Off-label use of intravitreal bevacizumab: A global conundrum.

Bevacizumab is a monoclonal, humanized, full-length antibody targeting vascular endothelial growth factor(VEGF-A), known for its anti-angiogenic properties. The off-label use of bevacizumab has stirred legal, financial, industrial, and ethical complexities. With its potential to treat diverse ocular conditions, this commentary delves into the multifaceted dimensions of bevacizumab's off-label utilization, encompassing clinical trials, regulatory frameworks, safety considerations, comparative effectiveness, and economic implications.

Efficacy, Safety and Immunogenicity of Sun's Ranibizumab Biosimilar in Neovascular Age-Related Macular Degeneration: A Phase 3, Double-Blind Comparative Study.

INTRODUCTION: The study aimed to evaluate comparability in terms of efficacy, safety and immunogenicity of Sun's ranibizumab biosimilar with reference ranibizumab in patients with neovascular age-related macular degeneration (nAMD). METHODS: This prospective, randomised, double-blind, two-group, parallel-arm, multicentre, phase 3 comparative study included patients with nAMD ≥ 50 years, randomised (in a 2:1 ratio) in a double-blind manner to receive 0.5 mg (0.05 mL) intravitreal injection of either Sun's ranibizumab or reference ranibizumab in the study eye every 4 weeks until week 16 (total of four doses). RESULTS: Primary endpoint results demonstrated equivalence in the proportion of patients who lost fewer than 15 letters from baseline best-corrected visual acuity (BCVA) to the end of week 16 (99% of patients in Sun's ranibizumab and 100% in reference ranibizumab; p > 0.9999), with the proportional difference (90% confidence interval) at -1% (-2.51, +0.61) lying within a pre-specified equivalence margin. Visual acuity improved by 15 or more letters in 43% of Sun's ranibizumab group and 37% of the reference ranibizumab group (p = 0.4267). The mean increase in BCVA was 15.7 letters in Sun's ranibizumab group and 14.6 letters in the reference ranibizumab group (p < 0.001 within both groups and p = 0.5275 between groups). The mean change in central macular thickness was comparable between groups (p = 0.7946). Anti-ranibizumab antibodies were found in one patient of the reference ranibizumab group, while neutralising antibodies were not found in any patients. Both products were well tolerated. CONCLUSION: Sun's ranibizumab biosimilar is found to be therapeutically equivalent to reference ranibizumab in patients with nAMD. There were no additional safety or immunogenicity concerns. TRIAL REGISTRATION: CTRI/2020/09/027629, registered on 07 September 2020.

Breast cancer: A comparative review for breast cancer detection using machine learning techniques.

Breast cancer is the most common cancer among women globally and presents a significant challenge due to its rising incidence and fatality rates. Factors such as cultural, socioeconomic, and educational barriers contribute to inadequate awareness and access to healthcare services, often leading to delayed diagnoses and poor patient outcomes. Furthermore, fostering a collaborative approach among healthcare providers, policymakers, and community leaders is crucial in addressing this critical women's health issue, reducing mortality rates, alleviating, and the overall burden of breast cancer. The main goal of this review is to explore various techniques of machine learning algorithms to examine high accuracy and early detection of breast cancer for the safe health of women.

Anti-Vegf therapy leads to an improvement in grade of retinal pigment epithelium alterations on single layer retinal pigment epithelium map in diabetic macular edema.

PURPOSE: In bovine retinal pigment epithelium (RPE) cells, increased secretion of vascular endothelial growth factor (VEGF) has a positive linear association with proliferation of RPE. Spectral domain optical coherence tomography (SD-OCT) based improvement in grades of topographic retinal pigment epithelium alterations (RPE-A), were evaluated after intravitreal anti-VEGF therapy, in diabetic macular edema (DME), for the first time. METHODS: A tertiary care center-based, prospective study. Forty-four consecutive patients, 40-65 years of age with type 2 diabetes mellitus (DM) with DME, were administered three doses of anti-VEGF therapy at monthly intervals. Pre- and post-intervention SD-OCT was done and central sub field thickness (CST), cube average thickness (CAT) and topographic grades of RPE-A were assessed using single layer RPE map (SL-RPE) as; Grade 0: No alterations, Grade 1: Alteration in two quadrants, Grade 2: Alteration in more than two quadrants. RESULTS: CST decreased from 354.2 ± 16.0 µm pre-intervention to 233.2 ± 7.9 µm post-intervention. CAT reduced from 340.6 ± 6.5 µm pre-intervention to 274.1 ± 5.1 µm post-intervention. Significant improvement in grades of RPE-A pre- v/s post-intervention were observed. (Grade 0: 0 v/s 39; Grade 1: 17 v/s 3; Grade 2: 27 v/s 2) (p < 0.001). CONCLUSION: Anti-VEGF therapy is associated with an improvement in grades of RPE-A in DME.The study was registered with the Clinical Trial Registry of India (CTRI/2019/03/018135).

E2F1-induced lncRNA, EMSLR regulates lncRNA LncPRESS1.

E2F1 induces hundreds of protein-coding genes influencing diverse signaling pathways but much less is known about its non-coding RNA targets. For identifying E2F1-dependent oncogenic long non-coding RNAs (lncRNAs), we carried out genome-wide transcriptome analysis and discovered an lncRNA, EMSLR, which is induced both in lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC). EMSLR depletion blocks the cells in G1 phase and inhibits the clonogenic ability indicating that it is essential for the tumor-related phenotypes. We discovered that EMSLR represses the promoter activity of another lncRNA, LncPRESS1, which is located 6.9 kb upstream of EMSLR and they display an inverse expression pattern in lung cancer cell lines. Depletion of C-MYC results in downregulation of EMSLR and simultaneous upregulation of EMSLR target LncPRESS1, exemplifying how C-MYC and E2F1 signal transduction pathways control the network of lncRNA genes to modulate cell proliferation and differentiation.

Higher Levels of Serum Ionic Calcium are Associated with Macular Edema in Patients with Diabetic Retinopathy.

BACKGROUND: The aim was to study the association of serum total calcium, ionic calcium, and magnesium with increases in macular thickness parameters and photoreceptor ellipsoid zone (EZ) disruption in diabetic macular edema (DME). METHODS: This study is a tertiary care center based observational cross-sectional study with sixty-six consecutive cases, divided into 3 groups of 22 cases each with no diabetic retinopathy (No DR), non-proliferative diabetic retinopathy (NPDR), and proliferative diabetic retinopathy (PDR) and a control group of 22 healthy controls. Best corrected visual acuity (BCVA) was measured on logMAR scale. Central subfield thickness (CST), cube average thickness (CAT), and EZ disruption were assessed using spectral-domain optical coherence tomography (SD-OCT). Serum total calcium, ionic calcium, and magnesium were measured using standard protocol. Data was analyzed statistically. RESULTS: Significant correlation was found between the increase in CST and the increase in serum total calcium and serum ionic calcium. Increase in CAT was significantly correlated with an increase in serum total calcium, serum ionic calcium, and a decrease in serum magnesium. Grades of EZ disruption and logMAR BCVA were also found to be significantly positively associated with serum total calcium and ionic calcium and negatively with serum magnesium. CONCLUSIONS: Increased levels of serum ionic calcium and decreased levels of serum magnesium are associated with an increase in macular thickness and EZ disruption in DME.

Analysis of biometric parameters of 2340 eyes measured with optical biometer Lenstar LS900 in a Caucasian population.

OBJECTIVES: To describe the pattern and mutual relationships between basic biometric characteristics of the eye in a Central European Caucasian population. METHODS: A single-centre retrospective study of 2340 patients (965 males, 1375 females) scheduled for cataract surgery between 2014 and 2016. Measurements using laser interferometry included AL (axial length), K (average corneal curvature), ACD (anterior chamber depth), LT (lens thickness), CCT (central corneal thickness), AST (astigmatism) and WTW (white to white). Subjects were stratified by gender and controlled for age. Descriptive, correlation and regression analyses were performed on the data. RESULTS: The mean AL was 23.33 ± 1.01 mm - higher in males (23.59 ± 0.99 mm), in comparison to females (23.15 ± 0.99 mm). The elderly had lower ACD and higher LT, while males had higher AL independent of age. Furthermore, LT and K decreased with AL, while ACD decreased with LT and increased with AL independent of age and gender. CONCLUSIONS: The estimates of the biometrics are obtained on a large sample of subjects and can serve as normative values for Lenstar LS900 in the Central European Caucasian population.

External limiting membrane and ellipsoid zone structural integrity in diabetic macular edema.

The external limiting membrane (ELM) and ellipsoid zone (EZ) can be observed exquisitely by SD-OCT. In diabetic macular edema (DME), dysfunction of mitochondria, represented by the EZ in the foveal photoreceptors results in reduced visual acuity (VA). An increase in VEGF was found to correlate with increased severity of DR, increased central subfield thickness (CST), and sequential disruption of ELM and EZ. The mechanism of ELM and EZ restoration after anti-VEGF therapy in DME has been discovered. The ELM restores first followed by EZ restoration. Thus, authors have discovered and established ELM as a novel retinal structural barrier.

Elevated advanced glycation end products are associated with subfoveal ellipsoid zone disruption in diabetic macular edema.

PURPOSE: Advanced glycation end products (AGEs), due to increased production and a slow turnover rate, serve as mediators of "metabolic memory" even after the resolution of hyperglycemia. A prospective study was undertaken to evaluate the association of AGEs with subfoveal ellipsoid zone (EZ) disruption in diabetic macular edema (DME). METHODS: A tertiary-care-center-based cross-sectional study included 40 consecutive cases with DME and 20 healthy controls in the age group of 40-65 years. All the study subjects underwent spectral-domain optical coherence tomography (SD-OCT) for cross-sectional imaging of the retina. The EZ was defined as a hyperreflective band below the external limiting membrane. The disruption of EZ was graded as intact EZ and disrupted EZ. Serum AGEs were assessed by assay of Nε-carboxymethyl-lysine (Nε-CML) using the standard protocol. Data were analyzed statistically. RESULTS: Subfoveal EZ disruption was noted in 80% (32/40) of the cases of DME. In the cases without EZ disruption, visual acuity (LogMAR VA) was 0.60 ± 0.52, whereas in cases with EZ disruption, LogMAR VA was 0.96 ± 0.56 (P < 0.001). In the cases without EZ disruption, Nε-CML was 94.31 ± 57 ng/mL, whereas in cases with EZ disruption Nε-CML was 120.64 ± 71.98 ng/mL (P < 0.001). CONCLUSION: In DME, increased levels of AGEs are significantly associated with EZ disruption on SD-OCT.

The lncRNA HMS recruits RNA-binding protein HuR to stabilize the 3'-UTR of HOXC10 mRNA.

Long noncoding RNAs (lncRNAs) have been reported to drive key cancer pathways but the functions of majority of lncRNAs are unknown making a case for comprehensive functional evaluation of lncRNAs. With an aim to identify lncRNAs dysregulated in human cancers, we analyzed the cancer patient database of lung adenocarcinoma (LUAD), which revealed an upregulated lncRNA, LINC02381 (renamed HOXC10mRNA stabilizing factor or HMS in this study), whose depletion results in proliferation defects and inhibition of colony formation of human cancer cells. In order to identify the binding targets of HMS, we screened for cis-genes and discovered that HOXC10, an oncogene, is downregulated in the absence of HMS. Depletion of HMS does not affect the HOXC10 promoter activity but inhibits the HOXC10 3'-UTR-linked luciferase reporter activity. Since lncRNAs have been known to associate with RNA-binding proteins (RBPs) to stabilize mRNA transcripts, we screened for different RBPs and discovered that HuR, an ELAV family protein, stabilizes HOXC10 mRNA. Using RNA pull-down and deletion mapping experiments, we show that HuR physically interacts with the cytosine-rich stretch of HMS and HOXC10 3'-UTR to stabilize HOXC10 mRNA. HOXC10 is overexpressed in many human cancers, and our discovery highlights that lncRNA HMS sustains the HOXC10 mRNA levels to maintain the invasive phenotypes of cancer cells.

Sequential restoration of external limiting membrane and ellipsoid zone after intravitreal anti-VEGF therapy in diabetic macular oedema.

BACKGROUND/OBJECTIVES: To study the mechanism of restoration of retinal photoreceptor ellipsoid zone (EZ), after intravitreal bevacizumab (IVB) therapy, in diabetic macular oedema (DMO). SUBJECTS/METHODS: Forty-four consecutive patients aged 40-65 years having type 2 diabetes mellitus (DM) with DMO were prospectively recruited for IVB therapy. It comprised of three doses (1.25 mg in 0.05 ml) of IVB at monthly intervals. Patients with other ocular and systemic diseases affecting retinal vessels and earlier ophthalmological interventions were excluded. Visual acuity (logMAR VA) was recorded. Spectral domain optical coherence tomography (SD-OCT) was performed pre and post intervention. Central sub-foveal thickness (CST) and grades of disorganization of retinal inner layers (DRIL), external limiting membrane (ELM) and EZ were assessed. Data were statistically analysed on SPSS software. Clinical trials registry: CTRI/2019/03/018135. RESULTS: Mean logMAR VA decreased after IVB therapy from 1.78 ± 0.07 pre-intervention to 0.42 ± 0.05 post intervention (p < 0.001). Similarly, CST reduced from 354.23 ± 15.0 µm pre-intervention to 233.18 ± 7.88 µm post intervention (p < 0.001). Among qualitative variables, DRIL decreased from 93.2% pre-intervention to 13.6% post intervention. Likewise, global ELM disruption reduced from 81.8 to 9.1% and global EZ disruption reduced from 79.5 to 11.4%. ELM restoration preceded EZ restoration. CONCLUSION: Anti-VEGF therapy restores the barrier effect of ELM. It causes ELM to restore first followed by EZ restoration in DMO.

Otological and Visual Implications of Diabetes Mellitus in North Indian Population.

The worldwide population of diabetic patients is increasing alarmingly with India claiming number one position. It causes irreversible damage to cochlear hair cells, vestibular apparatus, visual pathway, nephrons, nerves, if not checked in time. A total of 188 patients of diabetes mellitus were included in this prospective study. The patients underwent routine anamnesis, hearing handicap inventory and dizziness handicap inventory assessment along with clinical examination for audiological, vestibular, neurological and ophthalmological (fundoscopy) status. In our study a sensorineural hearing loss, retinopathy, neuropathy, vestibulopathy was seen in diabetic patients.

Connective tissue fibroblasts from highly regenerative mammals are refractory to ROS-induced cellular senescence.

A surveillance system in mammals constantly monitors cell activity to protect against aberrant proliferation in response to damage, injury and oncogenic stress. Here we isolate and culture connective tissue fibroblasts from highly regenerative mammals (Acomys and Oryctolagus) to determine how these cells interpret signals that normally induce cellular senescence in non-regenerating mammals (Mus and Rattus). While H2O2 exposure substantially decreases cell proliferation and increases p53, p21, p16, and p19 in cells from mice and rats, cells from spiny mice and rabbits are highly resistant to H2O2. Quantifying oxygen consumption and mitochondrial stability, we demonstrate that increased intracellular H2O2 is rapidly detoxified in regenerating species, but overwhelms antioxidant scavenging in cells from non-regenerative mammals. However, pretreatment with N-acetylcysteine (NAC) protects mouse and rat cells from ROS-induced cellular senescence. Collectively, our results show that intrinsic cellular differences in stress-sensing mechanisms partially explain interspecific variation in regenerative ability.

The microRNAs miR-449a and miR-424 suppress osteosarcoma by targeting cyclin A2 expression.

MicroRNAs of the miR-16 and miR-34 families have been reported to inhibit cell cycle progression, and their loss has been linked to oncogenic transformation. Utilizing a high-throughput, genome-wide screen for miRNAs and mRNAs that are differentially regulated in osteosarcoma (OS) cell lines, we report that miR-449a and miR-424, belonging to the miR-34 and miR-16 families, respectively, target the major S/G2 phase cyclin, cyclin A2 (CCNA2), in a bipartite manner. We found that the 3'-UTR of CCNA2 is recognized by miR-449a, whereas the CCNA2 coding region is targeted by miR-424. Of note, we observed loss of both miR-449a and miR-424 in OS, resulting in derepression of CCNA2 and appearance of aggressive cancer phenotypes. Ectopic expression of miR-449a and miR-424 significantly decreased cyclin A2 levels and inhibited proliferation rate, migratory potential, and colony-forming ability of OS cells. To further probe the roles of miR-449a and miR-424 in OS, we developed an OS mouse model by intraosseous injection of U2OS cells into the tibia bone of NOD-scid mice, which indicated that miR-449a and miR-424 co-expression suppresses tumor growth. On the basis of this discovery, we analyzed the gene expression of human OS biopsy samples, revealing that miR-449a and miR-424 are both down-regulated, whereas cyclin A2 is significantly up-regulated in these OS samples. In summary, the findings in our study highlight that cyclin A2 repression by miRNAs of the miR-16 and miR-34 families is lost in aggressive OS.

Prevalence of Age-Related Macular Degeneration in Slovakia and Associated Risk Factors: A Mobile Clinic-Based Cross-Sectional Epidemiological Survey.

PURPOSE: Age-related macular degeneration (AMD) is one of the leading causes of blindness in the elderly population. Although there are prevalence studies for AMD in Europe, data are scarce for the Slovakian population. METHODS: This was a prospective, multicenter, non-interventional, mobile clinic-based cross-sectional study that assessed age-specific prevalence of AMD in the Slovakian population and risk factors associated with AMD. The type of AMD was graded based on the international age-related maculopathy grading system; optical coherence tomography (OCT) was used for the differential diagnosis. Overall, 3,278 patients were screened; the fundus photographs, OCT scans, and self-reports were collected at the mobile clinic in a single visit. RESULTS: The prevalence of AMD in the study population was 8.99% (wet AMD 1.01%; dry AMD 7.85%), whereas the extrapolated estimate in the entire Slovakian population was 3.3% (wet AMD 0.3%; dry AMD 3.0%). Age, smoking, and hypertension were risk factors associated with AMD; however, contrary to reports in the literature, no gender-specific association was observed. CONCLUSION: Based on the results of this study, mobile clinics may be an effective way to extend health care access to a larger population. Early diagnosis of AMD will assist in early treatment and effective disease management of the population at risk.

MicroRNA-874-mediated inhibition of the major G1/S phase cyclin, CCNE1, is lost in osteosarcomas.

The tumor microenvironment is characterized by nutrient-deprived conditions in which the cancer cells have to adapt for survival. Serum starvation resembles the growth factor deprivation characteristic of the poorly vascularized tumor microenvironment and has aided in the discovery of key growth regulatory genes and microRNAs (miRNAs) that have a role in the oncogenic transformation. We report here that miR-874 down-regulates the major G1/S phase cyclin, cyclin E1 (CCNE1), during serum starvation. Because the adaptation of cancer cells to the tumor microenvironment is vital for subsequent oncogenesis, we tested for miR-874 and CCNE1 interdependence in osteosarcoma cells. We observed that miR-874 inhibits CCNE1 expression in primary osteoblasts, but in aggressive osteosarcomas, miR-874 is down-regulated, leading to elevated CCNE1 expression and appearance of cancer-associated phenotypes. We established that loss of miR-874-mediated control of cyclin E1 is a general feature of osteosarcomas. The down-regulation of CCNE1 by miR-874 is independent of E2F transcription factors. Restoration of miR-874 expression impeded S phase progression, suppressing aggressive growth phenotypes, such as cell invasion, migration, and xenograft tumors, in nude mice. In summary, we report that miR-874 inhibits CCNE1 expression during growth factor deprivation and that miR-874 down-regulation in osteosarcomas leads to CCNE1 up-regulation and more aggressive growth phenotypes.

The Impact of Hyperglycemia on VEGF Secretion in Retinal Endothelial Cells.

BACKGROUND: Diabetic retinopathy is a serious sight-threatening complication which is manifested by excessive angiogenesis in diabetic patients. AIM: We hypothesize that cultured Rhesus monkey retinal endothelial cells (RhRECs) respond to high glucose with a change in cell proliferation and vascular endothelial growth factor (VEGF) secretion. MATERIALS AND METHODS: In our study, 20 000 cells per well were treated without glucose or with 5.5 mM, 18.5 mM and 30 mM glucose for 24 hours. Viable cells were counted using trypan blue dye exclusion method. VEGF concentrations were measured in cell media by ELISA method. RESULTS: The number of viable cells incubated with 5.5 mM glucose increased significantly by 53.7% after 24 hours. In comparison, the number of viable cells decreased by 2.8% at 18.5 mM of glucose and by 20.4% at 30 mM of glucose after 24 hours of incubation. In contrast to this effect of glucose on the number of viable cells, a significant increase in VEGF levels (pg/mL) in the cell media with a glucose concentration of 0 mM compared to 5.5 mM of glucose was found. VEGF secretion in cell medium with 18.5 and 30 mM of glucose increased non-significantly in comparison with euglycemic levels. CONCLUSION: Our results show that viability of retinal endothelial cells and VEGF release are highly responsive to changes in glucose concentration. Such glucose-induced changes in retinal endothelial cells may negatively impact the integrity of the microvasculature in the diabetic retina leading to angiogenesis and microaneursym.

The phospho-caveolin-1 scaffolding domain dampens force fluctuations in focal adhesions and promotes cancer cell migration.

Caveolin-1 (Cav1), a major Src kinase substrate phosphorylated on tyrosine-14 (Y14), contains the highly conserved membrane-proximal caveolin scaffolding domain (CSD; amino acids 82-101). Here we show, using CSD mutants (F92A/V94A) and membrane-permeable CSD-competing peptides, that Src kinase-dependent pY14Cav1 regulation of focal adhesion protein stabilization, focal adhesion tension, and cancer cell migration is CSD dependent. Quantitative proteomic analysis of Cav1-GST (amino acids 1-101) pull downs showed sixfold-increased binding of vinculin and, to a lesser extent, α-actinin, talin, and filamin, to phosphomimetic Cav1Y14D relative to nonphosphorylatable Cav1Y14F. Consistently, pY14Cav1 enhanced CSD-dependent vinculin tension in focal adhesions, dampening force fluctuation and synchronously stabilizing cellular focal adhesions in a high-tension mode, paralleling effects of actin stabilization. This identifies pY14Cav1 as a molecular regulator of focal adhesion tension and suggests that functional interaction between Cav1 Y14 phosphorylation and the CSD promotes focal adhesion traction and, thereby, cancer cell motility.