Molecular genetic diagnostic techniques in choroideremia.

PURPOSE: To optimize and streamline molecular genetics techniques in diagnosing choroideremia (CHM). METHODS: PCR primers were designed for exons 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, and 15 of the CHM gene. Each PCR protocol was optimized so that all exons could be amplified with the same component ratio and PCR conditions. Sense and antisense primers were tested for their ability to be used as sequencing primers. Fibroblast cells were cultured, and an immunoblot analysis was performed to detect the presence or absence of Rab escort protein 1 (REP-1) in a suspected CHM patient sample when no mutation was detected with sequencing. Multiplex ligation-dependent probe amplification (MLPA) of the CHM gene was performed and used to detect deletions and duplications in affected males and female carriers. RNA analysis using cDNA was used to detect the presence or absence of the CHM transcript and to search for splice defects. RESULTS: The newly designed PCR primers allow for more efficient PCR preparation and sequencing to detect point mutations in affected males and female carriers. Immunoblot successfully detects the absence of REP-1 in a CHM patient. MLPA identifies deletions and duplications spanning multiple exons in the CHM gene. RNA analysis aids in detecting splice variants. CONCLUSIONS: The development of new molecular biology techniques and ongoing optimization of existing methods allows for an improved integrated approach to confirm CHM diagnosis and carrier status in consideration of patient family history and available patient sample materials. CHM can be confirmed with an immunoblot assay. To detect the molecular cause of CHM, an examination of the genomic DNA or the mRNA must be performed. Presymptomatic carriers with no identifiable fundus signs can be identified only through molecular analysis of genomic DNA or through quantitative assays.

¿Cuáles deben ser las líneas de actuación de la sociedad valenciana de medicina familiar y comunitaria en los próximos 4 años? / What lines of action should the valencia society of family and community medicine adopt over the next four years?

Objetivo. Identificar las áreas de actuación en que debe implicarse la junta directiva de la Sociedad Valenciana de Medicina Familiar y Comunitaria en los próximos 4 años. Diseño. Metodología cualitativa. Emplazamiento. Miembros de la SVMFyC. Pacientes u otros participantes. Participaron 27 expertos miembros de la SVMFyC. Intervenciones. Técnicas cualitativas de búsqueda de consenso. Se aseguró la fiabilidad y validez de la técnica mediante triangulación y la selección de los expertos entre los distintos grupos profesionales de la SVMFyC. Mediciones y resultados principales. Para determinar las líneas recomendadas se analizaron: productividad, representatividad espontánea, intensidad de la recomendación y grado de acuerdo. Las líneas prioritarias recomendadas fueron: defender la vía MIR, proponer reformas en los planes de estudios del pregrado, velar por la transparencia de las convocatorias y por la convocatoria anual de la OPE, impulsar la carrera profesional, creación de plazas de profesores del área de conocimiento de medicina familiar y comunitaria, definir el tamaño de la población adscrita, proponer un catálogo de incentivos y estudiar alternativas para que el salario no sea uniforme, sino que se vincule a objetivos. Conclusiones. Se establecen las que, a juicio de los expertos, son las líneas de actuación recomendadas (AU)

Astigmatism after penetrating keratoplasty. Role of the suture technique.

The Authors retrospectively evaluated the evolution of corneal astigmatism 6, 12 and 24 months after penetrating keratoplasty in 75 eyes by comparing 2 suture techniques: running suture (RS) in 14 cases vs interrupted suture (IS) in 61 cases. The mean keratometric astigmatism in patients with RS was 7.75 +/- 3.10 D at 6 months, 5.89 +/- 2.50 D at 12 months and 3.90 +/- 1.70 D at 24 months. In patients with IS these values were 4.82 +/- 4.00 D at 6 months, 2.81 +/- 1.60 D at 12 months and 2.77 +/- 1.34 D at 24 months. A significant lower astigmatism (P < 0.05) was noted after using IS at 6 and 12 months. At 24 months, however, this difference was no longer statistically significant (P > 0.3). The IS suture allowed a better control of the post-surgical astigmatism, and a more rapid recovery of the visual acuity.

[Functional evaluation of the corneal endothelium using anterior segment fluorophotometry]. / Evaluation fonctionnelle de l'endothélium cornéen par la fluorophotométrie du segment antérieur.

The AA have studied the permeability of the corneal endothelium by anterior segment fluorophotometry using instillation of fluorescein. The Fluorotron Master TM has been modified with the introduction of a new slit (58.4 microns) and of the voltage output (6.5 v) has also been increased. The resolution has been improved from 2.39 mm to 0.59 mm. Of this study, we have examined 20 patients submitted to extracapsular cataract extraction with intraocular lens implantation (ECCE + IOL). The patients have been divided in two groups, according to the surgical technic used: "can-open" and intracapsular. Each patient have been submitted before surgery and 1 week after to an ophthalmologic examination, paquimetry and anterior segment fluorophotometry. The permeability coefficient (PAC) increases with the duration of surgery and when using "can-opener" instead of endocapsular.

[Follow-up of early diabetic maculopathy by angiography and fluorophotometry]. / Surveillance de la maculopathie diabétique précoce par angiographie et fluorophotométrie.

We evaluated prospectively for 12 months 22 patients with late-onset diabetes mellitus and minimal retinopathy by fluorescein angiography and fluorophotometry. The study showed that early retinopathy changes were not permanent or invariably progressive, but may regress. Microaneurysm gradings worsened in 24%, improved in 14% and remained stabilized in 62%. Capillary closure worsened in 33%, improved in 10% and remained stabilized in 57%. The evolution of early diabetic maculopathy is slow and irregular, and this study has shown that vitreous fluorophotometry must be included when short follow-up periods of up to 12 months are considered. Finally, eyes with higher fluorophotometry values at entry showed significantly more deterioration during the 12-months follow-up period (p less than 0.05).