Changes in corneal astigmatism among patients with visually significant cataract.

OBJECTIVE: To determine the prevalence of corneal astigmatism, changes in astigmatism, and biometry measurements with age and to correlate fellow eye measurements of patients undergoing cataract surgery. DESIGN: Prospective, observational study. PARTICIPANTS: 3498 eyes of 1814 patients undergoing cataract surgery. METHODS: Setting was a single center teaching hospital. Preoperative biometry data measured by partial coherence interferometry (IOLMaster; Carl Zeiss Meditec, Jena, Germany) were collected and analyzed. RESULTS: Mean age of the cohort was 74.52 ± 10.23 years. Mean corneal astigmatism was 1.04 ± 0.78 D OD and 1.04 ± 0.79 D OS. About 19.7% and 4.9% of eyes had corneal astigmatism greater than 1.5 and 2.5 D, respectively. The prevalence of against-the-rule astigmatism significantly increased with age bilaterally. Paired fellow eye analysis found a positive correlation for corneal astigmatism, axial length, anterior chamber depth, and white-to-white distance (p < 0.001). About 33.33% of right eyes with corneal astigmatism greater than 2.5 D had more than 2.5 D of astigmatism in the fellow eye. This is in comparison with only 1.5% of right eyes with corneal astigmatism of less than 1.5 D having corneal astigmatism greater than 2.5 D in the fellow eye. CONCLUSIONS: Patients with high corneal astigmatism in 1 eye are more likely to have significant astigmatism in the fellow eye. This may necessitate the need for bilateral toric intraocular lens (IOL) implantation during cataract surgery. Against-the-rule astigmatism should be treated more aggressively during cataract surgery, because this is likely to worsen with age. Such information is useful when calculating toric IOL power at the time of surgery.

Visual outcomes following orbital biopsy.

BACKGROUND: The risks of orbital biopsy depend on the lesion's location and relationship with surrounding structures. Complications include reduced vision, although visual outcomes following orbital biopsy are not widely reported. AIMS: To determine visual outcomes following orbital biopsy in Gartnavel General Hospital's Oculoplastic and Oncology Service. METHODS: Case note review of 50 consecutive patients undergoing orbital incision or excision biopsy between January 2006 and December 2010. Data collected included preoperative clinical examination, radiological and histological features, preoperative and postoperative corrected distance visual acuity (CDVA) and surgical complications. The main outcome measure was change in CDVA. Mean follow-up duration was 1.32 years. RESULTS: Histological diagnoses following biopsy included idiopathic orbital inflammation (n = 13) and lymphoma (n = 9). Of the radiologically defined lesions, 86.7% were extraconal (13.3% intraconal). Extraconal lesions were anterior in 59.0% (41.0% posterior). Mean preoperative LogMAR CDVA was 0.10 which was maintained at day one post-biopsy, indicating the absence of immediate sight-threatening complications such as retrobulbar haemorrhage or optic nerve compression, and there was no significant reduction at one-year follow-up (p = 0.239). Further analysis of change in CDVA showed no difference between: anterior and posterior lesions; extraconal and intraconal lesions; incision and excision biopsies; anterior and lateral surgical approaches. CONCLUSIONS: CDVA is retained for one year following orbital biopsy. Significant visual loss is a very rare complication of this procedure.

Changes in ocular monochromatic higher-order aberrations in the aging eye.

PURPOSE: To characterize corneal, internal, and total ocular monochromatic higher-order aberration (MHOA) changes that occur in the aging eye. METHODS: Prospective observational case series including 300 eyes of 167 patients (mean age = 63.8 years) attending the ophthalmology service at University Hospital Ayr, Scotland. Corneal, internal, and total ocular aberrations were measured over a 6-mm dilated pupil. Zernike coefficients were obtained to the sixth order. Changes in MHOA between age groups and inter-eye correlations between right and left eyes were analyzed. RESULTS: A significant inter-eye correlation was found for refractive mean spherical equivalent and cylinder. A significant inter-eye correlation for the whole eye, corneal, and internal MHOA was found (p < 0.001). Right eye analysis found a significant positive correlation between age and the root mean square of whole eye MHOA (p = 0.012), with an increase from 0.517 µm in the fifth decade to 0.824 µm in the ninth. Total internal MHOA increased from 0.411 to 0.704 µm. A significant positive correlation was found between age and internal fourth- (p = 0.007), fifth- (p = 0.029), and sixth-order (p = 0.025) root mean square aberrations. There were no significant age-related changes in corneal MHOA or corneal spherical aberration. Overall mean (SD) corneal SA was 0.203 (0.082) µm. CONCLUSIONS: A strong correlation between the right and left eyes exists for MHOA. Whole eye MHOA increases with age. Such changes can be attributed to age-related changes in the internal optical quality of the eye. Such normative data are useful to the cataract surgeon when considering the use of an aspherical IOL to counteract corneal-induced SA during cataract surgery.

Long term visual outcomes, graft survival and complications of deep anterior lamellar keratoplasty in patients with herpes simplex related corneal scarring.

AIMS: To report long term visual outcomes, complications and graft survival of patients undergoing deep anterior lamellar keratoplasty (DALK) to treat corneal scarring secondary to herpes simplex virus (HSV) keratitis. METHODS: Retrospective, non-comparative case series. 18 patients who underwent DALK for HSV keratitis related corneal scarring between January 2004 and February 2007 were included. DALK was performed by Anwar's big bubble technique. Data collected for analysis included preoperative characteristics, intraoperative complications and postoperative acuity, complications and subsequent operations. RESULTS: Mean best corrected distance visual acuity (LogMAR) improved from 1.51 ± 0.90 preoperatively to 0.82 ± 0.85 at the last follow-up (p=0.05). 27% of patients with more than 4 years follow-up had a best corrected distance visual acuity of 6/12 or better and 64% were 6/24 or better. Six patients (33%) experienced a recurrence of HSV keratitis and 9 (50%) experienced an episode of graft rejection. There were five cases (28%) of graft failure, four of whom had had a previous episode of graft rejection. Logistic regression did not find an association with graft rejection, HSV recurrence, any other observed postoperative host corneal vascularisation and any postoperative complication. The majority of patients underwent a second operation with 50% requiring cataract surgery. CONCLUSIONS: DALK for the treatment of HSV related corneal scarring is associated with a high percentage of postoperative complications. DALK in this context is also associated with a large percentage of secondary operations. Patients should be aware of this when giving informed consent for DALK to treat HSV related corneal scars.

A novel technique of tangential, circumferential, scleral tunnel in 20-gauge transconjunctival sutureless vitrectomy: optical coherence tomography-aided analysis of wound integrity and clinical outcome.

AIM: To describe a novel technique of sclerotomy construction to facilitate 20-gauge transconjunctical sutureless vitrectomy (TSV) along with the evaluation of the wound integrity. METHODS: The surgical technique is described. One hundred consecutive patients who underwent TSV were evaluated for wound leaks, postoperative hypotony, endophthalmitis and any other complication related to surgery. The sclerotomies of eight patients (24 ports) were analysed by imaging with anterior segment optical coherence tomography immediately after surgery, and on the first postoperative day and after the first postoperative month. RESULTS: 104 eyes of 100 patients were evaluated with a mean follow-up of 9.6 months. All cases underwent surgery with standard 20-gauge instrumentation and vitrectomy techniques. Five sclerotomies were found to leak at the end of surgery, requiring a suture. Anterior segment optical coherence tomography images were obtained from eight eyes with good apposition of the tunnel noted in all the cases. Mean intraocular pressure was 18.7 mm Hg on the first postoperative day. One patient had hypotony without leak and this patient had pre-existing hypotony due to chronic panuveitis. There was no instance of postoperative endophthalmitis. CONCLUSIONS: This technique of 20-gauge TSV achieves good wound apposition with a low incidence of complications while using standard 20-gauge instrumentation and vitrectomy techniques.

Limbal stem cell failure secondary to advanced conjunctival squamous cell carcinoma: a clinicopathological case report.

A 67-year-old man with a history of multiple myeloma (treated with chemotherapy) was referred with a left hyperaemic conjunctival lesion covering almost 360° of the limbus and extending onto the corneal surface. Conjunctival biopsy revealed conjunctival intraepithelial neoplasia. Initial treatment consisted of topical and intralesional injections of interferon α-2b. The patient subsequently developed limbal stem cell deficiency resulting in a persistent non-healing corneal epithelial defect. This was successfully managed with total excisional biopsy of the lesion, combined with limbal stem cell autograft (from the fellow eye) and amniotic membrane transplantation. Histopathology revealed a conjunctival squamous cell carcinoma. The corneal epithelium completely healed postoperatively and there is no evidence of tumour recurrence at 1 year follow-up. This case highlights a rare case of advanced ocular surface neoplasia causing secondary limbal stem cell deficiency. Medical and surgical management of ocular surface neoplasia with limbal stem cell transplantation is effective in treating such cases.

Use of cryopreserved cells for enabling greater flexibility in compound profiling.

Measurement of intracellular calcium release following agonist challenge within cells expressing the relevant membrane protein is a commonly used format to derive structure-activity relationship (SAR) data within a compound profiling assay. The Fluorometric Imaging Plate Reader (FLIPR) has become the gold standard for this purpose. FLIPR traditionally uses cells that are maintained in continuous culture for compound profiling of iterative chemistry campaigns. This supply dictates that assays can only be run on 4 of 5 weekdays, or alternative cell culture machinery is required such that plating can occur remotely at the weekend. The data reported here demonstrate that high-quality compound profiling data can be generated from the use of cryopreserved cells and that these cells can also be plated at various densities to generate equivalent data between 24 and 72 h post-plating. Hence, the authors report a method that allows data generation throughout the week and without the requirement of highly automated cell culture or continuous culture.

Oncostatin M and leukemia inhibitory factor regulate the growth of normal human breast epithelial cells.

We have previously reported the inhibitory effects of oncostatin M (OSM) and leukemia inhibitory factor (LIF) on the proliferation of breast cancer cell lines. In this study, we examined the action of OSM and LIF on normal, non-malignant human breast epithelial cells (HBECs). We demonstrated expression of three components of the OSM receptor; gp130, the leukemia inhibitory factor receptor (LIFRbeta) and the OSM specific receptor (OSMRbeta). Treatment of the normal HBECs with OSM and LIF resulted in inhibition of proliferation, even in the presence of the breast mitogen, epidermal growth factor (EGF), which is required for HBEC growth. The inhibition was associated with a reduction of cells in the S-phase of the cell cycle and an accumulation of cells in G0/G1. These results suggest a previously unrecognised physiological role for these growth factors in the regulation of normal breast epithelium.

Oncostatin M induces the differentiation of breast cancer cells.

We have recently described the action of Oncostatin M (OSM) to inhibit the proliferation of breast cancer cells. In this study we examined the action of OSM on 2 breast cancer cell lines to further characterize the nature of OSM inhibition of cellular proliferation. Treatment with OSM for 6 days resulted in an approximately 2- to 5-fold decrease in cell number, which was independent of estrogen receptor status. Consistent with this, colony formation was reduced to approximately 50% when cells were exposed to OSM in primary agar cultures. Clonogenicity was further inhibited following 7 days treatment with OSM in monolayer cultures: the total number of clonogenic cells was suppressed approximately 10-fold. Analysis of cell cycle status in OSM-treated cells demonstrated a 40% reduction in the proportion of cells in S phase within 12 hr, with an increase in cells in G0/G1. After 6 days, there was a 10-fold reduction in the absolute number of cells in S phase in OSM-treated cultures. These changes were associated with striking changes in cellular morphology, including disruption of intercellular junctions and the production of lipid droplets. There was a 5-fold increase of c-fos and c-myc mRNA within 30 min of commencing treatment with OSM. In addition, in the ER positive cells there was a decrease in ER mRNA (evident within approximately 2 hr) and ER protein expression following treatment with OSM. Conversely, there was a 5-fold increase in epidermal growth factor receptor (EGFR) mRNA within 4 hr, and a 2.5-fold rise in mRNA for transforming growth factor alpha (TGF alpha). Thus, the inhibition of breast cancer cells by OSM was associated with decreased clonogenicity, a decrease in S phase cells and a variety of phenotypic changes, all consistent with the induction of differentiation.

Expression and function of members of the cytokine receptor superfamily on breast cancer cells.

Receptors for the cytokines leukemia inhibitory factor (LIF), interleukin-6 (IL-6), oncostatin M (OSM), ciliary neurotrophic factor (CNTF) and interleukin-11 (IL-11) are members of the structurally conserved hemopoietin receptor superfamily. In addition, they all share the transmembrane signalling protein gp130. In this paper the expression and function of this family of receptors in breast cancer cells was examined. RT-PCR analyses demonstrated that gp130 was expressed in 12/12 breast cell lines and the specific receptor alpha-chains for IL-6, LIF, IL-11 and CNTF were expressed in the majority of these cell lines. This was in contrast to other hemopoietin receptors. Examination of 50 clinical samples of malignant breast tissue by RT-PCR showed a similar pattern of expression of gp130 associated receptors. Treatment of breast cancer cell lines with OSM resulted in changes in cellular morphology. Cellular proliferation was inhibited following exposure to OSM (3/4 cell lines), IL-11 (2/4 cell lines), and by IL-6 and LIF (1/4 cell lines). Cell surface binding of LIF and OSM was also documented. The expression of these receptors in 12/12 cell lines and greater than 95% of clinical samples suggests that these molecules may be important in regulating the growth of breast cells.

Direct sequencing of double-stranded PCR products incorporating a chemiluminescent detection procedure.

A simple and reliable method is described for direct sequencing of material generated by the polymerase chain reaction (PCR). Sequencing reactions can be performed directly on PCR products without the need for purification of the template by removal of residual deoxyribonucleoside triphosphates or primers. The coupling of a chemiluminescent detection system with the use of the same primers in the initial and sequencing PCR's allows for sequencing of a number of PCR products on the one gel.

Purification of human leucocyte DNA: proteinase K is not necessary.

A rapid nontoxic method for the purification of DNA from human leucocytes is described. Preliminary experiments which tested different methods of DNA purification indicated that digestion of proteins with proteinase K was unnecessary. This led to the development of a simple procedure involving lysis of the cells in SDS followed by extraction with 6 M NaCl. The method described overcomes the requirement for lengthy incubations in the presence of expensive proteinase K and subsequent extraction with toxic chemicals.

Low calcium reperfusion of ischemic myocardium.

The mechanisms underlying protection of ischemic myocardium by reperfusion with solutions containing reduced concentrations of calcium (CA) were studied in isolated vascularly perfused rabbit interventricular septa at 37 degrees C. After 45 min of total ischemia adenosine triphosphate (ATP), and phosphocreatine (PC) contents fell to 6.8 +/- 0.82, and 12.4 +/- 2.0 micrometer/g dry tissue (dt) (+/- S.E.M.). After 5 min of reperfusion with 0.75 mM Ca these values had recovered significantly to 10.3 +/- 1.0, and 33.3 +/- 3.6, (micrometer/g dt), while reperfusion with 2.5 mM Ca produced 5.9 +/- 1.0 and 22.0 +/- 2.6 (micrometer/g dt). The significantly greater recovery of ATP and PC after 0.75 mM Ca reperfusion persisted after an additional 25 min of reperfusion with 2.5 mM Ca when compared to septa reperfused for the entire 30 min with 2.5 mM Ca. When mechanical work was reduced by cessation of stimulation during the first 5 min of reperfusion, ATP, PC and total creatine (TC) recovery were significantly improved in muscles exposed to 2.5 mM Ca but showed no additional improvement in muscles reperfused with 0.75 mM Ca. A further reduction to 100 micrometers Ca reperfusion in quiescent muscles did significantly improve ATP, PC and TC recovery. Quiescence achieved with 16.0 mM K impaired ATP, PC and TC recovery. This effect was reversed by 100 micrometers CA reperfusion. Measurements of mitochondrial oxygen consumption, respiratory control and ADP/O showed results parallel to the ATP, PC and TC determinations. Both mechanical work and CA itself influence mitochondrial respiration, ATP, PC and TC content after ischemia. As previously demonstrated for mechanical function, metabolic recovery can be improved by interventions limited to the first 5 min of reperfusion.