Antonio Bergua

Nitrergic and VIPergic neurons in the choroid and ciliary ganglion of the duck Anis carina.

Immunohistochemistry for neuronal nitric oxide synthase (nNOS) and vasoactive intestinal peptide (VIP), and NADPH diaphorase histochemistry, were applied to investigate neurons in the choroid and the ciliary ganglion of the muscovy duck Anis carina. Up to 1000 neurons in the choroid stained for NADPH diaphorase and showed virtually complete colocalization for nNOS immunoreactivity. Almost all of them co-stained for VIP, while about 90% of VIP immunoreactive cell bodies showed colocalization for nNOS. Two-thirds of the neurons were located, mostly singly, at nodes of a widemeshed nerve plexus in the suprachoroid and were only rarely grouped in ganglia of up to 3 neurons. Numerous varicose nNOS/NADPH-diaphorase-positive nerve fibers were seen around large arterial blood vessels. These fibers derived mainly from paravascular cell bodies that represented about one-third of all choroidal neurons and also displayed costaining for nitrergic markers and VIP. Colocalization of nNOS/NADPH-d and VIP could be demonstrated in most of the perivascular fibers, while slightly more VIP-positive axons in the suprachoroid plexus did not costain for nNOS/NADPH-d. Small-caliber blood vessels and those localized in the choriocapillaris were not endowed with VIP/nNOS/NADPH-diaphorase-positive fibers. A few reactive neuronal cell bodies were also found in ciliary nerves, while most ciliary axons were unstained. In the ciliary ganglion a small subpopulation of neurons showed VIP/nNOS/NADPH-diaphorase colocalization. There were also nNOS/ NADPH-d-positive cap-like terminals on ciliary ganglion cells. The presence of VIP/nNOS/NADPH-diaphorase positive neurons and nerve fibers in both the choroid and ciliary ganglion, and in the choroidal perivascular plexus, indicates peripheral nitrergic and VIPergic control of blood flow in the choroid of the duck.

Impaired Plasmacytoid Dendritic Cell Innate Immune Responses in Patients with Herpes Virus-Associated Acute Retinal Necrosis

Plasmacytoid dendritic cells (PDC), the main producers of type I IFNs in the blood, are important for the recognition and control of viral and bacterial infections. Because several viruses induce IFN-α production, severe courses of herpes virus infections in nonimmunocompromised patients may be related to numerical or functional PDC deficits. To evaluate this hypothesis, PBMC and PDC were repeatedly isolated from nine patients with acute retinal necrosis (ARN), caused by herpes simplex or varicella zoster virus. The patients experienced meningitis/encephalitis and frequent infections in childhood (n = 2), recurrent herpes virus infections at unusual localizations (n = 2), ocular surgery (n = 1), infections (n = 4), and stress around ARN (n = 6). The median percentage of isolated PDC was significantly lower in patients compared with 18 age-matched healthy controls (p < 0.001), confirmed by FACS analysis using peripheral blood, and was extremely low during acute disease. PDC counts dropped in five controls suffering from respiratory infections or diarrhea. IFN-α production in PDC and PBMC exposed to different stimuli was significantly lower in patients than in controls (p < 0.05). Anergy to these stimuli was observed on four occasions, in particular during acute disease. PDC of patients showed up-regulated IFN regulatory factor-7 mRNA levels and evidence of in vivo activation (CD80) and maturation (CD83) (p < 0.05). CD8+ cell responses were significantly lower in patients vs controls (p = 0.04). These data support a risk factor model in which numerical and functional deficits in PDC-mediated innate immune responses contribute to an impaired control of latent herpes virus infections and subsequent development of ARN.

Vasoactive intestinal and calcitonin gene-related peptides, tyrosine hydroxylase and nitrergic markers in the innervation of the rat central retinal artery

The vascular supply of the optic nerve has been studied with different methods including corrosion casts both in humans and in other mammals. In man, primates and some other mammals, such as the rat, a distinct central retinal artery accompanies the optic nerve, and runs through the lamina cribosa to reach the optic nerve head. Similarities between human and rat central retinal artery could serve to understand changes in the autonomic perivascular innervation in glaucoma using the rat as an animal model. Nitric oxide, calcitonin gene-related peptide, neuropeptide Y, substance P and vasoactive intestinal peptide have been identified around the monkey central retina artery. Innervation of the rat central artery, however, has not been described in detail. Using immuno- and histochemical methods, the present study investigates the peptidergic, adrenergic and nitrergic innervation of the rat posterior ciliary artery as well as the central retina artery. Numerous nitric oxide positive nerve fibers were visualized posterior and anterior to the lamina cribosa of the optic nerve. They colocalized with NADPH-diaphorase positive fibers, which could also be observed in two of six specimens studied at the level of the optic nerve head. Calcitonin gene-related peptide, tyrosine hydroxylase, and VIP positive fibers were also observed surrounding the vessels of the rat optic nerve. The presence of neuronal nitric oxide/NADPH-diaphorase and vasoactive intestinal peptide positive nerve fibers surrounding the posterior ciliary and central retinal arteries indicates a vasodilator effect in the rat optic nerve. Tyrosine hydroxylase positive innervation indicates the presence of sympathetic activity, and calcitonin gene-related peptide positive fibers indicate sensory innervation by trigeminal primary efferents.

Visual evoked potentials under luminance contrast and color contrast stimulation in glaucoma diagnosis.

Purpose: To evaluate the diagnostic value of visual evoked potential (VEP) assessment with luminance‐contrast and color‐contrast stimulation in the detection of glaucoma. Patients and Methods: The study included 59 patients (96 eyes) with glaucomatous changes of the optic disc and visual field defects and 58 control eyes of 29 healthy patients. Four types of pattern VEP stimulation (0.9 cycle/degree) were performed in all patients: achromatic, alternating sine‐wave stripe pattern: 6 reversals per second, contrast of 10% (activation of predominantly the magnocellular pathway); isoluminant, red‐green stripe pattern: 83.3 milliseconds onset, 83.3 milliseconds offset, contrast of 30% and 80% (activation of predominantly the parvocellular pathway); and blue grating with yellow background adaptation: 200 milliseconds onset, 500 milliseconds offset (activation of the blue‐sensitive pathway). Results: The glaucoma group and the control group differed significantly (P < 0.01) in the peak times of all chromatic VEP responses and to a lesser degree in the achromatic VEP. Considering the amplitudes, only the low‐contrast red‐green stimulus showed a statistically significant reduction in glaucoma. At a predefined specificity of 90%, in separating patients with glaucoma from healthy control subjects, the peak time of the blue‐yellow VEP had a high sensitivity (90%), whereas the sensitivity of the achromatic VEP was low (31%). The red‐green VEP showed a sensitivity of 73% using low contrast and 71% using high contrast. In a paired correlation analysis with visual field defects, all stimulations showed significant (P < 0.05) results. Correlation coefficients were highest (R = 0.79, P < 0.01) for the peak time of the blue‐yellow VEP. Conclusions: VEP measurements with presumable stimulation of single neuronal pathways can detect glaucomatous optic nerve damage in a considerable fraction of patients with visual field loss. Occipital responses to chromatic stimulation seem to be more sensitive to glaucoma damages than do responses to achromatic pattern reversal stimulation.

Topical Ranibizumab inhibits inflammatory corneal hem- and lymphangiogenesis.

Purpose:  Ranibizumab (Lucentis®) is a Fab‐Fragment of a recombinant, humanized, monoclonal VEGF (anti‐vascular endothelial growth factor) antibody. This study analyzed the ability of topical Ranibizumab to inhibit lymphangiogenesis in addition to hemangiogenesis after acute corneal inflammation in vivo. In addition, the effect of Ranibizumab on the proliferation of human lymphatic endothelial cells (LECs) and blood endothelial cells (BECs) in vitro was studied.

Self-Enucleation in Drug-Related Psychosis

Objective: To report on a patient who committed self-enucleation with his own fingers. Methods: Case report of a 28-year-old man who self-enucleated his right eye. After ophthalmological treatment, he was referred to the psychiatric department for further evaluation and treatment. Results: Reconstruction of the orbit was performed, and neither ophthalmological nor neurological complications were noted. The psychiatric history revealed continuous drug abuse (cannabis and amphetamines). The diagnosis of a drug-induced psychosis was established. Conclusions: Self-enucleation or ‘oedipism’ is a rare entity which requires, besides an operative reconstruction of the orbit, neurological monitoring to prevent possible intracranial complications. Long-term psychiatric therapy should be attempted to prevent further self-injurious behaviour.

Melatonin analogue agomelatine reduces rabbit's intraocular pressure in normotensive and hypertensive conditions.

In the search for new compounds to reduce intraocular pressure (IOP), with fewer side effects, we have found that agomelatine, a melatonin analogue, can reduce IOP being, therefore, interesting for the treatment of ocular hypertension and glaucoma. In normotensive conditions, agomelatine (10μl 100μM) reduced IOP by 20.8±1.4% (n=18) with a maximal effect 180min after the compound application and 68.8±5.7% (n=8) in a hypertensive condition. Concentration-response curve depicted a sigmoid behaviour presenting a pD2 value of 9.7±0.3 which was equivalent to an EC50 of 0.19nM. The effect of agomelatine was partially antagonized by 4PPDOT (MT2 antagonist receptor. 10μl 100μM) and prazosin (MT3 antagonist receptor. 10μl 100μM) (85.6±1.6% and 87.2±1.9%, N=18 respectively.) Agomelatine hypotensive effect in normotensive condition was comparable to latanoprost (40μl) and brimonidine (40μl) and it was no so effective as dorzolamide (40μl) or timolol (40μl). These results may suggest the use of this melatonin analogue for the treatment of those ocular conditions, which involve an abnormal raise of intraocular pressure.

Tele-transmission of Stereoscopic Images of the Optic Nerve Head in Glaucoma via Internet

The objective was to describe an inexpensive system to visualize stereoscopic photographs of the optic nerve head on computer displays and to transmit such images via the Internet for collaborative research or remote clinical diagnosis in glaucoma. Stereoscopic images of glaucoma patients were digitized and stored in a file format (joint photographic stereoimage [jps]) containing all three-dimensional information for both eyes on an Internet Web site (www.trizax.com). The size of jps files was between 0.4 to 1.4 MB (corresponding to a diagonal stereo image size between 900 and 1400 pixels) suitable for Internet protocols. A conventional personal computer system equipped with wireless stereoscopic LCD shutter glasses and a CRT-monitor with high refresh rate (120 Hz) can be used to obtain flicker-free stereo visualization of true-color images with high resolution. Modern thin-film transistor-LCD displays in combination with inexpensive red-cyan goggles achieve stereoscopic visualization with the same resolution but reduced color quality and contrast. The primary aim of our study was met to transmit stereoscopic images via the Internet. Additionally, we found that with both stereoscopic visualization techniques, cup depth, neuroretinal rim shape, and slope of the inner wall of the optic nerve head, can be qualitatively better perceived and interpreted than with monoscopic images. This study demonstrates high-quality and low-cost Internet transmission of stereoscopic images of the optic nerve head from glaucoma patients. The technique allows exchange of stereoscopic images and can be applied to tele-diagnostic and glaucoma research.

An early antecedent to modern random dot stereograms : The Secret Stereoscopic writing of Ramón y Cajal

The use of computerized random dot stimuli in modern neuroscience was introduced by Julesz in the 1960s. This method made it possible to study exclusively cortical processing of binocular information by disparity-sensitive neurons, and it has attained widespread use among neuroscientists and psychologists. It is now largely forgotten that in the last century, the famous neuroanatomist Ramón y Cajal had worked on random dot stereograms as a means of encoding written information. A brief note was finally published in a Spanish journal on photography in 1901. We present a translation of this text and summarize the early ideas on random dot stereograms, and we also supply a brief historical account on stereoscopic perception.

Innervation pattern of the preocular human central retinal artery.

The central retinal artery (CRA) is the main vessel for inner retinal oxygen and nutrition supply. While the intraocular branches lack autonomic innervation, the innervation pattern of the extra-ocular part of this vessel along its course within the optic nerve is poorly investigated. This part however is essential for maintenance of retinal blood supply, in physiological and pathological conditions. Therefore, the aim of this study was the characterization of the autonomic innervation of the preocular CRA in humans with morphological methods. Meeting the Declaration of Helsinki, eyes of body or cornea donors were processed for single or double immunohistochemistry against tyrosine hydroxilase (TH), dopamine-β-hydroxylase (DBH), choline acetyl-transferase (ChAT), vesicular acetylcholine transporter (VAChT), neuronal nitric oxide synthase (nNOS), calcitonin gene-related peptide (CGRP), substance P (SP), vasoactive intestinal polypeptide (VIP), and cytochemistry for NADPH-diaphorase (NADPH-d). For documentation, light-, fluorescence-, and confocal laser-scanning microscopy were used. TH and DBH immunoreactive nerve fibres were detected in the CRA vessel wall, although a distinct perivascular plexus was missing. Further, nerve fibres immunoreactive for ChAT and VAChT were found, while CGRP, SP, and VIP were not detected. NADPH-d staining revealed scattered nerve fibres in the adventitia of the CRA and in close vicinity; however, nNOS-immunostaining could not confirm this finding. The CRA receives adrenergic and cholinergic innervations, indicating sympathetic and parasympathetic components, respectively. Remarkably, a peptidergic primary afferent innervation was missing. Since clinical results suggest an autoregulation of intraretinal vessels, further studies are needed to clarify the impact of CRA innervation for retinal perfusion.