An in vitro assessment of the thermoreversible PLGA-PEG-PLGA copolymer: Implications for Descemet's membrane endothelial keratoplasty.

BACKGROUND: To explore the use of a thermoreversible copolymer gel coating to prevent donor tissue scrolling in Descemet's membrane endothelial keratoplasty (DMEK). METHODS: PLGA-PEG-PLGA triblock copolymer was synthesised via ring opening polymerisation. Two formulations were fabricated and gelation properties characterised using rheological analyses. Endothelial cytotoxicity of the copolymer was assessed using a Trypan Blue exclusion assay. Thickness of the copolymer gel coating on the endothelial surface was analysed using anterior segment optical coherence tomography (OCT) (RTVue-100, Optovue Inc.). Gold nanoparticles were added to the copolymer to aid visualisation using OCT. Prevention of Descemet membrane donor scrolling was represented via a novel, in vitro, immersion of copolymer coated donor graft material. RESULTS: Two different formulations of PLGA-PEG-PLGA copolymer were successfully fabricated and the desired peak gelling temperature of 24°C was achieved by polymer blending. Application of 20%, 30% and 40% (wt/vol) polymer concentrations resulted in a statistically significant increase in polymer thickness on the endothelium (p < 0.001). There was no detectable endothelial cytotoxicity. The polymer was easy to apply to the endothelium and prevented scrolling of the DMEK graft. CONCLUSION: This PLGA-PEG-PLGA thermoreversible copolymer gel could be exploited as a therapeutic aid for preventing DMEK graft scrolling.

Cataract risk stratification and prioritisation protocol in the COVID-19 era.

BACKGROUND: The COVID-19 pandemic halted non-emergency surgery across Scotland. Measures to mitigate the risks of transmitting COVID-19 are creating significant challenges to restarting all surgical services safely. We describe the development of a risk stratification tool to prioritise patients for cataract surgery taking account both specific risk factors for poor outcome from COVID-19 infection as well as surgical 'need'. In addition we report the demographics and comorbidities of patients on our waiting list. METHODS: A prospective case review of electronic records was performed. A risk stratification tool was developed based on review of available literature on systemic risk factors for poor outcome from COVID-19 infection as well as a surgical 'need' score. Scores derived from the tool were used to generate 6 risk profile groups to allow prioritised allocation of surgery. RESULTS: There were 744 patients awaiting cataract surgery of which 66 (8.9 %) patients were 'shielding'. One hundred and thirty-two (19.5 %) patients had no systemic comorbidities, 218 (32.1 %) patients had 1 relevant systemic comorbidity and 316 (46.5 %) patients had 2 or more comorbidities. Five hundred and ninety patients (88.7 %) did not have significant ocular comorbidities. Using the risk stratification tool, 171 (23 %) patients were allocated in the highest 3 priority stages. Given an aging cohort with associated increase in number of systemic comorbidities, the majority of patients were in the lower priority stages 4 to 6. CONCLUSIONS: COVID-19 has created an urgent challenge to deal safely with cataract surgery waiting lists. This has driven the need for a prompt and pragmatic change to the way we assess risks and benefits of a previously regarded as low-risk intervention. This is further complicated by the majority of patients awaiting cataract surgery being elderly with comorbidities and at higher risk of mortality related to COVID-19. We present a pragmatic method of risk stratifying patients on waiting lists, blending an evidence-based objective assessment of risk and patient need combined with an element of shared decision-making. This has facilitated safe and successful restarting of our cataract service.

An Evaluation for the Causes of Reduced Hb A2 and the Molecular Characterization of HBD Variants in Hong Kong.

Prenatal screening of ß-thalassemia (ß-thal) carriers is based on the hallmark phenotype of microcytosis and raised Hb A2. The unanticipated birth of ß-thal major (ß-TM) offspring to ß-thal carriers who were misdiagnosed during prenatal screening have been reported. A subset of these resulted from the masked phenotype due to the coinheritance of HBD variants. In a broader sense, the causes of reduced Hb A2 in thalassemia screening, the prevalence and spectrum of HBD variants in Hong Kong remain to be characterized. Over a 13-month period, a total of 2982 samples were referred for thalassemia screening. Surplus samples with reduced Hb A2 levels (2.0%) were evaluated. HBD variations were assessed by direct sequencing. Sixty-six samples were tested. Hb H disease, HBD variants, α-thalassemia (α-thal) trait and iron deficiency were detected in 40 (60.6%), 12 (18.2%), eight (12.1%) and seven (10.6%) samples, respectively. Seven samples carried more than one of the mentioned conditions. The cause remained elusive in seven samples. Thirteen HBD variants were detected and two were recurrent, including HBD: c.-127T>C [-77 (T>C)] and HBD: c.314G>A (Hb Chori-Burnaby). A novel nonsense variant HBD: c.262C>T [codon 87 (C>T)] was detected in cis with HBD: c.-127T>C. Overall, the prevalence of HBD variants was 0.4%. This study advanced our understanding of the causes of reduced Hb A2 in clinical practice and identified hereditary disorders of α- and δ-globin genes as the prevailing causes. It established the landscape of HBD variations in our locality and highlighted the pitfall of phenotypic screening of ß-thal carriers.

Distinct mutation spectrum, clinical outcome and therapeutic responses of typical complex/monosomy karyotype acute myeloid leukemia carrying TP53 mutations.

The present study aimed to define a subtype of complex/monosomal karyotype (CK/MK) acute myeloid leukemia (AML) by its distinct clinical features, p53 signaling and responses to p53 targeting agents. Ninety-eight young adults (range: 21-60 years; median: 49 years) with CK/MK AML were studied. They received standard induction, consolidation and allogeneic hematopoietic stem cell transplantation from siblings or matched unrelated donors if available. Chromosomal abnormalities most commonly affected chromosome 5 (30%), 7 (22%) and 17 (21%). Next generation sequencing of a 54-myeloid gene panel were available in 76 patients. Tumor protein 53 (TP53) mutations were most common (49%) and associated with the presence of -5/5q- (P < .001) and -17/17p- (P < .001), but not -7/7q- (P = .370). This "typical" CK/MK AML subtype was associated with significantly lower presenting white cell counts, higher number of karyotypic abnormalities, and inferior leukemia-free and overall survivals, compared with CK/MK AML without the typical features. Blood or bone marrow samples from typical CK/MK AML patients showed defective p53 signaling upon induction by etoposide. In vitro drug sensitivity analysis showed that they were sensitive to APR-246 that targeted mutant p53, but resistant to MDM2 antagonist MI-77301. Novel therapeutic strategies targeting TP53 mutations in CK/MK AML should be developed and tested in clinical trials.

The relationship between oestrogen receptor-alpha phosphorylation and the tumour microenvironment in patients with primary operable ductal breast cancer.

AIMS: Although the role of phosphorylation of oestrogen receptor (ER) at serines 118 (p-S118) and 167 (p-S167) has been studied, the relationship between p-S118, p-S167 and the tumour microenvironment in ER-positive primary operable ductal breast cancers have not been investigated. The aims of this study are to investigate (i) the relationship between p-S118/p-S167 and the tumour microenvironment, and (ii) the effect of p-S118/167 on survival and recurrence in ER-positive primary operable ductal breast cancers. METHODS AND RESULTS: Patients presenting at three Glasgow hospitals between 1995 and 1998 with invasive ductal ER-positive primary breast cancers were studied (n = 294). Immunohistochemical staining of p-S118 and p-S167 was performed and their association with clinicopathological characteristics, cancer-specific survival (CSS) and recurrence-free interval (RFI) were examined. In the whole cohort, tumour size (P < 0.05) and microvessel density (P < 0.05) were associated with high p-S118 while increased micovessel density (P < 0.05), apoptosis (P < 0.05), general inflammatory infiltrate measured using the Klintrup-Makinen score (P < 0.05) and macrophage infiltrate (P < 0.05) were found to be associated with high p-S167. Only high p-S167 was associated with shorter CSS (P < 0.005) and shorter RFI in the whole cohort (P = 0.001) and separately in the luminal A (P < 0.05) and B tumours (P < 0.05). CONCLUSIONS: This study showed that both p-S118 and p-S167 were associated with several microenvironmental factors, including increased microvessel density. In particular, p-S167 was associated with reduced RFI and CSS in the whole cohort and RFI in luminal A and B tumours and could possibly be employed to predict response to kinase inhibitors.

First Report of Hb Kent [ß37(C3)Trp→Cys (TGG>TGC) HBB: c.114G>C] in a Chinese Family.

We report a novel HBB: c.114G>C mutation in a Chinese family. This mutation resulted in a ß37(C3)Trp→Cys amino acid substitution and was synonymous with Hb Kent, a hemoglobin (Hb) variant that was reported exclusively in patients of European descent. Though Hb Kent has a normal oxygen affinity and molecular stability, it has a characteristic dual variant appearance on cellulose acetate electrophoresis (CAE) and high performance liquid chromatography (HPLC) caused by the posttranslational modification of cysteine. We also report the phenotypic expression of this variant when coinherited with the Southeast Asian (- -SEA) double α-globin gene deletion.

Statin usage, vascular diagnosis and vascular risk factors in Parkinson's disease.

Background and aims Vascular disease is a common comorbidity in Parkinson's disease patients. Statins are potentially neuroprotective for Parkinson's disease through non-vascular mechanisms. We investigated prevailing statin use in a Parkinson's disease cohort. Methods and results Data on diagnostic indication for statins, anti-Parkinson therapy, vascular risk factors, and statin prescription, were obtained from electronic medical record review for consecutive Parkinson's disease patients. The ASsessing cardiac risk using Scottish Intercollegiate Guidelines Network system was used to calculate future cardiovascular risk and identify those warranting statin use. Of 441 patients included, 59.9% were male, with a mean age of 68.9 years (standard deviation 10.3). One hundred and seventy-four (39.5%) patients had at least one diagnostic indication for statin use, of whom 136 (78.2%) were prescribed a statin. In the 267 (60.5%) cases without a diagnostic indication, 54 (20.2%) were excluded owing to age limitations defined in ASsessing cardiac risk using Scottish Intercollegiate Guidelines Network. Of the remaining 213, 62 (29.1%) had an ASsessing cardiac risk using Scottish Intercollegiate Guidelines Network score in the recommended range for statin therapy, of whom 15 (24.1%) were prescribed statins. Conclusion There is suboptimal implementation of statin therapy in Parkinson's disease patients. Given the possible neuroprotective effects of statins in Parkinson's disease in addition to reducing cardiovascular risk, reasons for suboptimal implementation warrant further investigation.

A Multi-locus Approach to Characterization of Major Quantitative Trait Loci Influencing Hb F Regulation in Chinese ß-thalassemia Carriers.

Genetic association studies showed that Hb F is under the influence of major quantitative trait loci (QTL) in ß-thalassemia (ß-thal) carriers. Single nucleotide polymorphisms (SNPs) at three major QTLs, BCL11A, HBS1L-MYB intergenic region and XmnI-HBG2 were individually validated in univariate models. However, their relative effect sizes on Hb F regulation are unknown. We genotyped 99 Chinese ß-thal carriers for the three major QTLs and performed genetic association studies using three different statistical models, including mass univariate analysis, multivariate linear regression and partial least square regression structural equation modeling (PLS-SEM). Performances of the three models were compared and effect sizes of the three QTLs in a multivariate model were assessed. Traditional mass univariate analysis and multivariate linear regression showed limited statistical power in our small cohort and the latter was constrained by multicollinearity. Partial least structural equation modeling showed significant positive associations of each QTL (p <0.05) with Hb F regulation, together explained 34.4% of variance. The HBS1L-MYB intergenic region polymorphism (HMIP) demonstrated the highest effect on Hb F prediction with effect size f2 0.294. PLS-SEM offered a statistically powerful multivariate model for multi-locus genetic association studies. We reproduced findings of previous studies with a much smaller cohort and demonstrated HMIP as the strongest regulator of Hb F in Chinese ß-thal carriers.

Adherence to topical glaucoma therapy in patients attending virtual clinics.

BACKGROUND/OBJECTIVES: Poor adherence to medical therapy is a major challenge to the effective treatment of chronic diseases including glaucoma. Potential factors influencing adherence include treatment complexity and patient understanding of disease and health beliefs. An increasing number of patients are seen in virtual clinics, where there is no face-to-face consultation, potentially reducing opportunities for patient education and reinforcement of the importance of treatment. The aim of this study was to examine adherence among patients attending a virtual glaucoma clinic. METHODS: 100 consecutive patients attending the virtual clinic were surveyed, with 78 using topical medications included in the analysis. All patients completed a validated adherence questionnaire with a score of >2 defined as poor adherence. The relationship between adherence and age, sex, duration since diagnosis, and disease severity was examined. RESULTS: The mean age was 73.1 ± 13.4 years, with an average mean deviation of -5.9 ± 5.5 dB and duration since first diagnosis of 9.0 ± 5.7 years. 93.6% reported self-instilling eye drops. Seventy-one patients (91.0 %) had good self-reported adherence. Multivariate logistic regression revealed those instilling eye drops independently had higher odds of good adherence. CONCLUSIONS: The level of medication adherence in the virtual glaucoma clinic was higher than adherence in previous studies examining patients attending face-to-face clinics. Virtual clinics should incorporate methods to ensure effective two-way communication with patients and strategies for patient education.

PACS plus criteria: a retrospective cohort review of 612 consecutive patients treated with bilateral YAG peripheral iridotomies.

INTRODUCTION: With an ageing population and better life expectancy, the prevalence of angle closure disease is expected to increase by 20% per decade. In 2022, the Royal College of Ophthalmologists (RCOphth) issued a guideline on managing angle closure disease. Hospital eye service (HES) referral and prophylactic treatment are recommended only for primary angle closure suspect (PACS) with "Plus" features only. We aimed to examine patients previously treated with YAG peripheral iridotomies (YAG PI) for the presence of "PACS Plus" features. METHODS: A retrospective cohort study of consecutive patients treated with YAG PI between 2015 and 2019 at a tertiary referral NHS eye centre was reviewed. Cases were examined to identify and classify patients into Primary Angle Closure (PAC), PACS, and Primary Angle Closure Glaucoma (PACG). Patients with PACS were studied for "Plus" features. RESULTS: Six hundred twelve patients with gonioscopy-confirmed angle closure (defined as a minimum 180 degrees iridotrabecular contact) treated with YAG PI from years 2015 to 2019 were included in the analysis. The mean age of patients presenting with angle closure disease was 68.5 years (SD 11.3). There were 390 (63.7%) patients with PACS, 102 (16.6%) with PAC and 120 (19.7%) with PACG. Of the PACS patients, 159(40.8%) patients had no "Plus" features. 181 (40.2%) patients had 1 "Plus" feature, 37 (9.5%) had 2 "Plus" features and 13 (3.3%) patients had 3 "Plus" features. CONCLUSION: In our cohort, a considerable proportion (40.8%) of PACS patients treated with YAG PI did not have Plus features and therefore that would not meet the proposed criteria for HES referral and YAG PI. With the proposed guidance, we expect a considerable reduction in HES referrals. Nonetheless, community optometry services should be supported and trained to provide monitoring for patients with PACS not referred to the HES.

Surgical management of aphakia.

Despite the safety and efficacy of cataract surgery, intraoperative complications can hamper the ability to place an intraocular lens in the capsular bag. With vast numbers of cataract surgeries performed daily, complications occur often enough that every ophthalmologist should be equipped with techniques to manage aphakia. Medical management of aphakia used to be commonplace but these techniques have their disadvantages including thick bulky lenses, poor cosmesis, and aniseikonia. Surgical management of aphakia overcomes these disadvantages and offers patients the possibility of a spectacle and contact lens-free lifestyle. This article reviews the various options of surgical management of aphakia and their advantages and disadvantages. Comparison of outcomes between techniques and a protocol for deciding between techniques is presented.

Is there an association of socioeconomic deprivation with acute primary angle closure?

BACKGROUND: Socioeconomic deprivation is known to increase the risk of late presentation of many diseases. This is the largest study in United Kingdom investigating the relationship between socioeconomic deprivation and acute primary angle closure (APAC). METHODS: A retrospective review of case notes was conducted of 718 consecutive patients who underwent laser peripheral iridotomy (LPI) in Edinburgh (Princess Alexandra Eye Pavilion) and Fife (Queen Margaret Hospital) between 2015 and 2019. Baseline demographics including sex, age, ethnicity, pre-existing diabetes, use of anti-depressants, and family history of glaucoma were collected. Deprivation was scored using the Scottish Index of Multiple Deprivation (SIMD) Index 2020v2. A lower rank and decile indicate higher degrees of deprivation. We investigated differences in characteristics between patients who were referred routinely versus patients who referred as APAC. RESULTS: The SIMD rank and deciles were consistently lower in patients who were referred urgently with APAC in both centres (P = <0.05) when compared to those referred routinely for LPI. On univariate and multivariate logistic regression, the presentation of APAC is negatively associated with SIMD Decile (OR = -0.101, 95% CI -0.178 to -0.026, P = 0.008) and family history of glaucoma (OR = -1.010, 95% CI -1.670 to -0.426, P = 0.001), and positively associated with age (OR = 0.029, 95% CI 0.009-0.049, P = 0.004). CONCLUSIONS: Socioeconomic deprivation is an important risk factors for patients presenting with APAC. Socioeconomic deprivation should be incorporated into the design of glaucoma services and considered when triaging patients for prophylactic and therapeutic LPI and cataract surgery.

Spotlight on the Disc-Damage Likelihood Scale (DDLS).

The disc damage likelihood scale (DDLS) is a tool for classifying glaucomatous structural changes to the optic disc based on the radial width of the neuroretinal rim at its thinnest location, or if no rim is present, the extent of absence of the rim. Unlike cup disc ratio (CDR), the DDLS also considers disc size. Twenty years after its first description, the aim of this review was to critically appraise evidence for the DDLS and evaluate its role in current practice. A literature search by two independent authors identified 33 relevant articles for inclusion. Five studies evaluated reproducibility, 5 diagnostic performance, and 2 studies examined ability to detect progression. Eleven studies evaluated correlation between DDLS and other markers of glaucoma. Despite the widespread availability of imaging devices such as optical coherence tomography (OCT), clinical examination of the optic disc remains an essential component of glaucoma diagnosis and monitoring. The DDLS provides a reliable method for semi-quantitative clinical grading of the optic disc in glaucoma, with higher reproducibility than methods such as CDR.

Macular vessel density, branching complexity and foveal avascular zone size in normal tension glaucoma.

The aim of this study was to investigate the relationship between glaucoma severity and perifoveal vessel density (pfVD), branching complexity, and foveal avascular zone (FAZ) size in normal tension glaucoma (NTG). 31 patients with NTG washed out of glaucoma medications were subjected to tests including; intraocular pressure measurement; standard automated perimetry; optical coherence tomography (OCT) measurement of macular ganglion cell complex (mGCC), inner macular thickness (IMT) and circumpapillary retinal nerve fibre layer (cpRNFL); and OCT angiography measurement of pfVD, FAZ perimeter and multispectral fractal dimensions (MSFD). Eyes with more severe glaucoma had significantly thinner mGCC and cpRNFL and lower pfVD. MD decreased by 0.4 dB (95% CI 0.1 to 0.6 dB, P = 0.007) for every 1% decrease in pfVD. Lower MSFD was observed in eyes with lower pfVD and in patients with systemic hypertension. Multivariable analysis, accounting for age and OCTA quality, found lower pfVD remained significantly associated with thinner IMT, thinner mGCC and worse MD but not with MSFD. pfVD was reduced in NTG and was diminished in eyes with worse MD. Macular vessel branching complexity was not related to severity of visual field loss but was lower in patients with systemic hypertension.

Predictive Biomarkers for Endocrine Therapy: Retrospective Study in Tamoxifen and Exemestane Adjuvant Multinational (TEAM) Trial.

Background: Aromatase inhibitors improve disease-free survival compared with tamoxifen in postmenopausal women with hormone receptor-positive breast cancer. The Tamoxifen and Exemestane Adjuvant Multinational (TEAM) trial compared exemestane monotherapy with sequential therapy of tamoxifen followed by exemestane. The trial failed to show a statistically significant difference between treatment arms. A robust translational program was established to investigate predictive biomarkers. Methods: A tissue microarray was retrospectively constructed using a subset of patient tissues (n = 4631) from the TEAM trial (n = 9766). Immunohistochemistry was performed for biomarkers, classed into three groups: MAPK pathway, NF-kappa B pathway, and estrogen receptor (ER) phosphorylation. Expression was analyzed for association with relapse-free survival (RFS) at 2.5 and 10 years and treatment regimen using Kaplan-Meier curves and log-rank analysis. All statistical tests were two-sided. Results: In univariate analysis, ER167 (hazard ratio [HR] = 0.71, 95% confidence interval [CI] = 0.59 to 0.85, P < .001), IKKα (HR = 0.74, 95% CI = 0.60 to 0.92, P = .005), Raf-1338 (HR = 0.64, 95% CI = 0.52 to 0.80, P < .001), and p44/42 MAPK202/204 (HR = 0.77, 95% CI = 0.64 to 0.92, P = .004) were statistically significantly associated with improved RFS at 10 years in patients receiving sequential therapy. Associations were strengthened when IKKα, Raf-1338, and ER167 were combined into a cumulative prognostic score (HR = 0.64, 95% CI = 0.52 to 0.77, P < .001). Patients with an all negative IKKα, Raf-1338, and ER167 score favored exemestane monotherapy (odds ratio = 0.56, 95% CI = 0.35 to 0.90). In multivariable analysis, the IKKα, Raf-1338, and ER167 score (P = .001) was an independent prognostic factor for RFS at 10 years in patients receiving sequential therapy. Conclusions: The IKKα, Raf-1338, and ER167 score is an independent predictive biomarker for lower recurrence on sequential therapy. Negative expression may further offer predictive value for exemestane monotherapy.

Clonal evolution of 8p11 stem cell syndrome in a 14-year-old Chinese boy: a review of literature of t(8;13) associated myeloproliferative diseases.

We describe a case of coexisting BCR-ABL negative myeloproliferative disorder and precursor T-cell lymphoblastic lymphoma associated with t(8;13) involving FGFR1 at 8p11 in a 14-year-old boy who presented with generalized lymphadenopathy and an abdominal mass. JAK2 mutation and FIP1L1-PDGFRalpha were not detected. RT-PCR revealed the ZNF198-FGFR1 fusion transcript in both the bone marrow (BM) and lymph node (LN) of the patient at diagnosis. Of interest, reciprocal FGFR1-ZNF198 fusion transcript was demonstrated in the BM but not LN. Also differential clonal TcRgamma gene rearrangements in the BM and LN samples were observed. These findings provide novel insights into the genetic pathogenesis.