Pharmacotherapies for glaucoma

Glaucoma is a group of progressive optic neuropathies in which the axons in the optic nerve are injured, retinal ganglion cell numbers are reduced and vision is gradually and permanently lost. The only approved and effective way to treat glaucoma is to reduce the intraocular pressure (IOP). This is usually accomplished by surgical and/or pharmacological means. Drugs designed to reduce IOP target one or more of the parameters that maintain it. These parameters (collectively known as aqueous humor dynamics) are the production rate of aqueous humor, the pressure in the episcleral veins and the drainage of aqueous humor through the trabecular or uveoscleral outflow pathways. Intraocular pressure lowering drugs can be classified as inflow or outflow depending on whether they reduce aqueous humor inflow into the anterior chamber or improve aqueous humor outflow from the anterior chamber. Inflow drugs, like β adrenergic antagonists and carbonic anhydrase inhibitors, reduce the rate of aqueous humor production. Outflow drugs, like prostaglandin analogs, cholinergic agonists and sympathomimetics, increase the rate of drainage through the uveoscleral outflow pathway and/or increase the facility of outflow through the trabecular meshwork. Some drugs have mixed inflow/outflow effects. This review summarizes the pharmacological treatments for glaucoma in use today and some new drugs showing potential for use in the future.     

Transplantation immunology of the anterior chamber of the eye. II. Immune response to allogeneic cells.

The mechanism by which the anterior chamber of the eye extends immunologic privilege to allogeneic donor tissues has been studied in inbred rats. Inoculation of allogeneic lymphoid cells into the anterior chamber demonstrated that although the site lacks a lymphatic drainage, the afferent limb of the immunologic reflex arc is intact because the recipient can recognize and mount a specific immune response. In addition, host immunity was able to express itself within the anterior chamber when induced systemically, indicating that the efferent limb of the reflex arc is also intact. Therefore, it is suggested that the unique immunologic features of the anterior chamber may result from the obligate intravenous presentation of graft antigen to the host's systemic immunologic apparatus, a route that prejudices the host's response in the direction of tolerance and/or enhancement rather than cell-mediated, tissue destructive immunity.     

Noninvasive high-resolution in vivo imaging of cell biology in the anterior chamber of the mouse eye

There is clearly a demand for an experimental platform that enables cell biology to be studied in intact vascularized and innervated tissue in vivo. This platform should allow observations of cells noninvasively and longitudinally at single-cell resolution. For this purpose, we use the anterior chamber of the mouse eye in combination with laser scanning microscopy (LSM). Tissue transplanted to the anterior chamber of the eye is rapidly vascularized, innervated and regains function. After transplantation, LSM through the cornea allows repetitive and noninvasive in vivo imaging at cellular resolution. Morphology, vascularization, cell function and cell survival are monitored longitudinally using fluorescent proteins and dyes. We have used this system to study pancreatic islets, but the platform can easily be adapted for studying a variety of tissues and additional biological parameters. Transplantation to the anterior chamber of the eye takes 25 min, and in vivo imaging 1-5 h, depending on the features monitored.     

Ultrastructure of the swim bladder of the goldfish,Carassius auratus

Summary The swim bladder of the cyprinid Carassius auratus (goldfish) is a two-chambered organ connected to the esophagus by a pneumatic duct. The anterior chamber is lined by a single type of squamous epithelial cell. Two types of epithelial cells are present in the posterior chamber. Flattened cells with differences in the electron density of the cytoplasm line most of the chamber. Darker cells generally contain large amounts of glycogen. Cuboidal epithelial cells also occur in the posterior chamber. A glandular layer external to the muscularis in the posterior chamber is composed of large cells containing little glycogen, an extensive Golgi apparatus, and numerous mitochondria with single large granules. Capillaries and nerves are present in large numbers in this layer. Blood vessels form micro-retia mirabilia in the submuscular layer external to the glandular layer. Vessels are of two distinct types with wide lumina and flattened endothelium characterizing the venous vessels. Arterial vessels have smaller lumina, thick endothelial cells with prominent pinocytotic vesicles, and surrounding pericytes. Collagen is present in three forms in this swim bladder — large tactoids in the tunica externa of the anterior chamber, smaller tactoids in the lamina propria of the posterior chamber, and small fibrils in all other areas.Supported by a Young Investigator Pulmonary Research Grant # 1 R23 HL 19593-01 and by HL 23338-01 from the National Institutes of Health     

Studies on water and ion transport in homopteran insects: ultrastructure and cytochemistry of the cicadoid and cercopoid Malpighian tubules and filter chamber

The filter chamber is a complex junction of anterior and posterior extremities of the midgut and Malpighian tubules. The sac-like anterior extremity, or filter chamber proper, comprises two cell types. These are large cuboidal cells which secrete a mucoprotein, and extremely thin cells which have regular tubular invaginations of the basal plasma membrane. The posterior extremity of the midgut and the internal Malpighian tubules coil round the filter chamber proper. They consist of thin epithelial cells identical in ultrastructure. The basal plasma membrane in these cells is formed into leaflets. A thin cellular sheath and thick muscle layers surround the filter chamber. The filter chamber proper is lined by the mucoprotein secretion of the cuboidal cells. This secretion appears to bind potassium ions. ATPase and alkaline phosphatase cannot be detected in the filter chamber epithelia. The structure and cytochemistry of the filter chamber suggests that water flows from filter chamber proper to midgut and Malpighian tubules by passive osmosis. This may be facilitated by ion binding in the filter chamber proper and by hydrostatic pressure engendered by contraction of the muscular coat. The Malpighian tubules appear to be structurally and chemically adapted for ion secretion by active transport and possibly for reabsorption in the Malpighian duct segment.     

THE MECHANISM OF AQUEOUS HUMOR FORMATION INFERRED FROM CHEMICAL STUDIES ON BLOOD-AQUEOUS HUMOR DYNAMICS

The importance of considering the effect of a possible flow out of the anterior chamber before inferring any mechanism of aqueous humor formation from the relative concentration of a substance in the aqueous humor and plasma under equilibrium conditions has been stressed. Several processes to account for the chemical equilibria between aqueous humor and blood based on the ultrafiltration and secretion hypotheses with a possible simultaneous loss of aqueous humor by flow have been outlined. On the basis of these processes, equations were formulated which would relate the rates of transfer into and out of the anterior chamber to the ratio of concentration of a substance in the aqueous to that in the blood at various intervals after its introduction into the blood. The explanation of equilibrium ratios above and below one for aqueous constituents is made apparent from the mathematical formulations. For each substance tested a determination was made of the best fit when the concentration in the aqueous humor is plotted against time. This fit was obtained by plotting the rate of transfer in against the rate of transfer out of the anterior chamber for all of the experimentally found concentration ratios on the basis of both the ultrafiltration and secretory hypotheses. Two sets of values were obtained from these calculations, one set for each hypothesis. The substantial agreement of all the experimental data with an assumed rate of leakage out of the anterior chamber of approximately 4 c. mm. per minute was shown to be compatible only with the idea that all the monovalent electrolytes tested entered the anterior chamber as a result of secretory process. It could not be decided from these chemical studies whether the non-electrolytes and the one multivalent electrolyte tested enter the anterior chamber by ultrafiltration or secretion. Experimental findings from other sources were cited which would suggest that non-electrolytes enter the anterior chamber as a result of ultrafiltration. The implications of the mechanism outlined in the paper with respect to intraocular pressure have been discussed. Supplementary evidence from the literature has been given in support of the conclusions presented here.     

Immune response to immunization via the anterior chamber of the eye. I. F. lymphocyte-induced immune deviation.

The immunizing abilities of alloantigens placed within the anterior chamber of the eye have been studied in inbred rats. Although intracameral inoculation of F1 hybrid lymphocytes into parental strain recipients elicited both cell- and antibody-mediated immunity, a delimited interval was identified postinoculation during which the systemic cell-mediated immune response was suppressed as indicated by prolonged acceptance of orthotopic skin allografts. The prompt appearance of hemagglutinating antibodies in the serum of immunized rats followed a time course which coincided with the suppression of cell-mediated immunity and suggested that the two events are casually related. Since exposure to allogeneic antigens on lymphoid cells via the anterior chamber elicits a transient suppression in cell-mediated immunity, where humoral immunity is preserved, the phenomenon resembles immune deviation.     

Mathematical model of flow in the vitreous humor induced by saccadic eye rotations: effect of geometry

Saccadic eye rotations induce a flow in the vitreous humor of the eye. Any such flow is likely to have a significant influence on the dispersion of drugs injected into the vitreous chamber. The shape of this chamber deviates from a perfect sphere by up to 10–20% of the radius, which is predominantly due to an indentation caused by the lens. In this paper we investigate theoretically the effect of the domain shape upon the flow field generated by saccades by considering an idealized model. The posterior chamber geometry is assumed to be a sphere with a small indentation, undergoing prescribed small-amplitude sinusoidal torsional oscillations, and, as an initial step towards understanding the problem, we treat the vitreous humor as a Newtonian fluid filling the chamber. The latter assumption applies best in the case of a liquefied vitreous or a tamponade fluid introduced in the vitreous chamber after vitrectomy. We find the flow field in terms of vector spherical harmonics, focusing on the deviation from the flow that would be obtained in a perfect sphere. The flow induced by the departure of the domain geometry from the spherical shape has an oscillating component at leading order and a smaller-amplitude steady streaming flow. The oscillating component includes a circulation cell formed every half-period, which migrates from the indentation towards the center of the domain where it disappears. The steady component has two counter-rotating circulations in the anterior part of the domain. These findings are in good qualitative agreement with the experimental results of Stocchino et al. (Phys Med Biol 52:2021–2034, 2007). Our results predict a significant reduction in the expected time for drug dispersal across the eye compared with the situation in which there is no fluid flow present.     

The gas bladder of Argentina silus L., with special reference to the ultrastructure of the gas gland cells and the countercurrent vascular bundles

Summary The orifice between the two chambers of the gas bladder in Argentina silus is surrounded by a sphincter muscle. Gas analyses of the gas bladder contents of fish from 400 meters depth give 0–1% carbon dioxide and 9–72% oxygen. Micro-retia mirabilia form a countercurrent vascular system, and the arterial component has peripherally a sphincter mechanism. The function of the glandular layer of the anterior chamber remains uncertain, but the structure indicates secretion into blood capillaries. The lining epithelium of the anterior chamber may secrete some substance into blood or directly into the lumen, which may be involved in a secretory mechanism. This conclusion is not supported by our histochemical tests. The posterior chamber has no micro-retia and the blood vessels have a different origin from those of the anterior chamber. The blood vessels form a plexus of capillaries or sinuses in contact with the flat lining epithelium, thus allowing gases to pass freely by diffusion. — The muscular layers of both chambers are innervated by catecholamine-containing nerve fibres.The investigation was supported by grants from the Swedish Natural Research Council (No. 99-35) and by the Faculty of Mathematics and Science, University of Lund.     

Pseudorhabdosynochus venus n. sp. (Monogenea: Diplectanidae) from Epinephelus howlandi (Perciformes: Serranidae) off New Caledonia

Pseudorhabdosynochus venus n. sp. is described from specimens collected from the gills of Epinephelus howlandi off Nouméa, New Caledonia, South Pacific. The male sclerotised quadriloculate organ of P. venus, 61-101 microm in internal length, has an anterior chamber with a thin anterior wall, a very short cone and a short posterior tube. The sclerotised vagina, 50-67 microm in total length, is composed of an anterior open trumpet, an S-shaped canal, a tear-shaped principal chamber and a spherical accessory chamber; all parts are heavily sclerotised. The two squamodiscs have 10-11 rows of separate rodlets and no central closed row of rodlets. P. venus is differentiated from all other species of Pseudorhabdosynochus by the spectacular morphology of its sclerotised vagina. It is the first diplectanid described from E. howlandi.     

Developmental studies on the loricate choanoflagellateStephanoeca diplocostata ellis

Summary Cells ofStephanoeca diplocostata comprise a colourless, flagellated, protoplast lodged in a lorica made of siliceous costae. The single anterior flagellum creates a water current from which bacteria and other food particles are filtered by the collar and ingested by linguiform pseudopodia that arise from the protoplast at the base of the collar. A waist divides the lorica into two chambers, the anterior of which contains three transverse and 17–20 longitudinal costae whereas the posterior chamber comprises two systems of spirally deflected costae and on some cells a pedicel at the hind end. Between 150–185 costal strips of similar length form the lorica. A thin investment covers the inner surface of the posterior chamber and lower part of the anterior chamber and joins with the protoplast at the level of the waist. Costal strips are produced within membrane-bounded vesicles in the peripheral cytoplasm and, although the origin of these vesicles is unknown, there is usually a close association with the Golgi apparatus. Once complete, strips are apparently released sideways through the plasmalemma into the cavity of the posterior lorica chamber. Later, bundles of strips are transferred to the top of the inner surface of the collar where they collectively form a horizontal ring. When sufficient strips to form a lorica have been accumulated at the top of the collar, cell division proceeds.     

The chemical composition and the osmotic pressure of the aqueous humor and plasma of the rabbit

Measurements were made of the osmotic pressure of plasma, and of aqueous humor taken from the anterior chamber of the right and left eyes and from the posterior chamber of unanesthetized rabbits. Aqueous humor from the anterior chamber was found to be hypertonic to the plasma by approximately 3 mM/liter equivalent of sodium chloride. The aqueous humor from the anterior and posterior chambers of the right and left eyes was isotonic. The concentration of chloride in the anterior and posterior chambers was the same. The concentration of all the major components of the aqueous humor and plasma has been determined by chemical analysis on fluid samples obtained from unanesthetized rabbits at approximately the same time. The calculated osmotic pressure of the total of these substances in terms of sodium chloride equivalent agrees to within better than 1 per cent of the total osmotic pressure as measured experimentally. The distribution of some individual anions and cations of the aqueous humor and plasma was determined. This distribution is widely different from that which would obtain at a state of equilibrium. The positive and negative charges carried by the ions in the aqueous humor were approximately equal. Sources of error in the experiments are discussed.     

Structural organization of the copulation site in the medfly Ceratitis capitata (Diptera: Tephritidae) and observations on sperm transfer and storage

The copulation site of the medfly Ceratitis capitata was investigated at anatomical and ultrastructural levels. It consists of the anterior vagina, with a ventral fertilization chamber and a dorsal insemination pocket into which the two spermathecal ducts open. The fertilization chamber is an organ comprised of a number of alveoli that in virgin females are filled with a filamentous secretion, whereas in mated females contain sperm bundles. Through study of the internal morphology of the aedeagus, its position in the anterior vagina, and the direct observation of sperm transfer and storage, we confirmed that sperm are ejaculated through two gonopores at the top of the distiphallus and another at the base of the genital rod. The sperm flow dorsally into the insemination pocket and ventrally into the fertilization chamber. During copulation, the two spermathecae and the fertilization chamber are progressively filled with spermatozoa.     

An intracameral injection of antigen induces in situ chemokines and cytokines required for the generation of circulating immunoregulatory monocytes

Anterior Chamber-Associated Immune Deviation (ACAID) induced by an intracameral injection of antigen generates antigen-specific regulatory splenic T cells that suppress specifically cell-mediated immunity specific for the injected antigen. Circulating F4/80(+) cells recovered from mice receiving an intracameral injection of antigen are thought to be ocular in origin and induce the development of thymic and splenic regulatory T cells. We have shown previously that after the intracameral injection of antigen there is a CCR2/CCL2-dependent infiltration of circulating F4/80(+) cells into the anterior chamber associated with the generation of circulating, ACAID-inducing F4/80(+) monocytes. Here we tested the hypothesis that the intracameral injection of antigen induces events in the anterior chamber that are associated with the induction of circulating immunoregulatory monocytes that induce the suppression of cell-mediated immunity. The intracameral injection of antigen resulted in aqueous humor (i) a time- dependent increase of CCL2 and CCL7, (ii) a transient increase in TNF-α, and (iii) an infiltration of CD11b(hi), Gr1(hi) and F4/80(+) as well as F4/80(-) and Gr1(hi) peripheral blood cells into the anterior chamber. Further characterization of these F4/80(+) cells revealed that they are Ly 6C(hi), LY6G(lo) or negative, 7/4 (LY6B)(hi), CD115(+), CD45(+), CD49B(+), and CD62 L(+). Antibody-mediated neutralization of TGF-β in situ in the anterior chamber prevented the induction of circulating, ACAID-inducing monocytes and ACAID. These cells did not increase in the irides of ACAID-refractory CCR2-/- and CCL2-/- mice that received an intracameral injection of antigen. Our results extend our suggestion that ACAID is initiated as the result of a mild proinflammatory response to intracameral injection that results in the infiltration of a CCR2(+) subset of monocytes into the anterior chamber where there is a TGF-β-dependent induction of an immunosuppressive phenotype in the infiltrated monocytes that recirculate to induce antigen-specific regulatory T cells.     

Lattice organs in y-cyprids of the Facetotecta and their significance in the phylogeny of the Crustacea Thecostraca

Scanning and transmission electron microscopy (SEM and TEM) were used to study lattice organs in facetotectan y‐cyprids from the White Sea and from Norwegian and Bahamian waters. The larvae represent at least four and possibly five different species of Facetotecta. Y‐cyprids have five pairs of lattice organs in the head shield (carapace) organized into two anterior pairs and three posterior pairs. Both groups of lattice organs are arranged around a large central pore. The facetotectan lattice organs are elongate areas with a longitudinal keel, just as in the Ascothoracida and some Cirripedia Acrothoracica. The terminal pore of the organs is situated posteriorly in all five pairs. TEM confirms that the organs have the same general morphology as in the Cirripedia and Ascothoracida, namely, a cuticular chamber into which project ciliary segments from the chemosensory cells. Unlike Cirripedia the cuticular roof of the chamber lacks any pores. We conclude that five pairs of lattice organs represent an autapomorphy for the Thecostraca, which supports the monophyly of this taxon. In the ground pattern the terminal pore is posterior in all five pairs. The anterior position of the pore in lattice organ pair 2 is apomorphic for the Cirripedia, while within this taxon an anterior position also in pair 1 is apomorphic for a monophylum comprising the Thoracica and the Rhizocephala. Minute pores in the roof of the organs is another apomorphy of the Cirripedia, but its elaboration into pores visible with SEM may have been subject to some homoplasy. Since lattice organs are omnipresent in the settling instar of the Thecostraca they probably serve a critical role for the function of these cypris or cypris‐like larvae.     

Morphogenesis of the eye of Siberian sturgeon

The most relevant changes in Acipenser baeri eye organization were detected between hatching and 5 days post hatch. At this age, the eye had an anterior chamber, lens, iris, choroid gland, scleral cartilage, cornea and a vitreous chamber lined by the retina (with two photoreceptors: rods and single cones).     

Transmission of 308 nm excimer laser radiation for ophthalmic microsurgery--medical, technical and safety aspects

A surgical system using 308 nm excimer laser radiation transmitted by quartz fibers is described. In-vitro surgery has been performed on the cornea, in the anterior chamber angle, on the lens and in the vitreous with a special application tip for each procedure. Possible damage of ocular structures by 308 nm radiation is discussed. In-vitro experiments showed that UV exposure of intraocular structures sensitive to 308 nm radiation can be reduced by appropriate design of the application tips and by the application of UV absorbing drugs.     

The ultrastructure of the swimbladder of the toadfish,Opsanus tau L.

Summary The anterior chamber of the swimbladder of the toadfish Opsanus tau L. is lined by a single layer of columnar gas gland cells, cuboidal cells that resemble gas gland cells but are located outside of the gas gland region, and squamous cells. Multilamellar bodies are numerous in the gas gland cells and the cuboidal cells and are present in smaller numbers in the squamous cells. Capillaries lie in the lamina propria directly below the epithelial lining. A thick continuous muscularis mucosae and a submucosa consisting of tightly packed cells, cell processes, and connective tissue may contribute to the impermeability to gases of the wall of the anterior chamber.The posterior chamber of the swimbladder is lined by a single type of squamous epithelial cell. Multilamellar bodies were occasionally observed in these cells also. Other types of cells frequently form a partial second layer between the epithelial lining and the basement lamina. A thin muscularis mucosae lies directly below the basement lamina and the capillaries of the posterior chamber are located in the submucosa. The tunica externa is a layer of dense connective tissue that surrounds both the anterior and posterior chambers. Collagen fibrils in the form of tactoids are present in this layer.Part of this work was submitted by S.M.M. in partial fulfillment of the requirements for the degree of Doctor of Philosophy, Biology Department, Boston University. S.M.M. is grateful for a National Science Foundation Traineeship.     

Morphology and ultrastructure of the alimentary canal of the cicada Platypleura kaempferi (Hemiptera: Cicadidae)

The alimentary canal of cicada Platypleura kaempferi is described. It comprises the oesophagus, filter chamber, external midgut section and hindgut. The elongate oesophagus expands posteriorly, with its posterior end constricting to become a bulb. The filter chamber consists of two parts: a very thin sheath and a filter organ. The filter organ is composed of the anterior and posterior ends of the midgut (internal midgut section), and the internal proximal ends of the Malpighian tubules. The external midgut section differentiates into a collapsed sac and a midgut loop. The latter is divided into three distinct segments. The hindgut contains a dilated rectum and a long narrow ileum. The distal portions of the four Malpighian tubules are enclosed in a peritoneal sheath together with the distal ileum before reaching to the rectum. Ultrastructurally, the oesophagus and the hindgut are lined with a cuticle. The filter chamber sheath consists of cells with large irregular nuclei. Filamentous substances coat the microvilli of the cells of the internal midgut section. The posterior end of the midgut comprises two types of cells, with the first type of cells containing many vesicles and scattered elements of rough endoplasmic reticulum. The anterior and posterior segments of the midgut loop cells have ferritin‐like granules. The ileum cells have well‐developed apical leaflets associated with mitochondria. Accumulations of virus‐like particles enclosed in the membrane are observed in the esophagus, conical segment, mid‐ and posterior segments of the midgut loop.     

In vivo antivascular endothelial growth factor treatment induces corneal endothelium apoptosis in rabbits through changes in p75NTR–proNGF pathway

Intravitreal injection (IVT) of antivascular endothelial growth factor (anti‐VEGF) agents is widely used for the treatment of retinal vascular diseases. Recently, the injection of anti‐VEGF agents in the ocular anterior chamber has been proposed for the treatment of neovascular glaucoma and potential side effects on the corneal structures have been investigated with contrasting results. Increasing evidence has demonstrated that VEGF inhibition is associated with cellular apoptotic changes and that this effect may be mediated by alterations in nerve growth factor (NGF) pathway. In this study, we demonstrated that anterior chamber injection (IC), but not IVT injection of two different anti‐VEGF agents, aflibercept and ranibizumab, affects rabbit corneal endothelium in terms of survival and apoptosis and is associated with changes in endothelial expression of NGF precursor (proNGF) and p75 neurotrophin receptor (p75NTR) receptor. We observed an increase in corneal endothelial cell incorporation of trypan blue and expression of cleaved‐caspase 3 (c‐Casp3), p75NTR, and RhoA after IC injection of both anti‐VEGF drugs when compared with the vehicle. Our results showed that apoptosis induction by aflibercept was more pronounced when compared with that of ranibizumab. Aflibercept also mediated a significant increase in endothelial expression of proNGF when compared with the vehicle. In line with these data, IC administration of both anti‐VEGF agents induced the activation of apoptotic signals in endothelial cells, including an increase in c‐Casp3, decrease in Bad Ser 112 phosphorylation, and unbalance of AKT phosphorylation. These results demonstrated that administration of anti‐VEGF in the anterior chamber of rabbit affects endothelial cell survival by inducing apoptosis through alteration of NGF pathway.