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Macular hole surgery without routine membrane excision or use of adjuvants.

PURPOSE: To present results of macular hole surgery without adjunctive treatment or routine membrane dissection, but with a long duration tamponade. METHODS: Fifty-one consecutive eyes underwent surgery with vitrectomy. Membrane peeling was performed only when traction was obvious. Pure perfluoropropane gas was injected as a tamponade followed by prone positioning for two weeks. All eyes were examined with scanning laser ophthalmoscopy (SLO) before and after surgery. RESULTS: The macular hole was closed successfully in 48 eyes (94%) after one procedure. Visual acuity improved to 0.50 or more in 18 eyes (35%). Improvement of two or more lines was achieved in 35 eyes (69%). Two eyes developed retinal detachments and two eyes showed post-operative visual field defects previously reported after macular hole surgery. CONCLUSION: In this series the results and complication rates compare favourably with other reports of macular hole surgery. However, in contrast to many previous studies, membrane peeling was not done. The extent and long duration of the gas tamponade was thought to be important for the surgical success.

Aged↗

Cystoid macular oedema following neodymium:YAG laser capsulotomy a prospective study.

Neodymium:YAG laser capsulotomy was performed in 65 eyes of 65 patients because of vision loss due to posterior capsular opacification or wrinkling following uneventful extracapsular cataract extraction with intraocular lens implantation. Eyes with pre-existing macular pathology were excluded from this study. In all of the eyes the posterior capsule was successfully opened. None developed clinical or angiographic cystoid macular oedema. One eye developed retinal detachment nine months after capsulotomy.

Aged↗

Positional cloning of the gene associated with X-linked juvenile retinoschisis.

X-linked juvenile retinoschisis(RS) is a recessively inherited vitreo-retinal degeneration characterized by macular pathology and intraretinal splitting of the retina. The RS gene has been localized to Xp22.2 to an approximately 1 Mb interval between DXS418 and DXS999/DXS7161. Mapping and expression analysis of expressed sequence tags have identified a novel transcript, designated XLRS1, within the centromeric RS locus that is exclusively expressed in retina. The predicted XLRS1 protein contains a highly conserved motif implicated in cell-cell interaction and thus may be active in cell adhesion processes during retinal development. Mutational analyses of XLRS1 in affected individuals from nine unrelated RS families revealed one nonsense, one frameshift, one splice acceptor and six missense mutations segregating with the disease phenotype in the respective families. These data provide strong evidence that the XLRS1 gene, when mutated, causes RS.

Amino Acid Sequence↗

Visual outcomes following ICG assisted ILM peel for Macular Hole.

PURPOSE: To evaluate the anatomical and visual outcome of indocyanine green (ICG)-assisted internal limiting membrane (ILM) peeling for macular hole surgery. A literature search identified potential factors that may affect the functional success of the procedure. METHODS: Retrospective case note review of 14 consecutive patients undergoing phaco-vitrectomy, gas and ICG-assisted ILM peeling for macular hole from July 2001 to July 2002. A 0.5% ICG solution (osmolarity 270 mOsm) was left in contact with the retina for 1-3 min. The outcome measures were hole closure, change in visual acuity, and macular pigmentary changes. RESULTS: Anatomical hole closure was achieved in 13 of 14 eyes (92.8%). The mean logMAR Snellen acuities were 0.80 (range 0.60-1.30) preoperatively and 0.77 (0.48-1.30) postoperatively. Seven eyes (50%) developed retinal pigmentary changes in the macular area. CONCLUSIONS: ICG-assisted ILM peeling for macular hole surgery achieves high rates of anatomical hole closure, but functional results are less encouraging. Previous studies suggest toxicity of the ICG to the retina, at the level of the RPE or inner retina. The results may be optimized by using a lower concentration, iso-osmolar, viscous solution, and by minimizing contact time of the solution and intensity, and duration of illumination.

Aged↗

QN1/KIAA1009: a new essential protein for chromosome segregation and mitotic spindle assembly.

We previously reported the involvement of QN1 (quail neuroretina 1) protein in cell cycle control during retinal development. We show here that QN1 is an ATPase conserved through evolution, from fugu to humans. We show that chicken/quail QN1 protein is orthologous to the KIAA1009 protein in humans, the function of which was not known. We demonstrate here for the first time that QN1/KIAA1009 protein is located at the spindle poles of the mitotic apparatus and at centrosomes during mitosis. The siRNA-mediated depletion of KIAA1009 led to abnormal mitosis with chromosome segregation defects and abnormal centrosome separation leading to the death of PC12 and MCF7 cells. Thus, QN1/KIAA1009 is a new microtubule-associated ATPase involved in cell division.

Adenosine Triphosphatases↗

Extreme altitude mountaineering and Type 1 diabetes; the Diabetes Federation of Ireland Kilimanjaro Expedition.

AIMS: To examine the effects of extreme altitude mountaineering on glycaemic control in Type 1 diabetes, and to establish whether diabetes predisposes to acute mountain sickness (AMS). METHODS: Fifteen people with Type 1 diabetes and 22 nondiabetic controls were studied during the Diabetes Federation of Ireland Expedition to Kilimanjaro. Daily insulin requirements, blood glucose estimations and hypoglycaemic attacks were recorded in diaries by the people with diabetes. The performance of blood glucose meters at altitude was assessed using standard glucose solutions. Symptoms of acute mountain sickness were recorded daily by people with diabetes and by the nondiabetic controls using the Lake Louise Scoring Charts. The expedition medical team recorded the incidence of complications of altitude and of diabetes. The final height attained for each individual was recorded by the expedition medical team and verified by the expedition guides. RESULTS: The final altitude ascended was lower in the diabetic than the nondiabetic group (5187 +/- 514 vs. 5654 +/- 307 m, P = 0.001). The mean daily insulin dose was reduced from 67.1 +/- 28.3-32.9 +/- 11.8 units (P < 0.001), but only 50% of recorded blood glucose readings were within the target range of 6-14 mmol/L. There were few hypoglycaemic attacks after the first two days of climbing. Both blood glucose meters tested showed readings as low as 60% of standard glucose concentrations at high altitude and low temperatures. The Lake Louise questionnaires showed that symptoms of AMS occurred equally in the diabetic and nondiabetic groups. There were two episodes of mild diabetic ketoacidosis; two of the diabetic group and three of the nondiabetic group developed retinal haemorrhages. CONCLUSIONS: People with Type 1 diabetes can participate in extreme altitude mountaineering. However, there are significant risks associated with this activity, including hypoglycaemia, ketoacidosis and retinal haemorrhage, with the additional difficulties in assessing glycaemic control due to meter inaccuracy at high altitude. People with Type 1 diabetes must be carefully counselled before attempting extreme altitude mountaineering.

Altitude↗

Targeted disruption of ATF4 discloses its essential role in the formation of eye lens fibres.

BACKGROUND: Activating transcription factor-4 (ATF4)--also termed CREB2, C/ATF, and TAXREB67--is a basic-leucine zipper (bZip) transcription factor that belongs to the ATF/CREB family. In addition to its own family members, ATF4 can also form heterodimers with other related but distinct bZIP proteins such as the C/EBP, AP-1 and Maf families, which may give rise to a variety of combinatorial diversity in gene regulation. In order to assess the in vivo essential role of ATF4, we have generated mice lacking ATF4 by gene targeting. RESULTS: ATF4-deficient mice exhibited severe microphthalmia. Although ATF4-deficient eyes revealed a normal gross lens structure up to embryonic day 14.5, later on the ATF4-deficient lens, degenerated due to apoptosis without the formation of lens secondary fibre cells. Retinal development was normal in the mutant mice. The lens-specific expression of ATF4 in the mutant mice led not only to the recovery of lens secondary fibres but also to the induction of hyperplasia of these fibres. CONCLUSION: These results demonstrated that ATF4 is essential for the later stages of lens fibre cell differentiation.

Activating Transcription Factor 4↗

[Early vitrectomy in the treatment of severe eye injuries].

Several authors regard early vitrectomy as the best guarantee of success in cases of severe ocular injury. The present authors reviewed their own group of 27 patients, which included 13 cases with intraocular foreign bodies, 6 cases of blunt trauma and 8 with perforating injuries. Aside from cases which later developed retinal complications, vitrectomy was performed in 25 patients, though rarely more than a month after injury. The mean duration of postoperative follow-up was 22 months (minimum 6 month, maximum 66 months). The results and the need for silicone oil injection are discussed and compared with the literature. The timing of vitrectomy is of major importance.

Adolescent↗

Peroxisomal straight-chain Acyl-CoA oxidase and D-bifunctional protein are essential for the retroconversion step in docosahexaenoic acid synthesis.

Docosahexaenoic acid (DHA, C22:6n-3) is essential for normal brain and retinal development. The nature and subcellular location of the terminal steps in DHA biosynthesis have been controversial. Rather than direct Delta4-desaturation of C22:5n-3, it has been proposed that this intermediate is elongated to C24:5n-3, desaturated to C24:6n-3, and "retroconverted" to DHA via peroxisomal beta-oxidation. However, this hypothesis has recently been challenged. The goal of this study was to determine the mechanism and specific enzymes required for the retroconversion step in human skin fibroblasts. Cells from patients with deficiencies of either acyl-CoA oxidase or D-bifunctional protein, the first two enzymes of the peroxisomal straight-chain fatty acid beta-oxidation pathway, exhibited impaired (5-20% of control) conversion of either [1-14C]18:3n-3 or [1-14C]22:5n-3 to DHA as did cells from peroxisome biogenesis disorder patients comprising eight distinct genotypes. In contrast, normal DHA synthesis was observed in cells from patients with rhizomelic chondrodysplasia punctata, Refsum disease, X-linked adrenoleukodystrophy, and deficiency of mitochondrial medium- or very long-chain acyl-CoA dehydrogenase. Acyl-CoA oxidase-deficient cells accumulated 2-5 times more radiolabeled C24:6n-3 than did controls. Our data are consistent with the retroconversion hypothesis and demonstrate that peroxisomal beta-oxidation enzymes acyl-CoA oxidase and D-bifunctional protein are essential for this process in human skin fibroblasts.

17-Hydroxysteroid Dehydrogenases↗

Spacrcan binding to hyaluronan and other glycosaminoglycans. Molecular and biochemical studies.

Photoreceptors project from the outer retinal surface into a specialized glycocalyx, the interphotoreceptor matrix (IPM), which contains hyaluronan (HA) and two novel proteoglycans, Spacr and Spacrcan. This matrix must be stable enough to function in the attachment of the retina to the outer eye wall yet porous enough to allow movement of metabolites between these tissues. How this matrix is organized is not known. HA is a potential candidate in IPM organization since biochemical studies show that these proteoglycans bind HA. RHAMM (receptor for HA-mediated motility)-type HA binding motifs (HABMs) are present in their deduced amino acid sequence and may be the sites of this HA interaction. To test this hypothesis, we subcloned three fragments of mouse Spacrcan that contain the putative HABMs. We found that each recombinant fragment binds HA. Binding decreased when residues in the HABMs were mutated. This provides direct evidence that the RHAMM-type HABMs in Spacrcan are involved in hyaluronan binding. Since chondroitin sulfate and heparan sulfate proteoglycans are important for retinal development and function, we also evaluated the binding of these recombinant proteins to heparin and chondroitin sulfates, the glycosaminoglycan side chain of these proteoglycans. We found that each recombinant protein bound to both heparin and chondroitin sulfates. Binding to chondroitin sulfates involved these HABMs, because mutagenesis reduced binding. Binding to heparin was probably not mediated through these HABMs since heparin binding persisted following their mutagenesis. These studies provide the first evidence defining the sites of protein-carbohydrate interaction of molecules present in the IPM.

Amino Acid Sequence↗

Retina-derived fetuin (RDF): analysis by immunocytochemistry, reverse transcriptase-polymerase chain reaction and Southern blot.

PURPOSE: This study was undertaken to determine the presence of retina-derived fetuin (RDF) protein and its message in retinal tissues and retinal pigment epithelial (RPE) cells. The techniques utilized in this study included light micros-copy, immunochemistry, Western blot, reverse transcriptase-polymerase chain reaction (RT-PCR) and Southern blot. METHODS: Retinal tissues and sections from embryonic, early postnatal and adult normal rats and retinal pigment epithe-lial (RPE) cells from postnatal rats were immunostained for fetuin with a polyclonal fetuin antibody and a peroxidase conjugated-secondary antibody using immunocytochemistry and Western blot analyses. The cDNA generated from RNA isolated from early postnatal rat retinas and RPE was probed with primers for rat fetuin, amplified by PCR and the PCR products were analyzed by Southern blot. RESULTS: Fetuin (RDF) was immunolocalized to cells of the neuroepithelium in retinas of early postnatal rats and most prominently in the nuclei and perinuclear region of cultured neonatal rat RPE cells. In adult retinas, ganglion cells, inner segments of photoreceptor cells, some components of the outer plexiform layer, ganglion cells and optic nerve processes were immunoreactive for the fetuin protein. As shown by Western blot, fetuin (RDF) was higher in embryonic and early postnatal retinas than in late postnatal retinas, indicating that this protein may be developmentally regulated. Using RT-PCR, the message for rat fetuin was demonstrated in the retina and RPE of normal postnatal rats. Southern blot confirmed that the PCR product from the retina and RPE was generated from rat fetuin mRNA as well as from rat liver, the primary source of fetuin. CONCLUSIONS: Fetuin, termed retina-derived fetuin (RDF), is reported for the first time in retinal tissues. Fetuin is a cysteine protease inhibitor that may play a role in support of neuronal cell survival during early retinal development and the maintenance of neuronal activity. RDF may interact with other growth factors and cytokines in providing trophic support for neurons and possibly other cells of the developing retina.

Animals↗

Morphologic evidence of photoreceptor differentiation of pinealocytes in the neonatal rat.

The pineal body and the retina of the neonatal Sprague-Dawley rat were studied by light and electron microscopy, and the morphologic differentiation of the parenchymal cells of the pineal body was compared with that of the developing photoreceptor cells of the retina. Between the ages of 4 and 12 days after birth, some of the developing pinealocytes were observed to become elongated and polarized, with their nuclei located at one pole. "Synaptic" ribbons were observed within the cell body. At the opposite pole the cells developed elongated cell processes that initially contained microtubules and ribosomes. These cell processes projected into luminal spaces and were attached by structures resembling zonulae adherentes to the adjacent cells. Extending from the tips of the cell processes, cilia with a 9 + 0 arrangement were observed. Lamellated and vesicular membranes were noted at the tips of the cilia. Such morphologic differentiation, however, could be observed only in rats younger than 17 days. Comparison of the morphologic features of the neonatal pinealocytes with those of the developing retinal photoreceptor cells showed much similarity. It is suggested that the pinealocytes of the neonatal rat undergo "photoreceptor-like" differentiation during a transient neonatal period. Such morphologic differentiation may provide an explanation for light-induced biochemical changes described in neonatal rats whose eyes had been enucleated.

Animals↗

Evolution of mutations conferring multidrug resistance during prophylaxis and therapy for cytomegalovirus disease.

In a human immunodeficiency virus-infected subject, cytomegalovirus (CMV) isolated 9 months after the patient began oral ganciclovir prophylaxis was resistant to ganciclovir and cidofovir and contained mutations in both UL97 and Pol coding regions. At 1 year, retinitis developed, which progressed despite intravenous ganciclovir followed by foscarnet and then cidofovir. A subsequent buffy coat virus isolate was resistant to all three drugs and contained new mutations in UL97 and Pol. By individually transferring the observed mutations to laboratory strain AD169, it was shown that a mutation at codon 603 of UL97 conferred resistance to ganciclovir, a mutation at codon 412 of Pol conferred resistance to both ganciclovir and cidofovir, and a mutation at codon 802 of Pol conferred resistance to ganciclovir and foscarnet. This case illustrates the development of multidrug resistance during prolonged exposure to antiviral therapy for CMV and cross-resistance arising from point mutations in the CMV Pol gene.

AIDS-Related Opportunistic Infections↗

Will dietary omega-3 fatty acids change the composition of human milk?

The most abundant long-chain polyunsaturated fatty acid in brain and retinal lipids is docosahexaenoic acid (DHA, C22:6 omega 3). It becomes incorporated into nerve tissues mostly in utero and during the 1st yr of life. DHA is derived in humans either performed in the diet or by hepatic synthesis from dietary linolenic acid (C18:3 omega 3). Since human milk contains DHA, this study was designed to see if increased dietary DHA would be reflected in a higher DHA content in human milk. Eight lactating women were given supplements of a fish oil concentrate rich in omega-3 fatty acids, including DHA (11% of fatty acids). Six women took 5 g/day of fish oil for 28 days; five women consumed 10 g/day for 14 days; and one woman consumed 47 g/day for 8 days. Each intake level of fish oil produced significant dose-dependent increases in the DHA content of milk and plasma. Base-line DHA levels in milk were 0.1 +/- 0.06% of total fatty acids. Five g/day of fish oil raised the levels to 0.5 +/- 0.1% (p less than 0.001); 10 g/day raised DHA levels to 0.8 +/- 0.1% (p less than 0.001); and 47 g/day produced DHA levels of 4.8%. The results of this study indicated that relatively low intakes of dietary DHA significantly elevated milk DHA content. This would clearly elevate the infant's DHA intake and might have implications for brain and retinal development.

Adult↗

Mutations in the human RAX homeobox gene in a patient with anophthalmia and sclerocornea.

Anophthalmia and microphthalmia are among the most common ocular birth defects and a significant cause of congenital blindness. The etiology of anophthalmia and microphthalmia is diverse, with multiple genetic mutations associated with each of these conditions, along with potential environmental causes. Based on findings that mutations in the Rx/Rax homeobox genes in mice and fish lead to defects in retinal development and result in animal models of anophthalmia, we screened 75 individuals with anophthalmia and/or microphthalmia for mutations in the human RAX gene. We identified a single proband from this population who is a compound heterozygote for mutations in the RAX gene. This individual carries a truncated allele (Q147X) and a missense mutation (R192Q), both within the DNA-binding homeodomain of the RAX protein, and we have characterized the biochemical properties of these mutations in vitro. Parents and grandparents of the proband were found to be carriers without visible ocular defects, consistent with an autosomal recessive inheritance pattern. This is the first report of genetic mutations in the human RAX gene.

Anophthalmos↗

Taurine: an essential nutrient for the cat.

Cats fed a purified diet containing purified casein as the source of protein develop retinal degeneration due to the lack of taurine in the diet. To test whether cats can synthesize this sulfur amino acid from sulfate or cystine, radioisotopes of these substances were injected into taurine-depleted and control cats. Sulfate did not serve as a precursor for taurine synthesis, whereas cystine underwent only a moderate conversion to taurine. This is in keeping with the low level of cysteinesulfinic acid (CSA) decarboxylase activity in cat liver. There was no difference between the activity of CSA decarboxylase in tissues from control cats and that in tissues from taurine-depleted cats. The pattern of tissue accumulation of [35S]taurine and from [35S]cystine also indicated that tissues from taurine-depleted cats do not synthesize [35S]taurine more rapidly than tissues from control cats. The data did not indicate a difference in taurine uptake by tissues of control and deficient cats, but progressive accumulation in deficient cats suggested that the turnover rate of taurine is decreased by the deficiency. Since supplementation of the purified diet with cysteine has been found previously to be inadequate to prevent progressive taurine depletion of the retina and its subsequent degeneration and since conversion of sulfur compounds to taurine in vivo is inadequate, taurine can be considered an essential nutrient for the cat.

Animals↗

Taurine deficiency in the kitten: exchange and turnover of [35S] taurine in brain, retina, and other tissues.

Kittens fed a purified diet containing partially-purified casein as the source of protein become taurine-deficient and develop retinal degeneration. The present studies report the exchange and turnover of taurine in different areas of brain, retina and other tissues and fluids of control and taurine-deficient kittens. The various tissues and fluids have different rates of exchange and different half-lives of taurine; taurine deficiency causes a range of changes in these parameters. Generally, tissues and fluids from taurine-deficient kittens accumulated more [35S] taurine and had a longer half-life of taurine than tissues and fluids from control kittens. Nine areas of brain were studied and, of these areas, olfactory bulb had the greatest concentration of taurine. Olfactory bulb resisted taurine depletion to a greater extent than other areas; and, in contrast to other areas of brain, in taurine-deficient kittens, it accumulated [35S] taurine for most of the experiment. Retina resisted taurine depletion and, in taurine-deficient kittens but not in control kittens, it accumulated [35S] taurine throughout the experiment. The amount of taurine conjugated to bile acids was unchanged by the taurine depletion, but the kinetic behavior was altered and was similar to that of retina. The results provide support for the suggestion that, in the kitten at least, taurine is most important for the functions of bile and retina and that taurine depletion affects retina before bile. Taurine may have special importance in olfactory bulb but not necessarily in other regions of brain.

Adrenal Glands↗

Factors affecting long-chain polyunsaturated fatty acid composition of plasma choline phosphoglycerides in preterm infants.

Long-chain polyunsaturated fatty acid (LCPUFA) composition of choline phosphoglycerides was measured in the plasma of 22 preterm infants at birth and at expected date of delivery (EDD). In a subgroup of 10 infants, data were also collected at regular intervals between birth and EDD. Levels at birth showed a positive correlation between arachidonic acid (AA) and birth weight, p less than 0.01, and between docosahexaenoic acid (DHA) and gestational age, p less than 0.01. Percentage compositions of both AA and DHA fell rapidly between preterm birth and expected date of delivery, at a time when they would remain high in utero. For AA, the mean value fell from 16.52 to 7.18%, and for DHA from 4.49 to 2.63%. Levels of DHA fell less in babies fed breast milk than in those fed formula milk, p less than 0.05, and levels of AA fell less in those requiring a large number of blood transfusions, p less than 0.05. The level of DHA fell more in those infants with intrauterine growth retardation, p less than 0.05. Although these nutrients share common metabolic pathways for their synthesis, they appear to be affected by different factors in both the fetus and the preterm infant. Low levels at this time may adversely affect brain and retinal development.

Age Factors↗