Hearing and Vision Screening Program for newborns in Italy.

Materials and Methods: Data were collected through a nationwide survey conducted in 2018. All maternity wards active in Italy in 2017 were included. Background: The aims of this paper were to present data on the implementation and coverage of simultane-ous Universal Neonatal Hearing and Vision Screening programmes and to evaluate the organization and management of these healthcare procedures in Italy. Conclusions: Our results show that implementation of simultaneous hearing and vision screening increased the coverage of both screening tests and is a new multi-disciplinary approach to sensorineural disability. The use of both ABR and TEOAE tests in the hearing screening decreases the number of newborns sent for audiological evaluation, with a notable reduction of costs. The consideration that ocular problems are two to three times more common in deaf and hearing impaired children than their in hearing peers, confirms the importance of establishing guidelines for simultaneous hearing and vision screening, that favors the formation of a multi-disciplinary team (pediatrician, audio-logist, ophthalmologist). Results: HEARING SCREENING. Overall 427,365 newborns out of 448,386 (95.3%) received the hearing screening test (first level) in 391 out of the 409 maternity hospitals (95.5%), with a national mean referral rate of 3.63% (SD ± 4.58). A statistically significant increase (p<0.034) of newborns sent to audiological examination was found in maternity hospitals with Neonatal Intensive Care Unit (NICU) with "TEOAE only" protocol (9.32% SD ± 7.57), compared to those with "TEOAE/AABR" (3.0% SD ± 3.29). VISION SCREENING. Overall 335,262 newborns out of 448,386 (74.7%) received vision screening (Red Reflex test) for vision impairment in 302 out of 409 maternity hospitals (73.8%), before nursery discharge. The mean referral rate, recorded in only 22 maternity hospitals out of 302 (7.2%), was 0.48% with a rate of lost to follow up of 0.75 %.

Degenerative events in retina and optic nerve induced by inhibition of carnitine transport.

Biochemical and pharmacological evidence supports the hypothesis that the mechanism of action of mildronate [3-(2,2,2-trimethylhydrazinium)propionate dihydrate] is based on its regulatory effect on carnitine concentration. The present study demonstrates that carnitine acts as a neuroprotective agent both in optic nerve head and in retinal ganglion cell (RGC) by means of antioxidant and antiradical activities. In fact, carnitine normalized the increase in caspase-3, cellular apoptosis susceptibility protein (CAS) and inducible nitric oxide synthase (iNOS) expression by stabilizing mitochondrial membranes, as assessed by quantitative and qualitative analysis. This research shows that the neuroprotective effects of carnitine result, at least partially, from anti-neurodegenerative (anti-apoptotic) and anti-inflammatory mechanisms. It is suggested that the molecular conformation of carnitine can facilitate its easy binding to mitochondria, and regulate the expression of different signal molecules, hence maintaining normal cellular signaling and survival by modulating caspase-3 activity.

New therapies in common ocular surface disorders.

Lacrimal dysfunction syndrome (LDS) and meibomian gland dysfunction (MGD) are common pathologies of the ocular surface both characterized by quantitative and/or qualitative tear film changings. They may result in symptoms of eye irritation, clinically apparent inflammation and ocular surface disease. Recent researches about the chronic inflammatory nature of dry eye, revealed the possibility to develop new treatments for LDS like new immunological drugs. MGD, alone or if it occurs contemporary to LDS, can be treated with a thermal pulsation system, a useful tool to alleviate symptoms.

Mechanisms of ocular neuroprotection by antioxidant molecules in animal models.

This work was conducted to evaluate the efficacy of a treatment on retinal ganglion cells (RGC) and on astrocytes of the optic nerve of glaucomatous eyes, using a combination of alpha-lipoic acid (ALA) and superoxide dismutase (SOD). Thirty-two male Wistar rats were fed with a diet supplemented with ALA, SOD, ALA and SOD or with no product for 8 weeks. Ocular hypertension was induced with 2% methylcellulose (MTC) and then rats were sacrificed. TUNEL assay showed a marked fluorescence in the ganglion cells and astrocytes of MTC-treated rats evidencing induction of apoptosis. In contrast, sections of eyes pretreated with ALA and SOD showed a lack of fluorescence quite similar to that of the controls. Similarly, eyes sections from rats pre-treated with ALA and SOD showed reduced differential expression of inducible nitric oxide synthase (iNOS) and of caspase-3 in compared to normally-fed/MTC-inoculated cases. An increase of ALA and SOD exerts an antiapoptotic effect and protects against oxidative stress and hence against the structural remodelling of the RGCs and astrocytes of the optic nerve in the presence of an ischemic and pressure stress.

Forskolin and rutin prevent intraocular pressure spikes after Nd:YAG laser iridotomy.

AIM: the purpose of this research was to evaluate whether an oral treatment with an association of forskolin and rutin can blunt the intraocular pressure (IOP) spikes and avoid the damage that may occur after laser iridotomy. METHODS: Ten patients underwent bilateral Neodymium:YAG (Nd:YAG) laser iridotomy (Visulas YAG III Laser, Zeiss), for the prevention of primary closed-angle glaucoma. IOP was measured in subjects before and after 7 days of pretreatment with placebo or forskolin and rutin by Goldman applanation tonometry. The IOP was measured before surgery and after surgery at 30-60-120 minutes, and 4-7 days. RESULTS: Analysis of variance indicated a significant increase of the postoperative values in patients receiving treatment with placebo (p < 0.001), but not in those who received treatment with the forskolin and rutin association. T test analysis confirmed that IOP still remained significantly elevated 7 days after laser intervention in placebo treated patients, whereas it stayed within normal values in forskolin/rutin treated patients. CONCLUSION: Forskolin and rutin can blunt the increase of IOP that occurs after Nd-YAG laser iridotomy. This can avoid serious risk to the optic nerve of the patients under laser treatment for iridotomy.

The effect of night vision goggles on the retinocortical bioelectrical activity and its improvement by food supplement.

AIM: To investigate the effect of luminance variations, as well as the oral administration of a food supplement, on the visual bioelectric response while using of Night Vision Goggles (NVG). METHODS: Two trials were performed, both enrolling healthy male aircrew members wearing NVG, and recording Visual Evoked Potentials (VEPs) from scalp electrodes. Both foveal and parafoveal response were evaluated. Latency and amplitude, P100 peak, were measured. In the first set of measurements, VEPs parameters were recorded during unaided photopic conditions and mesopic conditions while using 3rd generation plus NVG (ANVIS 9). In the second set of experiments, after the first basal electrophysiological investigation during mesopic conditions using NVG, patients started a 45 days oral treatment, during which they took 3 tablets per day of a food supplement. The tablets contained a mix of anthocyanosides, procyanidolic oligomers, lutein and vitamins A and E. At the end of this treatment, patients were tested again by pattern-reversal VEP investigation during aided vision condition (wearing NVG) in a mesopic environment. RESULTS: VEPs parameters, statistically evaluated using a two tailed paired t-test, showed that latency and amplitude were respectively increased (p < 0.001 and p < 0.01 for 15' and 60' minutes of arc) and decreased (p < 0.05) when measured using NVG with respect to unaided basal conditions. Furthermore, the VEP response in NVG aided vision was positively affected by the oral treatment with the food supplement, showing a significant (p < 0.05) decrease of latency and increase of amplitude. CONCLUSION: The use of NVG impairs the VEP response, and such effect is effectively counteracted by the oral treatment with a food supplement containing a combination of sight improving molecules that might enhance foveal selectivity, central photoreceptors sensitivity and magnocellular fibers effectiveness.

Keratoconjunctivitis by confocal microscopy after topical cyclosporine.

AIM: To describe the observations obtained by confocal microscopy concerning corneal stromal structure in the process of adenoviral epidemic keratoconjunctivitis, from the onset of the disease up to 20 weeks of follow-up, after topical cyclosporine. METHODS: A 16-years-old boy has developed subepithelial infiltrates and an epidemic keratoconjunctivitis from adenovirus in both eyes. Clinical case was analyzed by confocal microscopy (40x mode, Nidek ConfoScan 4) before and after administration of topical cyclosporine. RESULTS: Two weeks after the onset of the symptoms, corneal stroma showed the presence of cells with highly reflective area. These cells, comparable to keratocytes, had a typical fusiform shape and assumed a particular rosette disposition, never highlighted in the literature. Hyperreflective areas disappeared in both eyes after administration of topical cyclosporine 1% for 30 days. This outcome was also confirmed after five months. CONCLUSION: The confocal microscopy performed on the surface of the stroma before and after treatment with cyclosporine suggests the presence of a localized immune activation in subepithelial layer.

Retrospective study of glaucoma and closed-eyelid test: long-term outcomes in an Italian native population.

AIM: To establish a threshold value of intraocular pressure (IOP) increase after the closed-eyelid test (CET) that correlates with the highest probability of developing overt primary open-angle glaucoma (OAG) in an Italian native population from 1980 to 2010. METHODS: Retrospective analysis of data obtained from 161 patients with ocular hypertension who performed the CET in 1980, and were subsequently followed to see whether they developed OAG. CET was performed always in the morning Eyelids were closed by bandaging for 1 h in a quiet environment, with the patient seated and not sleeping. IOP was measured again 8 to 10s after opening the eyelids. RESULTS: Accurate statistical analysis of the obtained values indicated that 77% of the subjects showing an IOP increase after 1 hour of eyelid closure in a sitting position developed OAG in the following 30 years and that IOP increase values above 4 mmHg led to a subsequent diagnosis of glaucoma in more than 80% of the patients. CONCLUSION: Eyelid closure for 60 minutes results in a net elevation of IOP the extent of which depends on the balance between the increase of aqueous humour secretion and its outflow. Therefore, the CET may discriminate individuals with a normal outflow from individuals with a less functional outflow, which are evidently those at a higher risk of developing glaucoma.

Oxidative stress in the closed-eyelid test: management of glaucoma.

BACKGROUND AND OBJECTIVES: To evaluate the role of antioxidant drugs in the tonometric increase that follows the closed eyelid test (CET), a predicitive test for glaucoma, after administration of antioxidant substances was observed. MATERIALS AND METHODS: 30 subjects of 54.57+/=5.62 years, 13 males and 17 females, were examined by measuring the ocular pressure after 1 hour from the CET, both in normal conditions and after the administration of antioxidants such as: vitamin A (50,000 IU/die), vitamin E (600 mg/die), and vitamin C (1000 mg/die). The increases in temperature of the iridocorneal angle and of the iris were also measured in the same conditions with an infrared Thermo-Precision tonometer (Sola Electro-Optics, China) both before and after CET. RESULTS: The results showed increased pressure after CET and decreased pressure after the administration of each antioxidant substance, although vitamin A was found to be more effective and with statistically significant values compared to vitamins E and C. CONCLUSIONS: Considering the responses obtained after administration of antioxidant drugs, the ocular hypertension induced after CET could be a response to mixed stress, oxidative and thermic, with degenerative effects on the trabecular meshwork (TM). Besides, in light of these considerations the research results underline that the open angle glaucoma (OAG) should be considered a multifactorial degenerative disease.

Corneal haze in course of Fuchs' endothelial dystrophy.

This article describes the observations obtained with confocal microscopy (CM) on the corneal structure in course of corneal edema in a patient with Fuchs endothelial corneal dystrophy (FD). The patient was a 40 year old male, suffering from second stage FD, in course of corneal edema and bullous keratopathy. The tissue structure was analyzed with CM confoscan CS4 (Nidek Technologies(®), Birmingham, UK) using the 40x mode. The CM has shown the presence of gaps due to corneal edema and a diffuse stromal hyper reflectivity related to the alteration of the extracellular matrix. It has also showed the presence of binucleate cells, assimilable to keratocytes, in cytokinesis which presented a typical fusiform aspect with two highly reflective nuclei awaiting cell division. The total number of cells was much lower than that of healthy control subjects of similar age, sex and race. The CM in this case suggests a significantly lower number of cells, presumably keratocytes, compared to normal range, but mostly it shows the presence of cells undergoing cytokinesis, which witnesses the active processes of collagenogenesis and possible vasculogenesis that represent early stages of loss of the normal corneal transparency.

Topotecan hydrochloride effects on retinal vessels in newborn rats.

A physiological system, i.e. rodent retina during vessel formation and hierarchical organization, was utilised for assaying antiangiogenic properties of Topotecan, a topoisomerase I inhibitor, capable of inhibiting tumoral growth in animal models of retinoblastoma. In particular we analysed possible differences in effectiveness and side effects among different drug dosages and ways of administration. In the present research only qualitative analyses were undertaken. After preliminary experiments, in which suckling animals subcutaneously treated with Topotecan dosages comprised between 9 and 3 mg/kg underwent high lethality and extremely severe systemic damages, 7 day-old rats were subcutaneously, intravenously or peribulbary injected with a single dose of 1 mg/kg; retinal vessels were visualized in retinal fluorangio-graphies taken 1 and 2 weeks after treatment. The most important and frequent alterations were found to affect radial vessels, which showed non-perfused and/or regionally mislocated segments, together with abnormal branching and enlargements in retinal periphery; persistence of capillary-free periarteriolar regions, non-vascularised regions and spots of extravascular FITC were also detected. Despite the high individual variability the alterations were substantially similar among the different ways of drug administration, while they appeared milder in 21 day-old rats, with respect to younger ones. The extensive vascular remodelling found after Topotecan administration, besides demonstrating the antiangiogenic properties of this chemioterapic drug, confirms the rodent retina as a highly valuable model system for studying angiogenesis modulation.

Degenerative effects in rat eyes after experimental ocular hypertension.

This study was used to evaluate the degenerative effects on the retina and eye-cup sections after experimental induction of acute ocular hypertension on animal models. In particular, vascular events were directly focused in this research in order to assess the vascular remodeling after transient ocular hypertension on rat models. After local anaesthesia by administration of eye drops of 0.4% oxibuprocaine, 16 male adult Wistar rats were injected in the anterior chamber of the right eye with 15 µL of methylcellulose (MTC) 2% in physiological solution. The morphology and the vessels of the retina and eye-cup sections were examined in animals sacrificed 72 h after induction of ocular hypertension. In retinal fluorescein angiographies (FAGs), by means of fluorescein isothiocyanate-coniugated dextran (FITC), the radial venules showed enlargements and increased branching, while the arterioles appeared focally thickened. The length and size of actually perfused vessels appeared increased in the whole superficial plexus. In eye-cup sections of MTC-injected animals, in deep plexus and connecting layer there was a bigger increase of vessels than in controls. Moreover, the immunolocalization of astrocytic marker glial fibrillary acidic protein (GFAP) revealed its increased expression in internal limiting membrane and ganglion cell layer, as well as its presence in Müller cells. Finally, the pro-angiogenic factor vascular endothelial growth factor (VEGF) was found to be especially expressed by neurones of ganglion cell layer, both in control and in MTC-injected eyes. The data obtained in this experimental model on the interactions among glia, vessels and neurons should be useful to evaluate if also in glaucomatous patients the activation of vessel-adjacent glial cells might play key roles in following neuronal dysfunction.

First clinical case of effective medical treatment of the vitreoretinal traction with recovery of the visual acuity.

This case-report evaluates the effectiveness of the medical treatment on vitreomacular traction through the use of palmitoylethanolamide (PEA), orally administered. Our 69 year-old patient reported a significant eyesight reduction, metamorphopsia and photopsia. He underwent the following investigations, before and after medical treatment: visual acuity evaluation, dilated fundus examination, OCT. The treatment consisted of the administration of two tablets of PEA per day over 7 days. We observed that, while the visual acuity in the right eye was 2/10 compared to the initial 11/10, the dilated fundus examination highlighted a foveal hole. Moreover, the OCT confirmed the presence of a vitreomacular traction due to the incomplete posterior vitreous detachment. At the end of the treatment with PEA, the visual acuity was equal to 8/10 and the OCT showed a disappearance of the hyperreflective streak with recovery of the physiological retinal and foveal profile. The therapy was eventually carried on for 10 more days with a final visual acuity of 10/10. In conclusion, PEA orally administered, beyond a rare possibility of a spontaneous resolution of the vitreomacular traction, probably contributed, by anti inflammatory action, to the vitreolysis and thus to the disappearance of the vitreomacular traction and foveal hole with a subsequent recovery of the eyesight and of the metamorphopsia.

Oral administration of an association of forskolin, rutin and vitamins B1 and B2 potentiates the hypotonising effects of pharmacological treatments in POAG patients.

BACKGROUND/AIMS: Control of intraocular pressure is still the main strategy to treat glaucoma patients. Forskolin has already shown an ability to control intraocular pressure after topic administration, whereas rutin is known to improve ocular blood fl ow. Therefore, aim of this pilot study has been to observe whether administration of an association of oral forskolin and rutin to POAG patients under different regimens of medical therapy may contribute to their effects, further decreasing IOP values. MATERIALS AND METHODS: Forskolin (a natural compound present in the crude extract of the plant Coleus Forskohlii) and rutin are the main ingredients of a food supplement commercially available in Italy. In an open label pilot study, 16 patients with POAG under treatment with different topical drugs and with stable IOP were given additional treatment with the food supplement for 40 days, and their IOP values measured at enrolment, at the end of treatment and 40 days after treatment interruption. RESULTS: Further addition of forskolin and rutin to topical association treatments resulted in a further decrease of IOP by roughly 20% of the initial value. The effect was reversible upon suspension of the treatment. CONCLUSIONS: These data show for the fi rst time that forskolin and rutin given through the oral route appear to reach the ocular district, where they can act in synergy with topical pharmacological treatments, and contribute to the control of intraocular pressure.

An NMR spectroscopy study of bendaline-albumin interactions.

The complete assignment of the 1H and 13C NMR spectra of bendaline (BNDL) was performed by mono-dimensional and homo- and hetero-correlated two-dimensional NMR experiments. The interaction between bendaline and albumin was also studied by the analysis of the motional parameters spin-lattice relaxation times, allowing the motional state of the BNDL free and bound with albumin to be defined. In absence of albumin the indazolacetic and benzylic moieties are characterized by roughly the same mobility and by positive sigma (cross-relaxation rates) values. In the presence of the macromolecule, the indazolacetic and benzylic moieties and the lysine change their motional behaviour to different extents, as indicated by correlation times. Data obtained in absence and in presence of the protein show that the molecular moiety of the bendaline most involved in the binding with albumin is the fragment H-4 H-5. The binding constant was evaluated at 2.4x10(3)M(-1).

Age-related changes in rat optic nerve: morphological studies.

Age-related changes of the optic nerve were studied in 3-month-old (young), 12-month-old (adult) and 24-month-old (aged) male Sprague-Dawley rats. Cross sections of the intracranial portion of the optic nerves of animals of different age groups were stained with haematoxylin-eosin and examined under a light microscope at low and high magnification. Other sections were stained with crystal violet for demonstration of glial cells. A third group of sections were stained immunohistochemically to detect glial fibrillary acidic protein (GFAP) which is a marker for localizing and characterizing astrocytes. All morphological results were subjected to the quantitative analysis of images and to statistical analysis to identify significant morphometrical data. Tissue protein concentrations were determined on homogenized fragments of optic nerve. Our results demonstrate the following age-related changes: (1) increase of the optic nerve sheaths (meningeal membranes); (2) increased number of astrocytes; (3) increase of areal density of GFAP immunoreactivity; (4) increased diameter and area of the optic nerve; (5) decreased number of nerve fibres; (6) decreased-size of nerve fibres and (7) decrease of the nerve fibres/meningeal membrane ratio from 3:1 to 1:1. Moreover, the protein amount does not change with age. The rat optic nerve, therefore, appears sensitive to ageing processes and can be considered as a useful model for the studies on neuronal ageing.

A morphometric study of age changes in the rat optic nerve.

Age-related changes of the optic nerve fibres were studied in 3-month-old (young), 12-month-old (adult) and 24-month-old (aged) male Sprague-Dawley rats. The optic nerve was harvested with particular care from the intracranial portion. Cross sections from the optic nerve of animals of different age groups were stained with toluidine blue and examined under a light microscope at low and high magnification. Other sections were stained for the demonstration of glial cells using the method described by Holzer. A third group of sections were stained by the immunohistochemical method to detect glial fibrillary acidic protein, which is a marker for localising and characterising astrocytes. All these morphological results were subjected to the quantitative analysis of images and to statistical analysis of the values to identify significant morphometric data. Biochemical dosages of proteins were also performed on homogenised fragments of the optic nerve. Our results demonstrate that the following age-related changes can be observed: (1) an increase in meningeal membranes, (2) an increased number of astrocytes, (3) an increase in areal density of glial fibrillary acidic protein immunoreactivities, (4) an increased thickness of the entire optic nerve and an increased area of the nerve, (5) a decreased number of nerve fibres and (6) a decrease in the nerve fibre/meningeal membrane ratio from 3:1 to 1:1. Moreover, the amount of protein does not change with age. The rat optic nerve, therefore, appears sensitive to aging processes.

A murine cell culture model for post-trabeculectomy anfibrotic treatment: Induction of apoptosis by Cyclosporin.

PURPOSE: Experimental trials aimed at the research of selective antifibrotic agents are under development for the alternative treatment of glaucoma patients who are usually considered high-risk post-surgical individuals after trabeculectomy. Authors present here an in vitro model system for the treatment of post-trabeculectomy patients. The study is aimed at the evaluation of different drugs in a mouse fibroblast model. METHODS: The antifibrotic activity of Cyclosporin A, Interferon 2alpha, 5-Fluorouracyl was investigated on 3T6 cells in culture. Cell viability and proliferation was assessed after drug treatment. Molecular analysis of DNA degradation was evaluated by means of radioactive labeling and gel electrophoresis. RESULTS: The three drugs were shown to affect cell proliferation and viability in a differential fashion. However, only Cyclosporin A was able to control cell proliferation, inducing apoptosis. This phenomenon was reduced by supplementation of trolox, a compound known to inhibit programmed cell death. These results strongly suggest that this model system might be useful as a test of pharmacological functionality. CONCLUSION: A rapid and efficient model system is described for the assessment of cell viability and proliferation after treatment with agents of potential pharmacological use. Cyclosporin A induces a significant apoptosis. This is important for the negative control of fibrotic degeneration in post-trabeculectomy that is required for successful surgery in glaucoma patients. Therefore, Cyclosporin A might become a clinically interesting drug for the antifibrotic treatment of post-trabeculectomy.

Metabolic changes in rabbit lens induced by treatment with dexamethasone.

Metabolic changes in the rabbit lens have been studied by means of nuclear magnetic resonance spectroscopy. These changes have been induced by prolonged topical treatment with dexamethasone. Our results demonstrate an increase in sorbitol, sorbitol-3-phosphate, fructose-3-phosphate, glycerol-3-phosphate and glucose-6-phosphate levels and a decrease in glutathione sulphate (GSH) and myo-inositol levels, in agreement with what was observed in lenses from streptozocin-diabetic rats before lens opacity. The hyperglycaemia can only partially explain all these observed biochemical variations. The lack of increase in the intermediates of pentose cycle, such as sedoheptulose-7-phosphate, seems to support the hypothesis of an inhibition of glucose-6-phosphate dehydrogenase by dexamethasone treatment. Finally dexamethasone treatment induces a decrease in GSH. The decreasing or the loss of GSH has been suggested as a possible pathogenic mechanism in the cataract formation.