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The aim of this study was to evaluate the motor, sensory, functional, and head posture results of recession of the lateral rectus muscle contralateral to the involved eye in patients with exotropic Duane retraction syndrome (DRS) type 3. This was a retrospective, longitudinal, observational study of a consecutive clinical case series. Of the 11 patients with DRS type 3 operated on at a tertiary medical center from 1977 to 2012, 8 underwent recession of the lateral rectus muscle contralateral to the involved eye (with combined Y-splitting of ipsilateral lateral rectus muscle in 3 of them). Full ophthalmic, orthoptic, and neurological examination was performed before and after surgery. Main outcome measures included intragroup changes in motor misalignment, abnormal head turn, ocular upshoot, and stereopsis. Mean patient age was 8.75±3.1 years at surgery. Mean exodeviation for distance was −17.3±3.5 prism diopters (PD) preoperatively and −4.0±6.1 PD postoperatively; corresponding values for near were −23.1±7.2 PD and −5.9±8.7 PD. Motor deviation improved by 77% for distance (P=0.017) and 74.5% for near (P=0.01). In 7/8 patients, the postoperative residual exodeviation (distance and near) was

Objective To investigate the use of peripapillary optical coherence tomography for monitoring optic neuropathy in pediatric craniopharyngioma. Design Retrospective, consecutive-cohort, single-centre chart analysis. Participants Twenty children with craniopharyngioma treated at a pediatric medical centre from 1999 to 2011. Methods The medical files were reviewed for demographics and optic nerve function. Findings for visual acuity and visual fields were analyzed against repeated optical coherence tomography (OCT) measurements of peripapillary nerve fibre layer thickness (using either time-domain Stratus OCT or spectral-domain Cirrus OCT). Results Average age at diagnosis was 6.5 ± 3.88 years. The most common presenting symptom was headache; only 1 child complained of visual loss. Mean best corrected visual acuity (logMAR) was 0.036 ± 0.06 in the 17 healthy eyes and 1.05 ± 1.45 in the 23 eyes with optic neuropathy. Positive signs included relative afferent pupillary defect (8/20), visual acuity loss (7/20), temporal visual field loss (bilateral 4/15, unilateral 4/15), papilledema (3/20), and unilateral/bilateral optic disc pallor (14/20). RNFL thickness was significantly lower in eyes with optic neuropathy than in healthy eyes (65 ± 22 µm vs 86.2 ± 29 µm; p = 0.000) and correlated with visual acuity (r = –0.43 to –0.17, p = 0.0001) and presence or absence of a visual field defect (mean difference, 26.1 ± 5.8 µm, p = 0.003). Ten children showed no change in RNFL thickness over time (mean 18 ± 14.2 months). Conclusions A thinner RNFL on ocular coherence tomography is correlated with poorer visual acuity and visual field loss. Ocular coherence tomography may serve as an objective method to quantify axonal loss caused by craniopharyngioma. Further investigation is needed to determine its use for evaluating progressive axonal loss over time.

In pediatric brain tumours, dissemination of malignant cells within the central nervous system confers poor prognosis and determines treatment intensity, but is often undetectable by imaging or cytology. This study describes the use of fluorescence lifetime (FLT) imaging microscopy (FLIM), a novel diagnostic tool, for detection of metastatic spread. The study group included 15 children with medulloblastoma and 2 with atypical teratoid/rhabdoid tumour. Cells extracted from the tumour and the cerebrospinal fluid (CSF) 2 weeks postoperatively and repeatedly during chemo/radiotherapy were subjected to nuclear staining followed by FLT measurement and cytological study. Control CSF samples were collected from patients with infectious/inflammatory disease attending the same hospital. Median FLT was prolonged in tumour cells (4.27 ± 0.28 ns; P −16) and CSF metastatic cells obtained before chemo/radiotherapy (6.28 ± 0.22 ns; P −16); normal in inflammatory control cells (2.6 ± 0.04 ns) and cells from children without metastasis before chemo/radiotherapy (2.62 ± 0.23 ns; P = 0.858) and following treatment (2.62 ± 0.21 ns; P = 0.053); and short in CSF metastatic cells obtained after chemo/radiotherapy (2.40 ± 0.2 ns; P −16). FLIM is a simple test that can potentially identify CSF spread of brain tumours. FLT changes in accordance with treatment, with significant prolonged median values in tumours and metastases. More accurate detection of metastatic cells may guide personalised treatment and improve the therapeutic outcome.

MB-28. MicroRNA EXPRESSION IS HIGHLY SUBGROUP SPECIFIC IN MEDULLOBLASTOMA Sivan Gershanov1,2, Shalom Michowiz3,4, Orit Barinfeld2,3, Mali Salmon-Divon1, Nitza Goldenberg-Cohen3,4, and Helen Toledano3,4; Ariel University, Ariel, Israel; Felsenstein Medical Research Center, Petah-Tiqwa, Israel; Tel-Aviv University, Tel-Aviv, Israel; Schneider Children’s Medical Center of Israel, Petah-Tiqwa, Israel Medulloblastoma (MB), the commonest malignant brain tumor of childhood, is divided into four tumor subgroups representing distinct clinical and molecular entities. MicroRNAs (miRNAs) are involved in carcinogenesis and tumor progression by regulating post-transcriptional gene expression. However, our understanding of the miRNA-mRNA regulatory network is incomplete. The aim of the study is to identify novel miRNA subgroup biomarkers and their target mRNAs for rapid and specific diagnosis and future targeted therapy. With this aim, integrated whole transcriptome mRNA and miRNA expression analysis was performed on primary tumor samples collected from 10 MB patients. 867 mature miRNAs were identified in at least a single MB sample, of them 462 were common to all 4 subgroups. 25 (2.5%) of all expressed miRNAs appeared to be significantly differentially expressed between the MB subtypes (FDR , 0.1). Namely, upregulation of hsa-miR-224-5p and hsa-miR-449c-5p was found exclusively among WNT, while downregulation of hsa-miR-135b-5p characterized SHH. Among groups 3 and 4, hsa-miR-20a-5p was upregulated or downregulated, respectively. RNA-seq from the same tumor samples identified 500 genes that vary between subtypes (q value ,0.05), among which 69 (13.8%) have anticorrelated miRNA-mRNA interactions with the 25 detected miRNA biomarkers. The predicted mRNAs targets of these miRNAs are associated with different signaling pathways, known to have a role in MB biology. Our study demonstrates that miRNAs are readily detectible and are highly specific to distinct MB subgroups. Understanding the involvement of miRNAs and their targets in MB related signaling pathways may improve diagnosis and advance the development of targeted treatment for MB. Neuro-Oncology 18:iii97–iii122, 2016. doi:10.1093/neuonc/now076.26 #The Author(s) 2016. Published by Oxford University Press on behalf of the Society for Neuro-Oncology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Brain tumors are the second leading cause of cancer-related deaths in children, after leukemia. Patients with cancer in the central nervous system have a very low recovery rate. Today known imaging and cytology techniques are not always sensitive enough for an early detection of both tumor and its metastatic spread, moreover the detection is generally limited, reviewer dependent and takes a relatively long time. Medulloblastoma (MB) is the most common malignant brain tumor in children. The aim of our talk is to present the frequency domain fluorescence lifetime imaging microscopy system as a possible method for an early detection of MB and its metastatic spread in the cerebrospinal fluids within the pediatric population.

Purpose: To present the genetic cause of progressive deterioration in visual acuity and color vision in a child with high myopia and strabismus. Here we describe a novel x-linked mutation in the opsin 1 medium-wave-sensitive (OPN1MW) gene in a child, leading to cone rod dystrophy. Setting/Venue: Trio whole-exome sequencing (WES). Methods: We reviewed the clinical data and eye exams including family history since the patient's first visit 2008. Further evaluation included fundus photography, optical coherence tomography and electroretinography. The child was also referred to neurological assessment and magnetic resonance imaging was performed. Genetic evaluation included the extraction of DNA from peripheral blood leukocytes, trio WES and bioinformatics analysis using the Burrows-Wheeler Aligner (BWA) and the Genome Analysis Tool Kit (GATK) software. Results: A normal eye examination showed that the parents and brothers were healthy. The boy had bilateral impaired vision (best visual corrected 1/24), high myopia (BE -7.0 diopter) and esotropia. On fundus exam, normal fundus appearance was reported with only mild temporal pallor of the optic discs. Retinal thickness measured by optical coherence tomography was within normal limits. Electrophysiological studies were unspecified for cone rod dystrophy based on photopic and scotopic responses. The complete neurology examination and neuroimaging were normal. WES revealed no compound heterozygosity or recessive mutations. Conclusions: Here we described a boy with severely impaired vision, not explained by the high myopia or the strabismus, who also had a progressive course. The family history was negative. Genetic evaluation revealed a deletion of exon 5 in the OPN1MW gene. He was diagnosed with a blue cone monochromacy. This child is a representative case with the common symptoms that are sometimes under-diagnosed without genetic evaluation.

SummaryInherited optic neuropathies are a heterogeneous group of disorders characterized by mild to severe visual loss, colour vision deficit, central or paracentral visual field defects and optic disc pallor. Optic atrophies can be classified into isolated or non-syndromic and syndromic forms. While multiple modes of inheritance have been reported, autosomal dominant optic atrophy and mitochondrial inherited Leber's hereditary optic neuropathy are the most common forms. Optic atrophy type 1, caused by mutations in theOPA1gene is believed to be the most common hereditary optic neuropathy, and most patients inherit a mutation from an affected parent. In this study we used whole-exome sequencing to investigate the genetic aetiology in a patient affected with isolated optic atrophy. Since the proband was the only affected individual in his extended family, and was a product of consanguineous marriage, homozygosity mapping followed by whole-exome sequencing were pursued. Exome results identified a novelde novo OPA1mutation in the proband. We conclude, that thoughde novo OPA1mutations are uncommon, testing of common optic atrophy-associated genes such as mitochondrial mutations andOPA1gene sequencing should be performed first in single individuals presenting with optic neuropathy, even when dominant inheritance is not apparent.

Ischemic or mechanical injury to the optic nerve is an irreversible cause of vision loss, associated with limited regeneration and poor response to neuroprotective agents. The aim of this study was to assess the capacity of adult bone marrow cells to participate in retinal regeneration following the induction of anterior ischemic optic neuropathy (AION) and optic nerve crush (ONC) in a rodent model. The small-sized subset of cells isolated by elutriation and lineage depletion (Fr25lin(-)) was found to be negative for the neuroglial markers nestin and glial fibrillary acidic protein (GFAP). Syngeneic donor cells, identified by genomic marker in sex-mismatched transplants and green fluorescent protein, incorporated into the injured retina (AION and ONC) at a frequency of 0.35%-0.45% after intravenous infusion and 1.8%-2% after intravitreous implantation. Perivascular cells with astrocytic morphology expressing GFAP and vimentin were of the predominant lineage that engrafted after AION injury; 10%-18% of the donor cells incorporated in the retinal ganglion cell layer and expressed NeuN, Thy-1, neurofilament, and beta-tubulin III. The Fr25lin(-) cells displayed an excellent capacity to migrate to sites of tissue disruption and developed coordinated site-specific morphological and phenotypic neural and glial markers. In addition to cellular reconstitution of the injured retinal layers, these cells contributed to endothelial revascularization and apparently supported remodeling by secretion of insulin-like growth factor-1. These results suggest that elutriated autologous adult bone marrow-derived stem cells may serve as an accessible source for cellular reconstitution of the retina following injury.

Highlights ► We investigated the occurrence of BRAF, GNAQ, and GNA11 mutations in pediatric low-grade glioma. ► We found an increase in the copy number of BRAF. ► BRAF and GNAQ mutations are mutually exclusive. ► Understanding the molecular basis of low-grade glioma may increase treatment options., Fifty-two samples of pediatric low-grade glioma (48 primary, 4 recurrent) were analyzed for BRAF copy number variation (digital PCR analysis, CopyCaller) and point mutations of BRAF V600E, and exon 5 Q209 in GNAQ, and GNA11, using the MALDI-TOF mass spectrometer with validation by direct sequencing. An increased BRAF copy number was found in 18/47 primary samples tested; 15 of them (83.3%) were pilocytic astrocytomas. A BRAF mutation was found in 3/48 primary tumors, all with a normal BRAF copy number and no GNAQ mutation. One sample had a GNAQ209 mutation (Q209P626) with a normal BRAF gene; none of the tumors had a GNA11Q209 mutation. Recurrent or progressive tumors, analyzed in four patients, had the same molecular genotype as their primary. Increased BRAF copy number and activating BRAF mutations may be involved in the development of low-grade glioma via overactivation of the Ras/Raf pathway. This is the first report of a mutation in GNAQ209 in pediatric low-grade glioma. Understanding the molecular mechanisms underlying glioma initiation and growth may assist in the development of targeted therapies.

Bax expression is a prerequisite for retinal ganglion cell (RGC) apoptosis. Experimental studies have reported Bax protein upregulation following optic nerve transection. The stimuli that trigger apoptosis share a common executioner proteolysis cascade, including caspase-3 and poly-(adenosine diphosphate ribose) polymerase cleavage. This study sought to elucidate the role of the mitochondrial apoptotic pathway in RGCs using a Bax transgenic knockout mouse model.The right optic nerves of 26 C57BL mice, 7 Bax, 7 Bax, and 12 Bax, were subjected to crush injury and analyzed for apoptosis and neuronal cell loss on days 1, 3, and 21. Levels of Bax, Bcl-2, and caspase-3 messenger RNA expression were determined with real-time polymerase chain reaction.Multiple apoptotic cells were detected in the retinas of the Bax and Bax mice at days 1 and 3, but not in the Bax mice. The Bax/Bcl-2 ratio was higher in the Bax than in the Bax mice on day 1 (1.33 and 0.83, respectively), with a trend toward an increase on day 3 (1.47 and 1.66, respectively); Bax/Bcl-X showed the same elevation on day 1 in the wild-type mice (1.34) but decreased on day 3 (0.8). Bax gene expression was undetectable in the Bax mice. Caspase-3 gene expression was higher in the Bax than in the Bax mice on day 1 and dropped toward baseline on day 3. The opposite trend was noted in the Bax mice.The lack of apoptosis combined with the reduction in proapoptotic genes in the Bax mice after injury compared to the Bax and Bax mice suggests that Bax plays a crucial role in the induction of apoptosis. Suppression of Bax expression may reduce retinal cell loss.

Summary The phosphoinositide 3-kinase (PI3K)/v-akt murine thymoma (AKT) viral oncogene pathway is involved in regulating the signaling of multiple biological processes such as apoptosis, metabolism, cell proliferation, and cell growth. Mutations in the genes associated with the PI3K/AKT pathway including PI3K , AKT , RAS and PTEN , are infrequently found within head and neck squamous cell carcinoma and more specifically are rarely reported in oral squamous cell carcinoma (OSCC) cases. We aimed to investigate the frequency of mutations in AKT1 , PTEN , PIK3CA , and RAS ( K-RAS , N-RAS , H-RAS ) genes in 37 cases of oral squamous cell carcinoma (OSCC). Mutational analysis of PTEN , RAS , PIK3CA and AKT genes was performed using chip-based matrix-assisted laser desorption time-of-flight (MALDI-TOF) mass spectrometry and by direct sequencing. The only gene mutated in our series was the PIK3CA . Missense mutations of the PIK3CA gene were found in 4 of our cases (10.8%); no correlation has been found with oral location, stage and survival. The absence of mutations in AKT1 , PTEN , and RAS genes in the present study is in accordance with previous studies confirming that these genes are rarely mutated in OSCC. Our data confirm that PIK3CA is important to OSCC tumorigenesis and can contribute to oncogene activation of the PIK3CA / AKT pathway in OSCC. The knowledge of the PIK3CA’s involvement in OSCC is important because a specific kinase inhibitor could be considered as a future therapeutic option for OSCC patients with PIK3CA mutations.

Purpose To investigate whether acquired somatic mutations in the iron response element of the ferritin L-chain gene account for the age-related cataract. Methods The 15 most prevalent point mutations causing hereditary hyperferritinemia cataract syndrome (HHCS) were screened in patients with age-related cataract using MALDI-TOF Mass Spectrometry. DNA samples were obtained from the lens capsules of patients following cataract surgery, and subjected to PCR amplification. Products were analyzed by a Sequenom® mass spectrometer, and classified as a mutation or wild type according to molecular weight. For a positive control, L-ferritin G32T mutation detected by direct sequencing in 3 members of an Israeli family known to be affected by HHCS was used. Results DNA samples were isolated from the lens capsules of 90 patients, mean age 73.86, and screened for L-ferritin mutations. While the G32T mutation was detected in all 3 positive control cases, all other patients were negative for the 15 mutations. Conclusions Somatic mutations in the iron response elements (IRE) of the L-ferritin gene are infrequent in the age-related cataract. The role of L-ferritin genetic variations in the pathogenesis of age-related cataract is yet to be explored.

Purpose Werner syndrome is an autosomal recessive disease of premature aging caused by a polymorphic C1367T mutation in the Werner (WRN) gene. Although there are differences between the pathobiology of normal aging and the phenotype of Werner syndrome, the clinical age-related changes are similar. The aim of the study was to investigate the incidence of the C1367T (rs1346044) polymorphism in patients with age-related cataract. Methods The study group consisted of 81 patients with senile cataract undergoing cataract extraction surgery. Data on age, sex, and medical history of microvascular disease and cancer were obtained from the medical files. Anterior lens capsule material was collected during surgery. DNA was extracted, amplified by polymerase chain reaction, and screened for the C1367T polymorphism in WRN using restriction enzymes followed by sequencing. Results There were 33 male and 48 female patients of mean age 74.3±9 years. Genotypic frequencies were 67% for TT and 33% for TC. None of the patients had the CC genotype. Ten patients had a history of myocardial infarct, 8 cerebrovascular accident, and 8 various tumors. The distribution of these morbidities was similar in the two genotype groups. Conclusions The distribution of the C1367T WRN polymorphism in patients with senile cataract is similar to that in the normal population. Cataract formation in the elderly is not linked to a WRN mutation.

A percentage of sonic hedgehog (SHH) medulloblastomas will carry a germline mutation in PTCH1 (upstream) or SUFU (downstream) as part of Gorlin’s syndrome. It has been suggested that the tendency to develop medulloblastoma is greater in those with SUFU and that the syndromic features are different depending on the gene involved. We compared the gene and microRNA expression profile of the tumors of two patients with germline mutations - one in PTCH and one in SUFU. We performed RNA and microRNA sequencing from fresh tumor samples followed by ingenuity pathway analysis. We identified several differences in expression of genes and microRNA: the miRNAs miR-130a-3p and miR-181-5p showed lower expression in the SUFU sample with resultant increased expression of DICER1 either directly or indirectly. Other genes that exhibit decreased expression in the SUFU sample included Connexin involved in Gap junction signaling and Metalloproteinase involved in axonal guidance signaling. Genes that display higher expression in the SUFU sample included GABAR-A in the GABA receptor signaling pathway and Cahedrin in the beta-catenin signaling pathway. These findings need confirmation in a larger sample of tumors and may help to find treatable molecular targets for these types of medulloblastoma. This is vital since radiation - a standard component of treatment for medulloblastoma - is contraindicated in Gorlin’s due to the risk of basal cell carcinoma. In addition SHH inhibitors are ineffective for SUFU medulloblastoma as they inhibit upstream of the mutation.

Pain is the main cause of patient distress/dissatisfaction after strabismus surgery. The aim of the study was to evaluate the effect of sub-Tenon's block with ropivacaine at the end of strabismus surgery on post-operative pain.A prospective trial was conducted in 79 patients (age 1.0-65 years) scheduled for outpatient primary strabismus surgery with fixed sutures under general anesthesia (GA) at a major tertiary hospital. Half the patients were randomly allocated to receive sub-Tenon's block with ropivacaine 0.2% at conclusion of the operation. Primary outcome measures were visual analog scale (VAS) scores at arrival to the post-anesthesia care unit (PACU), at discharge 3 hr later, 12-16 hr post-operatively, and 24 hr post-operatively. Supplemental analgesia requirements and patient satisfaction were recorded as well. Data were presented as median (range). Mann-Whitney test, Pearson chi(2)-test or Fisher's exact test was used for statistical analysis; por= 0.05 was considered significant.There were no between-group differences in median VAS scores at arrival to the PACU and at discharge, with a borderline difference at 24 hr post-operatively (p = 0.06). At 12-16 hr post-operatively, the median score was 0.0 (range 0-5) in the study group and 4.0 (range 0-6) in the controls (p0.001). The lower VAS score in the study group was associated with a lower rate of supplemental analgesia use (21.9% versus 57.9%, p = 0.001), fewer doses of supplemental analgesia (10 doses versus 35, p = 0.03), and higher patient satisfaction (p0.001).Sub-Tenon's block with ropivacaine 0.2% at the completion of outpatient primary strabismus surgery with fixed sutures under GA reduces pain 12-16 hr post-operatively and analgesia requirements 4-23 hr post-operatively.

Purpose: To investigate the neuroprotective effect of brimonidine following induction of ischaemic optic neuropathy in rodents (rAION). Methods: Mice were treated with an intraperitoneal injection of brimonidine 48, 24 or 0 h before rAION induction or eye drops for 5 days after rAION induction. Retinal ganglion cell (RGC) loss and expression of genes involved in the angiogenesis (vascular endothelial growth factor [VEGF], pigment epithelium-derived factor [PEDF], The epidermal growth factor homology domains-2 [Tie-2]), ischaemia (haem oxygense-1 [HO-1], hypoxia-inducible factor 1α[HIF-1α], endothelial nitric oxide synthase [eNOS]) and oxidative stress (superoxide dismutase-1 [SOD-1], glutathione peroxidase-1 [GPX-1]) response to ischaemic damage were compared with sham or rAION-untreated mice. Results: No RGC loss was detected in the brimonidine-treated mice. Effect of post-rAION eye drops: day 1 – no decrease in retinal mRNA levels of angiogenesis-related genes, increase in ischaemia- and oxidative stress-related genes except HIF-1α; day 3 – baseline or higher levels of oxidative and ischaemia-related genes except HIF-1α, increase in VEGF, decrease in PEDF; day 21 – no change in angiogenesis-related genes. Effect of pre-rAION injection: baseline levels of angiogenesis-related genes with all injection schedules; increase in ischaemia-related genes with 48-h and 0-h pretreatment; decrease in HO-1 and eNOS with 24-h pretreatment; increase in oxidative-related genes except GPX-1. In optic nerve tissue, HO-1, HIF-1α and SOD-1 decreased on day 1 after topical administration and were still below baseline on day 3. Conclusions: The increase in HO-1 associated with rAION is mitigated with brimonidine treatment, especially when administered intraperitoneally. Topical brimonidine apparently reduces VEGF, Tie-2, HIF-1α and GPX-1 expression on day 21. These results agree with published data and may have therapeutic implications for patients diagnosed with AION in the acute phase.

The aim of this study was to characterize the molecular and histological changes that occur in the retina following central retinal artery occlusion (CRAO) in a mouse model. CRAO was induced in 60 mice by laser photoactivation of intravenously injected rose bengal. Mice were sacrificed at 3, 6, 12, and 24h and 7 and 21 days after CRAO induction for molecular analysis (5-13 mice/time point) and histological and apoptosis studies (3-4 mice/time point). Fundus examination and fluorescein angiography were also performed at various points. Retinal mRNA was analyzed for expression of T-cell antigen 1 (Thy-1), vascular endothelial growth factor (VEGF), heme oxygenase-1 (HO-1), and hypoxia-induced factor 1 alpha (HIF-1 alpha) using real-time polymerase chain reaction. The results showed that at 6-24h following CRAO induction, the retina was edematous, with interrupted blood perfusion. Fluorescein angiography showed reperfusion at 6h, and TdT-mediated dUTP nick end-labeling (TUNEL) assay revealed an increase in apoptotic cells in the first 24h. On histological sections, nuclear loss in the inner retinal layers was maximal on day 21. Thy-1 expression decreased to 30% of baseline (P

Patients on long-term hemodialysis undergo multiple interventions, including insertion of central catheters and arteriovenous anastomoses for creation of vascular access. The need for high-flow vessels to maintain hemodialysis efficiency leads to wear on the central veins and consequent stenosis and occlusion. In addition to local signs of impaired venous drainage, abnormal venous flow patterns involving the upper chest, face, and central nervous system might develop. We describe the first pediatric case of devastating intracranial hypertension presenting with visual loss in the eye contralateral to a high-flow vascular access in a patient on long-term hemodialysis. The literature on this rare complication of hemodialysis is reviewed.

Small-sized adult bone marrow cells isolated by counterflow centrifugal elutriation and depleted of lineage markers (Fr25lin(-)) have the capacity to differentiate into insulin-producing cells and stabilize glycemic control. This study assessed competitive migration of syngeneic stem cells to the bone marrow and islets in a murine model of chemical diabetes. VLA-4 is expressed in ∼ 25% of these cells, whereas CXCR4 is not detected, however, it is transcriptionally upregulated (6-fold). The possibility to enrich stem cells by a bone marrow homing (BM-H) functional assay was assessed in sequential transplants. Fr25lin(-) cells labeled with PKH26 were grafted into primary myeloablated recipients, and mitotically quiescent Fr25lin(-)PKH(bright) cells were sorted from the bone marrow after 2 days. The contribution of bone marrow-homed stem cells was remarkably higher in secondary recipients compared to freshly elutriated cells. The therapeutic efficacy was further increased by omission of irradiation in the secondary recipients, showing a 25-fold enrichment of islet-reconstituting cells by the bone marrow homing assay. Donor cells identified by the green fluorescent protein (GFP) and a genomic marker in sex-mismatched transplants upregulated PDX-1 and produced proinsulin, affirming the capacity of BM-H cells to convert in the injured islets. There was no evidence of transcriptional priming of freshly elutriated subsets to express PDX-1, insulin, and other markers of endocrine progenitors, indicating that the bone marrow harbors stem cells with versatile differentiation capacity. Affinity to the bone marrow can be used to enrich stem cells for pancreatic regeneration, and reciprocally, conditioning reduces the competitive incorporation in the injured islets.

The aim of the study was to investigate the incidence of nystagmus at diagnosis in children with optic pathway glioma involving the chiasm and hypothalamus.Twenty-two patients with a measurable optic pathway/hypothalamic glioma (without neurofibromatosis-1) were followed in our center from 2001 to 2013. The medical files were retrospectively reviewed for demographic and clinical findings, and the imaging scans, for tumor characteristics.There were 9 boys and 13 girls of mean age 3.5 ± 4.4 years at diagnosis; 15 were aged2 years. Tumor size ranged from 10 × 6 mm to 62 × 29 mm. Mean duration of follow-up was 8.3 ± 5.4 years. Nystagmus was detected at diagnosis in 10 children (45%), all2 years old (66.6% of the younger group); no child older than 2 years presented with nystagmus. Nystagmus, once present, did not resolve and continued throughout follow-up. There were no cases of new onset of nystagmus during follow-up in the children in whom it was not detected at diagnosis. Treatment consisted of partial resection/biopsy with/without shunting (n = 13) and chemotherapy (n = 19) with (n = 2) or without adjuvant radiation. Of the 22 children, 6 had a radiographic response to treatment, 8 remained stable, and 8 (all of whom received chemotherapy) showed disease progression despite treatment.In conclusion, monocular nystagmus is a more common presenting sign of optic pathway/hypothalamic glioma in children2 years old than previously estimated. Although subtle, nystagmus has a very narrow differential diagnosis, and its presence should raise suspicions of a chiasmal tumor with prompt referral for imaging. The visual prognosis is moderate to poor.

To examine the effects of hyperbaric oxygen (HBO) therapy and knockout of toll-like receptor 4 (TLR4) on the outcome of temporary middle cerebral artery occlusion (MCAO) in a mouse model.MCAO was induced in anesthetized male C57Bl/6 mice (WT) and TLR4 knockout mice (TLR4(-/-)) using an intra-arterial filament method. After 30 or 90 min, the filament was removed, and the mice were given either no treatment (WT and TLR4(-/-) groups) or HBO (WT only). Mice were euthanized 24 h after MCAO, and the brain infarct area was examined using 2,3,5-triphenyltetrazolium chloride (TTC) staining.In the WT group, without treatment, lesion volume was 120 ± 13 mm(3) in the mice subjected to 30 min' MCAO and 173 ± 23 mm(3) in the mice subjected to 90 min' MCAO. Respective values with HBO treatment were 66.5 ± 36.7 mm(3) and 53.2 ± 17.2 mm(3). The difference was significant only for 90-minute MCAO (p0.01, nonparametric test). In the TLR4(-/-) group (all untreated), lesion volume was 95.9 ± 17.9 after 90 min of MCAO, which was significantly lower than in the untreated WT animals (p0.05, nonparametric test).A single treatment of HBO immediately after MCAO followed by 24 h' reperfusion significantly reduces edema and may improve perfusion. TLR4 knockout protects mice from MCAO damage, but to a lesser extent than HBO treatment.

To describe a surgical procedure for loosening or tightening the superior oblique (SO) tendon that enables bedside adjustment following the surgery, with surgical outcome reported.A permanent suture separates the two cut ends of the SO tendon, with a sliding noose for adjustment. The noose is accessed by having the patient look up or straight ahead, not down, during adjustment at the bedside. Records of 17 patients who underwent this surgery between June 2000 and January 2003 were reviewed and analyzed for outcome.Seventeen patients, 18 eyes, mean age 43.7 years (range 5.9 to 71 years) had SO surgery with postoperatively adjustable sutures. Twelve eyes of 11 patients had a loosening procedure, and six eyes had a tightening procedure. Seven of the patients had had precious eye muscle surgery, four having had previous surgery on the same SO tendon. All but one patient returned for the follow-up examination, ranging from 1.5 to 7 months postoperatively. Torsional imbalances in 12 patients improved in all but two. In four patients with a preoperative A pattern, the A pattern improved from 16 PD to 0 PD on average. In eight patients with vertical misalignment in straight ahead gaze who had no other cyclovertical muscle surgery simultaneously, the mean reduction was 7 PD. All patients except one had improvement in preoperative symptoms.Both loosening and tightening procedures can be performed successfully using a postoperative adjustment technique for the SO tendon.

To investigate the role of inflammation in acute retinal artery occlusion (RAO).Levels of interleukin (IL)-6, IL-8, and tumor necrosis factor alpha (TNF-alpha) were measured in serum (n = 14) and aqueous humor (AqH) (n = 8) samples from patients with RAO. Findings were compared with 24 age- and disease-matched patients, 10 healthy subjects (serum), and 16 patients undergoing cataract surgery (AqH).Patients who arrived early (within 4-6 hours of occlusion) had higher serum IL-8 and IL-6 levels than controls; the IL-6 level in the AqH was lower than that of controls, while the IL-8 level was higher. In seven patients for whom both serum and AqH samples were available, serum IL-6 levels were higher than their corresponding AqH levels in most patients arriving within 10 hours of occlusion, and AqH IL-8 levels were higher than the corresponding serum levels in all but one. TNF-alpha levels were consistently higher in the serum than in the AqH at all time points.Serum IL-8 and IL-6 and AqH IL-8 are elevated immediately following acute RAO. The early local suppression of IL-6 may be related to ocular immune mechanisms.

Objective: To determine the long term visual and neurological outcome of patients diagnosed with cavernous sinus aneurysms (CCAs). Methods: Prospective follow up for at least five years or until death of 31 retrospectively recruited patients (27 women, 4 men) with treated and untreated CCAs. Results: There were 40 aneurysms in all. Mean age at diagnosis was 60.4 years (range 25 to 86; median 64). The most common symptoms were diplopia (61%), headache (53%), and facial or orbital pain (32%). Fifteen patients (48%) were diagnosed after they developed cranial nerve pareses, four (13%) after they developed carotid–cavernous sinus fistulas (CCFs), and 12 (39%) by neuroimaging studies done for unrelated symptoms. Twenty one patients (68%) had treatment to exclude the aneurysm from circulation, 10 shortly after diagnosis and 11 after worsening symptoms. Immediate complications of treatment occurred in six patients and included neurological impairment, acute ophthalmoparesis, and visual loss. Ten patients (32%) were observed without intervention. Over a mean (SD) follow up period of 11.8 (7.7) years, eight had improvement in symptoms, five remained stable, and eight deteriorated. Among the 10 patients followed without intervention, none improved spontaneously, three remained stable, and seven worsened. Conclusions: Most treated patients in this series improved or remained stable after treatment, but none improved without treatment. The long term prognosis for treated cases is relatively good, with most complications occurring immediately after the procedure. Endovascular surgery has decreased the morbidity and mortality of treatment so should be considered for any patient with a CCA.

Retrospective review to describe the features and visual outcome of traumatic optic neuropathy (TON) in children and adolescents.Interventional and observational case series of children18 years with TON.Forty children, 24 boys and 16 girls, were identified with 43 affected eyes. The mean age was 11.6 years (range, 2 to 18). The most common causes were motor vehicle accidents (62%) and sports injuries (22%). The trauma was blunt in 78% of cases and penetrating in 22%. Visual acuity at presentation for 27 patients whose visual acuity could be assessed ranged from no light perception to 20/80. Interventions included corticosteroids (n = 18), decompression of the optic canal (n = 3), and optic sheath nerve fenestration (n = 1). Visual outcome for 22 children with at least 1 month of follow-up was no light perception in 10 (45%),20/200 in 7(32%), 20/200 to20/80 in 1(4%), andor = 20/80 in 4 (18%). There was no difference in the rate or degree of improvement between treated and untreated patients.TON in children is caused by mechanisms similar to those that cause TON in adults. The severity of visual loss as well as the rate and degree of improvement are also similar. Our data do not indicate that treatment improves visual outcome.

This study aims to investigate the role of the inflammatory response following optic nerve crush (ONC) in knockout mice for the toll-like receptor-4 gene (TLR4-/-) compared to wild-type (WT) mice.ONC was induced in TLR4-/- and C57BL6 WT mice. Histological sections of the retina and optic nerve were analysed on days 1, 3 or 21 after injury. Molecular analysis with real-time quantitative polymerase chain reaction was used to study the expression of CD45, tumour necrosis-alpha (TNF-α) and glial fibrillary acidic protein, as well as retinal ganglion cell (RGC) markers THY-1 and Brn3b.There was a 25.5% and 38% loss in the RGC layer of the ONC-injured eyes of the TLR4-/- and the WT mice, respectively (with 27% and 9% of the remaining cells positive for Brn3a, respectively). Mean levels of Thy-1 and Brn3b were higher in the TLR4-/- mice. CD45 and Iba1 staining revealed infiltration of inflammatory cells into the injured nerve and retina in both groups. Molecular analysis of the optic nerve on day 1 showed increased TNF-α expression and reduced CD45 and GFAP expression; on day 3, CD45 reverted to baseline but GFAP remained low; on day 21, all 3 markers were at baseline in the TLR4-/- group and decreased in the WT group.Inflammation plays a major role in the response to ONC injury. Reduced levels of inflammation are associated with improved RGC preservation. The increase in TNF-α and reduction in CD45 in both TLR4-/- and WT mice may indicate the presence of an alternative pathway for induction of RGC death.

Parainfectious optic neuritis may appear at any age. The aim of our report was to compare the clinical manifestations and outcomes of this form of optic neuritis between children and adults.The study sample consisted of all patients diagnosed with parainfectious optic neuritis evaluated by 2 neuro-ophthalmology services between 2005 and 2012. Data were collected retrospectively from the medical files. Findings were compared between patients aged 0-18 years and 19 years or older.Ten children (50% female) and 8 adults (50% female) met the study criteria. Mean duration of follow-up was 29.4 months (range, 2-72 months) in the pediatric group and 14.2 months (range, 5-80 months) in the adult group. Respective rates of bilateral disease were 50% and 38%, and all patients had optic disc swelling. The associated pathogen was identified in 60% of the pediatric group, mainly Mycoplasma pneumoniae, and 75% of the adult group, in which no microorganism predominated. The interval from the febrile illness to symptom onset was 6 days (range, 1-14 days) in the pediatric group and 19.5 days (range, 14-30 days) in the adult group. Acute disseminated encephalomyelitis (ADEM) was diagnosed in 40% (4/10) of the children and none of the adults. Final visual outcome was 20/30 or better in all patients. There was a higher frequency of bilateral disease in prepubescent vs postpubescent children.Parainfectious optic neuritis is associated with a favorable visual prognosis regardless of age. Children tend to manifest visual symptoms sooner after the antecedent infectious illness and more often bilaterally and in conjunction with ADEM. The causative agent is isolated less frequently in children compared with adults.

We have recently reported that small-sized bone marrow cells (BMCs) isolated by counterflow centrifugal elutriation and depleted of lineage markers (Fr25lin-) have the capacity to differentiate and contribute to regeneration of injured islets. In this study, we assess some of the characteristics of these cells compared to elutriated hematopoietic progenitors (R/O) and whole BMCs in a murine model of streptozotocin-induced chemical diabetes. The GFPbrightCD45+ progeny of whole BMCs and R/O progenitors progressively infiltrate the pancreas with evolution of donor chimerism; are found at islet perimeter, vascular, and ductal walls; and have a modest impact on islet recovery from injury. In contrast, Fr25lin- cells incorporate in the islets, convert to GFPdimCD45-PDX-1+ phenotypes, produce proinsulin, and secrete insulin with significant contribution to stabilization of glucose homeostasis. The elutriated Fr25lin- cells express low levels of CD45 and are negative for SCA-1 and c-kit, as removal of cells expressing these markers did not impair conversion to produce insulin. BMCs mediate two synergistic mechanisms that contribute to islet recovery from injury: support of islet remodeling by hematopoietic cells and neogenesis of insulin-producing cells from stem cells.

Diabetic retinopathy (DR) results from vascular injury and leakage of blood. Genetic risk factors may play a role. Free hemoglobin is a potent oxidant protein. Haptoglobin protects from hemoglobin potential oxidative damage. Haptoglobin binds to hemoglobin during its transport and facilitates its removal from the extravascular compartment. Hp1-1, Hp2-1, and Hp2-2 differ markedly in their antioxidant ability. Patients who are Hp1-1 form dimeric haptoglobin molecules, whereas those who are Hp1-2 or Hp2-2 form multimers of two or more haptoglobin molecules. Haptoglobin multidimers are sterically hindered from binding to hemoglobin and have less access to the extravascular space. Hp2-2 genotype and diabetes mellitus may carry a higher risk of microvascular and macrovascular complications. Hp1-1 has a possible role in delaying the onset of retinopathy. Heterozygotes (Hp 2-1) are protected from both diabetes and DR. A few studies showed that haptoglobin genotype plays no role in the development or severity of proliferative retinopathy in type 2 diabetes. Type 2 diabetes may be independent of haptoglobin phenotype.

Background: Perfluorocarbon liquids are used as a vitreous substitute in the operative management of several ophthalmologic conditions. We examined the effects of residual perfluorocarbon droplets in the anterior chamber in patients after retinal detachment sur- gery. Methods: The study group consisted of seven patients (seven eyes) aged 28-74 years in whom droplets of perfluorocarbon appeared in the anterior chamber subsequent to retinal detachment surgery involving scleral buckling, perfluorodecalin injection, and per- fluorocarbon liquid-silicone oil exchange. Mean follow-up was 9.4 months. Results: From 1-15 droplets of perfluorocarbon liquid were found in the anterior cham- ber. There were no corneal complications or inflammatory reactions or blood vessel inva- sion into the corneal stroma. Some of the droplets appeared to be encapsulated in a membrane-like material. Perfluorocarbon liquid and silicone oil had to be removed in three patients at 20, 12, and 4 months postoperatively because of an increase in intraocular pressure due to blockage of the inferior iridectomy by a droplet of perfluorocarbon, emulsi- fication of the silicone oil, or residual perfluorocarbon liquid on the retina (one patient each). Increased intraocular pressure in a fourth patient was successfully treated pharma- cologically. Conclusion: Based on our experience, residual perfluorocarbon liquid droplets in the anterior chamber are well tolerated and do not induce corneal damage or ocular inflamma- tion. These patients should be closely followed, however, and the liquid removed if compli- cations develop.

PURPOSE Brain-derived neurotrophic factor (BDNF) and ciliary neurotrophic factor (CNTF) have limited and transient supportive effects on retinal recovery from ischemia. The aim of this study was to investigate their effect on engrafted adult bone marrow-derived stem cells in a rodent model of anterior ischemic optic neuropathy (rAION). METHODS Small cells were isolated from the bone marrow of green fluorescent protein expressing mice by counterflow centrifugal elutriation, depleted of cells expressing lineage markers, and grafted in conjunction with growth factors into the vitreous body of mice with unilateral rAION. Progenitors were mobilized with granulocyte macrophage colony-stimulating factor (GM-CSF) or stem cell factor (SCF). The contralateral eye served as a control. RESULTS At 4 weeks, the quantitative incorporation of donor cells in the injured retina was increased by BDNF (P < 0.01 versus control) and decreased by CNTF (P < 0.01 versus control), with no notable difference at 24 weeks. Both growth factors improved the short-term and long-term qualitative engraftment of cells adopting neural phenotypes in the retinal ganglion cell (RGC) layer and astrocyte phenotypes in the anterior vasculature. The RGC-engrafted cells formed extensions toward the inner nuclear layer. In the presence of growth factors, donor cells migrated to the optic nerve and contributed to repair by gliosis. Mobilization with GM-CSF restricted cell fate to microglia, whereas SCF was associated with limited neuroglial differentiation. CONCLUSIONS Both BDNF and CNTF enhance engraftment and neuroglial differentiation of adult bone marrow stem cells in injured retina, with BDNF having an early quantitative and qualitative advantage over CNTF. Mobilization with differentiation factors restricts cell fate in the injured retina.

PURPOSE To evaluate the effect of bevacizumab, a VEGF inhibitor, on optic nerve edema and retinal ganglion cell (RGC) loss in a mouse model of optic nerve crush (ONC). METHODS Two hundred C57BL/6 wild-type mice were anesthetized. Right ONC was induced in 150 mice, of which half (n = 75) received an intravitreal injection of bevacizumab immediately thereafter and half (n = 75) did not. The remaining 50 received only bevacizumab. The left eyes served as a control. Findings were analyzed by fluorescein angiography (days 0, 1, 3), histologic and immunohistochemical tests (days 1, 3, 4, 21), and quantitative real-time PCR. RESULTS Angiography revealed a reduction in postinjury disc leakage following bevacizumab injection (days 1, 3), confirmed with IgG staining. On PCR, expression of HO-1 and SOD-1 mRNA increased following ONC and further increased with bevacizumab. VEGF gene expression decreased following bevacizumab injection without ONC, remained at baseline after ONC, and increased slightly after ONC+bevacizumab. Histologically, there was a 38% RGC loss 21 days after ONC alone, which dropped to 14% with bevacizumab treatment; it was close to 15% with bevacizumab alone. Mean (SEM) microvascular perfusion in the optic nerve 4 days after ONC was significantly higher in the bevacizumab-treated (85% ± 10%) than the vehicle-treated (33% ± 13%) animals. CONCLUSIONS Bevacizumab treatment following ONC induction exerts a protective effect, manifested by reduced optic nerve head edema. The underlying mechanism probably involves a lesser interruption of axonal transport. Reduced expression of antioxidative and ischemic genes may contribute to RGC preservation.

Background/aim: Alterations of the immune system may have a role in thrombogenesis. Artery sites occluded with thrombi apparently release pro-inflammatory cytokines. Non-arteritic anterior ischaemic optic neuropathy (NAION) results from occlusion of the blood supply to the optic nerve. The aim of this study was to analyse levels of pro-inflammatory cytokines in patients with acute event of NAION. Methods: Study participants included 10 patients (12 eyes) with NAION and 20 age matched controls with the same risk factors for atherosclerosis disease. Peripheral blood samples were obtained immediately at the acute onset of NAION. Plasma interleukin 8 (IL-8), IL-6, and tumour necrosis factor alpha (TNF-α) levels were measured immediately following diagnosis and during the follow up intervals. Results: The plasma levels of IL-8 were significantly higher in NAION patients at the time of diagnosis in comparison to the control group (p = 0.002), and decreased during the follow up period (6–12 months) (p = 0.05). There were no differences in plasma levels of IL-6 and TNF-α between NAION patients and controls, either in the acute phase or during the follow up period. Conclusion: Plasma levels of IL-8 are elevated during the acute phase of NAION, but not IL-6 and TNF-α. These elevated levels are in accordance with other acute vascular thrombosis. The clinical significance of these findings should be further evaluated.

Purpose To describe the neuroimaging findings in a patient with an isolated abducens nerve palsy caused by a dolichoectatic basilar artery. Design Retrospective case report. Methods A 65-year-old man presented with a 3-year history of gradually worsening horizontal double vision. He subsequently underwent magnetic resonance imaging, magnetic resonance angiography, and computed tomographic angiography. Results Neuroimaging revealed dolichoectasia of the basilar artery. The enlarged, tortuous vessel appeared to be compressing the subarachnoid portion of the nerve at its exit from the brainstem. Conclusion Noninvasive neuroimaging studies are sufficient to establish a diagnosis of basilar artery dolichoectasia in patients with isolated cranial neuropathies.

Evaluation of potential tropic effects of vascular endothelial growth factor (VEGF) on the incorporation and differentiation of bone-marrow-derived stem cells (BMSCs) in a murine model of anterior ischemic optic neuropathy (AION).In the first approach, small-sized subset of BMCs were isolated from GFP donors mice by counterflow centrifugal elutriation and depleted of hematopoietic lineages (Fr25lin(-)). These cells were injected into a peripheral vein (1 × 10(6) in 0.2 ml) or inoculated intravitreally (2 × 10(5)) to syngeneic mice, with or without intravitreal injection of 5 μg/2μL VEGF, simultaneously with AION induction. In a second approach, hematopoietic cells were substituted by myelablative transplant of syngeseic GFP + bone marrow cells. After 3 months, progenitors were mobilized with granulocyte-macrophage colony-stimulating factor (GM-CSF) followed by VEGF inoculation into the vitreous body and AION induction . Engraftment and phenotype were examined by immunohistochemistry and FISH at 4 and 24 weeks post-transplantation, and VEGF receptors were determined by real time PCR.VEGF had no quantitative effect on incorporation of elutriated cells in the injured retina, yet it induced early expression of neuroal markers in cells incorporated in the RGC layer and promoted durable gliosis, most prominent perivascular astrocytes. These effects were mediated by VEGF-R1/Flt-1, which is constitutively expresses in the elutriated fraction of stem cells. Mobilization with GM-CSF limited the differentiation of bone marrow progenitors to microglia, which was also fostered by VEGF.VEGF signaling mediated by Flt-1 induces early neural and sustained astrocytic differentiation of stem cells elutriated from adult bone-marrow, with significant contribution to stabilization retinal architecture following ischemic injury.

Objective The genetic factors participating in oral melanoma evolution have not been studied extensively. We aimed to analyze the prevalence of BRAF and GNAQ mutations in a series of oral melanocytic tumors, nevi, and melanomas. Study Design The study group consisted of 4 melanomas and 10 nevi (6 intramucosal, 4 blue nevi). DNA was extracted from paraffin-embedded tissue sections, and mutations in GNAQ and BRAF were analyzed with the use of mass spectrometery. Results V600E point mutation was identified in the BRAF gene in 3 intramucosal nevi and in 2 melanomas. Only 1 blue nevus harbored the GNAQ209 mutation. None of the BRAF-positive samples harbored GNAQ mutations. Conclusions The finding of BRAF mutations in oral benign and malignant melanocytic lesions points to a potential initiating role of BRAF in malignant transformation, which may have important therapeutic implications as those with BRAF mutations may benefit from specific treatment using RAF inhibitors.

Brain ischemia has major consequences leading to the apoptosis of astrocytes and neurons. Glucose-regulated protein 78 (GRP78) known for its role in endoplasmic reticulum stress alleviation was discovered on several cell surfaces acting as a receptor for signaling pathways. We have previously described peptides that bind cell surface GRP78 on endothelial cells to induce angiogenesis. We have also reported that ADoPep1 binds cardiomyocytes to prevent apoptosis of ischemic heart cells. In this study we describe the effect of hypoxia on astrocytes and neurons cell surface GRP78. Under hypoxic conditions, there was an increase of more than fivefold in GRP78 on cell surface of neurons while astrocytes were not affected. The addition of the GRP78 binding peptide, ADoPep1, to neurons decreased the percentage of GRP78 positive cells and did not change the percent of astrocytes. However, a significant increase in early and late apoptosis of both astrocytes and neurons under hypoxia was attenuated in the presence of ADoPep1. Intravitreal administration of ADoPep1 to mice in a model of optic nerve crush significantly reduced retinal cell loss after 21 days compared to the crush-damaged eyes without treatment or by control saline vehicle injection. Histological staining demonstrated reduced GRP78 after ADoPep1 treatment. The mechanism of peptide neuroprotection was demonstrated by the inhibition of hypoxia induced caspase 3/7 activity, cytochrome c release and p38 phosphorylation. This study is the first report on hypoxic neuronal and astrocyte cell surface GRP78 and suggests a potential therapeutic target for neuroprotection.

To evaluate the optical and anatomical effects of oral propranolol treatment for infantile periocular capillary haemangioma. All children diagnosed with infantile capillary haemangioma in 2008–2010 at a tertiary paediatric medical centre underwent comprehensive evaluation, including imaging, by a multidisciplinary team followed by oral propranolol treatment. Clinical follow-up was performed regularly until the lesions disappeared. Main outcome measures included changes in anatomical extraocular extension, refractive sphere and cylindrical power, and spherical equivalent in the involved eye before and after treatment and between the two eyes. A total of 30 patients (8 male; mean age at diagnosis, 1.6±2.8 months) participated. The lesions affected the left eye in 53.3% and were located preseptally in 83.3%. Four patients (13.3%) received steroids before propranolol. A treatment dosage of 2 mg/kg per day was started at mean age 5.0±4.5 months, 3.3±4.3 months from disease onset. Side effects occurred in 11 patients and warranted a dose reduction (to 1 mg/kg per day) in 3 and treatment termination in 1. Findings were significant for mean reduction in involved extraocular area (P

Purpose To investigate the effect of hyperbaric oxygen (HBO) chamber treatment in mouse models of retinal ischemia. Methods Unilateral central retinal artery occlusion (CRAO) or optic nerve crush (ONC) was induced in 50 mice each, of which 30 were treated with 100% oxygen at 2 atm for 90 minutes immediately after injury and then daily for up to 14 days. Mice were euthanatized on days 1, 3, and 21 for histologic analysis, apoptosis assay, and quantitative real-time polymerase chain reaction test. Findings were analyzed by injury and by treatment. Results HBO treatment reduced cell loss from 58% to 30% in the CRAO model and from 52% to 32% in the ONC model. In both models, it was associated with significantly increased cell survival in the retinal ganglion cell layer. Expression levels of the proapoptosis genes (bax, caspase-3) decreased minimally in the HBO-treated CRAO mice on day 1, but this trend was reversed on day 3. In the ONC group, levels of caspase-3, bax, and bcl-x increased on day 1 and dropped below baseline on day 3. The pattern of changes in the expression levels of the ischemia- and oxidative-stress-related genes (HO-1, SOD-1, GPX-1, NOX-2) and the effectiveness of HBO treatment varied by model. Overall, however, gene expression levels that increased in the untreated mice increased further with HBO treatment and levels that decreased, decreased further with treatment. Conclusions HBO treatment protects injured neuronal cells from apoptosis. Response to treatment differs molecularly after ONC or CRAO. These results should prompt clinical trials of acute ischemic retinal damage.

Mutations in the LMNA gene encoding lamins A/C are responsible for Hutchinson-Gilford syndrome (HGS), a disorder of premature aging. Cataract is 1 of the main manifestations. The most prevalent mutation in Hutchinson-Gilford syndrome is C1824T, which activates a cryptic splice donor site to produce an abnormal lamin A protein. The purpose of this study was to investigate a possible association of the C1824T mutation with age-related cataract. Anterior lens capsule material was collected during cataract extraction surgery from 178 patients with senile cataract during 2007-2008. DNA and mRNA were extracted and sequenced for the LMNA gene. DNA and cDNA were screened for the C1824T mutation, which was not detected. Messenger RNA (mRNA) expression was normal, with no truncation. We found that human age-related nuclear cataract is not associated with LMNA gene mutations or truncation of lamin A.

Purpose There are 2 common alleles for Hp (Hp-1 and Hp-2) and 3 common Hp genotypes: Hp1-1, Hp2-1, and Hp2-2. The haptoglobin genotype may play a dual role in morbidities of diabetes: Hp1-1, protective and Hp2-2, provocative. This study investigated the possible association of haptoglobin genotypes with onset of retinopathy in Type 2 diabetes (DM2). Methods The sample included 98 consecutive adults with DM2 under routine outpatient follow-up from 2007 to 2009 who met the criteria for either no retinopathy at ≥10 years after diagnosis (Group 1) or proliferative retinopathy at ≤10 years after diagnosis (Group 2). Blood samples were collected for haptoglobin genotyping by polymerase chain reaction. Findings were compared between and within groups. Results Eighty-four patients had no retinopathy and 14 had early proliferative retinopathy. The distributions of the Hp genotypes were as follows: no-retinopathy group: 28.6% Hp1-1, 35.7% Hp2-1, and 35.7% Hp2-2 and proliferative retinopathy group: 22.6% Hp1-1, 27.4% Hp2-1, and 50% Hp2-2 (NS). On statistical analysis (limited to the larger no-retinopathy group), a predominance of Hp1-1 was noted in older patients; Hp2-2 was associated with an increased rate of stroke. Conclusion The Hp genotype apparently plays no role in the development or worsening of proliferative retinopathy in DM2. Hp1-1 may be involved in delaying the onset of diabetes. Hp2-2 may pose a microvascular risk.

An ongoing debate surrounds the existence of stem cells in the adult endowed with capacity to differentiate into multiple lineages. We examined the possibility that adult bone marrow cells participate in recovery from chemical diabetes through neogenesis of insulin-producing cells. Small-sized cells negative for lineage markers derived by counterflow centrifugal elutriation from the bone marrow were transplanted into mice made diabetic with streptozotocin and sublethal irradiation. These cells homed efficiently to the injured islets and contributed to tissue revascularization. Islet-homed CD45-negative donor cells identified by sex chromosomes downregulated GFP, expressed PDX-1 and proinsulin, and converted the hormone precursor to insulin. An estimated 7.6% contribution of newly formed insulin-producing cells to islet cellularity increased serum insulin and stabilized glycemic control starting at 5 weeks post-transplant and persisting for 20 weeks. Newly differentiated cells displayed normal diploid genotype and there was no evidence of fusion between the grafted stem cells or their myeloid progeny and injured β-cells. Considering the extensive functional incorporation of insulin-producing donor cells in the injured islets, we conclude that the adult bone marrow contains a subset of small cells endowed with plastic developmental capacity.

The contribution of stem cells derived from adult tissues to the recovery of pancreatic islets from chemical injury is controversial. Analysis of nonhematopoietic differentiation of bone marrow-derived cells has yielded positive and negative results under different experimental conditions. Using the smallest subset of bone marrow cells lacking immuno-hematopoietic lineage markers, we have detected incorporation and conversion into insulin-producing cells. Donor cells identified by genomic markers silence green fluorescent protein (GFP) expression as a feature of differentiation, in parallel to expressing PDX-1 and proinsulin. Here we elaborate potential experimental difficulties that might result in false-negative results. The use of GFP as a reporter protein is suboptimal for differentiation experiments: (a) the bone marrow of GFP donors partially expresses the reporter protein, (b) differentiating bone marrow cells silence GFP expression, and (c) the endocrine pancreas is constitutively negative for GFP. In addition, design of the experiments, data analysis, and interpretation encounter numerous objective and subjective difficulties. Rigorous evaluation under optimized experimental conditions confirms the capacity of adult bone marrow-derived stem cells to adopt endocrine developmental traits, and demonstrates that GFP downregulation and silencing is a feature of differentiation.

To determine the precise age of suppression of the oculocephalic reflex in infants and its relationship to specific clinical characteristics.The oculocephalic reflex was prospectively tested in 325 healthy full-term babies aged 1 to 32 weeks attending an orthopedic outpatient clinic. Two ophthalmologists raised the baby's head 30 degrees above horizontal and rapidly rotated it in the horizontal and vertical planes while watching the conjugate eye movement. Suppression of the reflex, by observer agreement, was analyzed in relation to gestational age, postpartum age, postconceptional age, birth weight, and current weight. The data were fitted to a logistic regression model to determine the probability of suppression of the reflex according to the clinical variables.The oculocephalic reflex was suppressed in 75% of babies by the age of 11.5 weeks and in more than 95% of babies aged 20 weeks. Although postpartum age had a greater influence than gestational age, both were significantly correlated with suppression of the reflex (p = 0.01 and p = 0.04, respectively; two-sided t-test). Postpartum age was the best single variable explaining absence of the reflex. On logistic regression with cross-validation, the model including postpartum age and current weight yielded the best results; both these factors were highly correlated with suppression of the reflex (r = 0.74).The oculocephalic reflex is suppressed in the vast majority of normal infants by age 11.5 weeks. The disappearance of the reflex occurs gradually and longitudinally and is part of the normal maturation of the visual system.

Purpose This study sought to characterize the expression of angiogenesis-related genes in a mouse model of corneal neovascularization, either untreated or after treatment with a single injection of bevacizumab by three different routes. In addition, the effectiveness of the treatment was compared to a rabbit model. Methods A chemical burn was induced in the mid-cornea of the right eye in 119 mice; 56 of them were untreated and 63 were bevacizumab-treated. Neovascularization was evaluated 2, 4, 8, 10, and 14 days later using digital photos, angiography and India ink perfusion. The relative area of new blood vessels was analyzed using slit-lamp examination in vivo and on histological and flat-mount sections. The levels of gene expression involved in the angiogenic process vascular endothelial growth factor [VEGF], insulin-like growth factor-1 [IGF-1], pigment epithelium derived factor [PEDF], and macrophage-inflammatory protein-2 [MIP-2]) were measured by a real-time polymerase chain reaction. Six rabbits underwent the same injury and treatment, and the response was compared to the mouse model. Results Neovascularization was first observed two days after injury. The affected section increased from 11.24% (±7.0) of the corneal area to 47.42% (±25.4) on day 8 and 50.62% (±24.7) on day 10. In the mice treated with bevacizumab, the relative area of neovascularization was significantly lower at the peak time points (p

PURPOSE. Many malignancies are known to be associated with abnormal activation of the PI3K-AKT pathway. Recently, a somatic mutation in the AKT1 gene (E17K) was identified in a small proportion of human tumors. This mutation activated AKT1 by means of abnormal membrane recruitment and stimulated downstream signaling. This study was designed to analyze AKT1 mutations in retinoblastoma and gain insights into the role PI3K-AKT pathway plays in the development of this tumor. METHODS. Twenty-four samples of retinoblastoma from children were analyzed for mutations in the AKT1, PTEN and K-RAS genes, using a chip-based matrix-assisted laser desorption-time-of-flight (NLkLDI-TOF) mass spectrometer. Mutations in the PIK3CA gene were analyzed in 16 retinoblastoma samples using direct sequencing. RESULTS. These results show that the mutation E17KlAKT1 was not detected in the 24 samples of retinoblastoma analyzed. K-RAS mutations were identified in two samples. There were no mutations in any of the other genes analyzed by a mass array system. On direct sequencing of 16 samples for the PIK3CA gene, one sample showed gain of function mutation in exon 9. In another sample, a genetic polymorphism of unknown significance (rs17849079) was detected in exon 20. CONCLUSIONS. Although the PI3K-AKT pathway is known to be dysregulated in retinoblastoma, the low frequency of oncogenic mutations in the AKT1, PIK3CA, and PTEN genes, suggests a different activating mechanism.

To evaluate the effect of combined intralesional and sub-Tenon's administration of corticosteroids for the treatment of refractory periocular and orbital capillary haemangioma in a retrospective, non-comparative study.Seven infants with resistant periorbital and orbital capillary haemangioma who attended our tertiary centre from 2000 to 2005 were treated with an intralesional injection of a mixture of betamethasone 6 mg/cc and triamcinolone 10 mg/cc, by body weight, together with sub-Tenon's infusion of betamethasone 6 mg/cc and triamcinolone 40 mg/cc of a volume of 1 cc, close to the orbital lesion, in the same session. Visual axis, proptosis, pre- and post-treatment changes in four refractive parameters, and parental satisfaction were evaluated.None of the babies had proptosis or visual axis obstruction after treatment. Mean spherical power decreased by 22.7% (P=0.06). Mean spherical equivalent decreased in 34%, but the difference was not statistically significant (P=0.09). No early or late ocular side effects were observed. Mean intraocular pressure results were normal before and immediately after treatment, and during the follow-up period. Parental satisfaction score during follow-up was 9/10. Mean time to improvement was 96.43+/-58.3 days.Combined local corticosteroid administration by posterior sub-Tenon's infusion and intralesional injection in babies with extensive capillary haemangioma is associated with a satisfactory anatomical and functional outcome and there were no ophthalmic side effects.

To evaluate the efficacy of topical lomefloxacin 0.3% versus chloramphenicol 0.2% with polymyxin B 2500 U/ml in the treatment of patients after strabismus surgery.Seventy-nine consecutive patients who underwent strabismus surgery from April through October 2006 were randomized to receive topical lomefloxacin 0.3% (n = 45, 58 eyes) or chloramphenicol 0.2% with polymyxin B 2500 U/ml (n = 34, 40 eyes) postoperatively, in addition to topical dexamethasone 0.1% and diclophenac 0.1%. The groups were compared for clinical signs of local cellulitis and findings on conjunctival cultures.From postoperative day 1 to day 28, rates of chemosis decreased from 96% to 9.5% in the lomefloxacin group and from 94% to zero in the chloramphenicol group; rates of hyperemia decreased from 100% to zero in both groups. Accordingly, rates of a cumulative clinical sign score (CCSS) decreased from 60% to zero in the lomefloxacin group and 50% to zero in the chloramphenicol group. Rates of positive bacterial cultures dropped from 59.6% preoperatively to 48.3% on day 14 in the lomefloxacin group and from 63% to 47% in the chloramphenicol group.Topical lomefloxacin is a potent alternative to topical chloramphenicol eye drops after strabismus surgery for the control of pain, infection and discharge, chemosis, hyperemia, and conjunctival hemorrhage.