RESUMEN

PURPOSE: Incisional hernia (IH) is one of the most frequent postoperative complications. Of all patients undergoing IH repair, a vast amount have a hernia which can be defined as a large incisional hernia (LIH). The aim of this study is to identify the preferred technique for LIH repair. METHODS: A systematic review of the literature was performed and studies describing patients with IH with a diameter of 10 cm or a surface of 100 cm2 or more were included. Recurrence hazards per year were calculated for all techniques using a generalized linear model. RESULTS: Fifty-five articles were included, containing 3,945 LIH repairs. Mesh reinforced techniques displayed better recurrence rates and hazards than techniques without mesh reinforcement. Of all the mesh techniques, sublay repair, sandwich technique with sublay mesh and aponeuroplasty with intraperitoneal mesh displayed the best results (recurrence rates of <3.6%, recurrence hazard <0.5% per year). Wound complications were frequent and most often seen after complex LIH repair. CONCLUSIONS: The use of mesh during LIH repair displayed the best recurrence rates and hazards. If possible mesh in sublay position should be used in cases of LIH repair.

Asunto(s)

Hernia Ventral/cirugía , Herniorrafia/métodos , Mallas Quirúrgicas , Hernia Ventral/etiología , Humanos , Implantación de Prótesis

RESUMEN

The superficial branch of the radial nerve (SBRN) is known for developing neuropathic pain syndromes after trauma. These pain syndromes can be hard to treat due to the involvement of other nerves in the forearm. When a nerve is cut, the Schwann cells, and also other cells in the distal segment of the transected nerve, produce the nerve growth factor (NGF) in the entire distal segment. If two nerves overlap anatomically, similar to the lateral antebrachial cutaneous nerve (LACN) and SBRN, the increase in secretion of NGF, which is mediated by the injured nerve, results in binding to the high-affinity NGF receptor, tyrosine kinase A (TrkA). This in turn leads to possible sprouting and morphological changes of uninjured fibers, which ultimately causes neuropathic pain. The aim of this study was to map the level of overlap between the SBRN and LACN. Twenty arms (five left and 15 right) were thoroughly dissected. Using a new analysis tool called CASAM (Computer Assisted Surgical Anatomy Mapping), the course of the SBRN and LACN could be compared visually. The distance between both nerves was measured at 5-mm increments, and the number of times they intersected was documented. In 81% of measurements, the distance between the nerves was >10 mm, and in 49% the distance was even <5 mm. In 95% of the dissected arms, the SBRN and LACN intersected. On average, they intersected 2.25 times. The close (anatomical) relationship between the LACN and the SBRN can be seen as a factor in the explanation of persistent neuropathic pain in patients with traumatic or iatrogenic lesion of the SBRN or the LACN.

Asunto(s)

Nervio Musculocutáneo/anatomía & histología , Neuralgia/etiología , Nervio Radial/anatomía & histología , Anciano , Anciano de 80 o más Años , Cadáver , Dolor Crónico/etiología , Femenino , Antebrazo/inervación , Humanos , Procesamiento de Imagen Asistido por Computador , Masculino , Persona de Mediana Edad

RESUMEN

STUDY QUESTION: Is in vitro fertilization treatment with or without intracytoplasmatic sperm injection (IVF/ICSI) associated with changes in first and second trimester embryonic and fetal growth trajectories and birthweight in singleton pregnancies? SUMMARY ANSWER: Embryonic and fetal growth trajectories and birthweight are not significantly different between pregnancies conceived with IVF/ICSI treatment and spontaneously conceived pregnancies with reliable pregnancy dating. WHAT IS KNOWN ALREADY: IVF/ICSI treatment has been associated with increased risks of preterm birth, fetal growth restriction and low birthweight. Decreased first-trimester crown-rump length (CRL) in the general population has been inversely associated with the same adverse pregnancy outcomes. STUDY DESIGN, SIZE, DURATION: In a prospective periconception birth cohort study conducted in a tertiary centre, 146 singleton pregnancies with reliable pregnancy dating and nonmalformed live borns were investigated, comprised of 88 spontaneous and 58 IVF/ICSI pregnancies. PARTICIPANTS/MATERIALS, SETTING, METHODS: Serial 3D ultrasound scans were performed from 6 to 12 weeks of gestation. As estimates of embryonic growth, CRL and embryonic volume (EV) were measured using the I-Space virtual reality system. General characteristics were obtained from self-administered questionnaires at enrolment. Fetal growth parameters at 20 weeks and birthweight were obtained from medical records. To assess associations between IVF/ICSI and embryonic growth trajectories, estimated fetal weight and birthweight, stepwise linear mixed model analyses and linear regression analyses were performed using square root transformed CRL and fourth root transformed EV. MAIN RESULTS AND THE ROLE OF CHANCE: In 146 pregnancies, 934 ultrasound scans were performed of which 849 (90.9%) CRLs and 549 (58.8%) EVs could be measured. Embryonic growth trajectories were comparable between IVF/ICSI pregnancies and spontaneously conceived pregnancies (CRL: ßIVF/ICSI = 0.10√mm; P = 0.10; EV: ßIVF/ICSI = 0.03(4)√cm³; P = 0.13). Estimated fetal weight and birthweight were also comparable between both groups (ßIVF/ICSI = 6 g; P = 0.36 and ßIVF/ICSI = 80 g; P = 0.24, respectively). LIMITATIONS, REASONS FOR CAUTION: Variations in embryonic growth trajectories of spontaneously conceived pregnancies with reliable pregnancy dating may partially be a result of less precise pregnancy dating and differences in endometrium receptivity compared with IVF/ICSI pregnancies. WIDER IMPLICATIONS OF THE FINDINGS: The absence of a significant difference in embryonic and fetal growth trajectories suggests safety of IVF/ICSI treatment with regard to early embryonic growth. However, further research is warranted to ascertain the influence of IVF/ICSI treatments in a larger study population, and to estimate the impact of the underlying causes of the subfertility and other periconceptional exposures on human embryonic and fetal growth trajectories. FUNDING STATEMENT: This study was supported by the Department of Obstetrics and Gynaecology of the Erasmus MC, University Medical Centre. CONFLICT OF INTEREST: No competing interests are declared.

Asunto(s)

Desarrollo Embrionario , Fertilización In Vitro/efectos adversos , Desarrollo Fetal , Adulto , Femenino , Humanos , Modelos Lineales , Embarazo , Resultado del Embarazo , Estudios Prospectivos , Inyecciones de Esperma Intracitoplasmáticas/efectos adversos , Ultrasonografía Prenatal

RESUMEN

OBJECTIVE: To investigate the association between periconception maternal folate status and embryonic size. DESIGN: Prospective periconception cohort study. SETTING: Erasmus University Medical Centre, Rotterdam, the Netherlands. POPULATION: Seventy-seven singleton pregnancies recruited in 2009 and 2010. METHODS: We recruited women before 8 weeks of gestation and performed weekly three-dimensional ultrasound scans from enrolment up to 13 weeks of gestation. As a measure of embryonic growth, crown-rump length (CRL) measurements were performed using V-Scope software in the BARCO I-Space. Maternal blood was collected to determine first-trimester long-term red blood cell (RBC) folate status. Non-malformed live births were included in the analysis. We calculated quartiles of RBC folate, square root-transformed CRL data and performed multivariable linear mixed model analyses. MAIN OUTCOME MEASURES: Serial first-trimester CRL measurements. RESULTS: In total, 484 ultrasound scans were performed in 77 women, in 440 (90.7%) of which CRLs could be measured. RBC folate in the third quartile (1513-1812 nmol/l) was significantly associated with an increased CRL compared with the first two quartiles (814-1512 nmol/l) and the upper quartile (1813-2936 nmol/l; P(overall) = 0.03; adjusted for gestational age, smoking, body mass index and fetal sex). Compared with the third quartile, embryos in the upper quartile were 24.2% smaller at 6(+0) weeks [4.1 mm (95% confidence interval 3.5, 4.7) versus 5.4 mm (95% confidence interval 4.8, 6.1)] and 7.6% smaller at 12(+0) weeks [55.1 mm (95% confidence interval 52.9, 57.3) versus 59.6 mm (95% confidence interval 57.4, 62.0)] of gestation. CONCLUSIONS: This study suggests that a very high maternal periconception folate status is associated with reduced embryonic size. Whether these effects are beneficial or harmful requires further investigation.

Asunto(s)

Largo Cráneo-Cadera , Ácido Fólico/sangre , Adulto , Femenino , Humanos , Países Bajos , Valor Predictivo de las Pruebas , Embarazo , Primer Trimestre del Embarazo , Estudios Prospectivos , Ultrasonografía Prenatal

RESUMEN

STUDY QUESTION: Can reliable size charts of human embryonic brain structures be created from three-dimensional ultrasound (3D-US) visualizations? SUMMARY ANSWER: Reliable size charts of human embryonic brain structures can be created from high-quality images. WHAT IS KNOWN ALREADY: Previous studies on the visualization of both the cavities and the walls of the brain compartments were performed using 2D-US, 3D-US or invasive intrauterine sonography. However, the walls of the diencephalon, mesencephalon and telencephalon have not been measured non-invasively before. Last-decade improvements in transvaginal ultrasound techniques allow a better visualization and offer the tools to measure these human embryonic brain structures with precision. STUDY DESIGN, SIZE, DURATION: This study is embedded in a prospective periconceptional cohort study. A total of 141 pregnancies were included before the sixth week of gestation and were monitored until delivery to assess complications and adverse outcomes. PARTICIPANTS/MATERIALS, SETTING, METHODS: For the analysis of embryonic growth, 596 3D-US scans encompassing the entire embryo were obtained from 106 singleton non-malformed live birth pregnancies between 7(+0) and 12(+6) weeks' gestational age (GA). Using 4D View (3D software) the measured embryonic brain structures comprised thickness of the diencephalon, mesencephalon and telencephalon, and the total diameter of the diencephalon and mesencephalon. MAIN RESULTS AND THE ROLE OF CHANCE: Of 596 3D scans, 161 (27%) high-quality scans of 79 pregnancies were eligible for analysis. The reliability of all embryonic brain structure measurements, based on the intra-class correlation coefficients (ICCs) (all above 0.98), was excellent. Bland-Altman plots showed moderate agreement for measurements of the telencephalon, but for all other measurements the agreement was good. Size charts were constructed according to crown-rump length (CRL). LIMITATIONS, REASONS FOR CAUTION: The percentage of high-quality scans suitable for analysis of these brain structures was low (27%). WIDER IMPLICATIONS OF THE FINDINGS:  The size charts of human embryonic brain structures can be used to study normal and abnormal development of brain development in future. Also, the effects of periconceptional maternal exposures, such as folic acid supplement use and smoking, on human embryonic brain development can be a topic of future research. STUDY FUNDING/COMPETING INTEREST(S): This study was supported by the Department of Obstetrics and Gynaecology of the Erasmus University Medical Center. M.G. was supported by an additional grant from the Sophia Foundation for Medical Research (SSWO grant number 644). No competing interests are declared.

Asunto(s)

Encéfalo/embriología , Tamaño de los Órganos , Primer Trimestre del Embarazo , Adulto , Encéfalo/patología , Largo Cráneo-Cadera , Femenino , Edad Gestacional , Humanos , Imagenología Tridimensional , Masculino , Análisis Multivariante , Embarazo , Estudios Prospectivos , Valores de Referencia , Reproducibilidad de los Resultados , Ultrasonografía Prenatal

RESUMEN

STUDY QUESTION: Are maternal characteristics and lifestyle factors associated with human embryonic growth trajectories? SUMMARY ANSWER: Periconception maternal age is associated with increased, and smoking and alcohol use with decreased embryonic growth trajectories, estimated with crown-rump length (CRL) measurements. WHAT IS KNOWN ALREADY: Fetal weight is associated with health and disease in later life. Maternal characteristics and lifestyle factors affect fetal growth in the second and third trimesters of pregnancy and at birth; however, little is known about the association of these characteristics with first trimester embryonic growth. STUDY DESIGN, SIZE, DURATION: In a tertiary centre, pregnant women were recruited and enrolled in a prospective periconception cohort study before 8 weeks of gestation. We selected 87 spontaneously conceived singleton pregnancies of women recruited in 2009 and 2010 that ended in non-malformed live births. PARTICIPANTS/MATERIALS, SETTING, METHODS: We performed weekly three-dimensional ultrasound scans from enrolment up to 13 weeks of gestation. At enrolment, a questionnaire was completed. Embryonic CRL measurements were performed using the V-Scope software in the BARCO I-Space. Associations between maternal characteristics and embryonic growth were assessed using square root transformed CRL as response in linear mixed model analyses, adjusted for potential confounders. MAIN RESULTS AND THE ROLE OF CHANCE: Four hundred and ninety-six scans from 87 pregnancies were included. In the multivariable analysis, maternal age was positively associated with first trimester CRL (difference per maternal year of age 0.024√mm (95% confidence interval (CI) 0.009, 0.040), P = 0.001). At 6 and 12 weeks of gestation, the CRL of an embryo from a 40-year-old mother was estimated 2.0 mm (61%) and 7.2 mm (14%) larger, respectively, compared with an embryo from a 20-year-old mother. Smoking of 10 or more cigarettes per day was negatively associated with CRL (difference -0.211√mm (95% CI -0.416, -0.006), P = 0.04), with embryos that were 0.9 mm (18.7%) and 3.1 mm (5.5%) smaller at 6 and 12 weeks, respectively, compared with non-smokers. Periconception alcohol use was negatively associated with CRL growth rate (difference -0.0025√mm (95% CI -0.0047, -0.0003)/day gestational age, P = 0.022), with embryos that were 0.2 mm (3%) and 1.1 mm (2%) smaller at 6 and 12 weeks, respectively, compared with non-alcohol users. Parity, BMI and moment of initiation of folic acid use were not significantly associated with embryonic CRL. LIMITATIONS, REASONS FOR CAUTION: Due to the selection of pregnancies in a tertiary centre and the small number of pregnancies, the external validity of the results has to be confirmed using larger sample sizes and other population-based periconception cohort studies. WIDER IMPLICATIONS OF THE FINDINGS: The association of maternal age and smoking with embryonic growth is in line with previous literature, whereas the association between embryonic growth and alcohol use is a new finding. However, concerning exposure to alcohol, the effect estimate was small and it is questionable whether this is of clinical value. More research is warranted to unravel underlying mechanisms and to assess the implications for preconception and early pregnancy care, such as the development and implementation of effective lifestyle interventions. STUDY FUNDING/COMPETING INTEREST(S): The work was funded by the Department of Obstetrics and Gynaecology, Erasmus MC, University Medical Centre, Rotterdam, The Netherlands. The authors declare no conflicts of interest.

Asunto(s)

Desarrollo Fetal , Edad Materna , Primer Trimestre del Embarazo , Ultrasonografía Prenatal , Adolescente , Adulto , Consumo de Bebidas Alcohólicas/efectos adversos , Largo Cráneo-Cadera , Femenino , Peso Fetal , Humanos , Estudios Longitudinales , Países Bajos , Embarazo , Estudios Prospectivos , Fumar/efectos adversos

RESUMEN

BACKGROUND: Tendon transfers are essential for reconstruction of hand function in tetraplegic patients. To transfer the extensor carpi radialis longus (ECRL), the extensor carpi radialis brevis (ECRB) has to be sufficiently strong. However, there is currently no reliable clinical test to individually analyse both muscles. In order to develop a reliable preoperative clinical test, the anatomy of the muscle (innervation) areas of ECRB, ECRL and brachio-radialis (BR) was examined. METHODS: In 20 arms, the ECRB, ECRL and BR were dissected and localised. Subsequently, muscle-innervation points were mapped and categorised. A novel method, computer-assisted surgical anatomy mapping (CASAM), was used to visualise muscle areas and innervation points in a computed arm with average dimensions. RESULTS: For both ECRL and ECRB a 100% area could be identified, a specific area in the computed average arm in which the muscle was present for all 20 arms. For the ECRL, this area was situated at 16% of the distance between the lateral epicondyle and the deltoid muscle insertion. The ECRB 100% area was 5 times bigger than that of the ECRL and was located at 40% of the distance between the lateral epicondyle and the radial styloid process. The ECRL and BR showed one to three innervation points, the ECRB one to four. In 47% of the cases, there was a combined nerve branch innervating both the ECRL and the ECRB. CONCLUSIONS: It is feasible to develop a preoperative test; the 100% areas can be used for needle electromyography (EMG) or local anaesthetic muscle injections.

Asunto(s)

Procesamiento de Imagen Asistido por Computador , Fuerza Muscular , Músculo Esquelético/fisiología , Transferencia Tendinosa , Estudios de Factibilidad , Pie/inervación , Humanos , Contracción Isométrica/fisiología , Músculo Esquelético/anatomía & histología , Músculo Esquelético/inervación , Periodo Preoperatorio

RESUMEN

Maternal smoking during pregnancy and a low socioeconomic status (SES) lead to increased risks of adverse pregnancy outcome. Maternal education is often used as proxy for SES. We explored the programming of the insulin pathway genes IGF2 DMR (insulin growth factor 2 differentially methylated region), IGF2R (insulin growth factor 2 receptor) and INSIGF [the overlapping region of IGF2 and insulin (INS)] in the child through any periconception maternal smoking and education level. In 120 children at 17 months of age, methylation of DNA derived from white blood cells was measured. Periconception smoking and low education were independently associated with INSIGF methylation and showed a relative increase in methylation of +1.3%; P = 0.043 and +1.6%; P = 0.021. Smoking and low education showed an additive effect on INSIGF methylation (+2.8%; P = 0.011). There were no associations with IGF2 DMR and IGF2R methylation. Our data suggest that periconception maternal smoking and low education are associated with epigenetic marks on INSIGF in the very young child, this warrants further study in additional populations.

Asunto(s)

Metilación de ADN , Proteínas Mutantes Quiméricas/genética , Efectos Tardíos de la Exposición Prenatal , Fumar/efectos adversos , Adulto , Escolaridad , Epigénesis Genética , Femenino , Genes Sobrepuestos , Humanos , Lactante , Insulina/genética , Insulina/metabolismo , Factor II del Crecimiento Similar a la Insulina/genética , Factor II del Crecimiento Similar a la Insulina/metabolismo , Modelos Lineales , Masculino , Proteínas Mutantes Quiméricas/metabolismo , Embarazo , Receptor IGF Tipo 2/genética , Receptor IGF Tipo 2/metabolismo , Factores Socioeconómicos

RESUMEN

BACKGROUND: We have recently demonstrated that expression profiling is a more accurate and objective method to classify gliomas than histology. Similar to most expression profiling studies, our experiments were performed using fresh frozen (FF) glioma samples whereas most archival samples are fixed in formalin and embedded in paraffin (FFPE). Identification of the same, expression-based intrinsic subtypes in FFPE-stored samples would enable validation of the prognostic value of these subtypes on these archival samples. In this study, we have therefore determined whether the intrinsic subtypes identified using FF material can be reproduced in FFPE-stored samples. METHODS: We have performed expression profiling on 55 paired FF-FFPE glioma samples using HU133 plus 2.0 arrays (FF) and Exon 1.0 ST arrays (FFPE). The median time in paraffin of the FFPE samples was 14.1 years (range 6.6-26.4 years). RESULTS: In general, the correlation between FF and FFPE expression in a single sample was poor. We then selected the most variable probe sets per gene (n=17,583), and of these, the 5000 most variable probe sets on FFPE expression profiles. This unsupervised selection resulted in a better concordance (R(2)=0.54) between expression of FF and FFPE samples. Importantly, this probe set selection resulted in a correct assignment of 87% of FFPE samples into one of seven intrinsic subtypes identified using FF samples. Assignment to the same molecular cluster as the paired FF tissue was not correlated to time in paraffin. CONCLUSION: We are the first to examine a large cohort of paired FF and FFPE samples. We show that expression data from FFPE material can be used to assign samples to intrinsic molecular subtypes identified using FF material. This assignment allows the use of archival material, including material derived from large-randomised clinical trials, to determine the predictive and/or prognostic value of 'intrinsic glioma subtypes' on Exon arrays. This would enable clinicians to provide patients with an objective and accurate diagnosis and prognosis, and a personalised treatment strategy.

Asunto(s)

Neoplasias Encefálicas/genética , Perfilación de la Expresión Génica/métodos , Glioma/genética , Análisis por Conglomerados , Fijadores , Formaldehído , Secciones por Congelación , Regulación Neoplásica de la Expresión Génica , Humanos , Adhesión en Parafina/métodos , Reproducibilidad de los Resultados , Fijación del Tejido/métodos

RESUMEN

Neurotransmission requires Ca(2+)-dependent release of secretory products through fusion pores that open and reclose (partial membrane distention) or open irreversibly (complete membrane distention). It has been challenging to distinguish between these release modes; however, in the work presented here, we were able to deduce different modes of depolarization-evoked exocytosis in neuroendocrine chromaffin and PC12 cells solely by analyzing amperometric recordings. After we determined the quantal size (Q), event half-width (t(50)), event amplitude (I(peak)), and event decay time constant (τ(decay)), we fitted scatter plots of log-transformed data with a mixture of one- and two-dimensional Gaussian distributions. Our analysis revealed three distinct and differently shaped clusters of secretory events, likely corresponding to different modes of exocytosis. Complete membrane distention, through fusion pores of widely varying conductances, accounted for 70% of the total amount of released catecholamine. Two different kinds of partial membrane distention (kiss-and-run and kiss-and-stay exocytosis), characterized by mode-specific fusion pores with unitary conductances, accounted for 20% and 10%, respectively. These results show that our novel one- and two-dimensional analysis of amperometric data reveals new release properties and enables one to distinguish at least three different modes of exocytosis solely by analyzing amperometric recordings.

Asunto(s)

Técnicas Electroquímicas/métodos , Exocitosis , Células Neuroendocrinas/citología , Potenciales de Acción/fisiología , Animales , Células Cromafines/citología , Células Cromafines/metabolismo , Ratones , Células Neuroendocrinas/metabolismo , Distribución Normal , Células PC12 , Ratas

RESUMEN

To evaluate the possibility to give a prediction of the future (disease-free) survival, given the fact that a patient with a history of early-stage cervical cancer has been disease free for a specific period after treatment. Between January 1984 and April 2005, 615 patients with cervical cancer stages I-IIA underwent radical hysterectomy with or without adjuvant radiotherapy. The Kaplan-Meier method was used to detect statistical significance and multistate risk models to estimate the influence of covariates and to generate predicted survival curves by simulation. Simulations were done for patients with positive lymph nodes (n= 123), patients with negative lymph nodes (n= 492), and 4 hypothetical patients. The 5-year cancer-specific survival and disease-free survival of the entire group was 84% and 76%, respectively. The probability of death of the two lymph node groups and the four hypothetical patients was demonstrated in predicted cumulative probability plots. It is possible with multistate risk models to give a detailed prediction of the future (disease-free) survival, given the fact that a patient has been disease free for a specific period after treatment. This possibility is an important step forward to improve the quality of cancer care.

Asunto(s)

Ganglios Linfáticos/patología , Recurrencia Local de Neoplasia/mortalidad , Neoplasias del Cuello Uterino/mortalidad , Neoplasias del Cuello Uterino/patología , Adulto , Anciano , Anciano de 80 o más Años , Estudios de Cohortes , Terapia Combinada , Supervivencia sin Enfermedad , Femenino , Estudios de Seguimiento , Humanos , Histerectomía/métodos , Inmunohistoquímica , Estimación de Kaplan-Meier , Persona de Mediana Edad , Análisis Multivariante , Invasividad Neoplásica/patología , Recurrencia Local de Neoplasia/patología , Estadificación de Neoplasias , Países Bajos , Valor Predictivo de las Pruebas , Modelos de Riesgos Proporcionales , Radioterapia Adyuvante , Estudios Retrospectivos , Análisis de Supervivencia , Resultado del Tratamiento , Neoplasias del Cuello Uterino/terapia

RESUMEN

OBJECTIVE: The immune system is thought to play a crucial role in regulating collateral circulation (arteriogenesis), a vital compensatory mechanism in patients with arterial obstructive disease. Here, we studied the role of lymphocytes in a murine model of hindlimb ischemia. METHODS AND RESULTS: Lymphocytes, detected with markers for NK1.1, CD3, and CD4, invaded the collateral vessel wall. Arteriogenesis was impaired in C57BL/6 mice depleted for Natural Killer (NK)-cells by anti-NK1.1 antibodies and in NK-cell-deficient transgenic mice. Arteriogenesis was, however, unaffected in J alpha281-knockout mice that lack NK1.1+ Natural Killer T (NKT)-cells, indicating that NK-cells, rather than NKT-cells, are involved in arteriogenesis. Furthermore, arteriogenesis was impaired in C57BL/6 mice depleted for CD4+ T-lymphocytes by anti-CD4 antibodies, and in major histocompatibility complex (MHC)-class-II-deficient mice that more selectively lack mature peripheral CD4+ T-lymphocytes. This impairment was even more profound in anti-NK1.1-treated MHC-class-II-deficient mice that lack both NK- and CD4+ T-lymphocytes. Finally, collateral growth was severely reduced in BALB/c as compared with C57BL/6 mice, 2 strains with different bias in immune responsiveness. CONCLUSIONS: These data show that both NK-cells and CD4+ T-cells modulate arteriogenesis. Promoting lymphocyte activation may represent a promising method to treat ischemic disease.

Asunto(s)

Arteriopatías Oclusivas/inmunología , Linfocitos T CD4-Positivos/inmunología , Circulación Colateral/inmunología , Células Asesinas Naturales/inmunología , Neovascularización Fisiológica/inmunología , Animales , Arteriopatías Oclusivas/fisiopatología , Linfocitos T CD4-Positivos/metabolismo , Modelos Animales de Enfermedad , Arteria Femoral/crecimiento & desarrollo , Miembro Posterior/irrigación sanguínea , Isquemia , Células Asesinas Naturales/metabolismo , Ratones , Ratones Endogámicos BALB C , Ratones Noqueados

RESUMEN

We propose a method to jointly detect influential biomarkers and estimate how they change with dose. The assessment is made in dose-ranging trials collecting multiple outcomes for efficacy, safety, pharmacokinetics or pharmacodynamics. We regress all these outcomes versus a non-parametric transformation of the dose. The regression coefficients and the parameters from the dose-response model are simultaneously estimated using a penalized alternating least-squares method. We illustrate the technique with a phase I clinical trial and a metabonomic experiment in rats.

Asunto(s)

Biomarcadores/análisis , Relación Dosis-Respuesta a Droga , Análisis de los Mínimos Cuadrados , Modelos Biológicos , Animales , Enfermedad Hepática Inducida por Sustancias y Drogas , Ensayos Clínicos como Asunto/métodos , Simulación por Computador , Dietilhexil Ftalato/farmacocinética , Dietilhexil Ftalato/toxicidad , Dietilhexil Ftalato/orina , Humanos , Hepatopatías/metabolismo , Ratas

RESUMEN

OBJECTIVE: The objective of this study was to systematically evaluate the molecular basis of the association between visceral fat mass and plasma plasminogen activator inhibitor-1 (PAI-1) levels in man. DESIGN: A comprehensive approach comprising observational, in vitro, and human intervention studies. MEASUREMENTS AND RESULTS: We confirmed an exclusive relationship between visceral fat and plasma PAI-1 levels (r=0.79, P<0.001) and corroborated preferential PAI-1 release from adipose tissue explants. Yet, messenger RNA analysis and in vivo measurement of PAI-1 release from visceral fat (AV-differences over the omentum) not only excluded visceral adipose tissue as a relevant source of circulating PAI-1, but also excluded visceral fat as a significant source of proinflammatory mediators such as tumor necrosis factor-alpha, IL-1 or transforming growth factor-beta that could induce PAI-1 expression in tissues other than visceral fat. Short-term interventions with acipimox and growth hormone (GH) as well as statistical evaluation excluded free fatty acids and GH as metabolic links. Further analysis of the metabolic data in a stepwise regression model indicated that plasma PAI-1 levels and visceral fat rather are co-correlates that both relate to impaired lipid handling. CONCLUSION: Our PAI-1 studies show that visceral fat mass and plasma PAI-1 levels are co-correlated rather than causatively related, with lipid load as common denominator.

Asunto(s)

Grasa Intraabdominal/metabolismo , Inhibidor 1 de Activador Plasminogénico/sangre , Adiposidad/fisiología , Adulto , Antropometría , Citocinas/biosíntesis , Femenino , Hormona de Crecimiento Humana/deficiencia , Humanos , Mediadores de Inflamación/metabolismo , Grasa Intraabdominal/anatomía & histología , Grasa Intraabdominal/patología , Metabolismo de los Lípidos , Persona de Mediana Edad , Obesidad/metabolismo , Obesidad/patología , Inhibidor 1 de Activador Plasminogénico/genética , ARN Mensajero/genética , Técnicas de Cultivo de Tejidos , Factor de Necrosis Tumoral alfa/biosíntesis

RESUMEN

Total mesorectal excision (TME) is the standard treatment for rectal cancer, while transanal endoscopic microsurgery (TEM) is a recently introduced surgical approach for the treatment of rectal adenomas. Incorrect preoperative staging before TEM is a problem. To identify genetic changes that might correlate with tumour stage and could lead to optimized treatment selection we performed a genome-wide chromosomal instability search in a homogeneous, clinical cohort of rectal tumours. 78 rectal tumours during different clinical stages were analysed with 10K single nucleotide polymorphism (SNP) arrays. Logistic regression was performed to build a quantitative model of specific chromosomal aberrations. Overall, most cases (95%) had one or more chromosomal aberrations. We observed a clear correlation between the total number of aberrations and the different tumour stages. Specifically, the chromosomal events: gain of 8q22-24, 13q and 20q, and loss of 17p and 18q12-22, were far more abundant in carcinoma than in adenoma. In adenoma fractions from cases with a carcinoma (infiltrating at least in the submucosa), twice the amount of such 'malignant aberrations' was observed, compared to pure adenomas. Furthermore, combined aberrations such as gain of 13q and loss of 18q were only found in adenomatous fractions of carcinomas and not in benign lesions. Based on these five genomic events associated with carcinoma, a clear distinction between adenoma and carcinoma tissue could be made. These data should be validated further in order that they may be used in preoperative staging of rectal tumours.

Asunto(s)

Adenoma/patología , Carcinoma/patología , Inestabilidad Cromosómica , Análisis de Secuencia por Matrices de Oligonucleótidos , Polimorfismo de Nucleótido Simple , Neoplasias del Recto/genética , Neoplasias del Recto/patología , Adenoma/genética , Análisis de Varianza , Carcinoma/genética , Diagnóstico Diferencial , Genoma , Humanos , Modelos Logísticos , Pérdida de Heterocigocidad , Estadificación de Neoplasias

RESUMEN

In this paper the performance of three alignment algorithms, correlation optimized warping, parametric time warping and semi-parametric time warping, is compared on real chromatograms. Among these, parametric time warping is the simplest and fastest; generally less than 1s is required to align two chromatograms. It does not require the optimization of input parameters and allows the alignment of peak shifts in only one direction, or non-complex peak shifts in both directions. With correlation optimized warping and semi-parametric time warping complex peak shifts in both directions can be corrected but at the expense of the optimization of two input parameters. Semi-parametric time warping requires the selection of the proper number of B-splines in the warping function and, if necessary, the optimization of the penalty parameter. Often the default values can be used to obtain aligned signals. The optimization of the input parameters for correlation optimized warping (section length, slack) is not easy and time-consuming. Moreover, dependent on the input parameters, the computation time of the correlation optimized warping algorithm can be twice as long as for semi-parametric time warping for which computation times up to 23 s are required. However, the performance of both algorithms is equally good considering the improvement of the precision of the peak retention times and correlation coefficients between the chromatograms, after alignment. For the data aligned in this study, the average retention time precision and the lowest correlation before warping were 14 and 0.17, and were improved to three and 0.83, and six and 0.87 after warping, with correlation optimized warping and semi-parametric time warping, respectively.

Asunto(s)

Algoritmos , Cromatografía Líquida de Alta Presión/métodos , Té/química

RESUMEN

Using genome-wide expression profiling of a panel of 27 human mammary cell lines with different mechanisms of E-cadherin inactivation, we evaluated the relationship between E-cadherin status and gene expression levels. Expression profiles of cell lines with E-cadherin (CDH1) promoter methylation were significantly different from those with CDH1 expression or, surprisingly, those with CDH1 truncating mutations. Furthermore, we found no significant differentially expressed genes between cell lines with wild-type and mutated CDH1. The expression profile complied with the fibroblastic morphology of the cell lines with promoter methylation, suggestive of epithelial-mesenchymal transition (EMT). All other lines, also the cases with CDH1 mutations, had epithelial features. Three non-tumorigenic mammary cell lines derived from normal breast epithelium also showed CDH1 promoter methylation, a fibroblastic phenotype and expression profile. We suggest that CDH1 promoter methylation, but not mutational inactivation, is part of an entire programme, resulting in EMT and increased invasiveness in breast cancer. The molecular events that are part of this programme can be inferred from the differentially expressed genes and include genes from the TGFbeta pathway, transcription factors involved in CDH1 regulation (i.e. ZFHX1B, SNAI2, but not SNAI1, TWIST), annexins, AP1/2 transcription factors and members of the actin and intermediate filament cytoskeleton organisation.

Asunto(s)

Neoplasias de la Mama/patología , Cadherinas/biosíntesis , Metilación de ADN , Perfilación de la Expresión Génica , Línea Celular Tumoral , Transformación Celular Neoplásica , Análisis Mutacional de ADN , Regulación hacia Abajo , Células Epiteliales , Femenino , Regulación Neoplásica de la Expresión Génica , Humanos , Mesodermo/citología , Invasividad Neoplásica , Reacción en Cadena de la Polimerasa , Regiones Promotoras Genéticas , Transcripción Genética , Factor de Crecimiento Transformador beta/fisiología

RESUMEN

To determine whether CCL2 mRNA expression is beneficial or detrimental for cervical cancer patients, the association between the expression of this molecule by cervical tumour cells, the number of tumour-associated macrophages, and clinicopathological parameters such as recurrence, relapse-free survival, and overall patient survival was investigated. In cervical cancer samples from 93 untreated cervical cancer patients, the CCL2 mRNA expression level was quantified using RNA in situ hybridization and verified using real-time quantitative RT-PCR. The number of tumour-associated macrophages was determined using immunohistochemistry. Furthermore, the study investigated whether lack of CCL2 expression was due to genetic alterations near the 17q11.2 (CCL2 genomic) region. CCL2 mRNA expression by cervical tumour cells was associated with the number of tumour-associated macrophages (p < 0.001). Lack of CCL2 mRNA expression (15 samples; 16%) was associated with increased cumulative relapse-free survival (log rank test, p = 0.030), increased cumulative overall survival (log rank test, p = 0.024), less post-operative surgery, reduced local and distant recurrence, reduced vascular invasion, and smaller tumour size (<40 mm). The absence of CCL2 mRNA expression corresponded with loss of heterozygosity (LOH) at 17q11.2 in five of six samples. The increased cumulative relapse-free survival and cumulative overall survival of cervical cancer patients lacking tumour cell-associated CCL2 mRNA suggest that the tumour-associated macrophages support tumour progression, presumably by promoting angiogenesis and production of growth factors.

Asunto(s)

Biomarcadores de Tumor/análisis , Carcinoma/genética , Quimiocina CCL2/genética , Recurrencia Local de Neoplasia/genética , ARN Mensajero/análisis , Neoplasias del Cuello Uterino/genética , Carcinoma/mortalidad , Carcinoma/patología , Supervivencia sin Enfermedad , Femenino , Expresión Génica , Humanos , Hibridación in Situ , Pérdida de Heterocigocidad , Macrófagos/patología , Recurrencia Local de Neoplasia/mortalidad , Recurrencia Local de Neoplasia/patología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Tasa de Supervivencia , Neoplasias del Cuello Uterino/mortalidad , Neoplasias del Cuello Uterino/patología

RESUMEN

Domoic acid (DA) is a naturally-occurring amino acid that causes a form of human intoxication called amnesic shellfish poisoning (ASP) following the consumption of shellfish. A rapid and sensitive HPLC-UV method has been developed for analysis of DA and analogues in shellfish without the need for SPE clean-up. Isocratic chromatographic separation of DA and its isomers from shellfish matrix interferences and from the prevalent amino acid, tryptophan, was achieved by careful control of the mobile phase pH. The optimised pH was found to be 2.5 when using a Luna(2) C18 column. Sample extraction was verified with control extracts from shellfish spiked at 5.0 and 10.0 microg/g of DA and with certified reference material. The average extraction efficiency was 98.5%. The calibration, based on mussel tissue spiked with DA standard, was linear in the range 0.05-5.0 microg/ml (r = 0.9999) and the detection limit (signal:noise 3:1) was better than 25 ng/ml. The DA assay achieved good precision; %RSD = 1.63 (intra-day, n = 6) and %RSD = 3.7 (inter-day, n = 8). This method was successfully applied to a variety of shellfish species, allowing the rapid screening of a large number of samples per day (20-30), without the need for SPE clean-up. Quantitative data were obtained for shellfish samples containing domoic acid in the concentration range 0.25-330 microg/g. Using the same chromatographic conditions, LC-MS3 was used to determine DA and its isomers, isodomoic acid D and epi-domoic acid, in scallop tissues.

Asunto(s)

Cromatografía Líquida de Alta Presión/métodos , Ácido Kaínico/análogos & derivados , Ácido Kaínico/análisis , Toxinas Marinas/análisis , Mariscos/análisis , Animales , Concentración de Iones de Hidrógeno , Estructura Molecular , Intoxicación por Mariscos , Espectrometría de Masa por Ionización de Electrospray/métodos

RESUMEN

SETTING: Human observers generally have a strong tendency to round analog measurements or estimates to 'nice' ending digits, such as 0, 5, or even numbers. This is known as digit preference; it is a well-known phenomenon in frequency distributions of indurations collected in tuberculin surveys. Digit preference can distort estimates of prevalence and other statistical parameters. METHODS: We have developed a statistical model that combines smoothing by penalized likelihood and the transfer of counts from non-preferred to preferred digits, to obtain estimates of: 1) the smooth underlying distribution and 2) the amount of digit preference. RESULTS: To illustrate the validity of the model, it was applied to data from several countries. CONCLUSION: With the proposed model, digit preference can be quantified and frequency distributions of indurations can be corrected for it.

Asunto(s)

Interpretación Estadística de Datos , Modelos Estadísticos , Prueba de Tuberculina/estadística & datos numéricos , Algoritmos , Sesgo , Conducta de Elección , Humanos , Funciones de Verosimilitud , Reproducibilidad de los Resultados