A genome-wide association study identifies novel gene associations with facial skin wrinkling and mole count in Latin Americans.

BACKGROUND: Genome-wide association studies (GWASs) have identified genes influencing skin ageing and mole count in Europeans, but little is known about the relevance of these (or other genes) in non-Europeans. OBJECTIVES: To conduct a GWAS for facial skin ageing and mole count in adults < 40 years old, of mixed European, Native American and African ancestry, recruited in Latin America. METHODS: Skin ageing and mole count scores were obtained from facial photographs of over 6000 individuals. After quality control checks, three wrinkling traits and mole count were retained for genetic analyses. DNA samples were genotyped with Illumina's HumanOmniExpress chip. Association testing was performed on around 8 703 729 single-nucleotide polymorphisms (SNPs) across the autosomal genome. RESULTS: Genome-wide significant association was observed at four genome regions: two were associated with wrinkling (in 1p13·3 and 21q21·2), one with mole count (in 1q32·3) and one with both wrinkling and mole count (in 5p13·2). Associated SNPs in 5p13·2 and in 1p13·3 are intronic within SLC45A2 and VAV3, respectively, while SNPs in 1q32·3 are near the SLC30A1 gene, and those in 21q21·2 occur in a gene desert. Analyses of SNPs in IRF4 and MC1R are consistent with a role of these genes in skin ageing. CONCLUSIONS: We replicate the association of wrinkling with variants in SLC45A2, IRF4 and MC1R reported in Europeans. We identify VAV3 and SLC30A1 as two novel candidate genes impacting on wrinkling and mole count, respectively. We provide the first evidence that SLC45A2 influences mole count, in addition to variants in this gene affecting melanoma risk in Europeans.

Relationship between tumor-associated immune infiltrate and p16 staining over clinicopathological features in acral lentiginous melanoma.

PURPOSE: This study aims to evaluate the association between composition of tumor-infiltrating lymphocytes (TIL) and expression of p16 in acral lentiginous melanoma (ALM), and their impact on prognosis. MATERIALS AND METHODS: A cohort of 148 surgical pathology specimens of ALM was studied. TIL were evaluated by immunohistochemical detection of CD3 and CD8, along with CD20, CD4, CD68, and CD163 in a subset of 43 cases. p16 protein expression was also investigated in all the cases. RESULTS: The median age was 66 years, median Breslow thickness was 6.0 mm, grade III TIL was found in 28.4% and lymph nodes were involved in 54.2%. Breslow thickness (p < 0.001), stage I-II (p < 0.001), negative lymph nodes (p < 0.001) and < 10% p16 (p = 0.01) were associated with longer survival. Grade III of TIL was associated with thinner Breslow thickness (p = 0.008) and lower mitosis (p = 0.047). A higher density of CD3 TIL was associated with male gender (p = 0.008), thinner Breslow thickness (p = 0.047), negative lymph node (p = 0.031), early stage (p = 0.046), and p16 nuclear expression of > 10% (p = 0.045). Higher CD8 TIL was associated with > p16 (p = 0.03). Survival analysis found that longer survival had a trend to be associated with high TIL (p = 0.090). Levels of CD3+ and CD8+ cells were correlated with those of CD4+, CD20+, CD68+ and CD163+ immune cells. CONCLUSIONS: Higher levels of TIL tend to be associated with better overall survival in ALM. Loss of expression of p16 is associated with lower levels of CD3+ and CD8+ TIL, indicating a probable relationship between p16 and TIL immune response in ALM .

Tumor infiltrating lymphocytes in acral lentiginous melanoma: a study of a large cohort of cases from Latin America.

PURPOSE: Acral lentiginous melanoma (ALM) is a poor prognosis subtype and is the most prevalent in non-Caucasian populations. The presence of tumor infiltrating lymphocytes (TILs) has been associated with poor prognosis in melanoma. A large cohort of ALM cases was studied to determine status of TIL and its association with outcome. METHODS: All patients with cutaneous melanoma presenting from 2005 to 2012 at Instituto Nacional de Enfermedades Neoplasicas in Peru were retrospectively identified. Clinicopathological information was obtained from the medical charts. A prospective evaluation of TIL was performed. Analysis of association between ALM and clinicopathological features including TIL as well as survival analysis compared the outcome of ALM to whole group and extremity NALM was performed. RESULTS: 537 ALM from a total of 824 cutaneous melanoma cases were studied. Older age (p = 0.022), higher Breslow (p = 0.008) and ulceration (p < 0.001) were found to be more frequent in ALM. Acral had worse overall survival (OS) compared with the whole group (p = 0.04). Clinical stage (CS) I-II patients had a median OS of 5.3 (95% CI 4.3-6.2) for ALM and 9.2 (95% CI 5.0-7.0) for extremity NALM (p = 0.016). Grade 0 (absence of TIL), I, II and III were found in 7.5, 34.5, 32.1, and 25.9%, respectively. Lower TIL grade was associated with larger tumor size (p = 0.003), higher Breslow (p = 0.001), higher Clark level (p = 0.007), higher CS (p = 0.002), extremity location (p = 0.048), histological subtype ALM (p = 0.024) and better OS (p = 0.001). CONCLUSIONS: ALM is highly prevalent in Peru and carries poor outcome. Lower TIL levels were associated with poor outcome and ALM.

Estudio clínico del primer molar permanente en niños de 6 años de edad de la comuna de calbuco, chile / Clinical study of the first permanent molar in children 6 years of age Calbuco commune, chile

Determinar la prevalencia de caries que presentan los Primeros Molares Permanentes (PMP) en niños chilenos de 6 años. Se realizó un estudio descriptivo, de corte transversal del estado de los PMP, en niños de seis años que asistieron al Hospital de Calbuco, X Región, Chile, durante el año 2008. Un operador calibrado examinó 166 niños de nivel socio-económico similar. La presencia de caries se estableció en base a los criterios propuestos por la OMS en el Oral Health Survey Basic Methods, para estudios epidemiológicos. El 65% (434/664) de los PMP se encontraban erupcionados, 47 (7%) semi- erupcionados y el 28% (183/664) aún no erupcionaban. De los PMP erupcionados y semierupcionados, el 69,02% (332/481) presenta caries. Al comparar la prevalencia de caries entre los PMP superiores y los inferiores se encontró una diferencia estadísticamente significativa (p<0,001). En los PMP superiores la mayor prevalencia de caries fue en la superficie oclusal, con un 56% (131/235), y en los PMP inferiores la superficie más afectadas fue la vestibular con un 68% (168/246).Conclusión: Los resultados demuestran la necesidad de incorporar estrategias de educativas y preventivas antes de la erupción del PMP

To determinate the prevalence of caries in first permanent molar (FPM) in six year-old Chilean children.Method: A Cross-sectional study was performed to state of FPM in six year-old children who attended in Calbuco Hospital, during 2008. A investigator previously calibrated to examined 166 Children belonging to the same socio-economic level. Caries diagnoses were carried out using standardized methods World Health Organization. 65% (434/664) of FPM were erupted and 47 (7%) partially erupted and 28% (183/664) were not erupted yet. From FPM erupted and partially erupted 69,02% (332/481) have caries. When comparing the prevalence of caries between upper and lower PFM, a significant statistical association was found (p<0,001). In the upper FPM the mayor prevalence of caries was found in the occlusal with 56% (131/235), in the lower FPM the most affected surface was the vestibular with a 68% (168/246). The results show the necessity of incorporating educative and preventive strategies before the eruption of FPM

Manejo del trauma penetrante cervical: serie de casos y revisión de literatura / Management of penetrating neck trauma: case series and review of the literature

Introduction: Penetrating neck trauma is a rare disease, but with high severity that affects the Emergency Room Services. Morbidity and mortality are due to injury of vascular structures and the aerodigestive tract. Conservative or surgical management depends on hemodynamic stability, the affected cervical zone and the presence of clinical signs of alert. Objective: To describe cervical penetrating trauma managed in the emergency unit of the Hospital Base Osorno, between January 2004 and April 2011 and review the literature. Material and Methods: We performed a retrospective review of all patients with penetrating cervical trauma seen at our center in this period. Results: Twelve patients were treated, nine were male. The average age was 29 years, ranging from 4 to 55 years. Zone II was the most affected with 9cases. Eight were stab injuries. The main surgical indications were active bleeding, subcutaneous emphysema and air exiting from the wound. The average length of hospital stay was 3,9 days. One patient died. Conclusions: In our center, penetrating neck trauma is a rare disease, which usually requires a surgical approach, forcing the emergency surgeon to be aware of warning signs to determine the need for surgical exploration or conservative management.

Central corneal thickness and intraocular pressure in Puerto Rican glaucoma patients

OBJECTIVE: To compare the variations in central corneal thickness and intraocular pressure measurements according to race, gender, and age. METHODS: A non-concurrent prospective study of 372 (744 eyes) glaucoma patients was conducted. Central corneal thickness was measured with ultrasound pachymeter and intraocular pressure with Goldmann tonometer. The relationship between CCT, race, gender, and age was evaluated using both descriptive and statistical analysis. RESULTS: The population age was 64 +/- 19.52 years. The mean central corneal thickness was 546 +/- 43.84 microm. The mean corrected intraocular pressure was 17 +/- 5.26 mm Hg. Central corneal thickness of male patients (549 +/- 43.43 pm) was thicker than that of female patients (546 +/- 41.83 microm). The mean corrected intraocular pressure in male patients (16 +/- 5.41 mm Hg) was less than that of female patients (17 +/- 5.18 mm Hg). The mean central corneal thickness in patients 0 to 9 years-of-age was 548 +/- 36.08 microm; 10 to 19 years-of-age was 606 +/- 82.30 microm; 20 to 29 years-of-age was 564 +/- 29.23 microm; 30 to 39 years-of-age was 579 +/- 15.32 microm; 40 to 49 years-of-age was 546 +/- 48.29 microm; 50 to 59 years-of-age was 550 +/- 38.12 mirom; 60 to 69 years-of-age was 545 +/- 40.22 microm; 70 to 79 years-of-age was 541 +/- 34.71 microm; 80 to 89 years-of-age was 541 +/- 34.05 microm; older than 90 years-of-age was 527 +/- 46.90 microm. CONCLUSIONS: Central corneal thickness of glaucoma patients in Puerto Rico was similar to that of Hispanics in the continental United States. However, the intraocular pressure and corrected intraocular pressure of glaucoma patients in Puerto Rico were statistically higher than that of Hispanics with glaucoma in the continental United States. In our study population, central corneal thickness and intraocular pressure were not affected by gender or age.

Early kinetics of amyloid fibril formation reveals conformational reorganisation of initial aggregates.

Understanding the initial steps of protein aggregation leading to the formation of amyloid fibrils remains a challenge. Here, the kinetics of such a process is determined for a misfolding protein model, ADA2h. The double nature of the very early kinetics suggests a step model of aggregation, where the denatured polypeptide folds into an aggregated beta-intermediate that subsequently reorganises into a more organised beta-sheet-richer structure that finally results in amyloid fibre formation. To determine the regions of the protein involved in amyloidosis, we have analysed a series of mutants previously made to study ADA2h folding. Using the algorithm TANGO, we have designed mutants that should enhance or decrease aggregation. Experimental analysis of the mutants shows that the C terminus of the molecule (comprising the last and edge beta-strand) is the major contributor to amyloid fibril formation, in good agreement with theoretical predictions. Comparison with proteins with similar topology reveals that family folds do not necessarily share the same principles of protein folding and/or aggregation.

Valoración del estado nutricional al ingreso de los pacientes internados en un hospital de niños / Evaluation of the nutritional status at admission in hospitalized patients in the pediatric hospital

Objetivos: conocer el estado nutricional de los pacientes que ingresan al hospital a través de un tamizaje nutricional. Determinar el día de inicio de la alimentación. Relacionar días medios de internación según estado nutricional y patología de base. Material y método: se estudiaron en forma prospectiva todos los pacientes ingresados en la salas de clínica médica (n:453) durante un período de tres meses. A través de una planilla se registraron los siguientes datos: fecha de ingreso y egreso; edad; sexo; diagnóstico de ingreso; peso; talla; día de inicio de la alimentación. Para evaluar el estado nutricional se tomó como patrón de referencia las tablas de NCHs/OMS y la clasificación del mismo se realizó en base a los criterios de Waterlow. Resultados: evaluaron 453 pacientes. El tamizaje nutricional mostró que el 47 por ciento de los pacientes al ingresar al hospital eran eutróficos y el 53 por ciento restante presentó algún grado de malnutrición: desnutrición aguda I° (15 por ciento), desnutrición aguda II° (8 por ciento), desnutrición aguda III° (3 por ciento); desnutrición crónica I° (9,5 por ciento)...

Protein engineering as a strategy to avoid formation of amyloid fibrils.

The activation domain of human procarboxypeptidase A2 (ADA2h) aggregates following thermal or chemical denaturation at acidic pH. The aggregated material contains well-defined ordered structures with all the characteristics of the fibrils associated with amyloidotic diseases. Variants of ADA2h containing a series of mutations designed to increase the local stability of each of the two helical regions of the protein have been found to have a substantially reduced propensity to form fibrils. This arises from a reduced tendency of the denatured species to aggregate rather than from a change in the overall stability of the native state. The reduction in aggregation propensity may result from an increase in the stability of local relative to longer range interactions within the polypeptide chain. These findings show that the intrinsic ability of a protein to form amyloid can be altered substantially by protein engineering methods without perturbing significantly its overall stability or activity. This suggests new strategies for combating diseases associated with the formation of aggregated proteins and for the design of novel protein or peptide therapeutics.

[Natural history of paroxysmal nocturnal hemoglobinuria in adolescents,adults, and children: the Mexican experience]. / Historia natural de la hemoglobinuria paroxística nocturna en adolescentes, adultos y en la edad pediátrica: la experiencia mexicana.

PURPOSE: To compare the clinical patterns and survival of young and adult (AP) versus paediatric (PP) patients with paroxysmal nocturnal haemoglobinuria (PNH). PATIENTS AND METHODS: The clinical records of 117 patients (82% AP, 18% PP) seen in four cities of the Mexican Republic were analysed, the clinical course and survival of both groups being compared. RESULTS: No sex difference was found in the two patient-groups: 51% and 52% males, 49% and 48% females in AP and PP, respectively. The onset of PNH had similar distribution for the two groups of patients: aplastic form, 45% in AP and 62% in PP; cytopenias, 24% in AP versus 27% in PP; haemolysis, 28% in AP and 9% in PP, and thrombosis, 3% in AP versus 0% in PP. The clinical features with significant difference were: anaemic+haemorrhagic syndrome (39 AP (40%) vs 14 PP (67%), p = 0.02), initial diagnosis of immunologic thrombocytopenic purpura (7 AP (7%) vs 7 PP (33%), p = 0.003), and death rate (17 AP (18%) vs 8 PP (38%), p = 0.04). The actuarial survival curves showed significant differences between both groups (p = 0.045, Cox-Mantel), with estimated 10-year survival of 81% for AP and 55% for PP, and 15-year survivals of 64% for AP and 55% for PP. CONCLUSIONS: Seemingly, PNH in paediatric age has poorer prognosis than in adults, which is associated to higher incidence of fatal haemorrhages due to thrombocytopenia.

Favourable native-like helical local interactions can accelerate protein folding.

BACKGROUND: Extensive studies of peptide conformation have provided reasonable knowledge of the rules determining helix stability. This knowledge can be used to stabilize proteins against chemical and thermal denaturation. This has been done in two proteins: the chemotactic protein from Escherichia coli, Che Y (a 129 aa alpha/beta parallel protein with five alpha-helices, which shows an accumulating intermediate during refolding) and the activation domain of human procarboxypeptidase A2, ADA2h (a 81 aa alpha + beta protein domain, with two alpha-helices, which follows a two-state mechanism). As the introduced stabilizing interactions are local in nature, the energy balance between the contribution of local and nonlocal interactions changes considerably. Recent theoretical analyses of protein folding using simplified models have indicated that optimization of folding speed requires this balance to be biased towards nonlocal interactions. To determine whether this is the case, we study here the folding kinetics of two ADA2h mutants in which alpha-helix 1 (mutant M1) or 2 (mutant M2) has been stabilized through local interactions, as well as the equilibrium and kinetic behaviour of a double mutant (DM) in which both helices have been stabilized. RESULTS: The stability of DM is considerably enhanced with respect to wild type (WI) and this mutant can be considered as a thermoresistant protein (Tm > 363 K). The thermodynamic parameters obtained by chemical denaturation (urea and GdnHCl) show that DM is approximately 2.6 kcal mol-1 more stable than WT. The effects on folding kinetics are different in each of the single mutants. M1 shows very little effect in refolding, while its unfolding is greatly decelerated with respect to WT. M2 shows, together with a deceleration in unfolding, a significant acceleration in refolding. As with equilibrium parameters, the kinetics of the double mutant can be explained by the simple addition of the effects found in each single mutant. Interestingly enough, the refolding slope mkf in mutants M2 and DM is smaller than in the wild-type and M1 mutant. CONCLUSIONS: Thermoresistance can be achieved, in some cases, by increasing favourable native local interactions. The balance between local and nonlocal interactions can be significantly changed in some proteins and still keep a cooperative unfolding transition similar to that of the wild type. The introduction of favourable local interactions by mutational redesign can also be used to increase the folding speed of certain proteins, showing that not all proteins in nature have been optimized for rapid folding, contrary to what has been theoretically indicated. This behaviour is probably also shared by other polypeptides with highly unstructured denatured states. All these phenomena have been shown experimentally in ADA2h by mutations that increase helix stability. However, the effects promoted for such an approach in proteins with residual structure and/or intermediates in the denatured ensemble could be different. This has been shown by experiments performed on CheY in which the cooperativity of the folding process was greatly affected.

Stabilization of proteins by rational design of alpha-helix stability using helix/coil transition theory.

BACKGROUND: Increasing protein stability is a major goal of protein engineering because of its potential industrial and pharmacological applications. Several different rule-of-thumb strategies have been employed for such a purpose, but a general rational method is still lacking. Recently, there has been significant progress in our understanding of the interactions responsible for helix stability in monomeric peptides and this information has been included in algorithms based on the helix/coil transition theory. We set out to investigate whether it is possible to use these algorithms to rationally increase protein stability. RESULTS: Using a helix/coil transition algorithm, AGADIRms, we have designed mutations affecting solvent-exposed residues which, as predicted, significantly increase the helical stability in aqueous solution of peptides corresponding to the two alpha-helices of the activation domain of procarboxipeptidase A. Introduction of the same mutations in the protein results in proteins more resistant to urea or temperature denaturation, and there is a qualitative agreement between the expected and observed increases in stability. CONCLUSIONS: In this work we demonstrate that by using a helix/coil algorithm to design helix-stabilizing mutations on the solvent-exposed face of helices, it is possible to rationally increase the stability of proteins.

Autoexamen de mamas, una práctica para el autocuidado en salud / Self-examination breasts, a practice for self care in health

El propósito de este trabajo es observar que factores favorecedores u obtaculizadores llevan a la realización del autoexamen de mama y como es la calidad de ésta en las 42 alumnas del cuarto año de la carrera enfermera matrona del primer semestre académico 1990. Para operacionalizar la variable calidad de la técnica se creó una escala compuesta por las siguientes categorías: tiempo de práctica; regularidad de la práctica; período del ciclo menstrual en que lo realiza y componentes de la técnica. La edad promedio de la población fue de 22.5 años, solteras y nulíparas. El 69% del grupo en estudio realiza el autoexamen de mamas. Los antecedentes mórbidos mamarios personales y familiares, no se demuestran como factores favorecedores ni obtaculizadores en la práctica del autoexamen de mamas, lo que difiere de la literatura revisada. El 95.2% se percibe capaz de realizar el autoexamen de mamas. El 100% del grupo en estudio considera la técnica útil, pero el 31% no la realiza por olvido. El 31% de la población percibe ser susceptible de desarrollar cáncer mamario, de este grupo 26% lo practica, lo que se podría considerar como un factor favorecedor para la realización de esta técnica. El 79% de los alumnos realiza el examen de mamas desde hace 6 meses o más lo que se podría asociar al aprendizaje de esta técnica en cursos atingentes al tema efectuados en semestres anteriores a esta investigación. En relación a la calidad de autoexamen de mamas el 80% de la población varía entre excelente y bueno. En cuanto a magnitud de la práctica del autoexamen de mamas el 69% de las alumnas lo realiza. Los hallazgos son preocupantes si se considera que son alumnas futuras enfermeras matronas, promotoras del autocuidado en salud

Analysis of the activation process of porcine procarboxypeptidase B and determination of the sequence of its activation segment.

The molecular events which lead to the proteolytic transformation of porcine procarboxypeptidase B (PCPB) in carboxypeptidase B (CPB) have been determined. Among pancreatic and other tested proteinases, trypsin is the only one capable of generating carboxypeptidase B activity from the zymogen, in vitro. In the first step of this process, trypsin produces cleavage at the boundary between the activation region and the CPB region. Subsequently, a definite sequence of cleavages occurs at the C-terminal end of the released activation segment of 95 residues, giving rise to characteristic intermediates and to a proteolytically resistant activation fragment of 81 residues. In this process, the newly formed CPB participates in the quick-trimming of the released activation peptides. Only a single CPB species is formed in the activation process. This fact and the inability of the released activation peptides to inhibit CPB--and, therefore, their inability to slow down the kinetics of appearance of CPB activity--are two important characteristics differentiating between the activation processes of procarboxypeptidases A and B. The sequence of the 95 residues (MW = 12,835) of the activation region of porcine PCPB has also been deduced, largely from the information obtained by Edman degradation of its fragments and in part by considerations of homology with the rat precursor. The porcine PCPB activation region contains a high percentage of acidic residues, lacks cysteines, methionines, and side-chain posttranslational modifications, and presents a low but significant homology (31%) with the corresponding sequence of porcine procarboxypeptidase A.

[Polychemotherapy in multiple myeloma. The COMPA protocol]. / Poliquimioterapia en mieloma múltiple. Esquema C.O.M.P.A.

Ninety three patients with multiple myeloma (MM) were treated with cyclophosphamide, vincristine, melphalan, prednisone and adriamycin (C.O.M.P.A.). Their median age was 60.9 years and sixty five were males. Seven patients were in stage I-A; 25 in II-A; 33 in III-A and 28 in III-B. Complete remission (CR) was achieved in 61 (65.6%), partial remission (PR) in 18 (19.3%) and no response in 14 (15%). At present, the mean survival of the CR group, is 32.3 months (10 to 78), and of the PR 11.2 months (6 to 18). The actuarial survival of the CR group is 37.9 months. A hemoglobin level lower than 8.5 g/dL, serum creatinine higher than 2.0 mg/dL, and stage III disease were factors that together negatively influenced in both response to treatment and survival. Proteinuria did not affect response, but it was a negative factor for survival. Thirty percent of deaths were due to infection, and 24.5% to myeloma activity associated with infection. We conclude that this five drug combination (C.O.M.P.A.) achieves a high percentage of complete remissions, but does not differ significantly from other reported schemes in the mean survival obtained for multiple myeloma.

Trombocitopenia y enfermedad tiroidea: presentación de 11 casos y revisión de la literatura / Thrombocytopenia and thyroid disease: report of 11 cases and review of the literature

Se informan once casos de trombocitopenia asociada a enfermedad tiroidea. Todos ellos mujeres. En 5 pacientes apareció la enfermedad tiroidea antes de la trombocitopenia, en 3 se trató de hipotiroidismo que se manifestó 13 años antes y en ellos se encontró el nivel más alto de inmunoglobulina "G" asociada a las plaquetas. En 4, se presentaron ambas enfermedades al mismo tiempo y en todas se trató de hipertiroidismo. En dos se resolvieron ambos problemas con tratamiento médico y en dos fue necesaria la esplenectomía. Tres recibieron Iodo radioactivo. En 2 pacientes la trombocitopenia precedió a la enfermedad tiroidea y tuvieron hipertiroidismo las dos, en una el cuadro clínico fue característico de tiroiditis de Hashimoto que se confirmó por histopatología después de cirugía de tiroides. No recibieron Iodo radioactivo y tienen el tiempo de observación más largo (8 años), sin evidencia de enfermedad autoinmune o linfoproliferativa. Se discuten los casos a la luz de los informes de la literatura y los mecanismos etiopatogénicos propuestos para tal asociación