Nitrosative stress and corneal transplant endothelial cell death during acute graft rejection.

BACKGROUND: Nitrosative stress takes place in endothelial cells (EC) during corneal acute graft rejection. The purpose of this study was to evaluate the potential role of peroxynitrite on corneal EC death. METHODS: The effect of peroxynitrite was evaluated in vivo. Fifty, 250, and 500 microM in 1.5 microL of the natural or denatured peroxynitrite in 50 microM NaOH, 50 microM NaOH alone, or balanced salt solution were injected into the anterior chamber of rat eyes (n=3/group). Corneal toxic signs after injection were assessed by slit-lamp, in vivo confocal imaging, pachymetry, and EC count. The effect of peroxynitrite was also evaluated on nitrotyrosine and leucocyte elastase inhibitor/LDNase II immunohistochemistry. Human corneas were incubated with peroxynitrite and the effect on EC viability was evaluated. A specific inducible nitric oxide synthase inhibitor (iNOS) was administered systemically in rats undergoing allogeneic corneal graft rejection and the effect on EC was evaluated by EC count. RESULTS: Rat eyes receiving as little as 50 microM peroxynitrite showed a specific dose-dependent toxicity on EC. We observed an intense nitrotyrosine staining of human and rat EC exposed to peroxynitrite associated with leucocyte elastase inhibitor nuclear translocation, a noncaspase dependent apoptosis reaction. Specific inhibition of iNOS generation prevented EC death and enhanced EC survival of the grafted corneas. However, inhibition of iNOS did not have a significant influence on the incidence of graft rejection. CONCLUSION: Nitrosative stress during acute corneal graft rejection in rat eyes induces a noncaspase dependent apoptotic death in EC. Inhibition of nitric oxide production during the corneal graft rejection has protective effects on the corneal EC survival.

Autosomal-Recessive Posterior Microphthalmos Is Caused by Mutations in PRSS56, a Gene Encoding a Trypsin-Like Serine Protease.

Posterior microphthalmos (MCOP) is a rare isolated developmental anomaly of the eye characterized by extreme hyperopia due to short axial length. The population of the Faroe Islands shows a high prevalence of an autosomal-recessive form (arMCOP) of the disease. Based on published linkage data, we refined the position of the disease locus (MCOP6) in an interval of 250 kb in chromosome 2q37.1 in two large Faroese families. We detected three different mutations in PRSS56. Patients of the Faroese families were either homozygous for c.926G>C (p.Trp309Ser) or compound heterozygous for c.926G>C and c.526C>G (p.Arg176Gly), whereas a homozygous 1 bp duplication (c.1066dupC) was identified in five patients with arMCOP from a consanguineous Tunisian family. In one patient with MCOP from the Faroe Islands and in another one from Turkey, no PRSS56 mutation was detected, suggesting nonallelic heterogeneity of the trait. Using RT-PCR, PRSS56 transcripts were detected in samples derived from the human adult retina, cornea, sclera, and optic nerve. The expression of the mouse ortholog could be first detected in the eye at E17 and was maintained into adulthood. The predicted PRSS56 protein is a 603 amino acid long secreted trypsin-like serine peptidase. The c.1066dupC is likely to result in a functional null allele, whereas the two point mutations predict the replacement of evolutionary conserved and functionally important residues. Molecular modeling of the p.Trp309Ser mutant suggests that both the affinity and reactivity of the enzyme toward in vivo protein substrates are likely to be substantially reduced.

Extent of Hypoattenuation On CT Angiography Source Images In Basilar Artery Occlusion: Prognostic Accuracy Of Posterior Circulation ASPECTS And The Pons-midbrain Score In The Basilar Artery International Cooperation Study

Background: The posterior circulation Acute Stroke Prognosis Early CT Score (pc-ASPECTS) and the combined Pons-midbrain score quantify the extent of early ischemic changes in the posterior circulation. We compared the prognostic accuracy of both scores if applied to CT angiography (CTA) source images (CTA-SI) of patients in the Basilar Artery International Cooperation Study (BASICS).Methods: BASICS was a prospective, observational, multi-centre, registry of consecutive patients who presented with acute symptomatic basilar artery occlusion (BAO). Functional outcome was assessed at 1 month. We applied pc-ASPECTS and the combined Pons-midbrain score to CTA-SI by 3-reader-consensus. Readers were blinded to clinical data. We performed multivariable logistic regression analysis, adjusting for thrombolysis, baseline NIHSS score and age, and used the output to derive ROC curves to compare the ability of both scores to discriminate patients with favourable (modified Rankin Scale [mRS] scores 0-3) from patients with unfavourable (mRS scores 4-6) functional outcome.Results: We reviewed CTAs of 158 patients (64% men, mean age 65 _ 15 years, median NIHSS score 25 [0-38], median GCS score 7 [3-15], median onset-to-CTA time 234 minutes [11-7380]). At 1 month, 40 (25%) patients had a favourable outcome, 49 (31%) had an unfavourable outcome (mRS score 4-5) and 69 (44%) were deceased. Both techniques of assessing CTA-SI hypoattenuation in the posterior circulation showed equally good discriminative value in predicting final outcome (C-statistics; area under ROC curve 0.74 versus 0.75, respectively; p_0.37). Pc-ASPECTS dichotomized at _6 versus _6 was an independent predictor of favourable functional outcome (RR _ 2.2; CI95 1.1-4.7; p _ 0.034).Conclusion: Compared to the combined Pons-midbrain score, the pc-ASPECTS score has similar prognostic accuracy to identify patients with a favourable functional outcome in BASICS. Dichotomized pc-ASPECTS (_6 versus _6) is an independent predictor of favourable functional outcome in this population. Author Disclosures: V. Puetz: None. A. Khomenko: None. M.D. Hill: None. I. Dzialowski: None. P. Michel: None. C. Weimar: None. C.A.C. Wijman: None. H. Mattle: None. K. Muir: None. T. Pfefferkorn: None. D. Tanne: None. S. Engelter: None. K. Szabo: None. A. Algra: None. A.M. Demchuk: None. W.J. Schonewille: None.

Estudio retrospectivo sobre cambios en la morfología corneal tras crosslinking

S'ha realitzat un estudi retrospectiu de 41 casos clínics de queratocon mesurant les variables abans i després de la intervenció (al primer mes postoperatori, als 3 mesos i als 6 mesos). El crosslinking corneal amb riboflavina i radiació ultraviolada sembla ser un procediment segur i efectiu, que ha assenyalat la parada a la progressió del queratocon, una reducció, encara que no estadísticament significativa de la curvatura corneal, de l'equivalent esfèric i de l'astigmatisme en ulleres a pacients que prèviament havien manifestat una progressió del queratocon. Conclusions que podem treure d'aquest estudi: 1) Es pot esperar una mínima millora de l'agudesa visual o una detecció la pèrdua d'aquesta en els casos de queratocon progressiu. Encara que aquesta millora no va ser estadísticament significativa. 2) No es demostra que el crosslinking freni la progressió del queratocon, però sí observem un augment de les mitjanes dels valors que ens indiquen el grau de duresa corneal comparant els valors abans de la intervenció, i als 3 i 6 mesos, encara que no hagi diferència estadísticament significativa. Podem constatar que el Crosslinking és una tècnica més que podem emprar en el tractament del queratocon. Possiblement el seu efecte en la visió no serà molt apreciable, però davant de casos amb alt risc de progressió pot ser factible el seu ús ja que no té efectes secundaris d'importància.

Modelo de caracterización cerebral del área anorrectal mediante técnicas de neuroimagen para su utilización posterior en pacientes con incontinecia fecal

Resumen: La incontinencia fecal es una patología con importantes implicaciones sociosanitarias, con un tratamiento complejo y no siempre satisfactorio, especialmente la incontinencia fecal idiopática. El sistema nervioso central regula los procesos de continencia y defecación. Los estudios de neuroimagen han demostrado ser útiles para caracterizar las áreas cerebrales que controlan el área anorrectal. A partir de un grupo de voluntarias sanas, se ha creado un modelo de caracterización de estas áreas cerebrales anorrectales, que podrá ser utilizado posteriormente para compararlo con un grupo de pacientes con incontinencia fecal idiopática, estudiando posibles diferencias y posibles opciones terapéuticas.

Relació de l’aspecte de la ferida quirúrgica en el postoperatori immediat d’una artroplàstia total de genoll amb l’evolució funcional posterior

Mitjançant un estudi observacional prospectiu, hem descrit i classificat la ferida quirúrgica en el postoperatori immediat de 159 pacients que se’ls hi va implantar una artroplàstia total de genoll, mitjançant una escala descriptiva fiable i reproduïble creada pels autors (estudi de validesa). Es va observar que l’aspecte de la ferida quirúrgica en el postoperatori immediat no tenia relació amb la funcionalitat de la pròtesis als 6 mesos de seguiment ni amb el dolor que patia el pacient en l’estància hospitalària. Si que vam observar correlació estadísticament significativa entre el pitjor aspecte de la ferida i l’aparició d’infeccions protèsiques profundes.

Studies on Cercariae from Kuwait Bay. XI. Description and surface topography of Cercaria kuwaitae XI sp.n. (Digenea: Echinostomatidae)

A new echinostome cercaria, Cercaria kuwaitae XI sp.n., from the prosobranch gastropod Cerithidea cingulata (Gmelin) from Kuwait Bay is described. The new cercaria is characterized by 23 collar spines and primary excretory tubules with distinct diverticula. The cercaria encysts in the snail host and is similar to those of Acanthoparyphium sp. The surface topography of the redia, cercaria and metacercarial cyst wall is studied by scanning electron microscopy. This is the first echinostome cercaria to be recorded in a gastropod from the Arabian Gulf region.

Repercusión visual posterior al explante de lente intraocular opacificada

INTRODUCCIÓ: Valorar els resultats funcionals i quirúrgics posteriors a l’explant de la lent intraocular H60M opacificada. MATERIAL I MÈTODES: Estudi retrospectiu de vint-i-cinc ulls sotmesos a intercanvi de H60M opacificada. Anàlisi dels resultats visuals i complicacions, abans, durant i després de l’explante. RESULTATS: Trobarem millors resultats visuals i una menor taxa de complicacions quan no associem vitrectomia anterior a la cirurgia, així como amb la col•locació de la lent secundaria a sulcus. CONCLUSIÓ: L’intercanvi de la LIO Hidroview opacificada suposa un risc quirúrgic addicional, que podem minimitzar durant el període intraoperatori intentant preservar les estructures oculars el més anatòmicament possible.

Posterior cortical atrophy: review of the recent literature.

Posterior cortical atrophy (PCA) is a group of neurodegenerative dementing disorders characterized by initial predominant visual complaints followed by progressive decline in cognitive functions. The visuospatial and visuoperceptual defects arise from the dysfunction of, respectively, the dorsal (occipito-parietal) and the ventral (occipito-temporal) streams. Clinical symptoms, results of neuropsychological examination, and findings of posterior cerebral atrophy and/or posterior hypoperfusion/hypometabolism contribute to the diagnosis. However, owing to the insidious onset of PCA and the non-specificity of initial symptoms, the diagnosis is often delayed. Specific etiologies include Alzheimer's disease, dementia with Lewy bodies, subcortical gliosis, corticobasal degeneration, and prion-associated diseases. Alzheimer's disease accounts for at least 80 % of PCA cases. Recent research has concentrated on better defining the clinical presentation of PCA, improving neuroimaging analysis, testing new neuroimaging techniques, and developing biological measurements. Selected recent papers on PCA are reviewed in this article.

Posterior spiracles of fourth instar larvae of four species of phlebotomine sand flies (Diptera: Psychodidae) under scanning electron microscopy

In the present study, posterior spiracles of laboratory-reared fourth instar larvae of Lutzomyia longipalpis, L. migonei, L. lenti, and L. whitmani (Diptera: Psychodidae) of the State of Ceará, Brazil, were examined under scanning electron microscopy. The number of papillae of spiracles examined varied according to the species examined, but no intraspecific differences were found. The importance of this structure to sand fly larva identification and phylogeny is commented.

Hyperglycemia-induced abnormalities in rat and human corneas: the potential of second harmonic generation microscopy.

BACKGROUND: Second Harmonic Generation (SHG) microscopy recently appeared as an efficient optical imaging technique to probe unstained collagen-rich tissues like cornea. Moreover, corneal remodeling occurs in many diseases and precise characterization requires overcoming the limitations of conventional techniques. In this work, we focus on diabetes, which affects hundreds of million people worldwide and most often leads to diabetic retinopathy, with no early diagnostic tool. This study then aims to establish the potential of SHG microscopy for in situ detection and characterization of hyperglycemia-induced abnormalities in the Descemet's membrane, in the posterior cornea. METHODOLOGY/PRINCIPAL FINDINGS: We studied corneas from age-matched control and Goto-Kakizaki rats, a spontaneous model of type 2 diabetes, and corneas from human donors with type 2 diabetes and without any diabetes. SHG imaging was compared to confocal microscopy, to histology characterization using conventional staining and transmitted light microscopy and to transmission electron microscopy. SHG imaging revealed collagen deposits in the Descemet's membrane of unstained corneas in a unique way compared to these gold standard techniques in ophthalmology. It provided background-free images of the three-dimensional interwoven distribution of the collagen deposits, with improved contrast compared to confocal microscopy. It also provided structural capability in intact corneas because of its high specificity to fibrillar collagen, with substantially larger field of view than transmission electron microscopy. Moreover, in vivo SHG imaging was demonstrated in Goto-Kakizaki rats. CONCLUSIONS/SIGNIFICANCE: Our study shows unambiguously the high potential of SHG microscopy for three-dimensional characterization of structural abnormalities in unstained corneas. Furthermore, our demonstration of in vivo SHG imaging opens the way to long-term dynamical studies. This method should be easily generalized to other structural remodeling of the cornea and SHG microscopy should prove to be invaluable for in vivo corneal pathological studies.

Expression of inducible nitric oxide synthase in bovine corneal endothelial cells and keratocytes in vitro after lipopolysaccharide and cytokines stimulation.

PURPOSE: To determine whether bovine corneal endothelial (BCE) cells and keratocytes express the inducible form of nitric oxide synthase (NOS) after exposure to cytokines and lipopolysaccharide (LPS), and to study the regulation of NOS by growth factors. METHODS: Cultures of bovine corneal endothelial cells and keratocytes were exposed to increasing concentrations of LPS, interferon-gamma (IFN-gamma), and tumor necrosis factor-alpha (TNF-alpha). At selected intervals after exposure, nitrite levels in the supernatants were evaluated by the Griess reaction. Total RNA was extracted from the cell cultures, and messenger RNA levels for inducible NOS (NOS-2) were measured by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: Exposure of BCE cells and keratocytes to LPS and IFN-gamma resulted in an increase of nitrite levels that was potentiate by the addition of TNF-alpha. Analysis by RT-PCR demonstrated that nitrite release was correlated to the expression of NOS-2 messenger RNA in BCE cells and keratocytes. Stereoselective inhibitors of NOS and cycloheximide inhibited LPS-IFN-gamma-induced nitrite release in both cells, whereas transforming growth factor-beta (TGF-beta) slightly potentiated it. Fibroblast growth factor-2 (FGF-2) inhibited LPS-IFN-gamma-induced nitrite release and NOS-2 messenger RNA accumulation in keratocytes but not in BCE cells. CONCLUSIONS: The results demonstrate that in vitro activation of keratocytes and BCE cells by LPS and cytokines induces NOS-2 expression and release of large amounts of NO. The high amounts of NO could be involved in inflammatory corneal diseases in vivo.

Investigar la relación entre la detección de glucosa en el fluido aspirado a través del catéter epidural y la aparición posterior de cefalea

En 33 mujeres en trabajo de parto se realiza analgesia epidural hasta el momento del parto. Una vez finalizado éste, se procede a la retirada del catéter epidural realizando al mismo tiempo una aspiración continua. El fluido obtenido es analizado con un glucómetro digital para averiguar si contiene glucosa, que indicaría su procedencia intrarraquídea. Se relaciona la presencia o no de glucosa con la aparición posterior de cefalea.