Correction: The replication initiator protein of a geminivirus interacts with host monoubiquitination machinery and stimulates transcription of the viral genome.

[This corrects the article DOI: 10.1371/journal.ppat.1006587.].

Eyelid sebaceous gland carcinoma, validation of AJCC 8th edition T staging- a retrospective study from North India.

PURPOSE: This study compares the 8th edition staging of AJCC for prognosis of eyelid Sebaceous Gland Carcinoma (SGC) patients with respect to the 7th edition. METHODS: A retrospective study was undertaken over a period of 100 months. Ninety-nine histopathologically proven cases of eyelid SGC available for follow-up were recruited. Patients were staged by both the 7th and 8th editions of AJCC and followed up at six monthly intervals after surgery. RESULTS: Of the 99 eyelid SGC patients recruited, recurrence and orbital invasion developed in 22%, lymph node metastasis in 21% and 4% had distant metastasis and also died. High-risk features include tumour size>20 mm, orbital invasion, exenteration and staging by both the 7th and 8th editions of AJCC. Cox regression analysis revealed that staging by AJCC 8th edition was associated with greater risk for local recurrence (HR 3.01,95% CI-1.65-5.51%, p < 0.01) lymph node metastasis (HR 8.26, 95% CI 3.96-17.19%, p < 0.01) and disease-free survival (HR 4.61, 95% CI 2.81-7.54). The Kaplan-Meir survival curves of eyelid SGC patients by the 8th edition AJCC staging were also significantly associated with lymph node metastasis (p < 0.01), tumour-related deaths (p < 0.01) and reduced disease-free survival (p = 0.07). The higher Harrell's values by the 8th edition signify better predictive value for lymph node metastasis and DFS (disease-free survival). The lower AIC values indicate a better monotonicity of gradients for lymph node metastasis, recurrence and DFS. CONCLUSION: Staging by the 8th AJCC edition is, therefore, recommended for eyelid SGC as it gives a better perspective about disease outcome. The orbital extension was the single most important predictor of lymph node metastasis, recurrence and death.

Health Care Organization Policies for Employee Safety and COVID-19 Pandemic Response: A Mixed-Methods Study.

OBJECTIVE: The coronavirus (COVID-19) pandemic impacted the well-being of health care workers. We examined the association between prepandemic perceptions of perceived organizational support for safety (using NOSACQ-50), safety hazards and the pandemic's impact on individual workers and institutions. METHODS: Questionnaires from health care staff of five public health care facilities were collected in 2018 ( n = 1059) and 2021 ( n = 1553). In 2021, 17 workers were interviewed from the same facilities. RESULTS: Interviewees reported that their organizations struggled to communicate due to changing guidelines, inadequate personal protective equipment, training, and infection control, early in the pandemic. Questionnaire reports of decreased staffing and increased workload during the pandemic were associated with lower baseline NOSACQ scores. CONCLUSION: Survey findings predicted some variation in organizational response to the pandemic. Better organizational policies for employee safety and pandemic planning could improve health care institutions' preparedness.

Association of kidney functions with a cascade of care for diabetes and hypertension in two geographically distinct Indian cohorts.

AIM: Type 2 diabetes (T2DM) and hypertension (HTN) are the main modifiable risk factors of chronic kidney disease (CKD), among the known traditional and non-traditional risk factors. METHODS: We determined the prevalence and care-cascade of these modifiable CKD-risk factors and their association with socioeconomic status in adjoining Lucknow and Puducherry cities of India. RESULTS: 439 participants reported no CKD were recruited. Serum analysis revealed an Estimated Glomerular Filtration Rate (eGFR) ≥ 90 ml/min/1.73 m2 in 60.36% of the population. Of them, 55.85% had HTN and/or T2DM as CKD-risk factors; however, less than half of this population was unaware of their CKD-risk status. Awareness and treatment were significantly higher in Puducherry and were associated with literacy, occupation, and residence place. Although the CKD-risk population was about two times higher in Puducherry than Lucknow, the populations with mild-low eGFR were comparable in the two regions. Moreover, in Lucknow, mild-low eGFR and low awareness were more prevalent among the younger participants (<30 years), relative to Puducherry. CONCLUSIONS: Despite a higher prevalence of CKD-risk factors in Puducherry, populations with mild-low eGFR were comparable to Lucknow. More heightened awareness and better care cascade for CKD-risk factors in Puducherry may prevent or delay eGFR reduction.

The Descemet Membrane in Primary Congenital Glaucoma.

PURPOSE: To evaluate Descemet membrane (DM) morphology in eyes with primary congenital glaucoma (PCG) in vivo using high-definition anterior segment optical coherence tomography (ASOCT) and on histopathology. METHODS: Corneal scans of patients with PCG (22 eyes of 15 patients) were evaluated for DM morphology and anterior chamber angle using ASOCT. The DM thickness in PCG eyes was compared with fellow eyes (8 eyes) of unilateral patients with PCG and healthy controls (12 eyes) on ASOCT. The DM morphology was also compared on the histopathology of corneal tissues (9) obtained from PCG eyes after keratoplasty and enucleated eyes of retinoblastoma (6 controls) on light microscopy with immunostaining for collagen IV. RESULTS: On ASOCT, all affected eyes showed the presence of either a thickened DM complex or a hyper-reflective double layer representing the thickened DM and pre-Descemet layer (PDL), unlike a single membrane in the controls and fellow eyes. On ASOCT, among patients with PCG, the DM showed significant thickening (32.0 ± 11.2 µm) versus fellow eyes (14.4 ± 3.3 µm) and controls (11.5 ± 1 µm) (P < 0.001; analysis of variance). The thickened DM complex continued peripherally into the trabecular meshwork as an abnormal membrane in 16/22 affected eyes. On histopathology, thickening of DM was also more among PCG eyes (median: 67.9 µm range: 27.2-214.9) versus controls (median: 27.7 µm, range: 22.1-36.1; P = 0.005) as also of PDL (median: 14 µm, range: 5.9-30.5) of PCG versus (median 3.5, range: 1.3-6.7 µm) in controls; P = 0.014. CONCLUSIONS: Thickening of DM and PDL occurs in eyes with PCG and is seen to have a peripheral extension upto the angle recess.

Vimentin overexpression as a novel poor prognostic biomarker in eyelid sebaceous gland carcinoma.

BACKGROUND: Vimentin is an intermediate-sized filament which is highly expressed in mesenchymal cells and is associated with epithelial-mesenchymal transition (EMT). EMT markers ZEB2 and Slug lead to Vimentin overexpression and E-cadherin loss, resulting in invasion and metastasis. However, the status of Vimentin remains unexplored in eyelid sebaceous gland carcinoma (SGC). The study aims to determine status of Vimentin in SGC and its association with EMT markers E-cadherin, ZEB2 and Slug. METHODS: Vimentin protein expression was undertaken in 66 cases with SGC by immunohistochemistry (IHC). Messenger RNA (mRNA) expression was determined in 42 fresh tissues by quantitative real-time PCR. Association of Vimentin with E-cadherin, ZEB2 and Slug was also analysed. Patients were followed up for 17-69 months (mean 34.02 ± 14.73 months). RESULTS: IHC revealed Vimentin overexpression in 37/66 (56%) cases. This overexpression showed significant association with lymph node metastasis (p=0.004) and pagetoid spread (p=0.05). Patients with high Vimentin expression also had poor disease-free survival (p=0.033). Univariate Cox regression model indicated that high Vimentin expression (p=0.043) and advanced tumour stage (p=0.002) were independent adverse prognostic factors. High Vimentin mRNA expression was seen in 16/42 (38%) cases and correlated significantly with lymph node metastasis (p=0.027), advanced tumour stage (p=0.002) and large tumour size (p=0.023). Vimentin expression overall showed a significant inverse association with E-cadherin and direct association with ZEB2 expression. CONCLUSIONS: Vimentin overexpression in SGC is associated with EMT and leads to poor clinical outcome. It also emerged as a novel predictor for lymph node metastasis and poor survival.

miRNA-200c and miRNA-141 as potential prognostic biomarkers and regulators of epithelial-mesenchymal transition in eyelid sebaceous gland carcinoma.

BACKGROUND: MicroRNA (miRNA)-200c and miRNA-141 are tumour suppressors, which regulate epithelial-mesenchymal transition (EMT), leading to tumour invasion and metastasis in various malignancies. miRNA-200c and miRNA-141 maintain the epithelial phenotype by post-transcriptionally inhibiting the E-cadherin repressors, zinc finger E-box binding homeobox (ZEB)1 and ZEB2. The present study was performed to determine the prognostic significance of miRNA-200c and miRNA-141, and their association with EMT markers ZEB1, ZEB2 and E-cadherin in eyelid sebaceous gland carcinoma (SGC). METHODS: Expression levels of miRNA-200c and miRNA-141 were determined in 42 eyelid SGC cases by quantitative real-time PCR (qPCR). Their association with ZEB1, ZEB2 and E-cadherin was determined by qPCR and immunohistochemistry. Kaplan-Meier plots and Spearman's rank correlation tests were applied to analyse the data. Patients were followed up for 7-44 months. RESULTS: Low expression levels of miRNA-200c and miRNA-141 were seen in 36/42 (86%) and 28/42 (67%) cases, respectively. Low miRNA-200c correlated significantly with large tumour size (p=0.03) and poor differentiation (p=0.03). Low miRNA-141 correlated significantly with large tumour size (p=0.02) and lymph node metastasis (p=0.04). Survival analysis revealed that patients with low miRNA-200c (p<0.05) and miRNA-141 expression (p=0.07) had shorter disease-free survival. There was a significant association of both miRNA-200c and miRNA-141 with E-cadherin and ZEB2 expression. CONCLUSIONS: Low levels of miRNA-200c and miRNA-141 in patients with eyelid SGC facilitates tumour progression by promoting EMT and miRNA-200c has emerged as a novel potential predictor of survival.

Chlamydia and ocular adnexal lymphomas: An Indian experience.

CHLAMYDIA AND OCULAR ADNEXAL LYMPHOMAS: AN INDIAN EXPERIENCE: Ocular adnexal lymphomas (OALs) are a heterogeneous group of malignancies, majority being extranodal mucosa-associated lymphoid tissue (MALT) type. Different geographical regions have reported association of Chlamydia with OALs (MALT type). In India, role of Chlamydia in OALs remains unexplored. The aim of this study was to detect Chlamydia and to correlate with clinicopathological features of OALs in India. The clinicopathological features of 41 OAL cases were studied prospectively. Chlamydia DNA was detected by genus specific PCR amplifying major outer membrane protein (MOMP) gene followed by DNA sequencing. Chlamydia immunoexpression was evaluated by immunofluorescence and immunohistochemistry. The results were correlated with clinicopathological features including follow-up and survival. Chlamydia genome was detected in 3/41 (7.3%) OAL cases by PCR. Direct sequencing revealed C. trachomatis in 3 positive cases. Immunofluorescence and immunohistochemistry showed Chlamydia antigen in 5/41 and 1/41 cases respectively. Immunofluorescence demonstrated higher sensitivity than immunohistochemistry. A significant association was observed between Chlamydia positivity and orbital location (P=0.05). Follow-up revealed relapse in 2 Chlamydia positive cases (P=0.056). Our results demonstrate for the first time presence of C. trachomatis genome in 7.3% OAL cases in India. As no other reports are documented, more detailed studies from different regions within India are needed to explore status of Chlamydia in OALs.

ZEB2/SIP1 as novel prognostic indicator in eyelid sebaceous gland carcinoma.

Epithelial-mesenchymal transition (EMT) plays a pivotal role in tumor invasion and metastasis in various malignancies. ZEB2/SIP1 is an important EMT regulator and down-regulates E-cadherin expression. The present study was planned to explore status of EMT-associated markers ZEB2/SIP1 and E-cadherin in eyelid sebaceous gland carcinoma (SGC) and to correlate with clinicopathological high-risk features. Expressions of ZEB2 and E-cadherin were evaluated by immunohistochemistry in 65 cases of histopathologically proven eyelid SGC. The results were correlated with clinicopathological high-risk features and survival of the patients to determine the prognostic significance of ZEB2, E-cadherin, and various high-risk features. Cytoplasmic overexpression of ZEB2 and membranous loss of E-cadherin were seen in 68% and 66% of cases of eyelid SGC, respectively. ZEB2 overexpression was significantly associated with E-cadherin loss (P = .002). Overexpression of ZEB2 also showed significant association with lymph node metastasis (P = .046), orbital invasion (P = .049), large tumor size (P = .018), and advanced tumor stages (P = .036). Survival analysis revealed that patients with ZEB2 overexpression had poor survival. ZEB2 overexpression and orbital invasion were found to be independent prognostic indicators (univariate analysis). However, multivariate analysis showed that ZEB2 (hazard ratio, 0.094; 95% confidence interval, 00.012-0.709; P = .022) was the best poor prognostic indicator of eyelid SGC. Our study demonstrates the role of both ZEB2 and E-cadherin in the promotion of EMT in eyelid SGC. The outcome of this study also points toward ZEB2 as an independent prognostic marker as well as a potential therapeutic target in eyelid SGC.