted handmade enucleation (CAHE) group vs polar body (PB) oriented handmade enucleation (OHE) group respectively]. After removal of the cumulus cells and partial digestion of the zona pellucida, oocytes with visible extrusion cones and/or polar bodies attached to the surface were subjected to oriented bisection. Putative cytoplasts without extrusion cones or PB were selected as recipients. Two cytoplasts were electrofused with one transgenic fibroblasts expressing green fluorescent protein (GFP), while non-transgenic fibroblasts were used as controls. Reconstructed embryos were cultured in Well of Wells (WOWs) with porcine zygote medium 3 (PZM-3) after activation. Cleavage and blastocyst rates were registered on day 2 and day 7 of in vitro culture respectively. Meanwhile, the total blastocyst cell number was counted on day 7. We found that the difference was only observed between blastocyst rates (38.6 +/- 2% vs 48.1 +/- 3%) of cloned embryos with GFP transgenic fibroblast cells after CAHE vs OHE. With adjusted time-lapse for zonae-free cloned embryos cultured in WOWs with PZM-3, it was obvious that in vitro developmental competence after CAHE was compromised when compared with the OHE method. OHE enucleation method seems to be a potential superior alternative method used for somatic cell nuclear transfer (SCNT) with transgenic fibroblast cells.

Comparison of topical travoprost eye drops given once daily and timolol 0.5% given twice daily in patients with open-angle glaucoma or ocular hypertension.

PURPOSE: This 9-month study compared the intraocular pressure (IOP)-lowering efficacy and safety of once-daily travoprost ophthalmic solutions (0.0015% and 0.004%) with twice-daily timolol 0.5%. PATIENTS AND METHODS: This study was conducted using a double-masked, randomized, parallel-group design; adult patients with open-angle glaucoma or ocular hypertension (IOP between 24 and 36 mm Hg, inclusive at 9 am and between 21 and 36 mm Hg, inclusive, at 11 am and 4 pm on two eligibility visits after an appropriate washout of previous treatments). In both eyes, the travoprost vehicle (placebo) was instilled at 9 am and travoprost (0.0015% or 0.004%) was instilled at 9 pm, or timolol 0.5% was instilled at both times. The primary efficacy variable was mean IOP measured at 9 am, 11 am, and 4 pm at baseline and follow-up visits. RESULTS: Five hundred seventy-three patients were randomized to the study treatments. Mean IOP, which was combined across study visits, was lower with travoprost 0.004% than with timolol 0.5% at 9 am (P = 0.0246), 11 am (P = 0.0039), and 4 pm (P = 0.0004). Intraocular pressure was lower with travoprost 0.004% than with travoprost 0.0015% at 11 am (P = 0.0314), the time of peak drug activity. Mean IOP was consistently lower with travoprost 0.0015% than with timolol 0.5%. Mean IOP reductions from baseline were significantly (P less than equal 0.0001) greater with travoprost 0.004% (8.0-8.9 mm Hg) than with timolol 0.5% (6.3-7.9 mm Hg). The most frequent related adverse events were hyperemia, pruritus, discomfort, pain, and iris pigmentation changes. The local tolerance was better in the timolol group compared with patients receiving travoprost. There were no serious unexpected treatment-related adverse events in any group. CONCLUSIONS: Travoprost 0.004% reduced diurnal mean intraocular pressure significantly more than timolol 0.5%. Both concentrations of travoprost were well tolerated and safe for use in patients with open-angle glaucoma or ocular hypertension.

The impact of glaucoma on the quality of life of patients in Norway. I. Results from a self-administered questionnaire.

PURPOSE: To evaluate the feelings and experiences of patients living with glaucoma. METHODS: A questionnaire was delivered to patients willing to take part, at a regular visit to their ophthalmologist, for filling in anonymously at home. A total of 589 questionnaires were returned. RESULTS: More than 80% reported negative emotions on learning that they had glaucoma, one-third were afraid of going blind. Half the patients had no visual problem at all, 14% complained of poor or very poor vision. This proportion increased with age. One-fourth of the patients on topical medication experienced adverse effects of moderate or high degree. About half the patients being treated with laser or surgery felt their situation had improved afterwards. Nine-tenths of the individuals were satisfied with the information and care given, although their knowledge about glaucoma was rather incomplete. One-fifth missed information, mainly on causes, treatment and prognosis of the disease. The younger patients were more anxious and inquiring, reported more side effects and were less satisfied than the older patients. The women were in general more dissatisfied than the men. CONCLUSION: Giving a patient a diagnosis of glaucoma influences his quality of life negatively. Only half of our patients experienced any visual difficulties, whereas one-fourth reported adverse reactions due to the therapy. Most of the patients were very satisfied with the information and care given. Ophthalmologists in private practice are quite central in the management and care of the glaucoma patients in a medical setting like ours.

The impact of glaucoma on the quality of life of patients in Norway. II. Patient response correlated to objective data.

PURPOSE: To elucidate the relationship between the visual difficulties reported by patients treated for glaucoma and their objective functional damage, and to evaluate the reliability of the patient responses. METHODS: Questionnaires concerning quality of life filled in at home by 589 patients treated for chronic open angle glaucoma were correlated to corresponding questionnaires returned from their ophthalmologists. RESULTS: Few of our patients had a visual field damage judged to be of functional significance. There was a weak to moderate association between both visual field defects and decreased visual acuity and self-reported visual difficulties. A high proportion of the patients had normal binocular visual field and a stable disease, raising the suspicion that some of them were treated for ocular hypertension. The agreement between the responses from the patients and the ophthalmologists concerning the topical treatment was good, regarding treatment duration and other diseases of the patients the agreement was moderate. CONCLUSION: The association between subjective visual disability and presence of visual field defects was weak to moderate in our patients treated for glaucoma, and this association was further weakened by adjusting for visual acuity. Some patients might be treated unnecessarily, and a favourable prognosis might be given to most of them. The reliability of the patients in general was good.