Efficacy of topical cobalt chelate CTC-96 against adenovirus in a cell culture model and against adenovirus keratoconjunctivitis in a rabbit model

BACKGROUND: Adenovirus (Ad), associated with significant morbidity, has no topical treatment. A leading CTC compound (CTC-96), a Co(III )chelate, was found to have potent in vitro and in vivo antiviral efficacy against herpes viruses. In this study CTC-96 is being tested for possible anti-Adenovirus activity. METHODS: The biological anti-adenovirus activity of CTC-96 in concentrations from 5 to 250 ug/ml, was evaluated initially by viral inactivation (viral exposure to CTC-96 followed by dilution and inoculation of cells), virucidal (viral exposure to CTC-96 and inoculation of cells without dilution) and antiviral (effect of CTC-96 on previously adsorbed virus) plaque assays on HeLa (human cervical carcinoma), A549 (human lung carcinoma) and SIRC (rabbit corneal) cells. After verifying the antiviral activity, New Zealand White rabbits were infected with Ad-5 into: 1) the anterior cul-de-sac scarifying the conjunctiva (Group "C+"); 2) the anterior cul-de-sac scarifying the conjunctiva and cornea (Group "CC+"); 3) the stroma (Group "CI+"). Controls were sham-infected ("C-", "CC-", "CI-"). Other rabbits, after "CC", were treated for 21 days with: 1) placebo, 9x/day ("-"); 2) CTC-96, 50 ug/ml, 9x/day ("50/9"); CTC-96, 50 ug/ml, 6x/day ("50/6"); CTC-96, 25 ug/ml, 6x/day ("25/6"). All animals were monitored via examination and plaque assays. RESULTS: In vitro viral inactivation, virucidal and antiviral assays all demonstrated CTC-96 to be effective against Adenvirus type 5 (ad-5). The in vivo model of Ad keratoconjunctivitis most similar to human disease and producing highest viral yield was "CC". All eyes (6/6) developed acute conjunctivitis. "CI" yielded more stromal involvement (1/6) and iritis (5/6), but lower clinical scores (area × severity). Infection via "C" was inconsistent (4/6). Fifty (50) ug/ml was effective against Ad-5 at 6x, 9x dosings while 25 ug/ml (6x) was only marginally effective. CONCLUSION: CTC-96 demonstrated virucidal activity against Ad5 in tissue culture with HeLa, A549 and SIRC cell lines. Animal Model Development: 1) "CC" produced conjunctival infection with occasional keratitis similar to human disease; "CI" yielded primarily stromal involvement; 2) "C" consistently produced neither conjunctivitis nor keratitis. CTC Testing: 1) Conjunctivitis in all eyes; 2) Resolution fastest in "50/9" ("50/9". "50/6" > "25/6" > "-"); 3) Efficacy in "50/6" was not statistically different than "50/9"; 4) Conjunctival severity was lower in treatment groups then controls; 5) Little corneal or intra-ocular changes were noted

Comparison of an immortalized human corneal epithelial cell line with Vero cells in the isolation of Herpes simplex virus-1 for the laboratory diagnosis of Herpes simplex keratitis

BACKGROUND: Herpes simplex keratitis (HSK) is a sight threatening ocular infection often requiring a specific and prompt laboratory diagnosis. Isolation of Herpes simplex virus (HSV-1) in culture provides the most reliable and specific method and is considered as the "Gold Standard" in the laboratory diagnosis of HSK in spite of its low sensitivity. Using "cell lines of corneal origin" for virus isolation may be beneficial under such circumstances, since these cells have been shown to be excellent substrates for the growth of HSV-1 isolated from the cornea. We report a comparative study of a novel human corneal epithelial cell line (HCE) and the Vero cell line in the isolation of HSV-1 from corneal scrapings employing a shell vial assay. METHODS: Corneal scrapings were obtained from 17 patients with a clinical diagnosis of HSK. All the cases were confirmed by virological investigations (PCR and viral antigen detection positive, n = 15, PCR positive, n = 1, Viral antigen positive, n = 1). Scrapings obtained from 10 patients with infectious keratitis of non-viral origin were included as controls. All the scrapings were simultaneously inoculated into shell vials of HCE and Vero cells. Cultures were terminated at 24 h post-infection. Isolation of HSV-1 was confirmed using an indirect immunofluorescence/ immunoperoxidase assay. RESULTS: Virus could be isolated using both or either of the cell lines in 10/17 (58.82%) patients with HSK. HSV-1 was isolated from 10/ 17 (58.82%) and 4/17(23.52%) specimens in HCE and Vero cells, respectively (P = 0.036). None of the controls yielded HSV-1. While all the 10 (100%) strains were isolated in HCE, Vero yielded only 4/10 (40%) strains in the shell vial culture (P = 0.014). CONCLUSIONS: HCE showed a statistically significant difference in the virus isolation rate with respect to Vero cells. HCE may be an excellent alternative cell line for the isolation of HSV-1, especially from corneal scrapings, for the laboratory diagnosis of HSK

Patient-Reported Ocular Disorders and Symptoms in Adults with Moderate-to-Severe Atopic Dermatitis: Screening and Baseline Survey Data from a Clinical Trial

Introduction: Patients with atopic dermatitis (AD) have a greater risk of conjunctivitis and other ocular surface disorders than the general population. We evaluated the burden of ocular surface disorders and related symptoms prior to treatment initiation in adults with moderate-tosevere AD. Methods: Patients were enrolled in a randomized, placebo-controlled, double-blinded, phase 3 trial of dupilumab administered with concomitant topical corticosteroids. At the beginning of the screening period, all enrolled patients completed a survey of ocular disorder diagnoses received in the past year; at baseline, patients completed a survey of frequency and severity of ocular symptoms (discomfort, itching, redness, and tearing) experienced in the past month. Results: A total of 712 of 740 patients enrolled in the trial provided responses to the survey. At screening, 286 of 740 patients (38.6%) reported having at least one ocular disorder in the past year. At baseline, 499 of 712 respondents (70.1%) reported having at least one symptom within the past month. Of these patients, 4.4%, 6.0%, 5.5%, and 4.4%, respectively, reported h

Cost of Dry Eye Treatment in an Asian Clinic Setting

10.1371/journal.pone.0037711PLoS ONE76

A study of family mediation during divorce in the Pakistani Muslim community in Bradford. Some observations on the implications for the theory and practice of conflict resolution.

Conflict resolution theory and practice have been increasingly criticised for ignoring the centrality of culture in their attempts to find theories and models that are applicable universally, not only across cultures but also across levels of society. Mediation is one form of conflict resolution, which has come to occupy a central position in the resolution of disputes both at international and local levels. At the level of family disputes, family mediation has failed to engage users from different ethnic groups in England and Wales. This thesis explores the hypothesis that culture and, in particular, culturally defined concepts of gender are the important factors determining the success or failure of mediation in divorce disputes.J. A. Clark Charitable Trus

Ring electrode for radio-frequency heating of the cornea: modelling and in vitro experiments

[EN] Radio-frequency thermokeratoplasty (RF-TKP) is a technique used to reshape the cornea curvature by means of thermal lesions using radio-frequency currents. This curvature change allows refractive disorders such as hyperopia to be corrected. A new electrode with ring geometry is proposed for RF-TKP. It was designed to create a single thermal lesion with a full-circle shape. Finite element models were developed, and the temperature distributions in the cornea were analysed for different ring electrode characteristics. The computer results indicated that the maximum temperature in the cornea was located in the vicinity of the ring electrode outer perimeter, and that the lesions had a semi-torus shape. The results also indicated that the electrode thickness, electrode radius and electrode thermal conductivity had a significant influence on the temperature distributions. In addition, in vitro experiments were performed on rabbit eyes. At 5 IN power the lesions were fully circular. Some lesions showed non-uniform characteristics along their circular path. Lesion depth depended on heating duration (60% of corneal thickness for 20s, and 30% for 10s). The results suggest that the critical shrinkage temperature (55-63degreesC) was reached at the central stroma and along the entire circular path in all the cases.Berjano, E.; Saiz Rodríguez, FJ.; Alió, J.; Ferrero, JM. (2003). Ring electrode for radio-frequency heating of the cornea: modelling and in vitro experiments. Medical & Biological Engineering & Computing. 41(6):630-639. https://doi.org/10.1007/BF02349970S630639416Alió, J. L., Ismail, M. M., Artola, A., andPérez-Santonja, J. J. (1997a): ‘Correction of hyperopia induced by photorefractive keratectomy using non-contact Ho: YAG laser thermal keratoplasty’,J. Refract. Surg.,13, pp. 13–16Alió, J. L., Ismail, M. M., andSanchez, J. L. (1997b): ‘Correction of hyperopia with non-contact Ho: YAG laser thermal keratoplasty’,J. Refract. Surg.,13, pp. 17–22Alió, J. 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(1999): ‘Temperature controlled and constant power radiofrequency ablation: what affects lesion growth?’,IEEE Trans. Biomed. Eng.,46, pp. 1405–1412Krasteva, V. Tz., andPapazov, S. P. (2002): ‘Estimation of current density distribution under electrodes for external defibrillation’,Biomed. Eng. OnLine,1, 7Labonté, S. (1992): ‘A theoretical study of radio-frequency ablation of the myocardium’,PhD dissertation, Department of Electrical Engineering, University of Ottawa, CanadaLabonté, S. (1994): ‘Numerical model for radio-frequency ablation of the endocardium and its experimental validation’,IEEE Trans. Biomed. Eng.,41, pp. 108–115Mannis, M. J., Segal, W. A., andDarlington, J. K. (2001): ‘Making sense of refractive surgery in 2001: Why, when, for whom, and by whom?’,Mayo Clin. Proc.,76, pp. 823–829McCally, R. L., Bargeron, R. A., andGreen, W. R. (1983): ‘Stromal damage in rabbit corneas exposed to CO2 laser radiation’,Exp. Eye Res.,37, pp. 543–550McDonald, M. B., Hersh, P. S., Manche, E. 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V., Arruda, M., Wang, X., Ohtomo, K., Beckman, K. J., McClelland, J. H., Lazzara, R., andJackman, W. M. (1995): ‘Comparison ofin vivo tissue temperature profile and lesion geometry for radiofrequency ablation with a saline-irrigated electrode versus temperature control in a canine thigh muscle preparation’,Circ.,91, pp. 2264–2273Panescu, D., Whayne, J. G., Fleischman, S. D., Mirotznik, M. S., Swanson, D. K., andWebster, J. G. (1995): ‘Three-dimensional finite element analysis of current density and temperature distributions during radio-frequency ablation’,IEEE Trans. Biomed. Eng.,42, pp. 879–890Plonsey, R., andHeppner, D. B. (1967): ‘Considerations of quasistationarity in electrophysiological systems’,Bull. Math. Biophys.,29, pp. 657–664Rowsey, J. J. (1987): ‘Electrosurgical keratoplasty: Update and retraction’,Invest. Ophthalmol. Vis. Sci.,28, p. 224Rutzen, A. R., Roberts, C. W., Driller, J., Gomez, D., Lucas, B. C., Lizzi, F. L., andColeman, D. J. 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Specific detection of fungal pathogens by 18S rRNA gene PCR in microbial keratitis

<p>Abstract</p> <p>Background</p> <p>The sensitivity and specificity of 18S rRNA polymerase chain reaction (PCR) in the detection of fungal aetiology of microbial keratitis was determined in thirty patients with clinical diagnosis of microbial keratitis.</p> <p>Methods</p> <p>Corneal scrapings from patients were used for Gram stain, culture and PCR analysis. PCR was performed with primer pairs targeted to the 18S rRNA gene. The result of the PCR was compared with conventional culture and Gram staining method. The PCR positive samples were identified by DNA sequencing of the internal transcribed spacer (ITS) region of the rRNA gene. Main outcome measures were sensitivity and specificity of PCR in the detection of fungus in corneal keratitis.</p> <p>Results</p> <p>Combination of microscopy and culture gave a positive result in 11 of 30 samples of microbial keratitis. PCR detected 10 of 11 samples that were positive by conventional method. One of the 19 samples that was negative by conventional method was positive by PCR. Statistical analysis revealed that the PCR to have a sensitivity of 90.9% and specificity of 94.7% in the detection of a fungal aetiology in microbial keratitis.</p> <p>Conclusion</p> <p>PCR is a rapid, sensitive and useful method to detect fungal aetiology in microbial keratitis.</p

Connexin Mediated Cataract Prevention in Mice

Cataracts, named for any opacity in the ocular lens, remain the leading cause of vision loss in the world. Non-surgical methods for cataract prevention are still elusive. We have genetically tested whether enhanced lens gap junction communication, provided by increased α3 connexin (Cx46) proteins expressed from α8(Kiα3) knock-in alleles in Gja8tm1(Gja3)Tww mice, could prevent nuclear cataracts caused by the γB-crystallin S11R mutation in CrygbS11R/S11R mice. Remarkably, homozygous knock-in α8(Kiα3/Kiα3) mice fully prevented nuclear cataracts, while single knock-in α8(Kiα3/−) allele mice showed variable suppression of nuclear opacities in CrygbS11R/S11R mutant mice. Cataract prevention was correlated with the suppression of many pathological processes, including crystallin degradation and fiber cell degeneration, as well as preservation of normal calcium levels and stable actin filaments in the lens. This work demonstrates that enhanced intercellular gap junction communication can effectively prevent or delay nuclear cataract formation and suggests that small metabolites transported through gap junction channels protect the stability of crystallin proteins and the cytoskeletal structures in the lens core. Thus, the use of an array of small molecules to promote lens homeostasis may become a feasible non-surgical approach for nuclear cataract prevention in the future

Método automático de clasificación de color en dientes humanos usando aprendizaje de máquina

Trabajo de InvestigaciónActualmente el proceso de identificación del color de los dientes para la fabricación de prótesis dentales es realizado manualmente por un experto que, utilizando un método de identificación visual, determina el color de las piezas dentales en la boca del paciente, usando guías de color como la VITA®. A pesar de que el método visual es el más utilizado para la identificación del color de dientes, este se ve afectado por distintas variables tales como: el cansancio del experto, la luminosidad en el ambiente, salud visual del especialista, entre otras que influyen en la identificación del color en los dientes. Los errores en la clasificación del color de los dientes pueden generar pérdidas de tiempo lo que implicaría en consecuencia sobrecostos que afectarían directamente al fabricante y la satisfacción final del cliente.1. Planteamiento del problema 2. Pregunta de investigación 3. Objetivos 4. Estado del arte 5. Marco de referencia 6. Alcances y limitaciones 7. Metodología 8. Diseño metodológico 9. Discusión y resultados 10. Conclusiones 11. Trabajos futuros 12. Bibliografía 13. ANEXOSPregradoIngeniero de Sistema