Dietary Lactobacillus casei can be used to influence intraepithelial lymphocyte migration and modulate mucosal immunity in chicks.

1. The role of probiotics in modulating intestinal mucosal immunity in chicks was investigated by measuring migration of intraepithelial lymphocytes (IEL) and cytokine signals in chicks fed on a diet supplemented with the Lactobacillus casei compared with those of chicks fed on an unsupplemented diet.2. Increased numbers of intraepithelial lymphocytes (IEL) were detected in the ileal epithelium at d 3 and d 7 after feeding a diet containing 108 CFU/g L. casei.3. Greater expression of chemokine genes for C-C motif chemokine ligand 3, C-X-C motif chemokine ligand 12, C-C motif chemokine receptor 5, and C-C motif chemokine receptor 9 were detected in the ileum on d 3, suggesting a greater number of IEL was associated with lymphocyte migration through the chemokine signalling pathway.4. After IEL migration, cell proliferation was evident in mucosal epithelial cells on d 14. Evidence of immune responses induced in the ileum from d 3-21 after feeding the diet containing L. casei was shown by the significant (P < 0.05) differences in transforming growth factor-ß, secretory immunoglobulin A, interferon-γ, tumour necrosis factor-α, interleukin-4, and interleukin-10.5. These results indicated that feeding L. casei helps guide IEL migration and modulates intestinal mucosal immunity.

Radiologic findings in infected and noninfected scleral buckles.

PURPOSE: To present the radiologic findings in scleral buckle infections and in the early postoperative period after scleral buckling. METHODS: Retrospective multicenter orbital computed tomography (CT) study of 14 patients and brain magnetic resonance (MR) in one patient with scleral buckle infections, some with the referring diagnosis of endophthalmitis, proliferative vitreoretinopathy, orbital cellulitis, or unilateral headache. The control population consisted of early postoperative prospective CT study of 38 consecutive patients with scleral buckle without clinical infection. RESULTS: Diffuse scleral thickening and preseptal soft tissue swelling were noted in acute scleral buckle infections. Scleral thickening decreased radiologically following prompt antibiotic therapy in five patients with acute infections. Silicone sponge had low attenuation without infection and high attenuation with infection. In chronically infected scleral buckle, the sclera was thickened around the buckle, with scleral melt under the buckle. MR showed increased signal intensity in the preseptal region in one patient with chronic fungal infection. In the controls, two had thickening of the sclera without soft tissue swelling. CONCLUSIONS: CT or MR can assist in the early diagnosis and management of scleral buckle infections.

Development of a safe and rapid neutralization assay using murine leukemia virus pseudotyped with HIV type 1 envelope glycoprotein lacking the cytoplasmic domain.

Neutralizing antibody (NAb) is a critical component of an immune system that can potentially provide sterilizing protection against human immunodeficiency virus type 1 (HIV-1). Therefore, an in vitro assay that can rapidly, safely, and accurately evaluate the NAb response vaccine candidates elicit, especially against a large number of HIV-1 variants, would be highly valuable. It has been demonstrated that HIV-1 envelope glycoprotein lacking the cytoplasmic domain can pseudotype murine leukemia virus encoding the beta-galactosidase gene and that this pseudovirus can specifically infect CD4(+) cells (Schnierle BS, Stitz J, Bosch V, et al.: Proc Natl Acad Sci USA 1997;94:8640-8645). Because the pseudovirus is not biohazardous and because the infection can be quantitatively determined within 2 days, we examined the feasibility of using the pseudovirus for high-throughput neutralization assays for HIV-1. We have generated viruses pseudotyped with gp140 of six different HIV-1 isolates (LAI, RF, Bal, AD8, 89.6, and DH12). All six pseudoviruses were infectious and exhibited expected coreceptor usage phenotype in HOS-CD4 cells expressing either CCR5 or CXCR4. More importantly, the neutralization sensitivity profile of these pseudoviruses was virtually identical to that observed from more conventional neutralization assays using either HIV-1 or SHIV. All pseudoviruses could be neutralized by broadly reactive human monoclonal antibody IgG1 b12. Our results indicate that the pseudoviruses are ideal for high-throughput evaluation of immune sera for their capacity to broadly neutralize a large number of HIV-1 isolates.

11-cis retinol dehydrogenase mutations as a major cause of the congenital night-blindness disorder known as fundus albipunctatus.

PURPOSE: Patients with fundus albipunctatus uniformly experience difficulty with vision at night. Their retinas are spotted with characteristic light yellow flecks of unknown composition that typically spare the macula. A defect in the transport or utilization of visual cycle retinoids is thought to underlie this recessive disorder with variable clinical expression. To elucidate the molecular defect we considered the genes for interphotoreceptor retinoid-binding protein (RBP3) and 11-cis retinol dehydrogenase (RDH5) as candidates for this disease. METHODS: We examined two unrelated families with fundus albipunctatus. The diagnosis was determined clinically and RBP3 and RDH5 were analyzed by molecular screening methods and direct genomic sequencing. RESULTS: Each family had two affected members with typical fundus albipunctatus. The affected members were siblings born to unaffected parents who were seventh cousins in the first family and unrelated in the second family. The probands from both families were clinically similar except for the fundus dots that were more extensive in the second family to the point of involving the parafoveal region. In the initial phase of genetic screening RBP3 defects were ruled-out as the cause of the disease in both families. In contrast, RDH5 mutations were found in the affected siblings in both families. The proband in one had a homozygotic Gly238Trp missense mutation (GGG -> TGG) involving exon 4 and in the other carried compound heterozygotic changes Arg280His (CGC -> CAC) and Ala294Pro (GCC -> CCC) in exon 5. The disease phenotype was only manifested in family members with two abnormal RDH5 alleles consistent with autosomal recessive inheritance in both pedigrees. CONCLUSIONS: These findings strongly implicate defects of RDH5 as the cause of fundus albipunctatus and point to a heterogeneity of RDH5 mutations in this form of congenital stationary night blindness with variable expressivity.

The new genotypic human calicivirus isolated in Seoul.

A new type of human calicivirus (HuCV) showing the classic cup-shaped surface morphology was identified in the stool sample from a child with symptoms of acute gastroenteritis in Seoul, Korea (SK virus). Genomic RNA was extracted directly from the stool sample, and the nucleotide sequence of 3.2 kb of the 3' end of SK virus was determined from cDNA. This region spanned sequences from the RNA-dependent RNA polymerase (RDRP) region in the open reading frame 1 (ORF1) to the 3' poly A tail. The non-structural and capsid protein coding sequences were fused in a single ORF as observed in Manchester type (Genogroup III). However, ORF2 of Manchester virus was missing in SK virus. In RDRP region, SK virus showed amino acid and nucleotide identities of 74-75% and 68-69% respectively, with those of Manchester virus, while showed 34-46% and 55-60% identities respectively with those of other human caliciviruses. However, capsid protein of SK virus showed a partial (29-46%) amino acid identity with those of other caliciviruses including Manchester type. The closest resemblance in amino acid (97-99%) and nucleotide sequence (85-86%) identities were found in RDRP region with Vanderbijlpark and Pretoria isolates recently found in South Africa. These results suggest that SK virus together with Vanderbijlpark and Pretoria isolates belong to a new type different from Manchester virus.

Electro-oculography in autosomal dominant vitreoretinochoroidopathy.

Thirteen members of a family presumed to be harboring the gene for autosomal dominant vitreoretinochoroidopathy were examined. In four affected members, electro-oculography demonstrated marked reduction of the Arden ratio (range, 1.1 to 1.5; normal, > or = 1.8), despite electroretinographic evidence of mildly affected rod function and normal cone function. These findings suggested that a diffuse disturbance of the photoreceptor-pigment epithelium complex may have been present prior to wide-spread loss of photoreceptor function in the affected members of this family. As in previously described families, the pattern of inheritance appeared consistent with autosomal dominance.

Experimental transconjunctival diode laser retinal photocoagulation through silicone scleral exoplants.

OBJECTIVE: To study the feasibility of inducing a chorioretinal lesion under a previously placed scleral buckle by experimental transconjunctival diode laser photocoagulation. METHODS: We performed transconjunctival diode laser photocoagulation in the peripheral retinas of seven pigmented rabbit eyes with a silicone exoplant (No. 42 band or No. 276 tire) and seven eyes without an exoplant. Each eye received burns with an intensity of grades 1 to 3 in different quadrants at varying power levels, with a 0.5-second duration and 650-micron spot size. Eyes were enucleated for histopathologic studies 1 day and 1 week after treatment. RESULTS: Although the irradiance emitted through the No. 42 band and the No. 276 tire was attenuated by 17% and 23%, respectively, the range of threshold powers needed to produce grades 1 to 3 burns was similar between eyes with and without a silicone exoplant. At 1 day, full-thickness coagulative necrosis was observed in all lesions, except that the ganglion cell layer and inner nuclear layer were preserved in two of four grade 1 burns and the ganglion cell layer was intact in one of six grade 2 burns. Inner scleral changes were noted acutely in three of five grade 3 lesions. At 1 week, burns of all intensity grades showed a full-thickness atrophic chorioretinal lesion with inner scleral changes. CONCLUSIONS: Experimental transconjunctival diode laser photocoagulation through hard silicone elements reproducibly created a chorioretinal lesion with histopathologic findings similar to those of lesions obtained without these elements. Although retinal photocoagulative effects were prominent, inner scleral abnormalities were also observed histologically.

A study of surgical approaches to retinal vascular occlusions.

OBJECTIVE: To develop a surgical approach to retinal vascular occlusive diseases. METHODS: Surgical manipulations were performed on the retinal vasculature to explore the feasibility of retinal vascular surgery. In a human cadaver eye model (25 procedures, 21 eyes), we performed (1) cannulations of retinal blood vessels with a flexible stylet and (2) arteriovenous sheathotomies. Histological findings were correlated with surgical outcomes. In an in vivo model (6 eyes, 5 animals), we examined the technical feasibility and anatomical outcome of surgical penetration of retinal blood vessels. RESULTS: Cannulations of branch retinal arterioles were successful in 7 of 9 procedures, cannulations of branch retinal venules were successful in 1 of 3 procedures, cannulations of central retinal arteries were successful in 0 of 2 procedures, and cannulations of central retinal veins were successful in 2 of 4 procedures. Arteriovenous sheathotomies were successful in 4 of 7 procedures. In the in vivo model, surgical penetration of retinal blood vessels was accomplished in 5 of 6 eyes. Immediately postoperatively, thrombus formation with obstruction of the retinal vasculature was observed. At 2 weeks postoperatively, the retinal vasculature was completely patent. CONCLUSIONS: Multiple surgical techniques aimed at assisting recanalization of occluded retinal vasculature have been evaluated. Retinal vascular surgery has become more feasible and deserves further investigation.

Echographic diagnosis of anterior hyaloidal fibrovascular proliferation.

High-resolution contact B-scan echographic imaging of the ciliary body and peripheral retina was performed on five eyes with anterior hyaloidal fibrovascular proliferation and media opacity by means of a wide (58 degrees) scanning arc. This technique determined the circumferential and anteroposterior extent of peripheral traction retinal detachment associated with anterior hyaloidal fibrovascular proliferation, which correlated highly with findings at subsequent vitrectomy. Final visual acuity of 20/400 or better was achieved in the two eyes (50%) with more limited peripheral traction detachment. In the presence of media opacity, anterior echographic imaging may allow early detection of traction retinal detachment associated with anterior hyaloidal fibrovascular proliferation and may be useful in characterizing the severity of this condition.

Surgical excision of the attached posterior hyaloid.

When vitrectomy is performed in eyes without posterior vitreous detachment, complete removal of the posterior hyaloid may be important to relieve existing vitreoretinal traction or to remove a surface for subsequent cellular proliferation. We used two alternative techniques of detaching the posterior hyaloid in an unselected consecutive series of ten patients; we used either a tapered extrusion needle or a microvitreoretinal blade to initiate separation of the posterior hyaloid at the optic disc margin. After surgical peeling of the posterior hyaloid from the retina, complete removal of the posterior vitreous and surgical relief of posterior vitreoretinal traction were facilitated.

Rebleeding in experimental traumatic hyphema treated with intraocular tissue plasminogen activator.

Tissue plasminogen activator has recently been shown to enhance the clearance of experimental nontraumatic hyphema in animals. However, hyphema in human eyes usually results from ocular trauma, and rebleeding is a serious complication. Hemorrhage is also a potential complication of fibrinolytic therapy. We assessed the incidence of rebleeding in an animal model of surgically induced traumatic hyphema after intracameral injection of tissue plasminogen activator (25 micrograms) or physiological saline. Eight eyes were each treated with tissue plasminogen activator or physiological saline at 10 minutes, 24 hours, 48 hours, or 72 hours after injury. Controls were 8 eyes with hyphema but no intracameral injection. No eyes treated with physiological saline (total, 32 eyes) or control eyes rebled. In contrast, the incidence of rebleeding from the injury site in eyes treated with tissue plasminogen activator was 88% (7/8) at 10 minutes, 75% (6/8) at 24 hours, 50% (4/8) at 48 hours, and 0% (0/8) at 72 hours after injury. Treatment of traumatic hyphema with tissue plasminogen activator prior to healing of damaged vascular endothelium may contribute to rebleeding.

Differentiation between recently resolved optic neuritis and central serous retinopathy. Use of tests of visual function.

A test battery was performed on 13 patients with resolved central serous retinopathy and on 13 patients with resolved optic neuritis to see whether the tests would help to distinguish between the two conditions. We found that the most useful discriminators were the relative afferent pupillary defect, followed by the visual evoked potential latency and the critical flicker frequency. The total error score on the Farnsworth-Munsell 100-Hue Test and the nature of the color vision defect were not helpful in separating the two diseases.

Ocular injuries caused by elastic cords.

Five patients with ocular trauma due to elastic cords with attached metal or plastic hooks were evaluated over a 13-month period. Injuries included hyphema, angle recession, lens subluxation, choroidal rupture, vitreous hemorrhage, retinal detachment, and scleral rupture. Two patients suffered penetrating ocular injuries, and three patients had final visual acuities of 20/200 or less. This series emphasizes the potential severity of ocular injury due to these commonly used devices and reiterates a modification in design that might decrease the likelihood of accidental trauma from elastic cords.

Combined phacoemulsification, pars plana vitrectomy, and posterior chamber intraocular lens insertion.

Eighteen eyes with coexisting cataract and vitreoretinal disease underwent combined phacoemulsification, pars plana vitrectomy, and posterior chamber lens implantation. Preoperative vitreoretinal disease included nonclearing vitreous hemorrhage (eight eyes), vitreous hemorrhage and tractional retinal detachment (three eyes), tractional retinal detachment (one eye), epiretinal membranes (three eyes), peripheral uveitis (two eyes), and a retained intraocular metallic foreign body (one eye). Postoperative visual acuity improved in each case; 14 eyes achieved visual acuity between 20/20 and 20/80 during an average postoperative period of 11 months (range, 3 to 39 months). Perioperative complications included an iatrogenic retinal break (one eye) and pupillary block glaucoma (one eye). Four eyes required YAG laser capsulotomy postoperatively. Phacoemulsification did not interfere with corneal clarity, allowed water-tight wound closure during vitrectomy, and preserved the capsular bag, allowing endocapsular fixation of the posterior chamber lens. Combining phacoemulsification, posterior chamber lens implantation, and pars plana vitrectomy allows rapid visual rehabilitation and functional unaided vision in these eyes.

Vitrectomy for traumatic retinal incarceration.

Traumatic retinal incarceration into a scleral wound may prevent successful surgical rehabilitation of eyes with severe posterior segment injury. We managed 15 consecutive eyes with traumatic retinal incarceration and associated retinal detachment with vitrectomy techniques. We based our approach on the anteroposterior location of the incarceration site and the amount of retina incarcerated into the wound. Despite successful anatomic reattachment in six of seven eyes with retinal incarceration posterior to the vortex vein ampullae, only two of seven eyes achieved visual acuity of 5/200 or better. In eyes with more peripheral retinal incarceration, anatomic reattachment was achieved in five of eight eyes and visual acuity of 5/200 or better was achieved in four eyes. Overall, a visual acuity of 5/200 or better was achieved in six (40%) of 15 eyes with a minimum follow-up of six months.

Histopathologic study of autosomal dominant vitreoretinochoroidopathy in a 26-year-old woman.

The clinicopathologic findings were obtained from enucleated eyes, obtained post mortem, of a 26-year-old woman with autosomal dominant vitreoretinochoroidopathy. Light microscopy demonstrated atrophic, disorganized peripheral retina with retinal blood vessels obscured by pigmented cells surrounding periodic acid-Schiff-positive deposits. Peripheral retinal pigment epithelial cells showed multilayering and pigmentation, with a thickened basal lamina. By electron microscopy, the peripheral retinal vessel endothelium was replaced by an arrangement of morphologically polarized pigmented cells of presumed retinal pigment epithelial origin oriented with their basal surfaces toward a fibrillar matrix occupying the vessel lumen. The similarity of the findings in this young patient to those of an aged patient described previously suggest that autosomal dominant vitreoretinochoroidopathy is an early-onset dystrophy of the peripheral retina with minimal subsequent progression, characterized by a retinal pigment epithelial response that includes marked intraretinal migration and extracellular matrix deposition.

Comparison of scleral tensile strength after transscleral retinal cryopexy, diathermy, and diode laser photocoagulation.

OBJECTIVES: To determine the effect of transscleral diode photocoagulation on the tensile strength of sclera in an experimental rabbit model and to compare it with that of transscleral cryotherapy and diathermy. METHODS: Twenty-four Dutch-belted rabbits received one of the following three treatment modalities in one eye: (1) transscleral cryotherapy, (2) transscleral diathermy, or (3) transscleral diode laser photocoagulation. The opposite eye served as a paired control. Tensile strength measurements of scleral strips excised from areas of treatment were obtained 6 weeks later. RESULTS: No statistically significant difference in mean tensile strength was observed between eyes receiving transscleral cryopexy (n = 7) or transscleral diode photocoagulation (n = 8) and their corresponding, paired, control eyes. In contrast, transscleral diathermy reduced mean scleral tensile strength to 26% of that of the paired control eyes (n = 8, P = .0001). CONCLUSIONS: In this rabbit model, scleral weakening is significant following transscleral diathermy while transscleral cryopexy or transscleral diode photocoagulation produces no significant weakening relative to paired, untreated controls.

Visual function abnormalities in central serous retinopathy.

Eighteen patients with central serous retinopathy (CSR) underwent a battery of visual function tests when first seen and after resolution of the subretinal fluid. Eyes with CSR had minimal relative afferent pupillary defects, reduced critical flicker-fusion thresholds, prolonged visual evoked potential (VEP) latencies, increased errors on the Farnsworth-Munsell 100-hue (FM 100) test, and depressed central visual fields (Octopus). The afferent pupillary defect and critical flicker-fusion thresholds were the first to improve after resolution of the subretinal fluid. Next in rate of improvement were the visual acuity, the VEP latency, and the FM 100 test results. The threshold of the central Octopus at fixation improved the slowest and was still abnormal during long-term follow-up, indicating a prolonged depression in the threshold of central field sensitivity after resolution of the CSR. Many of these abnormalities are also seen in patients with optic nerve disease.

Management of traumatic hemorrhagic retinal detachment with pars plana vitrectomy.

Traumatic hemorrhagic retinal detachment may prevent successful visual rehabilitation of eyes with severe posterior segment injury. We managed 19 consecutive cases of traumatic hemorrhagic retinal detachment with pars plana vitrectomy, scleral buckling, and fluid-gas exchange, with or without internal drainage of subretinal hemorrhage. We based our approach on the amount of subretinal hemorrhage present and the location of associated retinal breaks. Internal drainage of subretinal hemorrhage was performed in 16 eyes to allow adequate retinopexy to hemorrhagically elevated retinal breaks (9 eyes), to remove massive subretinal hemorrhage (4 eyes), and to allow intraoperative reattachment when the retina exhibited bullous retinal detachment (3 eyes). Overall, with a minimum of 6 months of follow-up, anatomic reattachment was achieved in 13 (68%) of 19 eyes, and functional success (visual acuity 5/200 or better) was achieved in 6 (32%) of 19 eyes. Anatomic failure resulted from proliferative vitreoretinopathy (4 eyes) and globe atrophy (2 eyes). Drainage of subretinal blood appeared to be beneficial for hemorrhagically elevated retinal tears to allow adequate retinopexy and may help to accomplish long-term anatomic attachment in eyes with massive subretinal hemorrhage or bullous retinal detachment.

A two-stage technique for intraoperative fluid-gas exchange following pars plana vitrectomy.

We describe herein a two-stage technique of intraoperative fluid-gas exchange following pars plana vitrectomy. The first stage is a complete fluid-air exchange using an air pump and intraocular linear suction. The air pump then maintains the intraocular pressure at the desired level during adjunctive procedures, such as laser endophotocoagulation. The second stage is an air-gas mixture exchange in which the desired gas, premixed in air to the desired final intraocular concentration, is manually flushed through the eye. This technique allows the attainment of an accurate concentration of intraocular gas if the air-gas mixture exchange is complete. In vitro and in vivo experiments in aphakic human eyes demonstrated that an effectively complete air-gas mixture exchange occurs with a 25-mL flush volume. This result compares favorably with the theoretical prediction derived from a "pharmacokinetic approximation" equation.