Cataract extraction in patients with primary congenital glaucoma.

OBJECTIVE: To assess incidence and risk factors of cataract extraction in patients with primary congenital glaucoma, surgical outcome, and complications. MATERIAL AND METHOD: Retrospective cohort study, in which 108 patients with primary congenital glaucoma were included. Data collected were need for cataract extraction and at what age, intraocular pressure at diagnosis of primary congenital glaucoma, required antiglaucomatous surgeries, possible mutation in the CYP1B1 gene, and final visual acuity. Among the patients who required cataract extraction were visual acuity, intraocular pressure, and complications, evaluated preoperatively and postoperatively. The data were analysed with STATA. RESULTS: A total of 198 eyes of 108 patients were included, with a median follow-up of 8 years (range: 5-53). In all, 32 eyes (16.2%) of 24 patients (22%) required cataract extraction. The median age for cataract extraction was 12.94 years (interquartile range: 2.42-22). The main identified risk factors associated with cataract extraction were antiglaucomatous surgeries (hazard ratio 1.48, p < 0.001) and valvular implant (hazard ratio 2.78, p < 0.001). Lens was implanted in 30/32 eyes and the main complications were intraocular pressure decontrol (n = 13), capsular fibrosis (n = 7), corneal decompensation (n = 4), lens subluxation (n = 4), and endophthalmitis (n = 1). Visual acuity improvement was observed after cataract extraction in 66.67% of eyes. CONCLUSIONS: There is a high incidence of cataract surgery in patients with primary congenital glaucoma, but generally outside of pediatric age. A greater risk of cataract extraction was identified in those patients with a greater number of antiglaucomatous surgeries, especially after valvular implantation. Despite the high rate of complications related to cataract extraction in primary congenital glaucoma, good visual improvement was observed after surgery.

Importance of clinical variables in the diagnosis of MODY2 and MODY3

La diabetes tipo MODY (del inglés maturity onset diabetes of the young) constituye un grupo de patologías bien definidas y caracterizadas por su aparición antes de los 25 años, herencia autosómica dominante y por el hecho de que no precisan un tratamiento con insulina (al menos, inicialmente) para evitar la formación de cuerpos cetónicos. A pesar de la importancia de una clasificación precisa del paciente diabético, no siempre resulta sencillo clasificar el diagnóstico de un paciente joven con diabetes, y los estudios genéticos, a menudo, se usan de forma inadecuada. Métodos Se describen las características clínicas de pacientes cuyo estudio para MODY2 y MODY3 resultó negativo, y se comparan con las características de pacientes con resultado de estudio positivo. Resultados Todos los pacientes con MODY3 habían sido diagnosticados antes de los 25 años de edad y requerían algún tratamiento farmacológico para controlar la glucemia. Los pacientes con MODY2 fueron diagnosticados a partir de la primera analítica realizada, bien de forma accidental o dentro de un contexto de cribado de diabetes gestacional. La descripción clínica de los 19 pacientes cuyo estudio para MODY2 y MODY3 resultó negativo, mostró que sólo dos pacientes presentaban un cuadro clínico compatible con MODY3 y solo un paciente con MODY2.ConclusionesLas características clínicas pueden ser utilizadas para excluir el diagnóstico de MODY2 y MODY3, y ello puede reducir la necesidad de estudios genéticos (AU)

MODY (maturity onset diabetes of the young) is a group of well-defined diseases clinically characterised by onset before age 25 years that does not require insulin treatment (at least initially)to prevent the formation of ketone bodies and autosomal dominant inheritance. Despite the importance of accurate classification, it is not always simple to catalogue the diagnosis of a young patient with diabetes, and genetic studies are often improperly used. Methods: We describe the clinical features of patients negative for MODY2 and MODY3 and compared them to patients positive for these subtypes. Results: All patients with MODY3 had been diagnosed before age 25 years and required drug therapy for blood glucose control. MODY2 patients were diagnosed at the first laboratory workup either incidentally or as part of gestational diabetes screening. The clinical description of the19 patients negative for MODY2 and MODY3 showed that only two patients presented a clinical picture consistent with MODY3 and one patient with MODY2.Conclusions: Clinical features can be used for early exclusion of a MODY2 or MODY3 diagnosis and may reduce the need for genetic testing (AU)

Importance of clinical variables in the diagnosis of MODY2 and MODY3.

AIMS: MODY (maturity onset diabetes of the young) is a group of well-defined diseases clinically characterised by onset before age 25 years that does not require insulin treatment (at least initially) to prevent the formation of ketone bodies and autosomal dominant inheritance. Despite the importance of accurate classification, it is not always simple to catalogue the diagnosis of a young patient with diabetes, and genetic studies are often improperly used. METHODS: We describe the clinical features of patients negative for MODY2 and MODY3 and compared them to patients positive for these subtypes. RESULTS: All patients with MODY3 had been diagnosed before age 25 years and required drug therapy for blood glucose control. MODY2 patients were diagnosed at the first laboratory workup either incidentally or as part of gestational diabetes screening. The clinical description of the 19 patients negative for MODY2 and MODY3 showed that only two patients presented a clinical picture consistent with MODY3 and one patient with MODY2. CONCLUSIONS: Clinical features can be used for early exclusion of a MODY2 or MODY3 diagnosis and may reduce the need for genetic testing.

Clinical differences between patients with MODY-3, MODY-2 and type 2 diabetes mellitus with I27L polymorphism in the HNF1alpha gene.

OBJECTIVE: The aim of our study was to describe and evaluate the clinical and metabolic characteristics of patients with MODY-3, MODY-2 or type 2 diabetes who presented I27L polymorphism in the HNF1alpha gene. METHODS: The study included 31 previously diagnosed subjects under follow-up for MODY-3 (10 subjects from 5 families), MODY-2 (15 subjects from 9 families), or type 2 diabetes (6 subjects) with I27L polymorphism in the HNF1alpha gene. The demographic, clinical, metabolic, and genetic characteristics of all patients were analyzed. RESULTS: No differences were observed in distribution according to sex, age of onset, or form of diagnosis. All patients with MODY-2 or MODY-3 had a family history of diabetes. In contrast, 33.3% of patients with type 2 diabetes mellitus and I27L polymorphism in the HNF1alpha gene had no family history of diabetes (p < 0.05). No differences were observed in body mass index, prevalence of hypertension, or microvascular or macrovascular complications. Drug therapy was required by 100% of MODY-3 patients, but not required by 100% of MODY-2 patients or 16.7% of patients with type 2 diabetes mellitus and I27L polymorphism in the HNF1alpha gene (p < 0.05). CONCLUSIONS: Occasional difficulties may be encountered when classifying patients with MODY-2, MODY-3 or type 2 diabetes of atypical characteristics, in this case patients who present I27L polymorphism in the HNF1alpha gene.

Implications of p53 gene mutations on patient survival in transitional cell carcinoma of the bladder: a long-term study.

OBJECTIVE: To determine the prognostic value of p53 gene mutations and P53 overexpression for predicting the incidence of recurrence, progression and long-term survival of patients with transitional cell carcinoma (TCC) of the bladder. METHODS: Prospective cohort study with 94 consecutive patients diagnosed and treated for TCC. DNA was obtained from tumor tissue to perform PCR-SSCP of p53 exons 5-9, with automatic sequencing of any mutated samples. Immunohistochemistry using anti-human P53 monoclonal antibody was also performed. Survival was analyzed and the survival curves compared (Mantel-Haenszel). Lastly, a Cox proportional hazards model was constructed. RESULTS: Mutations were found in 46.8% of samples, with 61.8% in infiltrating tumors. Exon 8 was involved in 42.3%. P53 overexpression (cutoff > or =20%) was found in 52.1%. Mean follow-up was 44.1 months; 43.6% had died by the end of this period. Mean survival was lower in patients with exon 8 mutations (38.4 months), compared with patients without this exon mutated (P = 0.016). There were no differences in patient survival based on positive or negative immunohistochemistry (cutoff > or =20%), although survival was lower in patients with a percentage higher than 50% of antibody-stained cells (P = 0.02). In the Cox analysis, tumor stage, pM stage, and interaction between stage > or =pT2 and mutated p53 gene were independent risk factors, with a 6.13-fold risk of death in these patients (P = 0.019). The number of tumors, nuclear grade, pTa stage, and the interaction between GI degree and nonmutated p53 gene remained in the Cox model for superficial tumors, such that these patients had a lower risk of recurrence or progression (P = 0.008). CONCLUSIONS: Alterations in the p53 gene may be indicative of poorer prognosis and greater recurrence in patients with urothelial bladder tumor, in particular, the presence of mutations in exon 8 and a greater percentage of stained cells in the immunohistochemistry. Nevertheless, the classic prognostic factors (primarily, pTNM stage) should still be considered the most useful factors for follow-up of these patients.

Sensitivity and specificity of p53 protein detection by immunohistochemistry in patients with urothelial bladder carcinoma.

OBJECTIVE: To analyze the correlation between the genotypic and phenotypic patterns of p53 in patients with transitional cell carcinoma (TCC) of the urinary bladder. MATERIALS AND METHODS: Cross-sectional study of 73 patients diagnosed with TCC. DNA was obtained from the tumor tissue to perform polymerase chain reaction-single strand conformational polymorphism (PCR-SSCP) of exons 5-9 of the p53 gene, with automatic sequencing done on any mutated samples. Immunohistochemistry (IHC) was also performed using anti-human P53 monoclonal antibody, and the diagnostic performance of this test was analyzed by a ROC curve, using the presence of p53 mutations found by PCR-SSCP as 'gold standard'. RESULTS: The cutoff point for defining immunopositivity was 20%. IHC had a specificity of 62.9%, and a sensitivity of 65.8%. The highest sensitivity values appeared in G3 tumors (75%) and infiltrating tumors (71.4%), and the highest specificity values were observed in G1 (77.7%) and G2 tumors (90%) and superficial tumors (66.6%). Mutations in exon 8 gave a positive result most frequently (73.7%) and were considered most relevant in terms of altering P53 function (60.9%). False negatives were documented in 28.5% of infiltrating tumors, and false positives in 33.4% of superficial tumors. CONCLUSIONS: There is a moderate correlation between p53 mutations and P53 protein overexpression, with this stronger in high-grade, infiltrating tumors, in exon 8 mutations, and when the mutation induces relevant changes in the protein structure. Although IHC is useful in routine clinical practice, the classic prognostic factors should still be considered the most important in the follow-up of these patients.

[Promoter hypermethylation status of the mismatch repair gene hMLH1 in patients with sporadic renal cell carcinoma]. / Hipermetilación del promotor del gen reparador hMLH1 en la patogenia del carcinoma de células renales esporádico.

BACKGROUND AND OBJECTIVE: Epigenetic inactivation is a gene function abnormality that produces no changes in the DNA sequence, with the most frequent epigenetic alteration being hypermethylation of CpG islands in the promoter regions of the genes. Based on recent indications of a potential relationship between mismatch repair genes and renal cell carcinoma (RCC), we were interested in investigating the existence of promoter hypermethylation of the hMLH1 gene in tumor DNA samples from patients with sporadic RCC. MATERIAL AND METHOD: Sixty-five tumor tissue specimens were collected consecutively. The DNA was first obtained and purified, then digested with the restriction enzymes Hpa II and Msp I, followed by polimerase chain reaction amplification of 3 promoter regions of the hMLH1 gene, agarose gel electrophoresis, and densitometric analysis of the images of the amplified bands. RESULTS: Mean patient age was 63.7 years. The most frequent cell type was clear cell carcinoma (67.7%). 73.9% of tumors were diagnosed in stages below pT2, 9.3% had gland involvement and 20%, distant metastasis. No somatic hypermethylation was detected in the promoter region of the hMLH1 gene in any of the patients studied. CONCLUSIONS: Our data indicate that promoter hypermethylation of the hMLH1 gene is not implicated in the pathogenesis of sporadic RCC, and therefore the existence of another type of mutation, microsatellite instability and/or loss of heterozygosity should be examined to determine the possible role of this gene in sporadic RCC.

Hipermetilación del promotor del gen reparador hMLH1 en la patogenia del carcinoma de células renales esporádico / Promoter hypermethylation status of the mismatch repair gene hMLH1 in patients with sporadic renal cell carcinoma

Fundamento y objetivo: La metilación de los islotes CpG (secuencias del genoma ricas en guanina y citosina) presentes en las regiones promotoras constituye la forma de inactivación epigenética más común y es una alteración en el mecanismo de regulación de los genes que no requiere cambios en la secuencia del ADN. Se ha descrito hipermetilación del promotor del gen hMLH1 en algunas líneas celulares procedentes de carcinoma de células renales (CCR). Nuestro objetivo ha sido determinar la existencia de hipermetilación en el promotor del gen hMLH1 en muestras de ADN tumoral de pacientes con CCR esporádico. Material y método: Se recogieron consecutivamente 65 muestras de tejido tumoral procedente de pacientes diagnosticados de CCR. Tras la obtención y purificación del ADN se procedió a su digestión con las enzimas de restricción HpaII y MspI, seguido de amplificación mediante reacción en cadena de la polimerasa de 3 regiones del promotor del gen hMLH1, electroforesis en gel de agarosa y análisis densitométrico de las bandas amplificadas. Resultados: La edad media de los pacientes fue de 63,7 años. El tipo celular más frecuente fue el carcinoma de células claras (67,7%). Un 73,9% de los tumores se diagnosticaron en estadios inferiores a pT2, un 9,3% tenían afectación ganglionar y un 20%, metástasis a distancia. No se detectó hipermetilación somática en la región promotora del gen hMLH1 en ninguno de los pacientes estudiados. Conclusiones: La hipermetilación del promotor del gen hMLH1 no parece intervenir en la patogenia del CCR esporádico, y es necesario analizar la existencia de otro tipo de mutaciones, inestabilidad de microsatélites y/o pérdida de heterocigosis para determinar el posible papel de este gen en la patogenia del CCR esporádico

Background and objective: Epigenetic inactivation is a gene function abnormality that produces no changes in the DNA sequence, with the most frequent epigenetic alteration being hypermethylation of CpG islands in the promoter regions of the genes. Based on recent indications of a potential relationship between mismatch repair genes and renal cell carcinoma (RCC), we were interested in investigating the existence of promoter hypermethylation of the hMLH1 gene in tumor DNA samples from patients with sporadic RCC. Material and method: Sixty-five tumor tissue specimens were collected consecutively. The DNA was first obtained and purified, then digested with the restriction enzymes Hpa II and Msp I, followed by polimerase chain reaction amplification of 3 promoter regions of the hMLH1 gene, agarose gel electrophoresis, and densitometric analysis of the images of the amplified bands. Results: Mean patient age was 63.7 years. The most frequent cell type was clear cell carcinoma (67.7%). 73.9% of tumors were diagnosed in stages below pT2, 9.3% had gland involvement and 20%, distant metastasis. No somatic hypermethylation was detected in the promoter region of the hMLH1 gene in any of the patients studied. Conclusions: Our data indicate that promoter hypermethylation of the hMLH1 gene is not implicated in the pathogenesis of sporadic RCC, and therefore the existence of another type of mutation, microsatellite instability and/or loss of heterozygosity should be examined to determine the possible role of this gene in sporadic RCC

Determination of vhl gene mutations in sporadic renal cell carcinoma.

OBJECTIVE: Characterization of the molecular status of the vhl gene and its relationship to the usual prognostic factors could help establish new diagnostic and therapeutic strategies. This study determines the presence of vhl gene mutations in tumor tissue from patients with sporadic renal cell carcinoma, the relationship between the mutations and classic prognostic factors, and the possible impact on protein function. MATERIALS AND METHOD: Cross-sectional analytical study investigating vhl gene mutations in tumor tissue by PCR testing of the gene exons and automatic sequencing in 96 patients with sporadic renal cell carcinoma. The characteristics of the mutations detected, as well as their relationship with various clinical and pathological parameters and the possible impact on protein function, were analyzed. RESULTS: Twenty-two mutations were found in 21 patients (21.9%); 68.2% were point mutations. The mutations were associated only with tumor histotype and were located in protein functional domains. Three mutations were detected in the intron position; 73.7% were considered relevant to protein function. CONCLUSION: The percentage of vhl gene mutations in sporadic renal cell carcinoma was relatively low, mutations were seen more often in clear cell carcinoma and were not related to the classic clinical prognostic factors. Mutations of the vhl gene produce important changes in the protein, causing it to lose its tumor suppressor function.

Relationship between the Arg72Pro polymorphism of p53 and outcome for patients with traumatic brain injury.

OBJECTIVE: To determine the relationship between the Arg72Pro polymorphism of p53 and the outcome after traumatic brain injury (TBI) in humans. DESIGN AND SETTING: A prospective study was carried out in a ten-bed surgical intensive care unit (SICU) of a university hospital. PATIENTS: The study included 90 caucasian patients who had experienced a severe TBI within the previous 24 h. The exclusion criterion was a previous deficit in the central nervous system. MEASUREMENTS AND RESULTS: The main recorded outcomes were values on the Glasgow Outcome Scale (GOS) at discharge from the SICU (GOS-0) and 6 months later (GOS-6). A blood sample was taken from each patient and all samples were analyzed by an allele-specific polymerase chain reaction for detection of Arg72Pro polymorphism of p53. These polymorphisms were tested for their association with values of GOS-0 and GOS-6. A two-tailed value of p<0.05 was considered statistically significant. The frequency of the argine/argine (Arg/Arg) genotype was greater among the patients who had a bad outcome at GOS-0 (69 vs 31% in the bad-outcome group; p=0.029). A multiple logistic regression analysis showed that patients with the Arg/Arg variation had a 2.9-fold greater risk of having a bad outcome at discharge from the SICU (95% confidence interval, 1.05-8.31; p=0.039 ). There was no similar relationship with respect to GOS-6. CONCLUSION: The Arg/Arg genotype of the Arg72Pro polymorphism in p53 is associated with increased likelihood of a bad outcome at discharge from the SICU.

[Mutational analysis of the vhl gene in the sporadic renal cell carcinoma]. / Análisis mutacional del gen vhl en el carcinoma de células renales de aparición esporádica.

OBJECTIVES: To assess and characterize mutations in the vhl gene in tumor tissue samples from 30 patients undergoing surgery for renal cell carcinoma in our department. METHODS: Descriptive, observational and analytical study of 30 patients undergoing surgery for RCC, analyzing the vhl gene sequence in tumor tissue, and using healthy renal tissue from the same patients as controls. Tissues were processed by DNA extraction, PCR amplification of the three exons that conform the gene, and ulterior automatic sequence analysis of the amplified exons between intronic primers previously designed. The sequence is compared with the corresponding exons included in the GeneBank. Alterations were checked by backwards sequence analysis. RESULTS: 9 mutations (30%) were found in the tumoral samples analyzed. 7 of them were punctual (one of them intronic); the other two were deletions. Mutations were distributed among the three exons: 3 in exon 1, 4 in exon 2, 1 in exon 3 and 1 intronic. One of the samples showed 2 mutations. Control tissue was free of mutations. CONCLUSIONS: Sporadic RCC shows mutations in the vhl gene which mainly appear in the clear cell subtype. Such alterations result in severe disturbances in the protein, disturbing its tumor suppressing function.