Investigation the effects of vitreous humor on proliferation and dedifferentiation of differentiated NTERA2 cells / Investigar os efeitos do humor vítreo na proliferação e desdiferenciação de células NTERA2 diferenciadas

Abstract Mammals have a limited capacity to regenerate their tissues and organs. One of the mechanisms associated with natural regeneration is dedifferentiation. Several small molecules such as vitamin C and growth factors could improve reprogramming efficiency. In this study, the NTERA2-D1 (NT2) cells were induced towards differentiation (NT2-RA) with 10-5 M retinoic acid (RA) for three days and then subjected to various amounts of vitreous humor (VH). Results show that the growth rate of these cells was reduced, while this rate was partly restored upon treatment with VH (NT2-RA-VH). Cell cycle analysis with PI method also showed that the numbers of cells at the S phase of the cell cycle in these cells were increased. The levels of SSEA3 and TRA-1-81 antigens in NT2-RA were dropped but they increased in NT2- RA-VH to a level similar to the NT2 cells. The level of SSEA1 had an opposite pattern. Expression of OCT4 gene dropped after RA treatment, but it was recovered in NT2-RA-VH cells. In conclusion, we suggest VH as a potent mixture for improving the cellular reprogramming leading to dedifferentiation.

Resumo Os mamíferos têm uma capacidade limitada de regenerar seus tecidos e órgãos. Um dos mecanismos associados à regeneração natural é a desdiferenciação. Várias moléculas pequenas, como vitamina C e fatores de crescimento, podem melhorar a eficiência da reprogramação. Neste estudo, as células NTERA2-D1 (NT2) foram induzidas à diferenciação (NT2-RA) com ácido retinóico (RA) 10-5 M por três dias e depois submetidas a várias quantidades de humor vítreo (VH). Os resultados mostram que a taxa de crescimento dessas células foi reduzida, enquanto essa taxa foi parcialmente restaurada após o tratamento com VH (NT2-RA-VH). A análise do ciclo celular com o método PI também mostrou que o número de células na fase S do ciclo celular nessas células estava aumentado. Os níveis de antígenos SSEA3 e TRA-1-81 em NT2-RA diminuíram, mas aumentaram em NT2-RA-VH a um nível semelhante ao das células NT2. O nível de SSEA1 teve um padrão oposto. A expressão do gene OCT4 diminuiu após o tratamento com AR, mas foi recuperado em células NT2-RA-VH. Em conclusão, sugerimos o VH como uma mistura potente para melhorar a reprogramação celular levando à desdiferenciação.

Research Progress on Estimation of Postmortem Interval Based on Ocular Tissues Structure / 法医学杂志

Estimation of postmortem interval (PMI) is one of the important research contents in forensic pathology, and it has always been the focus and hot spot of research work. In recent years, scholars at home and abroad have made some research progress in estimating PMI by using ocular tissue. After death, the changes of cornea, aqueous humor, iris, lens, vitreous humor and retina all show time sequence change rule highly related to PMI. This paper reviews the research progress of PMI estimation based on the morphological, biochemical, molecular and genetic material changes of different ocular tissue structures after death, and discusses the existing problems and development trends.

Changes of 1, 5-AG in Vitreous Humor of Rabbit Cadavers with Hyperglycemic Metabolism / 法医学杂志

OBJECTIVES@#To investigate the concentration and change characteristics of 1, 5-anhydroglucitol (1, 5-AG) in the vitreous humor of rabbit cadavers with hyperglycemic metabolism, and to explore the value of 1, 5-AG in forensic pathology identification of death caused by hyperglycemic metabolism disorders.@*METHODS@#A diabetic hyperglycemic rabbit model was established by using alloxan. Eighteen rabbits with fasting glucose concentration ≥13.80 mmol/L (experimental group) and 18 healthy rabbits with fasting glucose concentration ≤6.10 mmol/L (control group) were selected. After death from air embolism. The blood samples were collected immediately, and vitreous humor samples were collected at 0 h, 12 h, 24 h and 36 h after death. The concentration of 1, 5-AG in the blood and vitreous humor of rabbits was determined.@*RESULTS@#The blood glucose concentration in the experimental group was (25.10±3.14) mmol/L. At the time of death, there was no significant difference in the concentration of 1, 5-AG in the blood [(0.94±0.20) μg/mL] and in the vitreous humor (0.99±0.05 μg/mL, P>0.05). The concentration of 1, 5-AG in the vitreous humor of the experimental group was lower than that of the corresponding control group at all time points (P<0.05), and there was no significant difference betwwen 1, 5-AG concentration in vitreous humor between earch time point in the experimental group and the control group (P>0.05). Correlation analysis showed that the concentration of 1,5-AG in blood was negatively correlated with blood glucose in both control group and experimental group (control group: r=-0.79, P<0.05; experimental group: r=-0.97, P<0.05).@*CONCLUSIONS@#Vitreous humor can replace blood as an effective test sample for 1,5-AG detection. The concentration of 1, 5-AG in rabbit vitreous humor remains stable within 36 hours after death and is not affected by the change of postmortem interval. If the concentration of 1, 5-AG decreases significantly, it indicates the existence of hyperglycemia in rabbits before death.

Ocular cysticercosis in pediatric patient / Cisticercose ocular em paciente pediátrico

ABSTRACT Ocular cysticercosis is a parasitic infection caused by Taenia solium. Its early diagnosis and treatment decreases the possibility of visual morbidity. It can either compromise the anterior chamber or the posterior segment, which translates into an very variable and interspecific presentation that changes depending on the site of the infection. It is important to report this case due to its low presentation rate and the fact that a high suspicion index is required to make an assertive and timely diagnosis. This is especially important in geographical areas that are endemic to this parasite due to the direct relationship between an early diagnosis and treatment and better visual outcomes. In this case report, we will discuss the multidisciplinary interventions of a pediatric patient in a high complexity hospital.

RESUMO A cisticercose ocular é uma infecção parasitária causada pela Taenia solium. O diagnóstico e tratamento precoces diminuem a possibilidade de morbidade visual. Ela pode comprometer a câmara anterior ou o segmento posterior, o que se traduz em uma apresentação muito variável e interespecífica, que muda dependendo do local da infecção. É importante relatar esse caso devido à sua baixa taxa de apresentação e ao fato de que é necessário um alto índice de suspeita para fazer um diagnóstico assertivo e oportuno. Isso é especialmente importante em áreas geográficas endêmicas para esse parasita, devido à relação direta entre diagnóstico e tratamento precoces e melhores resultados visuais. Neste relato de caso, discutiremos as intervenções multidisciplinares de um paciente pediátrico em um hospital de alta complexidade.

Ranibizumab intravítreo para el tratamiento de oclusiones vasculares retinianas: análisis mediante tomografía de coherencia óptica / Intravitreal Ranibizumab for the Treatment of Retinal Vascular Occlusions: Analysis Using Optical Coherence Tomography

Introducción: El uso del medicamento ranibizumab intravítreo favorece la reducción del edema macular generador de las oclusiones vasculares retinianas causantes de la pérdida visual. Objetivo: Evaluar la eficacia y seguridad de la administración intravítreo de ranibizumab en el cambio de espesor macular central en las oclusiones vasculares retinianas analizado mediante tomografía de coherencia óptica. Material y métodos: Se desarrolló un estudio de tipo retrospectivo, analítico, correlacional y observacional de campo con diseño no experimental en 125 pacientes mayores de 30 años con oclusión vascular retiniana diagnosticados en la consulta de oftalmología del Hospital "Teodoro Maldonado Carbó" durante enero de 2017 a junio de 2018. La técnica ANOVA compara las medias para determinar mediante el proceso de contraste de hipótesis si existen diferencias estadísticamente significativas entre estas. Resultados: El análisis de la agudeza visual con escala logMAR demostró diferencias estadísticamente significativas entre los promedios obtenidos 3 meses antes y después de la aplicación del tratamiento (p=0,0001). Se encontró 28,8 por ciento de efectos adversos. Con frecuencia en aumento de presión intraocular (4 por ciento), sequedad ocular (16 por ciento) y hemorragia conjuntival (11,2 por ciento). Conclusiones: El ranibizumab en oclusiones vasculares retinianas proporciona una mejor agudeza visual corregida en relación con el grosor macular, favorece el desarrollo de nuevos vasos sanguíneos a partir de vasos preexistentes desde migración de células endoteliales(AU)

Introduction: The use of the intravitreal ranibizumab favors the reduction of the macular edema that generates retinal vascular occlusions that cause visual loss. Objective: To evaluate the efficacy and safety of the intravitreal administration of ranibizumab in the change in central macular thickness in retinal vascular occlusions analyzed by optical coherence tomography. Material and methods: A retrospective, analytical, correlational and observational field study with a non-experimental design was carried out on 125 patients over 30 years of age diagnosed with retinal vascular occlusion in the Ophthalmology Service of "Teodoro Maldonado Carbó" Hospital during the period between January 2017 and June 2018. The ANOVA technique was used to compare means in order to determine, through the hypothesis contrast process, if there are statistically significant differences between them. Results: Visual acuity analysis using the logMAR scale showed statistically significant differences between the averages obtained 3 months before and after the application of the treatment (p =0.0001). In addition, 28,8 percent of adverse effects were found. The most frequent ones included increased intraocular pressure (4 percent), dry eyes (16 percent), and conjunctival hemorrhage (11,2 percent). Conclusions: In retinal vascular occlusions, Ranibizumab provides a better corrected visual acuity in relation to macular thickness, favors the development of new blood vessels from pre-existing vessels from endothelial cell migration(AU)

Inferring Postmortem Submersion Interval in Rats Found in Water Based on Vitreous Humor Metabolites / 法医学杂志

OBJECTIVES@#The metabolomics technique of LC-MS/MS combined with data analysis was used to detect changes and differences in metabolic profiles in the vitreous humor of early rat carcasses found in water, and to explore the feasibility of its use for early postmortem submersion interval (PMSI) estimation and the cause of death determination.@*METHODS@#The experimental model was established in natural lake water with 100 SD rats were randomly divided into a drowning group (n=50) and a postmortem (CO2 suffocation) immediately submersion group (n=50). Vitreous humor was extracted from 10 rats in each group at 0, 6, 12, 18 and 24 h postmortem for metabolomics analyses, of which 8 were used as the training set to build the model, and 2 were used as test set. PCA and PLS multivariate statistical analysis were performed to explore the differences in metabolic profiles among PMSI and causes of death in the training set samples. Then random forest (RF) algorithm was used to screen several biomarkers to establish a model.@*RESULTS@#PCA and PLS analysis showed that the metabolic profiles had time regularity, but no differences were found among different causes of death. Thirteen small molecule biomarkers with good temporal correlation were selected by RF algorithm. A simple PMSI estimation model was constructed based on this indicator set, and the data of the test samples showed the mean absolute error (MAE) of the model was 0.847 h.@*CONCLUSIONS@#The 13 metabolic markers screened in the vitreous humor of rat corpses in water had good correlations with the early PMSI. The simplified PMSI estimation model constructed by RF can be used to estimate the PMSI. Additionally, the metabolic profiles of vitreous humor cannot be used for early identification of cause of death in water carcasses.

Ambliopia severa na infância relacionada à persistência da vasculatura fetal / Severe childhood amblyopia associated to persistence of fetal vasculature

RESUMO A persistência do vítreo primário hiperplásico, atualmente referida como persistência da vasculatura fetal, é uma anomalia congênita que resulta da não regressão do vítreo vascular primário e do sistema da artéria hialoide durante a embriogênese. Trata-se de uma anomalia unilateral na maioria dos casos, esporádica e comumente não associada a nenhum outro achado sistêmico. Clinicamente, essa condição pode ser classificada em persistência anterior e em persistência posterior da vasculatura fetal. A condição anterior está relacionada ao sistema da artéria ciliar, enquanto a persistência da vasculatura posterior associa-se à artéria hialoide e pode apresentar anormalidades, com desfecho visual desfavorável. A detecção da persistência do vítreo primário hiperplásico é de suma importância, visto que é um diagnóstico diferencial para retinoblastoma. O relato de caso a seguir descreve o acompanhamento ambulatorial em um Serviço de Oftalmologia de uma criança do sexo masculino com persistência da vasculatura fetal unilateral e sem alterações sistêmicas.

ABSTRACT Hyperplastic primary vitreous persistence, currently referred to as fetal vasculature persistence, is a congenital anomaly that results from non-regression of the primary vascular vitreous and hyaloid artery system during embryogenesis. It is a unilateral anomaly in the vast majority of cases, sporadic and commonly not associated with any other systemic finding. Clinically, this condition can be classified into anterior and posterior persistence of fetal vasculature. The anterior condition is related to the ciliary artery system, while the persistence of the posterior vasculature is associated with the hyaloid artery, which may present abnormalities with an unfavorable visual outcome. Detecting persistent hyperplastic primary vitreous is of paramount importance, as it is a differential diagnosis for retinoblastoma. The following case report describes the outpatient follow-up at the ophthalmology service of the Federal University of Triângulo Mineiro (UFTM) of a male child with persistent unilateral fetal vasculature and no systemic changes.

Correlación entre sodio, cloro y potasio del cuerpo vítreo y el intervalo postmortem sobre los 2. 850 msnm / Correlation between sodium, chlorine and potassium of the vitreous body and the postmortem interval above 2. 850 msnm

RESUMEN: El intervalo Postmortem (IPM) es un importante desafío por resolver en la patología forense, y consiste en poder determinar el tiempo transcurrido desde la muerte hasta el momento de la autopsia. Dada la poca confiabilidad de algunos métodos por la gran influencia de factores externos y propios del cadáver, la bioquímica forense ha recibido considerable atención por sus niveles de seguridad. La ciudad de Quito se ubica en la cordillera de Los Andes a 2850 msnm, sin embargo, en la literatura no existen reportes de medición de estos parámetros a una altura como ésta. El objetivo fue establecer una correlación entre sodio (Na+), cloro (Cl-) y potasio (K+) del humor vítreo del cuerpo vítreo y el IPM a la altura de la ciudad de Quito. Para el estudio se utilizaron 128 muestras de cuerpo vítreo provenientes de 16 autopsias practicadas en la Unidad de Medicina Legal Zona 9, en IPM de 6, 12, 24, 48, 72, 96, 120 y 144 horas. Para la obtención del humor vítreo se siguió el método convencional, se dividió para las 8 alícuotas y se llevó a congelación -20 °C hasta el momento de su análisis. La cuantificación de la concentración de electrolitos Na+, Cl- y Potasio+ se realizó mediante analizador ISE de Roche Cobas (Roche Diagnostics) C501, calibrado para uroanálisis, y no fue necesaria la dilución. Durante la calibración y cada tres muestras, se midió un estándar interno para corregir los efectos de la desviación sistemática en la calibración. El enfoque estadístico se basó en un análisis de correlación lineal, utilizando el coeficiente de correlación de Spearman. La correlación entre las horas postmortem y las concentraciones de los diferentes electrolitos, fueron estadísticamente significativas. Se pudo corroborar una correlación lineal significativa entre el IPM y el aumento del K+ en el HV.

SUMMARY: The postmortem interval (PMI) is an important challenge to be solved in forensic pathology, and it consists of being able to determine the time elapsed from death to the moment of autopsy. Given the unreliability of some methods due to the great influence of external factors and those specific to the corpse, forensic biochemistry has received considerable attention for its levels of safety. The city of Quito is located in the Andes mountain range at 2850 meters above sea level, however, in the literature there are no reports of measurement of these parameters at a height like this. The objective was to establish a correlation between sodium (Na+), chlorine (Cl-) and potassium (K+) of the vitreous humor and the MPI at a height of 2,850 masl. For the study, 128 samples of vitreous humor were used from 16 autopsies performed in the Zone 9 Legal Medicine Unit, in IPM of 6, 12, 24, 48, 72, 96, 120 and 144 hours. To obtain the vitreous humor, the conventional method was followed, it was divided for the 8 aliquots and it was frozen at -20 ° C until the moment of its analysis. The quantification of the concentration of electrolytes Na+, Cl- and K+ was carried out using an ISE analyzer from Roche Cobas (Roche Diagnostics) C501, calibrated for urinalysis, and no dilution was necessary. During calibration and every third sample, an internal standard was measured to correct for the effects of systematic deviation on the calibration. The statistical approach was based on a linear correlation analysis, using the Spearman correlation coefficient. The correlation between the postmortem hours and the concentrations of the different electrolytes were statistically significant. A significant linear correlation between the PMI and the increase in K+ in vitreous humor could be corroborated.

Vasculatura fetal persistente bilateral. Reporte de caso / Bilateral persistent fetal vasculature. A case report

Introducción: La vasculatura fetal persistente bilateral es una malformación ocular poco frecuente, generalmente unilateral, que suele presentarse como leucocoria. Objetivo: Presentar el caso de una paciente de 6 meses con leucocoria bilateral y diagnóstico de vasculatura fetal persistente bilateral. Resumen del caso: Reporte de caso por medio de recopilación de datos de la historia clínica, fotografías del segmento anterior de la paciente, ecografía ocular y resonancia magnética de cerebro. Conclusiones: La vasculatura fetal persistente bilateral es poco frecuente, pero hay que tenerla en cuenta en el momento del abordaje de un paciente con leucocoria bilateral, cuyo pronóstico visual depende de su reconocimiento y tratamiento tempranos.

Introduction: Bilateral persistent fetal vasculature is a rare developmental disorder of the eye, usually seen as leukocoria affecting only one eye. Objective: Report on a case of a 6 month old patient with bilateral leukocoria and diagnosis of bilateral persistent fetal vasculature. Case summary: Descriptive case report study with data from de medical history, anterior segment photographs, ocular ultrasonography and cerebral magnetic resonance imaging. Conclusions: Bilateral persistent fetal vasculature is an infrequent disease that has to be taken into account when approaching a patient with bilateral leukocoria, whose visual prognosis depends on the early detection and treatment

Usefulness of aqueous and vitreous humor analysis in infectious uveitis

OBJECTIVE: To evaluate the role of intraocular fluid analysis as a diagnostic aid for uveitis. METHODS: Twenty-eight samples (27 patients including 3 HIV-infected patients) with active (n=24) or non-active (n=4) uveitis were submitted to aqueous (AH; n=12) or vitreous humor (VH) analysis (n=16). All samples were analyzed by quantitative PCR for herpes simplex virus (HSV), varicella zoster virus (VZV), cytomegalovirus (CMV), Epstein-Barr virus (EBV) and Toxoplasma gondii. RESULTS: The positivity of the PCR in AH was 41.7% (5/12), with 50% (2/4) in immunocompetent and 67% (2/3) in HIV+ patients. The positivity of the PCR in VH was 31.2% (5/16), with 13% (1/8) in immunocompetent and 50% (4/8) in immunosuppressed HIV negative patients. The analysis was a determinant in the diagnostic definition in 58% of HA and 50% of VH. CONCLUSION: Even in posterior uveitis, initial AH analysis may be helpful. A careful formulation of possible clinical diagnosis seems to increase the chance of intraocular sample analysis being meaningful.

Facoanafilaxia por cristalino mergulhado na Síndrome de Marfan / Phacoanaphylactic by dislocation of the lens in the Marfan Syndrome

Resumo A síndrome de Marfan é uma doença de herança autossômica dominante e que afeta o tecido conjuntivo com manifestações fenotípicas que envolvem os sistemas esquelético, cardiovascular e ocular. As principais manifestações oculares são a subluxação do cristalino, a miopia e o descolamento da retina. O objetivo deste artigo foi relatar a conduta clínico-cirúrgica de um paciente portador da síndrome de Marfan com cristalino luxado para a cavidade vítrea e que evoluiu com severa reação facoanafilática caracterizada por um glaucoma secundário severo e descompensação corneana.

Abstract Marfan syndrome is an autosomal dominant inheritance disease that affects connective tissue with phenotypic manifestations involving the skeletal, cardiovascular and ocular systems. The main ocular manifestations are the subluxation of the lens, myopia and retinal detachment. The aim of this article was to report the clinical and surgical management of a patient with Marfan syndrome with luxated lens for the vitreous cavity and who developed a severe phacoanaphylactic reaction characterized by severe secondary glaucoma and corneal decompensation.

Ultrastructural changes detected using swept-source optical coherence tomography in severe vitreopapillary traction: a case report / Alterações ultraestruturais detectadas pela tomografia de coerência óptica de fonte de varredura em tração vitreopapilar grave: relato de caso

ABSTRACT Vitreopapillary traction is an uncommon condition characterized by strong adhesion and the traction of the posterior hyaloid onto the optic disc and peripapillary retina, leading to optic disc elevation and visual loss. An 85-year-old man presented with a 6-month history of slow, progressive visual loss in the left eye along with optic disc edema. Swept-source optical coherence tomography B-scans revealed circumpapillary anterior-posterior persistent traction of dense vitreous strands onto the optic disc. Visual field examination demonstrated mild, generalized, diffuse sensitivity loss and blind-spot enlargement. A 25-gauge posterior vitrectomy was performed with posterior hyaloid separation from the optic disc, resulting in significant anatomical and visual improvement. In conclusion, swept-source optical coherence tomography aids in understanding the mechanism underlying visual loss in vitreopapillary traction. Moreover, posterior vitrectomy can effectively promote anatomical and visual improvements in these cases.

RESUMO A tração vitreopapilar é uma condição incomum caracterizada por forte adesão e tração da hialoide posterior no disco óptico e retina peripapilar, levando à elevação do disco óptico e à perda visual. Um homem de 85 anos apresentou uma história de 6 meses de perda visual lenta e progressiva no olho esquerdo, juntamente com edema do disco óptico. A tomografia de coerência óptica por fonte de varredura revelou tração persistente ântero-posterior peripapilar com traves vítreas densas sobre o disco óptico. Exame de campo visual demonstrou perda de sensibilidade difusa, generalizada, leve e aumento do ponto cego. Uma vitrectomia posterior de calibre 25 foi realizada com separação hialóide posterior do disco óptico, resultando em melhora anatômica e visual significativa. Em conclusão, a tomografia de coerência óptica por fonte de varredura auxilia na compreensão do mecanismo subjacente à perda visual na síndrome de tração vitreopapilar. Além disso, a vitrectomia posterior pode efetivamente promover melhorias visuais e anatômicas nesses casos.

Ultrasonographic characteristics of active ocular toxoplasmosis / Achados ultrassonográficos em toxoplasmose ocular ativa

ABSTRACT Purpose: To evaluate ophthalmic ultrasonographic findings associated with active ocular toxoplasmosis. Methods: Forty-seven eyes with active ocular toxoplasmosis in 47 patients were subjected to ocular ultrasonography using the transpalpebral technique (10-MHz transducer) and fundus photography. Patient medical records were retrospectively reviewed. Results: Ocular ultrasonography revealed vitritis, posterior vitreous detachment, retinal wall thickening, and non-rhegmatogenous retinal detachment in 47 (100%), 36 [76.6%; partial in 12 (25.5%) and total in 23 (48.9%)], 12 (25.5%), and 5 eyes (10.6%). Thirty-five of the 36 eyes with posterior vitreous detachment (97.2%) exhibited posterior hyaloid thickening; moreover, adhesion to the exudative lesion and vitreoschisis were observed in 4 (11.1%) and 12 eyes (25.5%), respectively. Ultrasonography detected the location of the exudative focus in 12 eyes (25.5%). Conclusion: Ultrasonography is helpful for detecting important intraocular findings of acute ocular toxoplasmosis that can be hindered by medial opacity or posterior synechiae.

RESUMO Objetivo: Avaliar os achados da ultrassonografia na toxoplasmose ocular ativa. Métodos: Quarenta e sete olhos com toxoplasmose ocular ativa em 47 pacientes foram submetidos à ultrassonografia ocular pela técnica transpalpebral (transdutor de 10 MHz) e fundo de olho. Os prontuários médicos foram revistos retrospectivamente. Resultados: A ultrassonografia ocular revelou vitreíte, descolamento vítreo posterior, espessamento da parede da retina e descolamento de retina não regmatogênico em 47 (100%), 36 [76,6%; parcial em 12 (25,5%) e total em 23 (48,9%)], 12 (25,5%) e 5 olhos (10,6%). Trinta e cinco dos 36 olhos com descolamento vítreo posterior (97,2%) exibiram espessamento hialoide posterior; além disso, a adesão à lesão exsudativa e vitreosquise foi observada em 4 (11,1%) e 12 (25,5%), respectivamente. A ultrassonografia detectou a localização do foco exsudativo em 12 olhos (25,5%). Conclusão: A ultrassonografia é útil na detecção de importantes achados intra-oculares de toxoplasmose ocular aguda que podem ser prejudicados pela opacidade medial ou sinéquia posterior.

Intravitreal neurodegenerative and inflammatory mediators in proliferative diabetic retinopathy / Mediadores intravítreos da neurodegeneração e inflamação na retinopatia diabética proliferativa

ABSTRACT Purpose: To compare the intravitreal concentrations of cellular mediators involved in neurodegeneration, inflammation, and angiogenesis in patients with proliferative diabetic retinopathy and other vitreoretinal diseases. Methods: A multiplex bead immunoassay was used to measure vitreous levels of pigment epithelium-derived factor, serum amyloid P, C-reactive protein, complement C4, alpha-1 antitrypsin, vascular endothelial growth factor, platelet-derived growth factor-AA, platelet-derived growth factor-BB, interleukin-6, interleukin-8, interleukin-10, tumor necrosis factor alpha and beta in patients undergoing 23-gauge vitrectomy for proliferative diabetic retinopathy and other diagnoses (control group). Results: We evaluated 55 patients, of whom 24 had proliferative diabetic retinopathy and 31 had other diagnoses including vitreous hemorrhage, retinal detachment, macular hole, and epiretinal membrane. Patients with proliferative diabetic retinopathy demonstrated increased levels of serum amyloid P (85.49 vs. 31.38 ng/mL); C-reactive protein (59.89 vs. 41.75 ng/mL), vascular endothelial growth factor (2,330.11 vs. 554.25 pg/mL; p<0.001), platelet-derived growth factor A (127.32 vs. 39.11 pg/mL), platelet-derived growth factor B (29.37 vs. 7.12 pg/mL), interleukin-6 (69.37 vs. 33.58 pg/mL), interleukin-8 (175.25 vs. 59.71 pg/mL), and interleukin-10 (3.70 vs. 1.88 pg/mL); all p<0.004 when compared with the control group. Levels of pigment epithelium-derived factor (30.06 vs. 27.48 ng/mL; p=0.295), complement C4 (570.78 vs. 366.24 ng/mL; p=0.069), and alpha-1-antitrypsin (359.27 vs. 522.44 ng/mL; p=0.264) were not significantly different between the groups. Intravitreal levels of tumor necrosis factor-alpha and tumor necrosis factor-beta were undetectable. Serum Amyloid P, C-reactive protein, platelet-derived growth factor A, platelet-derived growth factor B, interleukin-6, and interleukin-8 were correlated positively with vascular endothelial growth factor. Conclusions: Cellular mediators involved in neurodegeneration and inflammation demonstrated increased levels in the vitreous humor of patients with proliferative diabetic retinopathy and may be part of the pathogenesis of diabetic retinopathy.

RESUMO Objetivo: Comparar as concentrações intravítreas de mediadores celulares envolvidos na neurodegeneração, inflamação e angiogênese em pacientes com retinopatia diabética proliferativa e outras doenças vítreo-retinianas. Métodos: Um ensaio imunomagnético foi utilizado para medir os níveis vítreos do fator derivado do epitélio pigmentar, amilóide P sérico, proteína-C-reativa, complemento C4, e alfa-1-antitripsina, fator de crescimento do endotélio vascular, fator de crescimento derivado das plaquetas AA, fator de crescimento derivado das plaquetas BB, interleucina-6, interleucina-8, interleucina-10, fator de necrose tumoral alfa e beta em pacientes submetidos à vitrectomia 23-gauge para retinopatia diabética proliferativa ou outros diagnósticos (grupo controle). Resultados: Foram avaliados 55 pacientes, dos quais 24 tinham retinopatia diabética proliferativa e 31 tinham outros diagnósticos, incluindo hemorragia vítrea, descolamento de retina, buraco macular e membrana epirretiniana. Pacientes com retinopatia diabética proliferativa demonstraram níveis aumentados de amilóide P sérico (85,49 vs 31,38 ng/mL), proteína-C-reativa (59,89 vs 41,75 ng/mL), fator de crescimento do endotélio vascular (2.330,11 vs 554,25 pg/mL, p<0.001), fator de crescimento derivado das plaquetas-A: (127,32 vs 39,11 pg/mL), fator de crescimento derivado das plaquetas-B (29,37 vs 7,12 pg/mL), interleucina-6 (69,37 vs 33,58 pg/mL), interleucina-8 (175,25 vs 59,71 pg/mL) e interleucina-10 (3,70 vs 1,88 pg/mL), todos com p<0,004 quando comparados ao grupo controle. Níveis de fator derivado do epitélio pigmentar (30,06 vs 27,48 ng/mL; p=0,295), complemento C4 (570,78 vs 366,24 ng/mL; p=0,069), alfa-1 antitripsina (359,27 vs 522,44 ng/mL; p=0,264) não foram significativamente diferente entre os grupos. Níveis intravítreos de fator de necrose tumoral alfa e fator de necrose tumoral beta foram indetectáveis. O amilóide P sérico, a proteína C-reativa, o fator de crescimento derivado das plaquetas A e B, a interleucina-6 e a interleucina-8 correlacionaram-se positivamente com o fator de crescimento do endotélio vascular. Conclusões: Os medidores celulares envolvidos na neurodegeneração e inflamação demonstraram níveis aumentados no humor vítreo de pacientes com retinopatia diabética proliferativa e podem ser parte da patogênese da retinopatia diabética.

Diagnostic value of pediatric blood culture bottles for acute postoperative endophthalmitis

OBJECTIVE: To report our experience using conventional culture methods (CM) and pediatric blood culture bottles (PBCBs) for vitreous sample culture of acute postoperative endophthalmitis. METHODS: A retrospective study was conducted at the Department of Ophthalmology, Hospital das Clinicas, HCFMUSP, Faculdade de Medicina, Universidade de Sao Paulo, Sao Paulo, BR, from January 2010 to December 2015, and it included 54 patients with clinically suspected acute postoperative endophthalmitis. Vitreous samples were obtained by vitreous tap or vitrectomy. Samples from January 2010 to December 2011 were cultivated in CM, whereas samples from January 2012 to December 2015 were inoculated in PBCBs. The measured outcome was the yield of positive cultures. RESULTS: Twenty cases were included in the CM group, and 34 cases were included in the PBCB group. The yield of positive cultures in PBCBs (64.7%) was significantly higher than that in conventional CM (35%, p=0.034). Staphylococcus epidermidis and Streptococcus viridans were the two most commonly found agents. CONCLUSION: PBCBs can be used successfully in clinically suspected endophthalmitis. The method showed a higher yield of positive cultures than the conventional method. This technique appears to have several advantages over the traditional method: it saves time, as only one medium needs to be inoculated; transportation to a laboratory is easier than in the traditional method, and there is no need to maintain a supply of fresh agar media. The use of PBCBs may be recommended as the primary method for microbiological diagnosis and is especially suitable for office settings and remote clinics.

Evaluation of Sodium and Chloride Biochemical Tests in Drowning Autopsy Cases / 대한법의학회지

There are several diagnostic findings required for confirming a postmortem diagnosis of drowning. However, postmortem diagnosis of drowning remains challenging for forensic pathologists. In previous reports, several biochemical tests using various body fluids have been studied for their potential use in the postmortem diagnosis of drowning. In this study, the concentration of sodium and chloride was tested in various postmortem body fluids (vitreous humor, sphenoid sinus fluid, pleural fluid, cerebrospinal fluid, etc.) and their results were interpreted for their potential use in postmortem diagnosis of drowning. We examined 67 autopsy cases (freshwater drowning, 12 cases; seawater drowning, 16 cases; control group, 39 cases). The sodium and chloride concentration in the vitreous humor, sphenoid sinus fluid, and pleural fluid significantly correlated with each other. Furthermore, the concentrations of sodium, chloride, and the sum of the concentrations of the two in the various postmortem body fluids were significantly different in the three groups, when compared with each other (generally the concentration being the highest in the seawater drowning group, followed by the control group and the freshwater drowning group). Biochemical tests using various postmortem body fluids may serve as useful indicators for the postmortem diagnosis of drowning and for the differential diagnosis between freshwater and seawater drowning.

Retinal changes in rabbit after intravitreal injection of sunitinib encapsulated into solid lipid nanoparticles and polymeric nanocapsules / Avaliação das alterações retinais no coelho após injeção intravítrea de sunitinib encapsulado em nanoparticulas lípidas sólidas e nanocapsulas polímeros

ABSTRACT Purpose: We aimed to evaluate the safety of single intravitreal injection of each of two concentrations of 0.1 ml of sunitinib (1 and 10 mg/ml), 0.1 ml of a drug-free dispersion containing solid lipid nanoparticles, and 0.1 ml of a drug-free dispersion containing polymeric nanocapsules for analyzing the possible toxic effects using electrophysiology and histology in albino rabbit retina. Methods: We conducted an experimental controlled study of 20 eyes of albino rabbits. Intravitreal injections of each specific agent were applied to one eye per rabbit in each 5-rabbit group, while the contralateral eyes received no treatment and were used as controls. Results: We noted no electroretinographic changes in the sunitinib (1 and 10 mg/ml) or in solid lipid nanoparticles groups. However, we observed significant abnormalities in ocular morphology and in the electroretinogram in the nanocapsules group. At the histological level, only the nanocapsules group demonstrated abnormal changes, including severe edema and cytoplasmic vacuole formation. Conclusions: While nanocapsules intravitreal injections indicated retinal toxic effects, sunitinib and solid lipid nanoparticles intravitreal injections were not toxic to the retina. Our results suggest that a sunitinib preparation with solid lipid nanoparticles for controlled release may offer a significant therapeutic approach for vasoproliferative ocular disease.

RESUMO Objetivos: O presente estudo teve por objetivo avaliar a segurança da injeção intravítrea de 0,1 ml de sunitinibe em duas concentrações (1 mg/ml e 10 mg/ml), 0,1 ml de dispersão contendo nanopartículas lipídicas sólidas sem droga e 0,1 ml de dispersão contendo nanocápsulas poliméricas livre de drogas analisando os possíveis efeitos tóxicos à retina de coelhos albinos detectados pela eletrofisiologia e histologia por microscopia óptica. Métodos: Um estudo controlado experimental foi rea­lizado com 20 olhos de coelhos albinos. Foram realizadas injeções intravítrea de duas concentrações diferentes de suniti­nibe, uma dispersão contendo nanopartículas lipídicas sólidas e uma dispersão contendo nanocápsulas. O olho contralateral não recebeu tratamento e foi utilizado como controle. Resultados: Não foram observadas alterações eletrorretinográficas nos grupos do sunitinibe (1 mg/ml e 10 mg/ml) e no grupo das nanopartículas lipídicas sólidas. No grupo das nanocápsulas, houve alterações significativas tanto na morfologia, quanto na amplitude e tempo das ondas do eletrorretinograma. Ao estudo histológico, somente o grupo das nanocápsulas apresentou al­terações degenerativas (núcleos tumefeitos) com acentuado edema e formação de vacúolos citoplasmáticos, sugerindo toxidade retiniana. Conclusões: As injeções intravítreas de sunitinibe e nanopartículas lipídicas sólidas não foram tóxicas para a retina. No entanto, nanocápsulas mostraram ser tóxicas para a retina. Sendo assim, a possibilidade de poder combinar o potencial de uma droga que possui a capacidade de inibir duas importantes vias da angiogênese, às vantagens de liberação controlada das nanopartículas lipídicas sólidas, pode ser um importante recurso terapêutico para doenças vasoproliferativas oculares.

Seleção de marcadores para a busca ativa de endoftalmite após cirurgia de catarata / Selection of markers for active search of endophthalmitis after cataract surgery / Selección de marcadores para la búsqueda activa de endoftalmitis tras cirugía de catarata

Objetivo: Identificar marcadores potenciais para auxiliar na busca ativa de endoftalmite após cirurgia de catarata. Método: Estudo retrospectivo, descritivo e longitudinal, realizado por meio da revisão de prontuários de pacientes submetidos à cirurgia de catarata. Resultados: A amostra incluiu 20 pacientes (grupo de estudo ­ GE) que desenvolveram endoftalmite pós-operatória e 309 pacientes (grupo controle ­ GC) que não apresentaram endoftalmite. Os dados foram analisados para identificar os marcadores clínicos e epidemiológicos com uma diferença percentual ≥30% entre os grupos. Em comparação com o GC, o GE teve frequência ≥30% em: sinais e sintomas pós-operatórios definidos (dor, reação da câmara anterior, hipópio, edema da córnea, hiperemia conjuntival e opacidade vítrea); mais de 4 retornos pós-operatórios; e realização de injeção de antibiótico intravítreo. Conclusão: Os indicadores selecionados são sugeridos para incorporação na busca ativa das infecções pós-operatórias de endoftalmite, visando à facilidade operacional do sistema de vigilância epidemiológica

Objective: To identify potential markers to assist in the active search of endophthalmitis after cataract surgery. Method: Retrospective, descriptive, and longitudinal study, conducted through review of medical records of patients who underwent cataract surgery. Results: The sample included 20 patients (study group - SG) who developed postoperative endophthalmitis and 309 patients (control group - CG) who did not have endophthalmitis. The data were analyzed to identify the clinical and epidemiological markers with a percentage difference ≥30% between groups. In comparison with the CG, the SG demonstrated frequency ≥30% in established postoperative signs and symptoms (pain, anterior chamber reaction, hypopyon, corneal edema, conjunctival hyperemia, and vitreous opacity); more than four postoperative appointments; and administration of an intravitreal antibiotic injection. Conclusion: The selected markers are suggested for incorporation into the active search for post-operative endophthalmitis, in order to facilitate the operation of the epidemiological surveillance system

Objetivo: Identificar marcadores potenciales para auxiliar en la búsqueda activa de endoftalmitis tras cirugía de facectomía. Método: Estudio retrospectivo, descriptivo y longitudinal, realizado por medio de la revisión de prontuarios de pacientes sometidos a la cirugía de catarata. Resultados: La muestra incluyó 20 pacientes (grupo de estudio ­ GE) que desarrollaron endoftalmitis pos-operatoria y 309 pacientes (grupo control ­ GC) que no presentaron endoftalmitis. Los datos fueron analizados para identificar los marcadores clínicos y epidemiológicos con una diferencia porcentual ≥30% entre los grupos. En comparación con el GC, el GE tuvo frecuencia ≥30% en: señales y síntomas pos-operatorios definidos (dolor, reacción de la cámara anterior, hipopión, edema de la córnea, hiperemia conjuntival y opacidad vítrea); más de 4 retornos pos-operatorios; y realización de inyección de antibiótico intravítreo. Conclusión: Los indicadores seleccionados son sugeridos para incorporación en la búsqueda activa de las infecciones pos-operatorias de endoftalmitis, buscando la facilidad operacional del sistema de vigilancia epidemiológica