Tacrolimus downregulates inflammation by regulating pro­/anti­inflammatory responses in LPS­induced keratitis.

Lipopolysaccharide (LPS)­induced keratitis is a progressive infectious ocular disease in which innate inflammatory responses often cause clinical tissue damage and vision loss. The present study aimed to investigate the effects of tacrolimus, an effective immunomodulator, on LPS­induced innate immune responses. The effects of tacrolimus on the apoptotic rate and viability of human corneal epithelial cells (HCECs), polymorphonuclear neutrophils (PMNs) and monocytes (THP­1 cells) were examined using flow cytome-try and MTT assays. Subsequently, the role of tacrolimus on LPS­induced inflammation in HCECs, PMNs and THP­1 cells was evaluated by detecting the expression levels of pro­inflammatory cytokines, including interleukin (IL)­1ß, IL­6 and matrix metallopeptidase 9; anti­inflammatory cytokines, including IL­10 and transforming growth factor­ß; and proangiogenic factors, including vascular endothelial growth factor and tumor necrosis factor­α using quantitative polymerase chain reaction. The results demonstrated that tacrolimus had good biocompatibility with HCECs, while promoting apoptosis and decreasing the viability of PMNs and THP­1 cells. Furthermore, tacrolimus effectively reduced the expression levels of pro­inflammatory cytokines and increased anti­inflammatory cytokines in LPS­induced keratitis in vitro. Notably, tacrolimus decreased the levels of proangiogenic factors, which are highly increased following LPS stimulation. Conclusively, tacrolimus appears to be a safe and effective treatment to suppress neutrophil and monocyte activity, modulate the balance of pro­/anti­inflammatory cytokines, and reduce the inflammatory response and angiogenic activity in LPS­induced bacterial keratitis.

Heat Shock Proteins 70kDa, Eosinophil Cationic Protein, and Nitric Oxide During Chronic Superficial Keratitis in Dogs.

The objective of the study is to determine the levels of eosinophil cationic protein (ECP), heat shock proteins 70, and nitric oxide ions measured as nitrite ions (Griess reaction) in dogs with chronic superficial keratitis (CSK). The study was conducted on 24 dogs with CSK. Blood sera from the animals were tested for concentrations of heat shock proteins 70, ECP, and nitrite ions before treatment and again 5 weeks and 6 months after treatment. Dogs with CSK were treated for 6 months with various regimes involving the use of ophthalmic drops containing dexamethasone, dimethyl sulfoxide, and cyclosporine. The control group consisted of 16 clinically healthy German Shepherds. The results obtained indicated a significant (P ≤ 0.05) elevation in the concentrations of heat shock proteins 70 and nitrite ions in dogs with CSK in comparison to healthy dogs and dogs after 5 weeks of therapy. After 6 months of treatment, concentrations of heat shock proteins 70, ECP, and nitrite ions had fallen below pretreatment values. Significant correlations were found between concentrations of heat shock proteins 70, ECP, and nitrite ions in healthy animals and animals with CSK. The elevated concentrations of heat shock proteins 70, ECP, and nitrite ions in dogs with CSK may indicate that the disease was both localized and systemic. The significant correlation between levels of heat shock proteins 70 and nitrite ions suggests that these parameters may be used as indirect indicators of CSK.

Queratitis por Absidia corymbifera en paciente inmunocompetente sin traumatismo corneal previo / Keratitis caused by Absidia corymbifera in an immunocompetent male with no corneal injuries

CASO CLÍNICO: Varón sano de 55 años sin antecedente de traumatismo corneal acude a urgencias por un infiltrado en el ojo izquierdo que responde parcialmente a tratamiento antibiótico tópico. Tras la introducción de corticoterapia tópica presenta un importante empeoramiento de la úlcera. El examen directo y la tinción de Gram permiten un rápido diagnóstico de las hifas de Absidia. Presenta buena respuesta al tratamiento combinado de anfotericina y posaconazol. DISCUSIÓN: Las queratitis por Zygomycetes son raras. Es rara la afectación de pacientes sanos sin antecedentes de traumatismo corneal. El tratamiento combinado de anfotericina y posaconazol ejerce un efecto sinergico contra hongos filamentosos

CASE REPORT: A healthy 55-years-old male went to emergency due to a white infiltrate in the left eye without corneal trauma which partially responds to antibiotic treatment. The infiltrate worsened by the use of topical steroids. Direct microscopic evaluation and Gram stain are a valuable diagnostic tool for the detection of Absidia filaments. There is a successful treatment with anphotericin and posaconazole. DISCUSSION: Keratitis caused by Zygomicetes are unusual. This is a rare condition in healthy patients with no corneal trauma. The treatment with amphotericin and posaconazole are synergistic against filamentous fungi

Interleukin 17 expression in peripheral blood neutrophils from fungal keratitis patients and healthy cohorts in southern India.

Interleukin 17A (IL-17) production by peripheral blood neutrophils was examined in patients with fungal keratitis and in uninfected individuals in southern India, which has high levels of airborne Aspergillus and Fusarium conidia. Il17a gene expression and intracellular IL-17 were detected in all groups, although levels were significantly elevated in neutrophils from patients with keratitis. There were no significant differences in plasma IL-17 and IL-23 between patients with keratitis and uninfected individuals; however, combined data from all groups showed a correlation between the percentage IL-17 producing neutrophils and plasma IL-23, and between plasma IL-17 and IL-6 and IL-23.

Application of umbilical cord serum eyedrops for the treatment of neurotrophic keratitis.

PURPOSE: To investigate the efficacy of umbilical cord serum eyedrops for the treatment of neurotrophic keratitis. DESIGN: Prospective noncomparative case series. PARTICIPANTS: Twenty-eight eyes of 28 patients with neurotrophic keratitis who were refractory to conventional treatment. METHODS: The patients with neurotrophic keraitis were treated with 20% umbilical cord serum eyedrops 6 to 10 times a day. Ophthalmic examinations including best-corrected visual acuity (VA) measurement, corneal sensitivity test, corneal fluorescein staining, and anterior segment photography were performed before and after the treatment. Concentrations of substance P, insulinlike growth factor 1 (IGF-1), and nerve growth factor (NGF) in umbilical cord serum, normal peripheral blood serum, and tears were measured. MAIN OUTCOME MEASURES: Epithelial healing time; changes of VA and corneal sensitivity after treatment; and levels of substance P, IGF-1, and NGF in umbilical cord serum, normal peripheral blood serum, and tears. RESULTS: The epithelial defect healed completely in all eyes, with a mean healing time of 4.4+/-4.0 weeks. The epithelial defect healed within 2 weeks in 8 eyes (28.6%), between 2 and 4 weeks in 14 eyes (50.0%), and after 4 weeks in 6 eyes (21.4%). After treatment, VA improved by >2 lines in 17 eyes (60.7%). Mean pretreatment corneal sensitivity was 21.1+/-10.5 mm, and mean posttreatment corneal sensitivity was 24.3+/-11.7 mm (P<0.01). Mean concentrations of substance P, IGF-1, and NGF were 245.3+/-53.9 pg/ml, 239.0+/-77.1 ng/ml, and 729.7+/-72.0 pg/ml in umbilical cord serum; 169.5+/-81.0 pg/ml, 375.5+/-51.3 ng/ml, and 401.7+/-98.1 pg/ml in peripheral blood serum; and 69.8+/-24.9 pg/ml, 75.7+/-50.5 ng/ml, and 107.5+/-70.9 pg/ml in tears, respectively. CONCLUSIONS: Umbilical cord serum contains many neurotrophic factors, and umbilical cord serum eyedrops appeared to be effective for the treatment of neurotrophic keratitis.

Onchocerca volvulus keratitis (river blindness) is exacerbated in BALB/c IL-4 gene knockout mice.

To determine the outcome of Onchocerca volvulus keratitis in IL-4(-/-) BALB/c mice, animals were immunized subcutaneously and injected into the corneal stroma with soluble O. volvulus antigens. IL-4(-/-) BALB/c mice had a deviated cellular response, with decreased serum IgE and IgG1 and elevated IgG2a compared with control BALB/c mice. In marked contrast to control BALB/c, C57BL/6, and IL-4(-/-) C57BL/6 mice, IL-4(-/-) BALB/c mice developed severe corneal opacification and neovascularization that was associated with a pronounced neutrophil infiltrate to the corneal stroma. STAT-6(-/-) BALB/c mice had the same phenotype as IL-4(-/-) BALB/c mice, and complement depletion had no effect on the severity of O. volvulus keratitis in these mice. These findings indicate that on a BALB/c background, IL-4 has a critical role in regulating neutrophil recruitment to the cornea and development of O. volvulus keratitis.

Tyrosinemia type II: nine cases of ocular signs and symptoms.

PURPOSE: To report nine cases of tyrosinemia type II, with ocular signs and symptoms. METHODS: Participants included nine patients (18 eyes) who were followed for a mean follow-up period of 6.5 years (range, 2 to 8 years). Intervention included dietary restriction of tyrosine and phenylalanine, which led to resolution of ocular and cutaneous lesions, improved behavior in one patient, and may have prevented developmental delay in others. The main outcome measures were visual acuity and serum tyrosine levels to determine the response to dietary therapy. Intelligence testing and developmental screening were performed when appropriate. RESULTS: All patients presented with ocular signs and symptoms as the primary manifestation of disease and serve to illustrate the typical ocular findings of this syndrome. Three patients presented with cutaneous manifestations, and one patient had mild mental impairment at the time of diagnosis. CONCLUSIONS: Recalcitrant pseudodendritic keratitis may be the presenting sign in tyrosinemia type II. Awareness of the presenting signs and symptoms may accelerate the diagnosis and dietary intervention. Initiation of a tyrosine-restricted and phenylalanine-restricted diet in infancy is most effective in preventing cognitive impairment.

[Analysis of nucleotide level of human erythrocytes in patients with liver-fire exuberance syndrome].

Level of erythrocytes in 30 patients with Liver-Fire Exuberance Syndrome (LFES) was observed, and that of 30 healthy persons was determined in the meantime as control by high-performance liquid chromatography. It was found that the adenosine triphosphate in erythrocytes of LFES patients was lower and level of adenosine monophosphate was higher than that of healthy persons. While, the levels of adenylate pool and energy load ratio were lower than that of control. These results showed the metabolism was exuberant, the energy consumption increased and the energy storage reduced in patients of LFES.

[Incurable keratitis and chronic palmoplantar hyperkeratosis with hypertyrosinemia. Cure using a tyrosine-restricted diet. Type II tyrosinemia]. / Kératite "inguérissable" et hyperkératose palmo-plantaire chronique avec hypertyrosinémie. Guérison par un régime pauvre en tyrosine. Tyrosinémie de type II.

One should henceforth systematically search for hypertyrosinemia which, too often, goes unrecognized for years, in patients presenting chronic keratitis associated with palmar and plantar hyperkeratosis. As a matter of fact, this highly crippling disease may be cured with an appropriate diet and the diagnosis, once suspected, is easily confirmed by simple investigations.

Polymorphonuclear leukocyte kinetics in experimentally induced keratitis.

The movement of polymorphonuclear leukocytes (PMNLs) into inflamed corneas was studied using a quantitative technique to measure PMNL chemotaxis in vivo. Our studies suggested that, in this model, most PMNLs enter the cornea through limbal vessels. A variety of bacterial agents, including viable bacteria, killed bacteria, culture filtrates, and endotoxin, were found to induce a significant corneal inflammatory response. Of the agents tested, viable Pseudomonas aeruginosa produced greatest inflammation. Host factors (serum, PMNLs) also induced movement of PMNLs into corneas, but only after preincubation with activating agents. Normal serum, resting PMNLs, and PMNL lysates derived from resting cells did not promote PMNL corneal ingress. These studies provide further insight into the movement of PMNLs into the inflamed cornea and information that may be of use in developing techniques to inhibit the corneal inflammatory response.

[Value of the immunoenzyme method (ELISA) in determining serum and aqueous humor antiherpes immunoglobulin levels (apropos of 155 cases)]. / Intérêt de la méthode immuno-enzymatique (ELISA) pour le dosage des immunoglobulines antiherpès dans l'humeur aqueuse et le sérum (à propos de 155 cas).

Immuno-enzymatic assay of aqueous humor and serum antiherpetic antibodies was performed in 155 patients with all types of uveitis and with keratitis. Globulin levels were also determined in the two fluids, employing an original nephelometry-laser technique. This immuno-enzymatic technique, which possesses greater reliability, reproducibility, and sensitivity than passive hemagglutination, appears suitable for microassay in aqueous humor, and demonstrates the production of anti-herpes antibodies in this fluid to the exclusion of other specificity. Applying limits of at least 1/40e for H.A. antibody levels, and at least 10 for the immunity load coefficient, these antibodies could be demonstrated in half of the cases of clinically confirmed herpes, in one-third of clinically suspected cases, and in two intermediary cases of uveitis where no predictive signs of herpes were present. In contrast, specific antibodies were never detected in 70 cases of anterior or total uveitis. Measuring anti-herpes antibodies in aqueous humor and serum by the ELISA method, in association with immunity load coefficients determination in the two fluides, appears to be a very useful method for the future etiological diagnosis of anterior and intermediary uveitis when the etiology is uncertain but clinical signs suggest a possible herpetic origin.

Experimental bacterial keratitis in neutropenic guinea pigs: polymorphonuclear leukocytes in corneal host defense.

Quantitative techniques were used to determine the relative concentrations of viable bacteria and polymorphonuclear leukocytes (PMNs) in the corneas of neutropenic and non-neutropenic guinea pigs with experimental bacterial keratitis induced with three strains of Pseudomonas aeruginosa. Neutropenia was produced by whole-body X-irradiation 1 week before infection. Significantly greater numbers of bacteria were present in the cornea of neutropenic animals 48 h after infection than were present in the corneas of non-neutropenic animals. The same was true 24 and 48 h after infecting animals with Staphylococcus aureus. Examination of histological sections showed that fewer PMNs were present in the corneas of infected neutropenic animals than in the corneas of infected non-neutropenic animals. Radiolabeling of PMNs confirmed a significant reduction in PMN concentration in the corneas of infected neutropenic animals. Tears and the corneal epithelium appear to be the most important elements protecting the cornea against local invasion by bacteria. However, once bacterial keratitis is established, PMNs play a role in limiting bacterial multiplication.

Tyrosinemia without liver or renal damage with plantar and palmar keratosis and keratitis (hypertyrosinemia type II).

A boy of 3 2/12 years of age with Richner-Hanhart syndrome (plantar and palmar keratosis and chronic keratitis) was found to have hypertyrosinemia and to excrete the hydroxyacids derived from tyrosine. A diet poor in phenylalanine and tyrosine cured the skin and corneal lesions. Clinical and biochemical observations are reported.

Anti-inflammatory effectiveness of topically administered corticosteroids in the cornea without epithelium.

The present studies demonstrate that modification of the derivative of a given steriod base alters its anti-inflammatory potential as measured by suppression of leukocyte invasion of the cornea. A comparison of each drug's corneal bioavailability with its anti-inflammatory effectiveness shows the acetate derivative of prednisolone to be a more potent anti-inflammatory agent than the phosphate derivative. Similarly, the free alcohol derivative of dexamethasone proved to be more potent than the phosphate derivative. Increasing the concentration of prednisolone acetate from 0.125 per cent to 1.0 per cent results in a significant increase in its anti-inflammatory effectiveness in the cornea following topical administration. The same increase in prednisolone phosphate concentration does not produce a significant increase in its ability to suppress polymorphonuclear leukocyte infiltration of the cornea. When the epithelium of the inflamed cornea is intact, prednisolone acetate, 1.0 per cent ophthalmic suspension, is the most effective of the corticosteroid preparations studied. In the absence of an intact epithelium, prednisolone acetate, 1.0 per cent ophthalmic suspension, again produces the greatest mean reduction in polymorphonuclear leukocytic infiltration of the cornea although here one cannot demonstrate a statistically significant difference from the anti-inflammatory effect produced by prednisolone phosphate, 1.0 per cent ophthalmic solution, or dexamethasone alcohol, 0.1 per cent ophthalmic suspension. Overall, therefore, prednisolone acetate 1.0 per cent is the most effective of the topical agents studied for suppression of corneal inflammation.