Search

The aim of this study is to evaluate molecules involved in oxidative stress (OS), inflammation, angiogenesis, and apoptosis, and discern which of these are more likely to be implicated in proliferative diabetic retinopathy (PDR) and diabetic macular edema (DME) by investigating the correlation between them in the plasma (PLS) and vitreous body (VIT), as well as examining data obtained from ophthalmological examinations. Type 2 diabetic (T2DM) patients with PDR/DME (PDRG/DMEG; n = 112) and non-DM subjects as the surrogate controls (SCG n = 48) were selected according to the inclusion/exclusion criteria and programming for vitrectomy, either due to having PDR/DME or macular hole (MH)/epiretinal membrane (ERM)/rhegmatogenous retinal detachment. Blood samples were collected and processed to determine the glycemic profile, total cholesterol, and C reactive protein, as well as the malondialdehyde (MDA), 4-hydroxynonenal (4HNE), superoxide dismutase (SOD), and catalase (CAT) levels and total antioxidant capacity (TAC). In addition, interleukin 6 (IL6), vascular endothelial growth factor (VEGF), and caspase 3 (CAS3) were assayed. The VITs were collected and processed to measure the expression levels of all the abovementioned molecules. Statistical analyses were conducted using the R Core Team (2022) program, including group comparisons and correlation analyses. Compared with the SCG, our findings support the presence of molecules involved in OS, inflammation, angiogenesis, and apoptosis in the PLS and VIT samples from T2DM. In PLS from PDRG, there was a decrease in the antioxidant load (p < 0.001) and an increase in pro-angiogenic molecules (p < 0.001), but an increase in pro-oxidants (p < 0.001) and a decline in antioxidants (p < 0.001) intravitreally. In PLS from DMEG, pro-oxidants and pro-inflammatory molecules were augmented (p < 0.001) and the antioxidant capacity diminished (p < 0.001), but the pro-oxidants increased (p < 0.001) and antioxidants decreased (p < 0.001) intravitreally. Furthermore, we found a positive correlation between the PLS-CAT and the VIT-SOD levels (rho = 0.5; p < 0.01) in PDRG, and a negative correlation between the PSD-4HNE and the VIT-TAC levels (rho = 0.5; p < 0.01) in DMEG. Integrative data of retinal imaging variables showed a positive correlation between the central subfield foveal thickness (CSFT) and the VIT-SOD levels (rho = 0.5; p < 0.01), and a negative correlation between the CSFT and the VIT-4HNE levels (rho = 0.4; p < 0.01) in PDRG. In DMEG, the CSFT displayed a negative correlation with the VIT-CAT (rho = 0.5; p < 0.01). Exploring the relationship of the abovementioned potential biomarkers between PLS and VIT may help detecting early molecular changes in PDR/DME, which can be used to identify patients at high risk of progression, as well as to monitor therapeutic outcomes in the diabetic retina.

STUDY QUESTIONCan risks associated with novelties in assisted reproduction technologies (ARTs) be assessed in a systematic and structured way?SUMMARY ANSWERAn ART-specific risk assessment tool has been developed to assess the risks associated with the development of novelties in ART (EuroGTP II-ART).WHAT IS KNOWN ALREADYHow to implement new technologies in ART is well-described in the literature. The successive steps should include testing in animal models, executing pre-clinical studies using supernumerary gametes or embryos, prospective clinical trials and finally, short- and long-term follow-up studies on the health of the offspring. A framework categorizing treatments from experimental through innovative to established according to the extent of the studies conducted has been devised. However, a systematic and standardized methodology to facilitate risk evaluation before innovations are performed in a clinical setting is lacking.STUDY DESIGN, SIZE, DURATIONThe EuroGTP II-ART risk assessment tool was developed on the basis of a generic risk assessment algorithm developed for tissue and cell therapies and products (TCTPs) in the context of the project ‘Good Practices for demonstrating safety and quality through recipient follow-up European Good Tissue and cells Practices II (EuroGTP II)’. For this purpose, a series of four meetings was held in which eight ART experts participated. In addition, several tests and simulations were undertaken to fine-tune the final tool.PARTICIPANTS/MATERIALS, SETTING, METHODSThe three steps comprising the EuroGTP II methodology were evaluated against its usefulness and applicability in ART. Ways to improve and adapt the methodology into ART risk assessment were agreed and implemented.MAIN RESULTS AND THE ROLE OF CHANCEAssessment of the novelty (Step 1), consisting of seven questions, is the same as for other TCTPs. Practical examples were included for better understanding. Identification of potential risks and consequences (Step 2), consisting of a series of risks and risk consequences to consider during risk assessment, was adapted from the generic methodology, adding more potential risks for processes involving gonadic tissues. The algorithm to score risks was also adapted, giving a specific range of highest possible risk scores. A list of strategies for risk reduction and definition of extended studies required to ensure effectiveness and safety (Step 3) was also produced by the ART experts, based on generic EuroGTP II methodology. Several explanations and examples were provided for each of the steps for better understanding within this field.LIMITATIONS, REASONS FOR CAUTIONA multidisciplinary team is needed to perform risk assessment, to interpret results and to determine risk mitigation strategies and/or next steps required to ensure the safety in the clinical use of novelties.WIDER IMPLICATIONS OF THE FINDINGSThis is a dynamic tool whose value goes beyond assessment of risk before implementing a novel ART in clinical practice, to re-evaluate risks based on information collected during the process.STUDY FUNDING / COMPETING INTEREST(S)This study was called EUROGTP II and was funded by the European Commission (Grant agreement number 709567). The authors declare no competing interests concerning the results of this study.

Peroxisomal biogenesis disorders (PBDs) are a heterogeneous group of genetic diseases. Multiple peroxisomal pathways are impaired, and very long chain fatty acids (VLCFA) are the first line biomarkers for the diagnosis. The clinical presentation of PBDs may range from severe, lethal multisystemic disorders to milder, late-onset disease. The vast majority of PBDs belong to Zellweger Spectrum Disordes (ZSDs) and represents a continuum of overlapping clinical symptoms, with Zellweger syndrome being the most severe and Heimler syndrome the less severe disease. Mild clinical conditions frequently present normal or slight biochemical alterations, making the diagnosis of these patients challenging. In the present study we used a combined WES and RNA-seq strategy to diagnose a patient presenting with retinal dystrophy as the main clinical symptom. Results showed the patient was compound heterozygous for mutations in PEX1. VLCFA were normal, but retrospective analysis of lysosphosphatidylcholines (LPC) containing C22:0–C26:0 species was altered. This simple test could avoid the diagnostic odyssey of patients with mild phenotype, such as the individual described here, who was diagnosed very late in adult life. We provide functional data in cell line models that may explain the mild phenotype of the patient by demonstrating the hypomorphic nature of a deep intronic variant altering PEX1 mRNA processing.

To investigate the macular and circumpapillary retinal microvasculature across all stages of Leber hereditary optic neuropathy (LHON) using swept source optical coherence tomography angiography (OCTA).This prospective, multicentre, cross-sectional, observational study analysed a total of 119 eyes of 60 patients with molecularly confirmed LHON across all stages and 120 eyes of 60 control subjects. Optical coherence tomography angiography maps centred on the fovea and optic disc were obtained to measure vessel densities (VDs) in the macular superficial (SCP) and deep (DCP) capillary plexuses, and the radial peripapillary capillary plexus (RPCP) respectively.In asymptomatic eyes, only the SCP showed significant changes on average (B coefficient = -0.72, 95% CI = -1.34 to -0.10, p = 0.022) or in sectors representing the papillomacular bundle (PMB) (B coefficient = -1.17, 95% CI = -2.23 to -0.11, p = 0.031). However, in chronic eyes, the greatest magnitude of change was found in the temporal sector of the RPCP (B coefficient = -12.36, 95% CI = -14.49 to -10.23, p 0.001). The RPCP showed the strongest correlations with visual acuity (VA, logarithm of the minimum angle of resolution; R = -0.677, p 0.001) and structural parameters (R = 0.747, p 0.001).Retinal VD changes in the circumpapillary region of the PMB appear later than in the macula but end up being more profound and correlate better with VA and structural parameters. Further studies are needed to assess the clinical utility of retinal VDs as potential LHON biomarkers.

Subtalar instability remains a topic of debate, and its precise cause is still unknown. The mechanism of injury and clinical symptoms of ankle and subtalar instabilities largely overlap, resulting in many cases of isolated or combined subtalar instability that are often misdiagnosed. Neglecting the subtalar instability may lead to failure of conservative or surgical treatment and result in chronic ankle instability. Understanding the accurate anatomy and biomechanics of the subtalar joint, their interplay, and the contributions of the different subtalar soft tissue structures is fundamental to correctly diagnose and manage subtalar instability. An accurate diagnosis is crucial to correctly identify those patients with instability who may require conservative or surgical treatment. Many different nonsurgical and surgical approaches have been proposed to manage combined or isolated subtalar instability, and the clinician should be aware of available treatment options to make an informed decision. In this current concepts narrative review, we provide a comprehensive overview of the current knowledge on the anatomy, biomechanics, clinical and imaging diagnosis, nonsurgical and surgical treatment options, and outcomes after subtalar instability treatment.

BACKGROUND: Current treatments for several corneal lesions show limited efficacy. Here we report the clinical evaluation of the efficacy of a novel eye drop preparation produced in a public cord blood (CB) bank. MATERIALS AND METHODS: In a multicentre, retrospective, consecutive case study we evaluated 33 patients (46 eyes) unresponsive to conventional treatments who required urgent intervention. The patients were given allogeneic eye drops obtained from cord blood platelet lysate (CBED) to treat severe ocular surface lesions under a compassionate use protocol. The CBED were prepared from CB units donated for haematopoietic stem cell transplantation that did not contain the minimum stem cell dose required for this use. Patients were grouped by acute conditions (neurotrophic ulcers: group I; other corneal ulcers: group II; corneal burns: group III), and chronic conditions (ocular graft-versus-host disease: group IV; severe dry eye syndrome: group V). The patients received one or two drops of the product to the affected eye four to six times per day for 19 days. A further 19-day cycle of treatment could be repeated according to the initial clinical response. RESULTS: Patients received a median of 19 CBED vials (interquartile range 19–57, range 19–442) to complete the therapy. Group I–II–III patients showed full and partial ulcer recovery in 25 (78%) and six (19%) eyes respectively. One eye (3%) did not respond to treatment. For groups IV–V improvement was reported for 12 (85%) eyes and lesions worsened on treatment in both eyes (15%) of one patient. No severe adverse events were directly attributed to CBED. DISCUSSION: Promptly available CBED resulted in a well-tolerated allogeneic treatment that showed evidence of efficacy in this cohort of patients. These positive results support further studies on CBED from platelet lysate as a novel product of CB banks. A prospective clinical trial in neurotrophic keratitis (NCT03084861) is ongoing to confirm these preliminary data.

Proliferative diabetic retinopathy (PDR) is characterized by intraocular formation of fibroglial vascularized tissue by active vasoproliferative mechanisms. Using immunocytochemistry, we have studied changes in the distribution pattern of fibronectin (FN), laminin (LN), vitronectin (VN) and their receptors in the newly-formed capillaries of PDR. In intraocular vascularized tissue of PDR patients, FN was present on both luminal and basal surfaces of endothelial cells, and was diffusely distributed in the interpericyte space. LN was also associated with the interpericyte space. VN was occasionally detected on the luminal capillary side, but was frequent in the basal aspect of the endothelium in the interpericyte space, where it was colocalized with FN. Beta-1 subunit complex receptors were detected on the luminal side, while alpha v beta 3 integrin was identified on both sides, more so in the luminal than in the basal endothelial domain. By slot-blotting techniques and densitometric analysis, increased concentrations of intravitreous FN and VN were found in PDR in comparison with normal samples. These results suggest that FN, VN and LN have a key role in the structural arrangement of newly formed capillaries in PDR, and that receptor expression could be involved in events of endothelial cell adhesion and proliferation.

Our goal was to analyze the epidemiological and clinical findings, as well as the diagnostic approach in a group of patients in whom uveitis was the first manifestation of sarcoidosis.Retrospective study (between March 1998 and July 2002) including 31 patients diagnosed with sarcoidosis after an episode of uveitis. The group consisted of 19 females (61%) and 12 males (39%). Mean follow-up was 34.5 months. Age, sex, ocular and systemic clinical findings were recorded. Angiotensin converting enzyme (ACE) levels were measured and radiological studies of the thorax were conducted including simple radiography (Rx), computerized axial tomography (CT) and gallium scan (67Ga). Biopsy was carried out in 14 patients.Mean age was 59.6 years. Six different clinical presentations of uveitic sarcoidosis were observed, the most common being bilateral panuveitis in 13 patients (42%) and chronic anterior bilateral uveitis in 11 (35%). Gallium scan, thoracic CT and chest radiography were positive in 88%, 77% and 33% cases, respectively. ACE levels were raised in 7 of the 27 patients analyzed (26%). Histological confirmation of disease was obtained in all 14 patients biopsied with a specificity and sensitivity of 100%.Uveitis may be the presenting manifestation of sarcoidosis, especially in women over 60 years of age. Bilateral panuveitis and chronic bilateral anterior uveitis are the most common clinical presentations. The sensitivity of gallium scan and thoracic CT is greater than simple chest radiography in diagnosing pulmonary and mediastinal lesions of sarcoidosis. The need for biopsy should be individualized in each case depending on complementary exams, age and potential morbidity of the procedure.

Carnitine octanoyltransferase (COT), which facilitates the transport of shortened fatty acyl-CoAs from peroxisomes to mitochondria, is expressed in the intestinal mucosa of suckling rats; its mRNA levels increase rapidly after birth, remain steady until day 15, and decrease until weaning, when basal, adult values are established, which remain unchanged thereafter. The process seems to be controlled at the transcriptional level since the developmental pattern of mRNA coincides with that of pre-mRNA values. Dam's milk may influence the intestinal expression of COT, since mRNA levels at birth are low and increase after the first lactation. Moreover, mRNA levels decrease in rats weaned on day 18 or 21. COT is also expressed in the liver of suckling rats. Hepatic COT mRNA is maximal at day 3, remains constant until day 9, and decreases thereafter; this pattern is also similar to that of pre-mRNA values. The profile of expression of COT in intestine and liver strongly resembles that of mitochondrial 3-hydroxy 3-methylglutaryl-coenzyme A synthase and carnitine palmitoyltransferase I, suggesting that analogous transcription factors modulate ketogenesis and mitochondrial and peroxisomal fatty acid oxidation.

RESUMO La interacción entre la matriz extracelular y los componentes celulares es la principal responsable de la diferenciación celular y dei desarrollo tisular. Mediante técnicas inmunocitoquímicas para microscopía óptica y electrónica, hemos estudiado el receptor de membrana celular dei subgrupo β1 de la familia de las integrinas, la laminina (LN) y la fibronectina (FN) en membranas epirretinianas (MER) de pacientes sometidos a vitrectomía portadores de una vitreorretinopatía proliferativa (VRP) como complicación de un desprendimiento de retina, y en membranas vítreorretinianas fibrovasculares de la retinopatía diabética proliferativa (RDP). Los resultados obtenidos indican que las MER presentan el complejo β1 en disposición pericelular, unido a la' membrana plasmática. La LN y la FN se hallan en la matriz extracelular en estrecha relación con la integrina. El receptor parece ser observado más frecuentemente en las MER de hasta 2 meses de evolución cuando comparadas a las de mayor tiempo de evolución. Por otro lado, dicha integrina se encuentra localizada en los capilares de neoformación de las membranas fibrovasculares obtenidas de pacientes diabéticos. Estos hallazgos aportan nuevos datos para una mejor comprensión de los mecanismos fisiopatológicos de la interacción célula-sustrato en el proceso proliferativo intraocular. SUMMARY Cell to extracellular matrix interaction is the main event in cellular differentiation and tissue development. By immunoelectron microscopy and fluorescence immunocytochemistry, we have identified the integrin-related receptor membrane complex, laminin (LN) and fibronectin (FN) in epiretinal membranes (ERM) from patients with proliferative vitreoretinopathy (PVR) after retinal detachment and in fibrovascular membranes from proliferative diabetic retinopathy. The results showed β1-receptor complex in a pericellular pattern joined with cell membrane in ERM. LN and FN were found in extracellular matrix correlated with this integrin. Receptor seems to be more frequently observed in ERM just up to two months evolution than others with more time evolution. Moreover we have also detected the integrin receptor in vessels of diabetic fibrovascular membranes. These data provide more accurate physiopathological knowledge about cell-matrix interaction mechanisms in proliferative intraocular process.

RESUMEN La Retinopatia Diabetica Proliferativa (RDP) se caracteriza por una proliferacion fibrovascular intraocular con la formacion de membranas prerretinianas que predisponen a Desprendimientos de Retina (DR) traccionales y hemorragias en el vitreo. Aunque sean considerables los estudios morfologicos acerca de estas membranas, su fisiopatogenesis es apenas conocida. Recientemente se ha descrito la posible participacion de factores humorales y de la matriz extracelular en la angiogenesis y en su formacion. Con el objetivo de identificar la distribucion de algunos de estos posibles factores, hemos realizado um estudio inmunocitoquimico para la deteccion de la Laminina (LN) y de la Fibronectina (FN), en una serie de 18 membranas fibrovasculares, obtenidas de ojos portadores de RDP y sometidos a vitrectomia. Ambas glucoproteinas presentam una localizacion vascular; la FN en disposicion luminal, y la LN en disposicion basal con respecto a las celulas endoteliales de los capilares. La inmunolocalizacion tambien ha confirmado la presencia de macrofagos, sugiriendo que el proceso proliferativo intraocular puede ser considerado como una forma modificada de formacion cicatricial.

Fluoropyrimidines may be effective in preventing proliferative vitreoretinopathy after repair of complicated retinal detachments. Liposome encapsulation of these antiproliferative drugs may extend the intravitreal half-life and increase their efficacy.The current study evaluated the pharmacokinetic behavior of intravitreally injected 5-fluorouridine (5-FUR), free and encapsulated in liposomes, either conventionally or coated with collagen into 25 New Zealand rabbits. Additionally, we investigated the retinal toxicity of intravitreal injections of 100, 250 and 500 microg as well as 1 mg 5-FUR as free drug or encapsulated in liposomes in the rabbit eye.The half-life of free 5-FUR after liposome injection into the vitreous cavity was 18.17+/-2.43 h, considerably longer than the half-life of free 5-FUR (0.82 h). Electrophysiologic tests did not show any changes in latency and a-wave amplitude and minimal changes in the b-wave amplitude. Histopathologic studies revealed integrity of the inner limiting membrane, and mild vacuolization in the outer retina.Encapsulation of 5-FUR within liposomes markedly increases its intravitreal half-life. Our study suggests that liposome-encapsulated 5-FUR is not toxic to the retina even at doses of 1 mg.