Loss of the DNA Repair Gene RNase H2 Identifies a Unique Subset of DDR-Deficient Leiomyosarcomas.

Targeting the DNA damage response (DDR) pathway is an emerging therapeutic approach for leiomyosarcoma (LMS), and loss of RNase H2, a DDR pathway member, is a potentially actionable alteration for DDR targeted treatments. Therefore, we designed a protein and genomic based RNase H2 screening assay to determine its prevalence and prognostic significance. Using a selective RNase H2 antibody on a pan-tumor tissue microarray (TMA), RNase H2 loss was more common in LMS (11.5%, 9/78) than across all tumors (3.8%, 32/843). In a separate LMS cohort, RNase H2 deficiency was confirmed in uterine LMS (U-LMS, 21%, 23/108) and soft-tissue LMS (ST-LMS) (30%, 39/102). In the TCGA database, RNASEH2B homozygous deletions (HomDels) were found in 6% (5/80) of LMS cases, with a higher proportion in U-LMS (15%; 4/27) compared to ST-LMS (2%; 1/53). Using the SNiPDx targeted-NGS sequencing assay to detect biallelic loss of function in select DDR related genes, we found RNASEH2B HomDels in 54% (19/35) of U-LMS cases with RNase H2 loss by IHC, and 7% (3/43) HomDels in RNase H2 intact cases. No RNASEH2B HomDels were detected in ST-LMS. In U-LMS patient cohort (n = 109), no significant overall survival difference was seen in patients with RNase H2 loss versus intact, or RNASEH2B HomDel (n=12) vs Non-HomDel (n=37). The overall diagnostic accuracy, sensitivity, and specificity of RNase H2 IHC for detecting RNASEH2B HomDels in U-LMS was 76%, 93% and 71% respectively, and it is being developed for future predictive biomarker driven clinical trials targeting DDR in U-LMS.

Influence of unilateral eyelid spasms and botulinum toxin treatment on intraocular pressure measured by transpalpebral tonometer.

PURPOSE: Eyelid spasms might be associated with elevated intraocular pressure (IOP) in hemifacial spasm (HFS) patients. IOP assessment using a Goldmann applanation tonometer (GAT) is often compromised by eyelid spasms. This study aimed to assess the effect of HFS on IOP measurements using the transpalpebral tonometer Diaton® before and after treatment with botulinum toxin type A (BTX-A) and compared Diaton® and GAT measurements after treatment with BTX-A. METHODS: IOP measurements were obtained with Diaton® in 27 patients with moderate-to-severe HFS before and after treatment with BTX-A. After treatment, the IOP was also measured using GAT and the results were compared with the ones measured with a Diaton®. The patients underwent automated perimetry, OCT, and pachymetry for screening to glaucoma. RESULTS: Mean IOP with Diaton® was 11 ± 3.42 mmHg before treatment in the affected eye and 9 ± 2.98 mmHg in the contralateral eye. This difference was statistically significant (P = 0.012). However, after treatment with BTX-A, no interocular difference was found in IOP obtained with Diaton® (P = 0.204) or GAT (P = 0.971). Comparison between GAT and Diaton® measurements showed no significant differences after BTX-A treatment between the affected (P = 0.212) and contralateral eye (P = 0.971). CONCLUSIONS: A significant reduction in IOP measurements on the affected side of HFS patients was observed after treatment with BTX-A, demonstrating that eyelid spasms may increase the IOP. No significant difference was observed between Diaton® and GAT measurements after the application of BTX-A. No differences were found in automated perimetry, OCT, and CCT when comparing affected eyes with contralateral eyes.

Trends in the epidemiological and microbiological profiles of infectious keratitis in southeastern Brazil / Perfil epidemiológico e microbiológico das ceratites infecciosas no sudeste do Brasil

ABSTRACT Purpose: To investigate the antibiotic susceptibility as well as the clinical, epidemiological, and microbiological profiles of microbial keratitis. Methods: This was a longitudinal retrospective study, and we retrospectively reviewed medical and laboratory records from 2015 to 2019. Results: In total, 380 pathogens (321 bacteria and 59 fungi) were isolated from the corneas of 352 patients. Staphylococcus species (45%) were most abundant within the organisms that were isolated, followed by Pseudomonas (18.4%), fungi (15.5%), Streptococcus (7.9%), and Serratia species (3.2%). The isolated gram-positive bacteria were not resistant to amikacin or vancomycin, although 14.8% of the gram-positive isolates were resistant to ciprofloxacin (p<0.05). All the gram-negative isolates were susceptible to amikacin. Male patients represented 62.8% of the 129 cases with accessible clinical data. The mean age of the patients was 53.17 ± 21 years. The time to presentation (from onset of symptoms) was 14.9 ± 19.4 days (median: 7 days). Large ulcers (>5 mm in any dimension) were present in 49.6% (64 eyes) of the cases. The duration of treatment was 49 ± 45.9 days (median: 38 days). Direct ocular trauma was reported by 48 (37.2%) patients, and 15 patients (11.6%) reported using contact lenses. For 72 (55.8%) patients, topical treatment had been previously prescribed, and 16 (12.4%) patients reported using other classes of drugs. Hospitalizations were required for 79 (61.2%) patients, and in terms of major complications, 53 (41.1%) patients had corneal perforations. A total of 40 patients (31%) underwent tectonic penetrating keratoplasty, and 28 (21.7%) developed secondary glaucoma. A progression to endophthalmitis occurred in 8 (6.2%) patients, with 50% of those patients' (3.1% of the total) endophthalmitis evolving to evisceration. The patients' microbial keratitis was largely treated empirically, with 94 (72.9%) patients prescribed moxifloxacin and 56 (43.4%) prescribed ciprofloxacin before receiving their culture results. Conclusions: For the most part, our hospital treated patients with severe microbial keratitis. Despite identifying gram-positive bacteria in most of the isolates, we also frequently identified gram-negative rods and fungi. Our susceptibility results support prescribing a combination of vancomycin and amikacin as an effective empirical therapeutic regimen to treat microbial keratitis.

RESUMO Objetivo: Investigar a susceptibilidade a antibió­ticos, o perfil clínico, epidemiológico e microbiológico das ce­ratites infecciosas. Métodos: Estudo retrospectivo longitu­dinal. Registros médicos e laboratoriais de 2015 a 2019 fo­ram revisados retrospectivamente. Resultados: Trezentos e oitenta patógenos (321 bactérias e 59 fungos) foram isolados das córneas de 352 pacientes. As espécies de Staphylococcus foram os microorganismos mais isolados (45%), seguidos de Pseudomonas (18,4%), fungos (15,5%), Streptococcus (7,9%) e Serratia (3,2%). Não houve resistência das bactérias Gram-positivas à amicacina ou vancomicina, enquanto 14,8% isolados Gram-positivos foram resistentes à ciprofloxacina (p<0,05). Todos os organismos Gram-negativos eram suscetíveis à amicacina. Pacientes do sexo masculino representaram 62,8% de 129 casos com dados clínicos acessíveis. A média de idade foi 53,17 ± 21 anos. O tempo até a apresentação (desde o início dos sintomas) foi de 14,9 ± 19,4 dias (mediana: 7 dias). Úlceras grandes (>5mm em qualquer extensão) representaram 49,6% (64 olhos) dos casos. A duração do tratamento foi de 49 ± 45,9 dias (mediana: 38 dias). Trauma ocular direto foi relatado por 48 (37,2%) pacientes e uso de lentes de contato por 15 (11,6%) pacientes. Foi prescrito tratamento prévio para 72 (55.8%) pacientes. Outras classes de medicamentos foram prescritas para 16 (12.4%). Setenta e nove (61,2%) pacientes tiveram que ser hospitalizados. Como complicações maiores, 53 (41,1%) pacientes apresentaram perfuração corneana, 40 pacientes (31%) foram submetidos à ceratoplastia penetrante tectônica e 28 (21,7%) desenvolveram glaucoma secundário. Oito pacientes (6,2%) evoluíram para endoftalmite. O tratamento empírico da ceratite microbiana foi amplamente empregado, com 94 (72,9%) pacientes em uso de moxifloxacina e 56 (43,4%) em uso de ciprofloxacina antes do resultado da cultura. Conclusões: Nosso hospital tratou predominantemente de pacientes com úlceras microbianas graves. Embora bactérias Gram-po­sitivas constituíssem a maioria dos isolados, bacilos e fungos Gram-negativos também foram frequentemente identificados nas ceratites microbianas. Os resultados de suscetibilidade sugerem a combinação de vancomicina e amicacina como um regime terapêutico empírico eficaz para essa condição grave com risco de perda visual permanente.

Traboulsi syndrome without features of Marfan syndrome caused by a novel homozygous ASPH variant associated with a heterozygous FBN1 variant.

BACKGROUND: Traboulsi syndrome is a rare disease clinically characterized by facial dysmorphism, abnormal spontaneous filtering blebs, ectopia lentis (EL) and multiple anterior segment abnormalities. MATERIAL AND METHODS: An 18-year-old female was referred to the Emergency Service of Hospital São Geraldo (HSG) claiming decreased right eye (RE) visual acuity associated with ocular pain that was noticed approximately 2 months earlier. She underwent a complete ophthalmic and physical examination including hands, ankle, wrist and chest X-ray, abdominal ultrasound, echocardiogram and genetic analysis (whole-exome sequencing). RESULTS: The ophthalmic examination revealed a high myopia with spherical equivalent of - 9.50 D and best corrected visual acuity (BCVA) of 20/60 in RE and - 9.25 D with BCVA of 20/30 in the left eye (LE). Slit-lamp examination showed normal conjunctiva in both eyes (BE) and a superior-temporal cystic lesion in RE and nasal in LE; the flat anterior chamber in BE with the transparent crystalline lens touches the central corneal endothelium in the RE. Fundoscopy suggested glaucoma as the cup/disc ratio was 0.7, although the intraocular pressure (IOP) was 10 mmHg in BE without medication. Validation of data from whole exome demonstrated a novel splicing homozygous pathogenic variant (PV) (c.1765-1G>A) of the ASPH gene as well as a heterozygous variant of unknown significance (VUS) of the FBN1 gene (c.6832C>T). CONCLUSION: We here report a novel splice-affecting homozygous pathogenic variant in the ASPH gene that was detected in a Brazilian patient with clinical features of Traboulsi syndrome.

Detection of Biallelic Loss of DNA Repair Genes in Formalin-Fixed, Paraffin-Embedded Tumor Samples Using a Novel Tumor-Only Sequencing Panel.

Patient selection for synthetic lethal-based cancer therapy may be improved by assessment of gene-specific loss of heterozygosity (LOH) and biallelic loss of function (LOF). This report describes SyNthetic lethal Interactions for Precision Diagnostics (SNiPDx), a targeted next-generation sequencing (NGS) panel for detection of LOH and biallelic LOF alterations in 26 target genes focused on DNA damage response pathways, in tumor-only formalin-fixed, paraffin-embedded (FFPE) samples. NGS was performed across all exons of these 26 genes and encompassed a total of 7632 genome-wide single-nucleotide polymorphisms on genomic DNA from 80 FFPE solid tumor samples. The Fraction and Allele-Specific Copy Number Estimates from Tumor Sequencing algorithm was optimized to assess tumor purity and copy number based on heterozygous single-nucleotide polymorphisms. SNiPDx demonstrated high sensitivity (95%) and specificity (91%) for LOH detection compared with whole genome sequencing. Positive agreement with local NGS-based testing in the detection of genetic alterations was 95%. SNiPDx detected 93% of biallelic ATM LOF mutations, 100% of ATM single-nucleotide variants and small insertions/deletions, and 100% of all ATM LOH status events identified by orthogonal NGS-based testing. SNiPDx is a novel, clinically feasible test for analysis of allelic status in FFPE tumor samples, which demonstrated high accuracy when compared with other NGS-based approaches in clinical use.

Outcomes of early versus delayed trabeculotomy for primary congenital glaucoma.

PURPOSE: In patients with primary congenital glaucoma (PCG), elevated intraocular pressure (IOP) causes abnormal eye growth. This study compared the outcomes of children with PCG who underwent ab externo trabeculotomy (TROC) at age ≤ 6 months (early TROC) and of those who underwent TROC at age > 6 months (delayed TROC). METHODS: Intraocular pressure, horizontal corneal diameter (HCD), central corneal thickness (CCT) and axial length (AL) were compared before TROC and at 1-, 3-, 6- and 12-month follow-up visits between the groups of children who underwent TROC until or after 6 months of age. The ALs of these groups were also compared with the ALs of healthy age-matched eyes examined under the same conditions. RESULTS: Trabeculotomy was performed in 43 children: 18 (33 eyes) aged 6 months (group 1) and 25 (37 eyes) aged >6 months (group 2); the mean ages were 86.56 ± 53.64 and 504.48 ± 448.14 days, respectively. The mean pre- and 12-month postoperative IOP values were 15.97 ± 4.78/16.62 ± 4.85 and 9.77 ± 2.88/10.93 ± 4.83 mmHg, respectively. Delayed TROC was associated with abnormal AL in 31 (88.6%) out of 37 eyes, while after early TROC, only 13 (41.9%) out of 33 eyes had abnormal AL (chi-square, 8.00; p = 0.03). In multivariable analysis, each 1-mmHg increase in preoperative IOP was associated with a 0.25-mmHg increase at 12 months (p = 0.04). On average, the mean IOP of the delayed TROC group was higher than that of the early TROC group by 3.72 mmHg at postoperative month 12 (95% CI = 0.44-6.99; p = 0.02). CONCLUSION: Compared with delayed TROC, early TROC is associated with reduced IOP and substantially reduced incidence of abnormal AL at postoperative month 12.

Trends in the epidemiological and microbiological profiles of infectious keratitis in southeastern Brazil.

PURPOSE: To investigate the antibiotic susceptibility as well as the clinical, epidemiological, and microbiological profiles of microbial keratitis. METHODS: This was a longitudinal retrospective study, and we retrospectively reviewed medical and laboratory records from 2015 to 2019. RESULTS: In total, 380 pathogens (321 bacteria and 59 fungi) were isolated from the corneas of 352 patients. Staphylococcus species (45%) were most abundant within the organisms that were isolated, followed by Pseudomonas (18.4%), fungi (15.5%), Streptococcus (7.9%), and Serratia species (3.2%). The isolated gram-positive bacteria were not resistant to amikacin or vancomycin, although 14.8% of the gram-positive isolates were resistant to ciprofloxacin (p<0.05). All the gram-negative isolates were susceptible to amikacin. Male patients represented 62.8% of the 129 cases with accessible clinical data. The mean age of the patients was 53.17 ± 21 years. The time to presentation (from onset of symptoms) was 14.9 ± 19.4 days (median: 7 days). Large ulcers (>5 mm in any dimension) were present in 49.6% (64 eyes) of the cases. The duration of treatment was 49 ± 45.9 days (median: 38 days). Direct ocular trauma was reported by 48 (37.2%) patients, and 15 patients (11.6%) reported using contact lenses. For 72 (55.8%) patients, topical treatment had been previously prescribed, and 16 (12.4%) patients reported using other classes of drugs. Hospitalizations were required for 79 (61.2%) patients, and in terms of major complications, 53 (41.1%) patients had corneal perforations. A total of 40 patients (31%) underwent tectonic penetrating keratoplasty, and 28 (21.7%) developed secondary glaucoma. A progression to endophthalmitis occurred in 8 (6.2%) patients, with 50% of those patients' (3.1% of the total) endophthalmitis evolving to evisceration. The patients' microbial keratitis was largely treated empirically, with 94 (72.9%) patients prescribed moxifloxacin and 56 (43.4%) prescribed ciprofloxacin before receiving their culture results. CONCLUSIONS: For the most part, our hospital treated patients with severe microbial keratitis. Despite identifying gram-positive bacteria in most of the isolates, we also frequently identified gram-negative rods and fungi. Our susceptibility results support prescribing a combination of vancomycin and amikacin as an effective empirical therapeutic regimen to treat microbial keratitis.

Guiding ATR and PARP inhibitor combinationswith chemogenomic screens.

Combinations of ataxia telangiectasia- and Rad3-related kinase inhibitors (ATRis) and poly(ADP-ribose) polymerase inhibitors (PARPis) synergistically kill tumor cells through modulation of complementary DNA repair pathways, but their tolerability is limited by hematological toxicities. To address this, we performed a genome-wide CRISPR-Cas9 screen to identify genetic alterations that hypersensitize cells to a combination of the ATRi RP-3500 with PARPi, including deficiency in RNase H2, RAD51 paralog mutations, or the "alternative lengthening of telomeres" telomere maintenance mechanism. We show that RP-3500 and PARPi combinations kill cells carrying these genetic alterations at doses sub-therapeutic as single agents. We also demonstrate the mechanism of combination hypersensitivity in RNase H2-deficient cells, where we observe an irreversible replication catastrophe, allowing us to design a highly efficacious and tolerable in vivo dosing schedule. We present a comprehensive dataset to inform development of ATRi and PARPi combinations and an experimental framework applicable to other drug combination strategies.

CCNE1 amplification is synthetic lethal with PKMYT1 kinase inhibition.

Amplification of the CCNE1 locus on chromosome 19q12 is prevalent in multiple tumour types, particularly in high-grade serous ovarian cancer, uterine tumours and gastro-oesophageal cancers, where high cyclin E levels are associated with genome instability, whole-genome doubling and resistance to cytotoxic and targeted therapies1-4. To uncover therapeutic targets for tumours with CCNE1 amplification, we undertook genome-scale CRISPR-Cas9-based synthetic lethality screens in cellular models of CCNE1 amplification. Here we report that increasing CCNE1 dosage engenders a vulnerability to the inhibition of the PKMYT1 kinase, a negative regulator of CDK1. To inhibit PKMYT1, we developed RP-6306, an orally bioavailable and selective inhibitor that shows single-agent activity and durable tumour regressions when combined with gemcitabine in models of CCNE1 amplification. RP-6306 treatment causes unscheduled activation of CDK1 selectively in CCNE1-overexpressing cells, promoting early mitosis in cells undergoing DNA synthesis. CCNE1 overexpression disrupts CDK1 homeostasis at least in part through an early activation of the MMB-FOXM1 mitotic transcriptional program. We conclude that PKMYT1 inhibition is a promising therapeutic strategy for CCNE1-amplified cancers.

Twenty-four hours intraocular pressure in keratoconic eyes assessed by applanation tonometry and Tono-Pen AVIA.

AIM: To assess intraocular pressure (IOP) during the daily curve of intraocular pressure (DCPo) in keratoconic eyes and compare Goldmann applanation tonometer (GAT), without and with astigmatism correction (nGAT and cGAT) and Tono-Pen AVIA (TPA) assessment methods. METHODS: Thirty-nine keratoconic eyes of 24 patients were assessed. DCPo was evaluated with five IOP measurements; four were performed with a GAT (nGAT and cGAT), and a Tono-Pen AVIA (TPA) at various times throughout the day. RESULTS: Mean IOP DCPo values (mm Hg) were: nGAT, 9.9±2.6; cGAT, 11.3±2.6; TPA 12.3±3.1. Mean IOP DCPo differences (mm Hg) and Spearman's correlation coefficients were as follows: cGATc-nGAT, 1.32±1.31, r s=0.879 (P<0.01); cGAT-TPA, -1.02±2.08, r s=0.723 (P<0.01); and nGAT-TPA, -2.35±2.23, r s=0.730 (P<0.01). Bland-Altman analysis for agreement between cGAT-TPA and nGAT-TPA mean IOP DCPo measurements revealed a mean difference of 1.02 (95%CI, 0.35-1.70) and 2.35 (95%CI, 1.62-3.07) mm Hg, respectively. Regression analysis yielded the following equation: TPA IOP=5.49+0.775×cGAT-0.015×ACD-0.299×corneal astig matism, which allowed us to infer TPA IOP values from other parameters. CONCLUSION: In keratoconic eyes, IOP peaks of DCPo measurements are identified at 6 a.m., independent of the tonometer. The mean DCPo values are: TPA>cGAT>nGAT. IOP TPA measures are predictive of cGAT values, adjusted according to anterior chamber depth and corneal astigmatism.

RP-3500: A Novel, Potent, and Selective ATR Inhibitor that is Effective in Preclinical Models as a Monotherapy and in Combination with PARP Inhibitors.

Ataxia telangiectasia and Rad3-related (ATR) kinase protects genome integrity during DNA replication. RP-3500 is a novel, orally bioavailable clinical-stage ATR kinase inhibitor (NCT04497116). RP-3500 is highly potent with IC50 values of 1.0 and 0.33 nmol/L in biochemical and cell-based assays, respectively. RP-3500 is highly selective for ATR with 30-fold selectivity over mammalian target of rapamycin (mTOR) and more than 2,000-fold selectivity over ataxia telangiectasia mutated (ATM), DNA-dependent protein kinase (DNA-PK), and phosphatidylinositol 3-kinase alpha (PI3Kα) kinases. In vivo, RP-3500 treatment results in potent single-agent efficacy and/or tumor regression in multiple xenograft models at minimum effective doses (MED) of 5 to 7 mg/kg once daily. Pharmacodynamic assessments validate target engagement, with dose-proportional tumor inhibition of phosphorylated checkpoint kinase 1 (pCHK1) (IC80 = 18.6 nmol/L) and induction of phosphorylated H2A.X variant histone (γH2AX), phosphorylated DNA-PK catalytic subunit (pDNA-PKcs), and phosphorylated KRAB-associated protein 1 (pKAP1). RP-3500 exposure at MED indicates that circulating free plasma levels above the in vivo tumor IC80 for 10 to 12 hours are sufficient for efficacy on a continuous schedule. However, short-duration intermittent (weekly 3 days on/4 days off) dosing schedules as monotherapy or given concomitantly with reduced doses of olaparib or niraparib, maximize tumor growth inhibition while minimizing the impact on red blood cell depletion, emphasizing the reversible nature of erythroid toxicity with RP-3500 and demonstrating superior efficacy compared with sequential treatment. These results provide a strong preclinical rationale to support ongoing clinical investigation of the novel ATR inhibitor, RP-3500, on an intermittent schedule as a monotherapy and in combination with PARP inhibitors as a potential means of maximizing clinical benefit.

Avaliação da relação entre a espessura da camada de fibras nervosas e variação da pressão intraocular em suspeitos de glaucoma e em pacientes com glaucoma primário de ângulo aberto / Evaluation of the relationship between nerve fiber layer thickness and intraocular pressure variation in glaucoma suspects and in patients with primary open-angle glaucoma

Introdução: Glaucoma é uma neuropatia óptica caracterizada pela perda irreversível das células ganglionares retinianas, sendo a pressão intraocular (Po) o principal fator de risco para a doença. Grandes variações da pressão intraocular durante um determinado período de tempo têm atraído a atenção como um potencial fator de risco para o desenvolvimento e progressão do glaucoma. Objetivo: Avaliar a relação entre a flutuação de 24 horas da pressão intraocular (∆Po) e a espessura da camada de fibras nervosas da retina (CFNR) medida por tomografia de coerência óptica de domínio espectral (TCO-DE). Método: O estudo incluiu 125 olhos de 65 pacientes. Sessenta e três olhos (50,4%) tiveram o diagnóstico de glaucoma primário de ângulo aberto (GPAA) e 62 (49,6%) foram considerados suspeitos de glaucoma (SG). Todos os olhos foram submetidos a imagens usando TCO-DE, juntamente com medição da Po de 24 horas e perimetria computadorizada acromática. Apenas pacientes com ∆Po anormal (>6 mmHg) foram incluídos. Correlação e modelos mistos lineares generalizados foram usados para investigar a relação entre ∆Po e a espessura da CFNR ajustando para potenciais fatores de confusão, como idade, diagnóstico de glaucoma, espessura da córnea, Po média durante 24 horas e gravidade da doença. Resultados: A idade média foi semelhante entre os grupos SG e GPAA (62,2 ± 15,6 vs. 64,6 ± 12,0, P = 0,50), enquanto o desvio médio do campo visual apresentou diferença entre SG e GPAA (0,41 ± 1,33 vs. -3,09 ± 3,23, P <0,001). A Po média também foi semelhante entre os grupos SG e GPAA (15,6 ± 3,47 vs. 15,6 ± 2,83 mmHg, P = 0,90), assim como o pico da Po às 6h (21,7 ± 3,85 vs. 21,3 ± 3,80 mmHg, P = 0,68). Correlações negativas estatisticamente significativas foram encontradas no grupo GPAA entre a Po às 6h e a espessura da CFNR global (rs = 0,543; P < 0,001), quadrantes inferior (rs = -0,540; P < 0,001), superior (rs = -0,405; P = 0,009) e nasal (rs = −0,561; P < 0,001). Correlações negativas também foram encontradas entre ∆Po e a espessura da CFNR global (rs = −0,591; P < 0,001), e todos os demais setores (P < 0,05). No SG a Po às 6h correlacionou-se apenas com a espessura da CFNR no quadrante inferior (rs = −0,307; P = 0,047). Cada 1 mmHg maior na ∆Po foi associado à afilamento de -1,44 µm na espessura global da CFNR (IC 95%: -2,77 a -0,11, P = 0,03). Além disso, cada incremento de 10 anos na idade foi associado à afilamento de -3,56 µm na espessura global da CFNR (IC 95%: -6,06 a -1,05, P = 0,006). O GPAA teve uma afilamento média de -10,91 µm na CFNR global em relação aos SG (IC 95%: -20,21 a -1,62, P = 0,02). A Po média não foi associada à espessura global da CFNR (IC 95%: -1,37 a 0,91, P = 0,69). Conclusão: Maior ∆Po foi associado à menor espessura global da CFNR medida pelo TCO-DE, em pacientes com ∆Po >6 mmHg. Esse achado corrobora um papel potencial da flutuação da Po como fator de risco para perda estrutural no glaucoma.

Introduction: Glaucoma is an optic neuropathy characterized by irreversible loss of retinal ganglion cells, with intraocular pressure (IOP) being the main risk factor for the disease. Large variations in IOP over a period of time have attracted attention as a potential risk factor for the development and progression of glaucoma. Purpose: To evaluate the relationship between 24-hour fluctuation of intraocular pressure (∆IOP) and retinal nerve fiber layer (RNFL) thickness measured by spectraldomain optical coherence tomography (SD-OCT). Material and Methods: The study included 125 eyes of 65 patients. Sixty-three eyes (50.4%) had a diagnosis of primary open angle glaucoma (POAG) and 62 (49.6%) were considered glaucoma suspects (GS). All eyes underwent imaging using SDOCT, along with 24-hour IOP measurement and standard automated perimetry. Only patients with abnormal ∆IOP (>6 mmHg) were included. Correlation and generalized linear mixed models were used to investigate the relationship between ∆IOP and RNFL thickness adjusting for potential confounding factors such as age, glaucoma diagnosis, corneal thickness, mean IOP during 24 hours, and disease severity. Results: Mean age was similar between the SG and POAG groups (62.2 ± 15.6 vs. 64.6 ± 12.0, P = 0.50), while the mean deviation of visual field showed a difference between SG and POAG (0.41 ± 1.33 vs. -3.09 ± 3.23, P < 0.001). The mean Po was also similar between GS and POAG groups (15.6 ± 3.47 vs 15.6 ± 2.83 mmHg, P = 0.90) as was IOP peak at 6 AM (21.7 ± 3.85 vs. 21.3 ± 3.80 mmHg, P = 0.68). Statistically significant negative correlations were found in POAG group between IOP at 6 AM and RNFL thickness in global (rs = −0.543; P < 0.001), inferior (rs = −0.540; P < 0.001), superior (rs = −0.405; P = 0.009), and nasal quadrants (rs = −0.561; P < 0.001). Negative correlations were also found between ∆IOP and RNFL thickness in global (rs = −0.591; P < 0.001), and all other sectors (P < 0.05). In GS IOP at 6 AM correlated only with RNFL thickness in the inferior quadrant (rs = −0.307; P = 0.047). Each 1 mmHg higher in ∆IOP was associated with thinning of -1.44 µm in global RNFL thickness (95% CI: -2.77 to -0.11, P = 0.03). Also, each 10-years increment in age was associated with thinning of -3.56 µm in global RNFL thickness (95% CI: 6.06 to -1.05, P = 0.006). POAG had on average -10.91 µm thinning in global RNFL than glaucoma suspects (95% CI: -20.21 to -1.62, P = 0.02). Mean IOP was not associated with global RNFL thickness (95% CI: -1.37 to 0.91, P = 0.69). Conclusion: Higher ∆IOP was associated with lower global RNFL thickness measured by SD-OCT, in patients with ∆IOP > 6 mmHg. This finding corroborates a potential role for IOP fluctuation as a risk factor for structural loss in glaucoma.

Use of machine learning to predict the risk of early morning intraocular pressure peaks in glaucoma patients and suspects / Uso de aprendizado de máquina para predizer o risco de picos de pressão intraocular no início da manhã em pacientes com glaucoma e suspeitos

ABSTRACT Purpose: To use machine learning to predict the risk of intraocular pressure peaks at 6 a.m. in primary open-angle glaucoma patients and suspects. Methods: This cross-sectional observational study included 98 eyes of 98 patients who underwent a 24-hour intraocular pressure curve (including the intraocular pressure measurements at 6 a.m.). The diurnal intraocular pressure curve was defined as a series of three measurements at 8 a.m., 9 a.m., and 11 a.m. from the 24-hour intraocular pressure curve. Two new variables were introduced: slope and concavity. The slope of the curve was calculated as the difference between intraocular pressure measurements at 9 a.m. and 8 a.m. and reflected the intraocular pressure change in the first hour. The concavity of the curve was calculated as the difference between the slopes at 9 a.m. and 8 a.m. and indicated if the curve was bent upward or downward. A classification tree was used to determine a multivariate algorithm from the measurements of the diurnal intraocular pressure curve to predict the risk of elevated intraocular pressure at 6 a.m. Results: Forty-nine (50%) eyes had intraocular pressure measurements at 6 a.m. >21 mmHg, and the median intraocular pressure peak in these eyes at 6 a.m. was 26 mmHg. The best predictors of intraocular pressure measurements >21 mmHg at 6 a.m. were the intraocular pressure measurements at 8 a.m. and concavity. The proposed model achieved a sensitivity of 100% and a specificity of 86%, resulting in an accuracy of 93%. Conclusions: The machine learning approach was able to predict the risk of intraocular pressure peaks at 6 a.m. with good accuracy. This new approach to the diurnal intraocular pressure curve may become a widely used tool in daily practice and the indication of a 24-hour intraocular pressure curve could be rationalized according to risk stratification.

RESUMO Objetivo: Utilizar aprendizado de máquina para predizer o risco de picos de pressão intraocular às 6 AM em pacientes com glaucoma primário de ângulo aberto e suspeitos. Métodos: Esse estudo observacional transversal incluiu 98 olhos de 98 pacientes submetidos à curva de 24 horas de pressão intraocular (incluindo as medidas às 6 AM). A curva diurna de pressão intraocular foi definida como uma série de três medidas da curva de 24 horas de pressão intraocular às 8 AM, às 9 AM e às 11 AM. Duas novas variáveis foram apresentadas: inclinação e concavidade. A inclinação da curva às 8 AM foi calculada como a diferença entre pressão intraocular às 9 AM e 8 AM e reflete a variação da pressão intraocular na primeira hora. A concavidade da curva foi calculada como a diferença entre as inclinações às 9 AM e às 8 AM e pode ser para cima ou para baixo. Uma árvore de classificação foi usada para determinar um algoritmo multivariado a partir das medidas da curva diurna para prever o risco de pressão intraocular elevada às 6 AM. Resultados: Quarenta e nove (50%) olhos apresentaram pressão intraocular às 6 AM >21 mmHg e a mediana do pico de pressão intraocularPIO foi 26 mmHg. Os melhores preditores de pressão intraocular às 6 AM >21 mmHg foram a pressão intraocular às 8 AM e a concavidade. O modelo proposto apresentou uma sensibilidade de 100% e uma especificidade de 86%, com uma acurácia de 93%. Conclusões: A abordagem de aprendizado de máquina foi capaz de prever o risco de picos de pressão intraocular às 6 AM com uma boa acurácia. Essa nova abordagem para a curva diurna de pressão intraocular pode se tornar uma ferramenta amplamente utilizada na prática clínica e a indicação da curva de 24 horas de pressão intraocular pode ser racionalizada de acordo com a estratificação de risco.

Use of machine learning to predict the risk of early morning intraocular pressure peaks in glaucoma patients and suspects.

PURPOSE: To use machine learning to predict the risk of intraocular pressure peaks at 6 a.m. in primary open-angle glaucoma patients and suspects. METHODS: This cross-sectional observational study included 98 eyes of 98 patients who underwent a 24-hour intraocular pressure curve (including the intraocular pressure measurements at 6 a.m.). The diurnal intraocular pressure curve was defined as a series of three measurements at 8 a.m., 9 a.m., and 11 a.m. from the 24-hour intraocular pressure curve. Two new variables were introduced: slope and concavity. The slope of the curve was calculated as the difference between intraocular pressure measurements at 9 a.m. and 8 a.m. and reflected the intraocular pressure change in the first hour. The concavity of the curve was calculated as the difference between the slopes at 9 a.m. and 8 a.m. and indicated if the curve was bent upward or downward. A classification tree was used to determine a multivariate algorithm from the measurements of the diurnal intraocular pressure curve to predict the risk of elevated intraocular pressure at 6 a.m. RESULTS: Forty-nine (50%) eyes had intraocular pressure measurements at 6 a.m. >21 mmHg, and the median intraocular pressure peak in these eyes at 6 a.m. was 26 mmHg. The best predictors of intraocular pressure measurements >21 mmHg at 6 a.m. were the intraocular pressure measurements at 8 a.m. and concavity. The proposed model achieved a sensitivity of 100% and a specificity of 86%, resulting in an accuracy of 93%. CONCLUSIONS: The machine learning approach was able to predict the risk of intraocular pressure peaks at 6 a.m. with good accuracy. This new approach to the diurnal intraocular pressure curve may become a widely used tool in daily practice and the indication of a 24-hour intraocular pressure curve could be rationalized according to risk stratification.

Comparison of Tono-Pen Avia and Handheld Applanation Tonometry in Primary Congenital Glaucoma.

PRCIS: Tono-Pen AVIA (TPA) intraocular pressure (IOP) values are different from those taken with handheld Goldmann applanation tonometer (GAT) in primary congenital glaucoma (PCG). These differences indicate both tonometers cannot be used interchangeably for measuring IOP in PCG. PURPOSE: The aim was to compare IOP measurements obtained using TPA and a handheld version of GAT in children with PCG. MATERIALS AND METHODS: Forty-two eyes from 23 patients were evaluated for central corneal thickness (CCT), axial length, biomicroscopy and IOP measurement with TPA and a handheld GAT under inhalation anesthesia. After 1 eye from each patient was randomized, paired the Student t-test and the Pearson correlation were used for analysis. Generalized linear mixed model was used to estimate the difference between tonometers. RESULTS: Mean age of children was 28.3±20.5 months. Mean axial length was 24.89±3.33 mm and mean CCT was 605.9±81.0 µm. Mean IOP was 22.1±9.6 for TPA and 14.0±4.5 mm Hg for GAT. There was a significant difference of 8.1±6.9 mm Hg between TPA IOP and GAT IOP (P<0.001). Each 6 months increase in age was associated with 1.32 mm Hg reduction in the difference between tonometers (P=0.002) and each 1 mm Hg higher of mean GAT IOP was associated with -0.73 mm Hg in the difference between TPA and GAT (P=0.002). Also, for every 20 µm increase in CCT an increase of 1.16 mm Hg in the difference between both devices was expected (P=0.003), after adjustment for potentially confounding variables. CONCLUSION: There is a significant difference between TPA IOP and GAT IOP in PCG. The difference between TPA and GAT in PCG is influenced by CCT, age and GAT IOP value.

The CIP2A-TOPBP1 axis safeguards chromosome stability and is a synthetic lethal target for BRCA-mutated cancer.

BRCA1/2-mutated cancer cells adapt to the genome instability caused by their deficiency in homologous recombination (HR). Identification of these adaptive mechanisms may provide therapeutic strategies to target tumors caused by the loss of these genes. In the present study, we report genome-scale CRISPR-Cas9 synthetic lethality screens in isogenic pairs of BRCA1- and BRCA2-deficient cells and identify CIP2A as an essential gene in BRCA1- and BRCA2-mutated cells. CIP2A is cytoplasmic in interphase but, in mitosis, accumulates at DNA lesions as part of a complex with TOPBP1, a multifunctional genome stability factor. Unlike PARP inhibition, CIP2A deficiency does not cause accumulation of replication-associated DNA lesions that require HR for their repair. In BRCA-deficient cells, the CIP2A-TOPBP1 complex prevents lethal mis-segregation of acentric chromosomes that arises from impaired DNA synthesis. Finally, physical disruption of the CIP2A-TOPBP1 complex is highly deleterious in BRCA-deficient tumors, indicating that CIP2A represents an attractive synthetic lethal therapeutic target for BRCA1- and BRCA2-mutated cancers.

Correlation between retinal nerve fiber layer thickness and IOP variation in glaucoma suspects and patients with primary open-angle glaucoma.

PURPOSE: To analyze the relationship between retinal nerve fiber layer thickness (RNFLT) and intraocular pressure (IOP) variation in glaucoma suspects (GS) and patients with primary open-angle glaucoma (POAG). METHODS: Thirty-one GS and 34 POAG patients underwent ophthalmologic examination and 24-h IOP measurements. GS had IOPs ranging from 19 to 24 mmHg and/or suspicious appearance of the optic nerve. POAG patients had reproducible abnormal visual fields. We only included patients who presented with short-term IOP fluctuation >6 mm Hg (∆IOP). Only one eye per patient was included through a randomized process. Peripapillary RNFLT was assessed by spectral-domain optical coherence tomography. We correlated RNFLT with IOP parameters. RESULTS: Mean IOP was similar between GS and POAG groups (15.6 ± 3.47 vs 15.6 ± 2.83 mmHg, p = 0.90) as was IOP peak at 6 AM (21.7 ± 3.85 vs 21.3 ± 3.80 mmHg, p = 0.68). Statistically significant negative correlations were found in POAG group between IOP at 6 AM and RNFLT in global (rs = -0.543; p < 0.001), inferior (rs = -0.540; p < 0.001), superior (rs = -0.405; p = 0.009), and nasal quadrants (rs = -0.561; p < 0.001). Negative correlations were also found between ∆IOP and RNFLT in global (rs = -0.591; p < 0.001), and all other sectors (p < 0.05). In GS IOP at 6 AM correlated only with inferior quadrant (rs = -0.307; p = 0.047). CONCLUSION: IOP at 6 AM and ∆IOP had negative correlations with RNFLT quadrants in POAG. In GS this correlation occurred between IOP at 6 AM and inferior quadrant. These findings may indicate potential risk factors for glaucoma progression.

Transcriptome analysis of cadmium exposure in kidney fibroblast cells of the North Atlantic Right Whale (Eubalaena glacialis).

An 8X15k oligonucleotide microarray was developed consisting of 2334 Eubalaena glacialis probes and 2166 Tursiops truncatus probes and used to measure the effects, at transcriptomic level, of cadmium exposure in right whale kidney fibroblast cells. Cells were exposed to three concentrations (1 µM, 0.1 µM, and 0.01 µM) of cadmium chloride (CdCl2) for three exposure times (1, 4, and 24 h). Cells exposed to 1 µM CdCl2 for 4 h and 24 h showed upregulated genes involved in protection from metal toxicity and oxidative stress, protein renaturation, apoptosis inhibition, as well as several regulators of cellular processes. Downregulated genes represented a suite of functions including cell proliferation, transcription regulation, actin polymerization, and stress fiber synthesis. The collection of differentially expressed genes in this study support proposed mechanisms of cadmium-induced apoptosis such as ubiquitin proteasome system disruption, Ca2+ homeostasis interference, mitochondrial membrane potential collapse, reactive oxygen species (ROS) production, and cell cycle arrest. The results also have confirmed the right whale microarray as a reproducible tool in measuring differentiated gene expression that could be a valuable asset for transcriptome analysis of other baleen whales and potential health assessment protocols.

Femtosecond laser-assisted anterior lamellar keratoplasty with suboptimal outcomes: a case series / Ceratoplastia lamelar anterior assistida por laser de femtosegundo com resultados subótimos: uma série de casos

ABSTRACT Objective To identify preoperative clinical characteristics of patients undergoing femtosecond laser-assisted anterior lamellar keratoplasty who failed to achieve optimal postoperative visual outcomes. Methods In this single-center, retrospective case series, patients who underwent femtosecond laser-assisted anterior lamellar keratoplasty between 2013 and 2018 were included if they required graft revision, subsequent corneal procedure, or additional postoperative visits for a femtosecond laser-assisted anterior lamellar keratoplasty-related issue. Visual outcomes assessed included best-corrected visual acuities and postoperative corneal astigmatism. Results Eight eyes of eight patients meeting the above criteria were included. Mean patient age was 64.5 years (range, 21 to 89 years). Mean included preoperative best-corrected visual acuities was one logarithm of the minimum angle of resolution (range, 0.3 logarithm of the minimum angle of resolution to counting fingers). Indications for femtosecond laser-assisted anterior lamellar keratoplasty included anterior stromal scarring due to viral keratitis (two cases), bacterial keratitis (one case), chronic epithelial defect (one case), Avellino dystrophy (one case), trauma (one case), and chronic endothelial failure (two cases). Six patients had history of prior intraocular surgeries including phacoemulsification (four cases), pars plana vitrectomy (one case), endothelial keratoplasty (two cases), and trabeculectomy (one case). Mean included best-corrected visual acuities at most recent follow-up was one logarithm of the minimum angle of resolution (range zero logarithm of the minimum angle of resolution to hand movements) representing improvement or stability in six of eight patients. Visually significant corneal astigmatism was present in four of eight patients. Post-femtosecond laser-assisted anterior lamellar keratoplasty procedures included graft repositioning, arcuate keratotomy, phacoemulsification, and regraft. Conclusion While femtosecond laser-assisted anterior lamellar keratoplasty offers a less-invasive treatment option compared to penetrating keratoplasty, intraoperative and postoperative management can be complex. Femtosecond laser-assisted anterior lamellar keratoplasty in patients with history of prior endothelial keratoplasty or ongoing ocular comorbidities should be pursued with caution.

RESUMO Objetivo Identificar as características clínicas pré-operatórias de pacientes submetidos à ceratoplastia lamelar anterior assistida por laser de femtossegundo que não alcançaram resultados visuais pós-operatórios ideais. Métodos Nesta série de casos retrospectiva em um único centro, os pacientes submetidos à ceratoplastia lamelar anterior assistida por laser de femtossegundo entre 2013 e 2018 foram incluídos se precisassem de revisão do enxerto, procedimento corneano subsequente ou visitas pós-operatórias adicionais por uma intercorrência relacionada à ceratoplastia lamelar anterior assistida por laser de femtossegundo. Os resultados visuais avaliados incluíram melhor acuidade visual corrigida e astigmatismo pós-operatório da córnea. Resultados Oito olhos de oito pacientes que atenderam aos critérios descritos foram incluídos. A idade média dos pacientes foi de 64,5 anos (variação de 21 a 89). A melhor acuidade visual corrigida pré-operatória média foi de um logaritmo do mínimo ângulo de resolução (variação de 0,3 logaritmo do mínimo ângulo de resolução para contagem de dedos). As indicações para ceratoplastia lamelar anterior assistida por laser de femtossegundo incluíram cicatriz do estroma anterior devido à ceratite viral (dois casos), ceratite bacteriana (um caso), defeito epitelial crônico (um caso), distrofia de Avellino (um caso), trauma (um caso) e insuficiência endotelial crônica (dois casos). Seis pacientes tinham história de cirurgias intraoculares anteriores, incluindo facoemulsificação (quatro casos), vitrectomia via pars plana (um caso), ceratoplastia endotelial (dois casos) e trabeculectomia (um caso). O mínimo ângulo de resolução médio no acompanhamento mais recente foi de um logaritmo do mínimo ângulo de resolução (variação de zero logaritmo do mínimo ângulo de resolução para movimentos das mãos), representando melhora ou estabilidade em seis de oito pacientes. Astigmatismo corneano visualmente significativo estava presente em quatro de oito pacientes. Os procedimentos pós-ceratoplastia lamelar anterior assistida por laser de femtossegundo incluíram reposicionamento do enxerto, ceratotomia arqueada, facoemulsificação e enxerto. Conclusão Embora a ceratoplastia lamelar anterior assistida por laser de femtossegundo ofereça uma opção de tratamento menos invasiva em comparação com a ceratoplastia penetrante, o manejo intra e pós-operatório pode ser complexo. A ceratoplastia lamelar anterior assistida por laser de femtossegundo em pacientes com história de ceratoplastia endotelial anterior ou comorbidades oculares correntes deve ser avaliada com cautela.

Author Correction: Characterization of Nigerian breast cancer reveals prevalent homologous recombination deficiency and aggressive molecular features.

The original version of this Article contained an error in the author affiliations. The affiliation of Kevin P. White with Tempus Labs, Inc. Chicago, IL, USA was inadvertently omitted. This has now been corrected in both the PDF and HTML versions of the Article.