Delayed Outgrowth in Response to the BDNF and Altered Synaptic Proteins in Neurons From SHR Rats.

Attention deficit hyperactivity disorder (ADHD) is a neurodevelopmental disorder characterized by inattention, hyperactivity, and impulsivity symptoms. Neuroimaging studies have revealed a delayed cortical and subcortical development pattern in children diagnosed with ADHD. This study followed up on the development in vitro of frontal cortical neurons from Spontaneously hypertensive rats (SHR), an ADHD rat model, and Wistar-Kyoto rats (WKY), control strain, over their time in culture, and in response to BDNF treatment at two different days in vitro (DIV). These neurons were also evaluated for synaptic proteins, brain-derived neurotrophic factor (BDNF), and related protein levels. Frontal cortical neurons from the ADHD rat model exhibited shorter dendrites and less dendritic branching over their time in culture. While pro- and mature BDNF levels were not altered, the cAMP-response element-binding (CREB) decreased at 1 DIV and SNAP-25 decreased at 5 DIV. Different from control cultures, exogenous BDNF promoted less dendritic branching in neurons from the ADHD model. Our data revealed that neurons from the ADHD model showed decreased levels of an important transcription factor at the beginning of their development, and their delayed outgrowth and maturation had consequences in the levels of SNAP-25 and may be associated with less response to BDNF. These findings provide an alternative tool for studies on synaptic dysfunctions in ADHD. They may also offer a valuable tool for investigating drug effects and new treatment opportunities.

Sex differences in maternal odor preferences and brain levels of GAP-43 and sonic hedgehog proteins in infant SHR and Wistar Kyoto rats.

Attention Deficit Hyperactivity Disorder (ADHD) is a neurodevelopmental disorder that presents sex differences in the severity and presentation of symptoms, whose neurobiological basis is still unknown. Both Growth-associated Protein 43 (GAP-43) and Sonic hedgehog (Shh) are considered essential proteins for the appropriate brain development, but their participation in ADHD neurobiology have not been investigated yet. In this study, we hypothesized that alterations in these proteins could be related to behavioral traits to ADHD phenotype. Thus, both sexes of infant Spontaneously hypertensive rats (SHR, used as ADHD animal model) were evaluated for developmental milestones, locomotor activity, olfactory and recognition memory. Both GAP-43 and Shh were assessed in the olfactory bulb, frontal cortex and hippocampus in early and late infancy. During early infancy, SHR reached three developmental milestones later, and females showed olfactory memory impairment accompanied by increased levels of Shh in the olfactory bulb. In later infancy, hyperlocomotion, impaired recognition memory, and decreased Shh in the hippocampus were observed in SHR from both sexes. While in early infancy GAP-43 was not altered, it was decreased in the frontal cortex and hippocampus of female SHR in late infancy. Therefore, both Shh and GAP-43 are involved in the sex-dependent behavioral alterations showed by infant SHR. Despite the disorder's complexity and heterogeneity, our findings reveal important developmental parameters during SHR development and also emphasizes the relevance of studying sex differences in the ADHD context.

Caffeine and adenosine A2A receptors rescue neuronal development in vitro of frontal cortical neurons in a rat model of attention deficit and hyperactivity disorder.

Although some studies have supported the effects of caffeine for treatment of Attention deficit and hyperactivity disorder (ADHD), there were no evidences about its effects at the neuronal level. In this study, we sought to find morphological alterations during in vitro development of frontal cortical neurons from Spontaneoulsy hypertensive rats (SHR, an ADHD rat model) and Wistar-Kyoto rats (WKY, control strain). Further, we investigated the effects of caffeine and adenosine A1 and A2A receptors (A1R and A2AR) signaling. Cultured cortical neurons from WKY and SHR were analyzed by immunostaining of microtubule-associated protein 2 (MAP-2) and tau protein after treatment with either caffeine, or A1R and A2AR agonists or antagonists. Besides, the involvement of PI3K and not PKA signaling was also assessed. Neurons from ADHD model displayed less neurite branching, shorter maximal neurite length and decreased axonal outgrowth. While caffeine recovered neurite branching and elongation from ADHD neurons via both PKA and PI3K signaling, A2AR agonist (CGS 21680) promoted more neurite branching via PKA signaling. The selective A2AR antagonist (SCH 58261) was efficient in recovering axonal outgrowth from ADHD neurons through PI3K and not PKA signaling. For the first time, frontal cortical neurons were isolated from ADHD model and they presented disturbances in the differentiation and outgrowth. By showing that caffeine and A2AR may act at neuronal level rescuing ADHD neurons outgrowth, our findings strengthen the potential of caffeine and A2AR receptors as an adjuvant for ADHD treatment.

Aged mice receiving caffeine since adulthood show distinct patterns of anxiety-related behavior.

Caffeine is the psychostimulant most consumed worldwide. Anxiogenic effects of caffeine have been described in adult animals with controversial findings about its anxiogenic potential. Besides, the effects of caffeine on anxiety with aging are still poorly known. In this study, adult mice (6months old) started to receive caffeine (0.3 and 1.0mg/mL, drinking water) during 12-14months only in the light cycle and at weekdays. The open field (OF) and elevated plus maze (EPM) testing were used to determine the effects of caffeine on anxiety-related behavior in adult and aged mice (18-20months old). Because aging alters synaptic proteins, we also evaluated SNAP-25 (as a nerve terminals marker), GFAP (as an astrocyte marker) and adenosine A1 and A2A receptors levels in the cortex. According to the OF analysis, caffeine did not change both hypolocomotion and anxiety with aging. However, aged mice showed less anxiety behavior in the EPM, but after receiving caffeine (0.3mg/mL) during adulthood they were anxious as adult mice. While SNAP-25 and adenosine A2A receptors increased with aging, both GFAP and adenosine A1 receptors were not affected. Caffeine at moderate dose prevented the age-related increase of the SNAP-25, with no effect on adenosine A2A receptors. The absence of effect for the highest dose suggests that tolerance to caffeine may have developed over time. Aged mice showed high responsiveness to the OF, being difficult to achieve any effect of caffeine. On the other hand this substance sustained the adult anxious behavior over time in a less stressful paradigm, and this effect was coincident with changes in the SNAP-25, suggesting the involvement of this synaptic protein in the ability of caffeine to preserve changes related to emotionality with aging.

Prenatal caffeine intake differently affects synaptic proteins during fetal brain development.

Caffeine is the psychostimulant most consumed worldwide. However, little is known about its effects during fetal brain development. In this study, adult female Wistar rats received caffeine in drinking water (0.1, 0.3 and 1.0 g/L) during the active cycle in weekdays, two weeks before mating and throughout pregnancy. Cerebral cortex and hippocampus from embryonic stages 18 or 20 (E18 or E20, respectively) were collected for immunodetection of the following synaptic proteins: brain-derived neurotrophic factor (BDNF), TrkB receptor, Sonic Hedgehog (Shh), Growth Associated Protein 43 (GAP-43) and Synaptosomal-associated Protein 25 (SNAP-25). Besides, the estimation of NeuN-stained nuclei (mature neurons) and non-neuronal nuclei was verified in both brain regions and embryonic periods. Caffeine (1.0 g/L) decreased the body weight of embryos at E20. Cortical BDNF at E18 was decreased by caffeine (1.0 g/L), while it increased at E20, with no major effects on TrkB receptors. In the hippocampus, caffeine decreased TrkB receptor only at E18, with no effects on BDNF. Moderate and high doses of caffeine promoted an increase in Shh in both brain regions at E18, and in the hippocampus at E20. Caffeine (0.3g/L) decreased GAP-43 only in the hippocampus at E18. The NeuN-stained nuclei increased in the cortex at E20 by lower dose and in the hippocampus at E18 by moderate dose. Our data revealed that caffeine transitorily affect synaptic proteins during fetal brain development. The increased number of NeuN-stained nuclei by prenatal caffeine suggests a possible acceleration of the telencephalon maturation. Although some modifications in the synaptic proteins were transient, our data suggest that caffeine even in lower doses may alter the fetal brain development.

New technique to demonstrate corneal magnification using trypan blue in cataract surgery / Nova técnica para demonstrar a magnificação corneana utilizando azul de tripan na cirurgia de catarata

PURPOSE: To demonstrate the corneal magnification using trypan blue in cataract surgery. METHODS: Eight eyes of eight patients undergoing phacoemulsification with an intraocular lens implantation were enrolled in this study. After staining the anterior capsule with Trypan Blue 0.1 percent and performing the capsulorhexis, the excised anterior capsule was placed on the corneal surface. By observing and measuring the relationship between the border of the excised anterior capsule and the intracameral capsulorhexis opening, the effect of corneal magnification was clearly demonstrated and calculated by linear method. RESULTS: The average magnification of the cornea was 20.88 percent using linear method with an average area magnification of 47.53 percent. CONCLUSION: The capsulorhexis stained by trypan blue is useful to demonstrate the magnification provided by the cornea helping to design an intended opening size.

OBJETIVO: Demonstrar a magnificação corneana utilizando o azul de tripan na cirurgia de catarata. MÉTODOS: Oito olhos de oito pacientes submetidos à facoemulsificação com implante de lente intraocular foram envolvidos neste estudo. Após corar a cápsula anterior do cristalino com azul de tripan 0,1 por cento e realizar a capsulotomia circular contínua, a cápsula excisada foi colocada sobre a superfície corneana. Após observar e aferir a relação entre a borda da cápsula anterior excisada e a abertura da cápsula intracameral, o efeito da magnificação corneana foi claramente demonstrada e calculada através do método linear. RESULTADOS: A média estimada da magnificação corneana foi de 20,88 por cento com uma média de magnificação de área de 47,53 por cento. CONCLUSÃO: A capsulorrexis corada pelo azul de tripan é útil na demonstração da magnificação corneana, auxiliando assim na correta confecção do seu tamanho.

Longitudinal study of intraocular lens exchange.

PURPOSE: To analyze the indications for intraocular lens (IOL) exchange, interval between the first IOL implantation and the exchange, type and mix of IOLs used, effect on vision, and frequency of complications. SETTING: Cincinnati Eye Institute-Cincinnati-Ohio-USA. METHODS: This retrospective study comprised 49 eyes of 49 adult patients who had IOL exchange between 1986 and 2002 performed by the same surgeon. The mean age was 70 years old, and 55% were women. The mean interval between surgeries was 53.8 months and the mean follow-up, 35.6 months. The patients were divided into 2 groups according to the type of IOL originally implanted: anterior chamber (AC) or posterior chamber (PC). RESULTS: There were 15 eyes with an AC IOL and 34 eyes with a PC IOL. The difference in mean age and follow-up were not statistically significant between groups. The mean interval between the primary surgery and IOL explantation was 82.3 months in the AC IOL group and 37.9 months in the PC IOL group. The main reason for IOL exchange was inflammation (53.34%) and dislocation/decentration (85.30%), respectively. The preoperative best corrected visual acuity was similar in both groups, and visual acuity was maintained or improved in 80%. Vitreous prolapse was the main intraoperative complication. CONCLUSIONS: The primary indication for IOL exchange was intraocular inflammation in patients with an AC IOL and IOL malposition in patients with a PC IOL. The results confirm the safety and positive visual outcome in this complex group of patients.

Fate of anterior capsule tears during cataract surgery.

PURPOSE: To determine the incidence of anterior capsule tears, at what stage of surgery they occurred, and their intraoperative behavior. SETTING: Ambulatory surgery center, Cincinnati Eye Institute, Cincinnati, Ohio, USA. METHODS: This 5-year retrospective study was of patients having phacoemulsification with posterior chamber intraocular lens (IOL) implantation complicated by unplanned peripheral extension of the capsulorhexis tear or a radial anterior capsule tear. The operative notes and a videotape of the surgery were reviewed. The stage at which the tear was initially observed and when it extended were identified, as was whether the tear extended to the posterior capsule. Anterior vitrectomy and the design and location of the IOL implanted were also analyzed. RESULTS: A discontinuous anterior capsulorhexis or a break in the anterior capsule rim was observed in 21 eyes of 2646 cases, for an overall incidence of 0.79%. Anterior capsule tears were identified during ophthalmic viscosurgical device injection in 1 eye, capsulorhexis in 13 eyes, hydrodissection in 2 eyes, phacoemulsification in 3 eyes, irrigation/aspiration (I/A) in 1 eye, and implantation of a prosthetic iris device in 1 eye. Seven of the 13 tears identified during the capsulorhexis were managed by redirecting the second edge of the "safety" capsulorhexis to incorporate the tear. In 14 eyes, the tear in the anterior capsule extended into the zonules; 4 of these tears were limited. Ten tears extended around the equator and through the posterior capsule, occurring during the hydrodissection in 1 eye, phacoemulsification in 2 eyes, I/A in 1 eye, and IOL implantation in 6 eyes. An anterior vitrectomy was required in 4 eyes that had posterior capsule involvement. Endocapsular fixation of a 1-piece acrylic IOL was achieved in 18 eyes. Three eyes required implantation of a 3-piece acrylic IOL in the ciliary sulcus. CONCLUSIONS: Extension of an anterior capsule tear can complicate cataract surgery at any stage. Extension of the tear through the posterior capsule occurred in almost half the eyes with an anterior capsule tear, often requiring an anterior vitrectomy. Managing an anterior capsule tear can be challenging yet compatible with implantation of a posterior chamber IOL.

[Unintentional staining of lens posterior capsule with trypan blue during extracapsular cataract extraction: case report]. / Impregnação inadvertida da cápsula posterior do cristalino com azul de tripano durante a facectomia extracapsular: relato de caso.

Report of a complication related to the use of 0.1% trypan blue during capsular staining of the anterior crystalline lens capsule in an extracapsular cataract extraction with intraocular lens implantation. The capsular dye was injected using an air-bubble technique, inadvertently, it was done using high pressure dispersing the dye through the zonules leading to a complete lost of red reflex, the following steps of the procedure becoming more difficult after the cataract extraction and causing a temporary disturbance of visual acuity in the postoperative period. In order to avoid this complication, the authors also describe the use of "Modified air-bubble technique" where dispersive viscoelastic is placed into the iridolenticular space 360 degrees to create a protection barrier.

Impregnacão inadvertida da cápsula posterior do cristalino com azul de tripano durante a facectomia extracapsular: relato de caso / Unintentional staining of lens posterior capsule with trypan blue during extracapsular cataract extraction: case report

Relato de complicacão relacionada ao uso do azul de tripano 0,1 por cento ocorrida ao corar a cápsula anterior do cristalino em facectomia extracapsular com implante de lente intra-ocular. O corante foi injetado sob alta pressão utilizando a técnica de bolha de ar ocasionando a migracão deste através da zônula e conseqüente perda do reflexo vermelho no peroperatório dificultando os passos subseqüentes à extracão da catarata e comprometendo temporariamente a acuidade visual no pós-operatório. Tendo como objetivo evitar esta complicacão, os autores descrevem ainda, o uso da "Técnica de bolha de ar modificada" no qual o viscoelástico dispersivo é injetado no espaco iridolenticular 360º para compor uma barreira de protecão.

Early uncorrected visual acuity as a measurement of the visual outcomes of contemporary cataract surgery.

PURPOSE: To determine the uncorrected visual acuity (UCVA) on the first postoperative day and the fifth week after routine slow-motion phacoemulsification with posterior chamber intraocular lens (IOL) implantation. SETTING: Cincinnati Eye Institute, Cincinnati, Ohio, USA. METHODS: This retrospective chart review performed by 3 research fellows analyzed the UCVA 1 day and 5 weeks postoperatively in 100 consecutive best-case scenario eyes of 99 patients who had routine slow-motion phacoemulsification with implantation of an AcrySof single-piece IOL (Alcon). Reasons for UCVAs worse than 20/40 were sought. The stability of the visual result was analyzed. RESULTS: The UCVA was 20/40 or better in 98% of eyes at 1 day. Ninety-seven percent had a UCVA of at least 20/40 by 5 weeks, confirming stability of acuity. The percentage of patients with a UCVA of 20/20 or 20/25 increased from 49% at 1 day to 77% at 5 weeks. CONCLUSIONS: The UCVA 1 day after slow-motion phacoemulsification was 20/40 or better in 98% of eyes and remained stable to the last measurement at 5 weeks. Early UCVA can serve as a more sensitive measurement than best corrected visual acuity for assessing new surgical technologies and techniques.

Subtle signs of zonular damage.

PURPOSE: To describe 5 subtle signs of zonular damage. SETTING: Cincinnati Eye Institute, Cincinnati, Ohio, USA. METHODS: Three patients with a history of ocular trauma and 1 patient with Marfan's syndrome are described. During the preoperative examination, subtle signs of zonular damage were detected. As a result of the early recognition, critical modifications in cataract surgery technique and technology were made. RESULTS: The signs of zonular damage identified in the patients were visibility of the lens equator during eccentric gaze, decentered nucleus in primary position, iridolenticular gap, changes in the contour of the lens periphery, and focal iridodonesis. CONCLUSION: Identification of subtle but important signs of zonular damage will better prepare the surgeon for a challenging surgical procedure.

Empty-bag technique for acrylic intraocular lens implantation using Healon5.

PURPOSE: To determine how often the empty-bag technique (EBT) for injecting an acrylic foldable intraocular lens (IOL) into the capsular bag using Healon5 (sodium hyaluronate 2.3%) will require a 2-compartment technique to remove the ophthalmic viscosurgical device (OVD) and compare this with that of a standard injection method using Healon5. SETTING: Cincinnati Eye Institute and Ambulatory Surgical Center, Cincinnati, Ohio, USA. METHODS: Forty-one eyes of 40 patients were randomly assigned to 1 of 2 groups. In Group A, standard capsular bag expansion was performed with Healon5 before implantation of an acrylic posterior chamber (PC) IOL. In Group B, Healon5 was placed in the anterior chamber followed by balanced salt solution expansion of the capsular bag before acrylic PC IOL implantation. In all patients, complete OVD removal was attempted with the automated aspiration tip anterior to the optic (rock 'n roll technique). The number of patients in each group who required placement of the aspiration tip posterior to the optic for complete removal was compared. Early postoperative intraocular pressure (IOP) was also compared. RESULTS: In Group A, 15 (75%) of 20 eyes required placement of the aspiration tip posterior to the optic for complete OVD removal and in Group B, 1 (0.05%) of 20 eyes. The postoperative IOP was similar in both groups. CONCLUSIONS: The EBT decreased the likelihood that a 2-compartment OVD removal method would be necessary when using Healon5 for acrylic PC IOL implantation. The limitations of this new technique must be fully understood before it is used.

Three-step technique for staining the anterior lens capsule with indocyanine green or trypan blue.

A 3-step technique for staining the anterior lens capsule was developed to optimize the use of a capsule dye (trypan blue or indocyanine green) during capsulorhexis. After the injection of a viscoadaptive ophthalmic viscosurgical device (OVD) (sodium hyaluronate 2.3% [Healon 5]), which fills the anterior chamber, balanced salt solution (BSS) is gently injected onto the anterior capsule, creating a wafer-thin, fluid-filled space beneath the OVD. The dye is slowly injected through a 27-gauge cannula (Duckworth & Kent), selectively mixing with the BSS and allowing relatively accurate painting of the anterior capsule, approximating the size and shape of the intended capsulorhexis. This 3-step technique avoids the use of air as well as the uncontrolled dispersion and excessive accumulation of dye within the OVD.

Intraocular lens exchange assisted by preoperative neodymium:YAG laser haptic fracture.

We present 3 cases of intraocular lens (IOL) exchange in which a neodymium:YAG (Nd:YAG) laser was used before surgery to fracture a haptic and make intraocular manipulations easier and safer. Strategic placement of the fracture facilitates maneuvering to explant the IOL segments. Delaying pupil dilation until after the Nd:YAG laser is used and prudent positioning of the patient's head minimize the risk that freed IOL segments will damage the cornea or subluxate posteriorly into the vitreous cavity. Neodymium:YAG laser IOL fracture before explantation/ exchange is a viable combined procedure.

Focal multistriae: postoperative finding in posterior lenticonus patients.

A brother and sister presented with a history of difficulty reading fine print and an intolerance to glare. Family history was positive for bilateral posterior lenticonus in the mother. Biomicroscopy confirmed that each sibling had bilateral posterior lenticonus that was more easily diagnosed in the brother. Uneventful phacoemulsification was performed in each eye with a foldable acrylic posterior chamber intraocular lens (IOL) placed vertically within the capsular bag in 3 eyes and horizontally in 1 eye. On the day after surgery, biomicroscopy revealed vertical wrinkles limited to the central 2.0 mm of the posterior capsule corresponding to the cone in the eyes in which the IOLs had been vertically placed. We hypothesize that these striae occurred within the cone as a result of redistributing the usual mechanical forces associated with capsule stretch between the most distal contact points of the haptics. We believe that the appearance of these multiple striae limited to the central posterior capsule represents an unreported postoperative finding associated with posterior lenticonus.

Modified capsular tension ring for patients with congenital loss of zonular support.

PURPOSE: To evaluate the results of implantation of a modified capsular tension ring (MCTR) and a posterior chamber intraocular lens (PC IOL) in patients with congenitally subluxated crystalline lenses. SETTING: Cincinnati Eye Institute, Cincinnati, Ohio, USA. METHODS: Ninety eyes of 57 patients with congenital loss of zonular support (Weill-Marchesani syndrome, idiopathic ectopia lentis, and Marfan's syndrome) had phacoemulsification with PC IOL and MCTR implantation. The preoperative examination included best corrected visual acuity (BCVA) and the presence or absence of phacodonesis, lens decentration, and vitreous prolapse. The postoperative evaluation included BCVA and the presence or absence of pseudophacodonesis, PC IOL centration, and posterior capsule opacification (PCO). RESULTS: At the last postoperative examination, the BCVA was 20/40 or better in 80 eyes (88.9%); 1 eye (1.1%) lost 1 line of acuity. Preoperatively, 18 eyes (20%) had phacodonesis; 1 eye had postoperative pseudophacodonesis. Decentration before surgery was present in 86 eyes (95.6%); 6 eyes (6.7%) developed late symptomatic PC IOL decentration a median of 17.84 months +/- 10.73 (SD) after surgery. Other complications were increased intraocular pressure (2.2%), persistent iritis (3.3%), broken suture (10.0%), retinal detachment (1.1%), and PCO (20.0%). CONCLUSIONS: Use of the MCTR resulted in centration of the capsular bag and PC IOL in 90 eyes with congenitally subluxated crystalline lenses. Fixation of a 9-0 polypropylene suture is recommended to decrease the risk for late suture breakage.

Techniques for managing common complications of cataract surgery.

Cataract surgery has evolved dramatically over the last two decades, largely as a result of technological advances. As a result, visual outcomes and patient convalescence have improved significantly. A second consequence of increased instrumentation and technology, however, is increased complexity of cataract surgery and the advent of complications unique to these advances. Cataract surgeons must be aware and capable of managing the many possible adverse events that can occur during cataract surgery. This review identifies many of the common complications of cataract surgery, describes methods to avoid these complications and discusses techniques to address complications that do occur. It is the ability to avoid, quickly identify, and properly manage complications that defines the accomplished cataract surgeon.