RESUMEN
BACKGROUND: Gastric and colorectal cancer (CRC) are both one of the most common cancers worldwide. In many countries fecal immunochemical tests (FIT)-based CRC screening has been implemented. We investigated if FIT can also be applied for detection of H. pylori, the main risk factor for gastric cancer. METHODS: This prospective study included participants over 18 years of age referred for urea breath test (UBT). Patients were excluded if they had used antibiotics/bismuth in the past 4 weeks, or a proton pomp inhibitor (PPI) in the past 2 weeks. Participants underwent UBT, ELISA stool antigen test in standard feces tube (SAT), ELISA stool antigen test in FIT tube (Hp-FIT), and blood sampling, and completed a questionnaire on user friendliness. UBT results were used as reference. RESULTS: A total of 182 patients were included (37.4% male, median age 52.4 years (IQR 22.4)). Of these, 60 (33.0%) tested H. pylori positive. SAT and Hp-FIT showed comparable overall accuracy 71.1% (95%CI 63.2-78.3) vs. 77.6% (95%CI 70.4-83.8), respectively (p = 0.97). Sensitivity of SAT was 91.8% (95%CI 80.4-97.7) versus 94.2% (95%CI 84.1-98.9) of Hp-FIT (p = 0.98). Serology scored low with an overall accuracy of 49.7% (95%CI 41.7-57.7). Hp-FIT showed the highest overall user convenience. CONCLUSIONS: FIT can be used with high accuracy and sensitivity for diagnosis of H. pylori and is rated as the most convenient test. Non-invasive Hp-FIT test is highly promising for combined upper and lower gastrointestinal (pre-) cancerous screening. Further research should investigate the clinical implications, benefits and cost-effectiveness of such an approach.
Asunto(s)
Infecciones por Helicobacter , Helicobacter pylori , Neoplasias Gástricas , Adolescente , Adulto , Heces , Femenino , Infecciones por Helicobacter/diagnóstico , Humanos , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Sensibilidad y EspecificidadRESUMEN
Autophagy is a cellular degradation mechanism, which is triggered by the bacterium Helicobacter pylori. A single nucleotide polymorphism (SNP) in the autophagy gene ATG16L1 (rs2241880, G-allele) has been shown to dysregulate autophagy and increase intestinal endoplasmic reticulum (ER) stress. Here, we investigate the role of this SNP in H.pylori-mediated gastric carcinogenesis and its molecular pathways. ATG16L1 rs2241880 was genotyped in subjects from different ethnic cohorts (Dutch and Australian) presenting with gastric (pre)malignant lesions of various severity. Expression of GRP78 (a marker for ER stress) was assessed in gastric tissues. The effect of ATG16L1 rs2241880 on H.pylori-mediated ER stress and pro-inflammatory cytokine induction was investigated in organoids and CRISPR/Cas9 modified cell lines. Development of gastric cancer was associated with the ATG16L1 rs2241880 G-allele. Intestinal metaplastic cells in gastric tissue of patients showed increased levels of ER-stress. In vitro models showed that H.pylori increases autophagy while reducing ER stress, which appeared partly mediated by the ATG16L1 rs2241880 genotype. H.pylori-induced IL-8 production was increased while TNF-α production was decreased, in cells homozygous for the G-allele. The ATG16L1 rs2241880 G-allele is associated with progression of gastric premalignant lesions and cancer. Modulation of H.pylori-induced ER stress pathways and pro-inflammatory mediators by ATG16L1 rs2441880 may underlie this increased risk.
Asunto(s)
Autofagia , Estrés del Retículo Endoplásmico , Infecciones por Helicobacter/fisiopatología , Helicobacter pylori/fisiología , Neoplasias Gástricas/fisiopatología , Adulto , Anciano , Australia , Proteínas Relacionadas con la Autofagia/genética , Proteínas Relacionadas con la Autofagia/metabolismo , Estudios de Cohortes , Progresión de la Enfermedad , Femenino , Microbioma Gastrointestinal , Infecciones por Helicobacter/genética , Infecciones por Helicobacter/metabolismo , Infecciones por Helicobacter/microbiología , Humanos , Masculino , Persona de Mediana Edad , Países Bajos , Polimorfismo de Nucleótido Simple , Neoplasias Gástricas/genética , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/microbiologíaRESUMEN
Background and study aims Gastric cancer (GC) is usually preceded by premalignant gastric lesions (GPLs) such as gastric intestinal metaplasia (GIM). Information on risk factors associated with neoplastic progression of GIM are scarce. This study aimed to identify predictors for progression of GIM in areas with low GC incidence. Patients and methods The Progression and Regression of Precancerous Gastric Lesions (PROREGAL) study includes patients with GPL. Patients underwent at least two upper endoscopies with random biopsy sampling. Progression of GIM means an increase in severity according to OLGIM (operative link on gastric intestinal metaplasia) during follow-up (FU). Family history and lifestyle factors were determined through questionnaires. Serum Helicobacter pylori infection, pepsinogens (PG), gastrin-17 and GC-associated single nucleotide polymorphisms (SNPs) were determined. Cox regression was performed for risk analysis and a chi-squared test for analysis of single nucleotide polymorphisms. Results Three hundred and eight patients (median age at inclusion 61 years, interquartile range (IQR: 17; male 48.4â%; median FU 48 months, IQR: 24) were included. During FU, 116 patients (37.7â%) showed progression of IM and six patients (1.9â%) developed high-grade dysplasia or GC. The minor allele (C) on TLR4 (rs11536889) was inversely associated with progression of GIM (OR 0.6; 95â%CI 0.4-1.0). Family history (HR 1.5; 95â%CI 0.9-2.4) and smoking (HR 1.6; 95â%CI 0.9-2.7) showed trends towards progression of GIM. Alcohol use, body mass index, history of H. pylori infection, and serological markers were not associated with progression. Conclusions Family history and smoking appear to be related to an increased risk of GIM progression in low GC incidence countries. TLR4 (rs11536889) showed a significant inverse association, suggesting that genetic information may play a role in GIM progression.
RESUMEN
BACKGROUND: In an earlier study published in this journal (Berger e.a. 2002) it was shown that the cognitive styles 'weak central coherence' and 'poor cognitive shifting' are common in autism spectrum disorders, but tests have revealed that the styles do not apply to every member of the patient group. This finding could have consequences for the course of treatment. AIM: To find out if the cognitive styles of autistic patients can predict whether their social functioning will improve after three years of treatment we conducted a follow-up study in which we examined 44 non-retarded adolescents with an autism spectrum disorder who were receiving residential treatment. METHOD: On the basis of factor scores awarded in an extensive battery of neuropsychological tests, we formed subgroups of patients with weak versus strong central coherence and cognitive shifting. Then analyses of variance were used to discover whether the subgroups were predictors of changes in three aspects of social functioning: autistic symptoms, social intelligence and social competence. RESULTS: We found a small but significant gain in all the social domains. However, there were clear individual differences in the degree of improvement. Cognitive shifting was found to be a predictor of a clinically meaningful improvement in social competence. CONCLUSION: The correlation found between cognitive shifting and social competence indicates that patients with an autism spectrum disorder should be given different forms of treatment that take differences in cognitive style into account.
Asunto(s)
Atención , Trastorno Autístico/psicología , Cognición , Inteligencia , Conducta Social , Adolescente , Adulto , Análisis de Varianza , Trastorno Autístico/fisiopatología , Trastorno Autístico/terapia , Formación de Concepto , Análisis Factorial , Femenino , Humanos , Control Interno-Externo , Masculino , Pruebas Neuropsicológicas , Valor Predictivo de las Pruebas , Ajuste SocialRESUMEN
Platelet suspensions, that secreted about 50% of their dense granule contents upon stimulation with alpha-thrombin, showed a dose-dependent increase in secretion after 30 min preincubation with 0.5-3.0 g low density lipoprotein (LDL) protein/1. A 1-5 min preincubation had no effect. The enhancement by LDL only occurred at about 20% secretion or more, indicating that a minimal degree of activation was required for LDL to become effective. Lysine-modified LDL was equally effective as native LDL. The effect of LDL on secretion was accompanied by enhanced thromboxane B2 formation caused by stimulation of the liberation of arachidonate from phosphatidylcholine and/or phosphatidylinositol. However, when thromboxane formation was inhibited or the prostaglandin H2-thromboxane A2-receptor was blocked, LDL remained a potent stimulator of the secretion response. Thus, LDL enhances platelet secretion by a thromboxane A2-dependent and a thromboxane A2-independent mechanism via an effect that is independent of specific binding sites on the platelet.
Asunto(s)
Plaquetas/fisiología , Lipoproteínas LDL/fisiología , Activación Plaquetaria , Ácidos Araquidónicos/metabolismo , Humanos , Cinética , Trombina/fisiología , Tromboxano B2/metabolismoRESUMEN
The relationship between metabolic energy and platelet aggregation and secretion was investigated by sudden exhaustion of the cell energy content after these platelet responses had been initiated. In normal platelets, optical aggregation was at any stage susceptible to energy exhaustion, whereas single platelet disappearance and secretion were hardly affected. Prelowering the platelet energy content, while preserving the adenylate energy charge, made both optical aggregation and the secretion from dense, alpha- and acid hydrolase-containing granules susceptible to energy exhaustion, but single platelet disappearance was not affected. Complete arrest of secretion occurred when the energy content had fallen below 3-3.5 mumol ATP equivalents (ATPeq)/10(11) platelets, while optical aggregation was interrupted below 2-2.5 mumol ATPeq/10(11) platelets. At any stage of optical aggregation and the three secretion responses, the dependence on energy remained the same, indicating a tight coupling between these functions and metabolic energy, which held during the entire responses.
Asunto(s)
Plaquetas/fisiología , Agregación Plaquetaria , Adenosina Trifosfato/metabolismo , Desoxiglucosa/farmacología , Metabolismo Energético/efectos de los fármacos , Humanos , Peróxido de Hidrógeno/farmacología , Serotonina/metabolismoRESUMEN
Studies with isolated lipoproteins and washed platelets suggest that lipoproteins may affect platelet functions. We investigated platelet-rich plasma (PRP) from a patient with abetalipoproteinemia (ABL), whose plasma lacks apo-B containing lipoproteins (VLDL, LDL and chylomicrons). ABL-PRP aggregated poorly with different agonists and failed to respond to arachidonate. Thromboxane B2 (TxB2) formation was severely impaired. After gel-filtration most of the aggregation defects persisted in agreement with reduced metabolism of endogenous arachidonate. However, arachidonate-induced aggregation and TxB2 production partially normalized. Normal platelets suspended in ABL-plasma showed similar defects in aggregation and TxB2 production but arachidonate-induced aggregation was much lower than expected on the basis of TxB2. We conclude that the abnormal platelet functions in ABL-PRP are caused by (i) an intrinsic platelet abnormality due to reduced arachidonate mobilization and (ii) a property in ABL plasma that inhibits aggregation partially by trapping the arachidonate and partially by an unidentified mechanism. The latter properties may be the result of the abnormal lipid composition of ABL-plasma.
Asunto(s)
Abetalipoproteinemia/sangre , Plaquetas/fisiología , Adenosina Trifosfato/metabolismo , Adulto , Ácido Araquidónico , Ácidos Araquidónicos/sangre , Plaquetas/metabolismo , Femenino , Citometría de Flujo , Humanos , Agregación Plaquetaria/fisiologíaRESUMEN
When platelets are treated with H2O2 the metabolic ATP content decreases sharply (Holmsen, H., and Robkin, L. (1977) J. Biol. Chem. 252, 1752-1757). Here we report that the loss of metabolic energy is fully recovered in phosphorylated glycolytic intermediates. A mixture of antimycin A/2-deoxy-D-glucose/D-gluconic acid-1,5-lactone blocks mitochondrial ATP resynthesis and prevents the entry of sugars into the glycolytic sequence. The energy-rich phosphates in the adenylate and the glycolytic pool are then consumed in a specific order. First, the glycolytic pool is consumed at a rate of 4.5 mumol of ATP equivalents/min/10(11) cells, and metabolic ATP and ADP are kept stable; then the consumption of the glycolytic pool decreases and metabolic ATP and ADP are consumed, together keeping up with the same rate of energy consumption. Thrombin stimulation increases the energy consumption to about 17 mumol of ATPeq/min/10(11) cells which is now furnished by both the glycolytic and the adenylate pool, again with a preferential consumption of the former. The results show that H2O2 triggers a shift of energy-rich phosphates from the adenylate to the glycolytic pool and that the latter remains rapidly accessible to energy consumption thereby stabilizing the level of metabolic ATP. The adenylate energy charge is independent of the distribution of energy among the two pools, which extends its importance to the regulation of energy supply and demand beyond the adenylate pool.
Asunto(s)
Nucleótidos de Adenina/metabolismo , Plaquetas/fisiología , Metabolismo Energético , Glucólisis , Adenosina Difosfato/metabolismo , Adenosina Trifosfato/metabolismo , Humanos , Peróxido de Hidrógeno/farmacología , Trombina/farmacologíaRESUMEN
Platelet-activating factor 1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine (PAF-acether) triggers exposure of fibrinogen binding sites on platelets via binding to specific receptors. Comparison of [3H]PAF-acether binding with 125I-fibrinogen binding shows that the rate with which PAF-acether binds to a number of receptors and not the degree of receptor occupancy determines how much fibrinogen binds. At low concentrations of PAF-acether (0.1-1.0 nM) binding site exposure is incomplete and parallels the rate of formation of the PAF-acether-receptor complex. Fibrinogen binding then primarily depends on the concentration of PAF-acether. At a high concentration of PAF-acether (500 nM) binding site exposure is complete within 2-5 min. Fibrinogen binding then depends on the concentration of fibrinogen. Exposure of binding sites in the absence of fibrinogen leads to disappearance of accessible binding sites. At 500 nM PAF-acether, this disappearance is exponential in nature and shows the same characteristics after 5-15 min incubation with fibrinogen as after 60 min. Exposure of binding sites is then complete within 5 min and their disappearance is not disturbed by other processes. At 0.5 nM PAF-acether, the same characteristics are found after 60 min incubation with fibrinogen, but shorter incubation times reveal an ongoing binding site exposure that interferes with the disappearance process. These results demonstrate close coupling between the PAF-acether receptors and fibrinogen binding sites and indicate that the rate of formation of the PAF-acether-receptor complex is a major factor in the regulation of binding site exposure.
Asunto(s)
Plaquetas/metabolismo , Fibrinógeno/metabolismo , Factor de Activación Plaquetaria/farmacología , Receptores Acoplados a Proteínas G , Sitios de Unión , Humanos , Cinética , Matemática , Glicoproteínas de Membrana Plaquetaria , Receptores de Superficie Celular/metabolismo , Factores de TiempoRESUMEN
The correlation between energy consumption and platelet responses induced by collagen, A23187 and ADP was investigated and compared with the energetics of thrombin-stimulated platelets established in earlier work. Aggregation, measured as single-platelet disappearance, and secretion correlated quantitatively with the increment but not with the total consumption of energy, suggesting that the former reflects the energy cost of these responses. The cost of complete aggregation was 2-3 mumol of ATP equivalents/10(11) platelets with collagen, ADP and thrombin as the stimulus. The cost of complete dense-granule secretion was 0.5-0.8 mumol of ATP equivalents/10(11) platelets with all agonists tested. The cost of combined secretion of alpha-granule and acid hydrolase granule contents was 5-7 mumol of ATP equivalents/10(11) platelets with thrombin and collagen. However, in the presence of A23187 much more energy was consumed during aggregation and secretion. Also ADP triggered more energy consumption during secretion than was seen with the other inducers. The effect of inhibitors of aggregation and secretion was investigated in thrombin-stimulated platelets. Raising the cellular cyclic AMP content sharply decreased the increment in energy consumption as well as aggregation and secretion. The cytoskeleton-disrupting agents cytochalasin B and colchicine left the increment in energy consumption intact, but decreased the basal consumption seen in unstimulated platelets. This was accompanied by normal (cytochalasin B) or diminished (colchicine) aggregation and secretion. Apart from the latter exception, all inhibitors decreased secretion and incremental energy consumption in parallel, thereby preserving the energy-versus-secretion relationship established in earlier work. In contrast, aggregation and energy consumption varied independently, suggesting that the coupling with energy consumption is much weaker for this response.
Asunto(s)
Plaquetas/efectos de los fármacos , Adenosina Difosfato/farmacología , Antimetabolitos/farmacología , Plaquetas/metabolismo , Calcimicina/farmacología , Colágeno/farmacología , Metabolismo Energético/efectos de los fármacos , Humanos , Técnicas In Vitro , Agregación Plaquetaria/efectos de los fármacos , Trombina/farmacologíaRESUMEN
Abrupt arrest of ATP resynthesis in blood platelets induces a rapid decline in metabolic ATP-ADP. This decline is biexponential with a 7-fold difference in the rate-constants of the two components. Stimulation with thrombin increases both rate-constants, and raises the relative contribution of the rapid component from 60 to 90% of total. The initial decline can be approximated by a single exponential term, yielding the rate-constant for initial ATP hydrolysis. Since this initial decline reflects energy consumption of undisturbed platelets, this approach offers a sensitive means to determine energy consumption and ATP turnover within short time intervals.
Asunto(s)
Adenosina Difosfato/sangre , Adenosina Trifosfato/sangre , Antimicina A/farmacología , Plaquetas/metabolismo , Metabolismo Energético , Gluconatos/farmacología , Plaquetas/efectos de los fármacos , Humanos , Cinética , Lactonas , Trombina/fisiologíaRESUMEN
The involvement of metabolic energy in platelet responses was investigated by measuring the energy consumption during aggregation and secretion from dense, alpha- and acid-hydrolase-containing granules. Gel-filtered human platelets were stimulated with different amounts of thrombin (0.05-5.0 units X ml-1). At various stages during aggregation and secretion the energy consumption was measured from the changes in metabolic ATP and ADP following abrupt arrest of ATP resynthesis. Stimulation with 5 units of thrombin X ml-1 increased the energy consumption from 6.2 +/- 0.9 to 17.8 +/- 0.4 mumol of ATPeq. X min-1 X (10(11) platelets)-1 during the first 15 s. It decreased thereafter and returned to values found in resting cells after about 30 s. With 0.05 unit of thrombin X ml-1, the energy consumption accelerated more slowly and took at least 3 min before it normalized. A strong positive correlation was found between the velocities of the three secretion responses and the concurrent energy consumption (a) at different stages of the responses induced by a given dose of thrombin, and (b) at different secretion velocities initiated by different amounts of thrombin. When, at different stages of the responses, the extent of secretion was compared with the amount of energy that had been consumed, a strong linear correlation was found with the increment in energy consumption but not with the total energy consumption. This correlation was independent of the concentration of thrombin and indicated that complete secretion from dense, alpha- and acid-hydrolase-containing granules was paralleled by an increment of 4.0, 6.5 and 6.7 mumol of ATPeq. X (10(11) platelets)-1, respectively. An energy cost of 0.7 mumol of ATPeq. X (10(11) platelets)-1 was calculated for separate dense-granule secretion, whereas the combined alpha- and acid-hydrolase granule secretion required 5.3 mumol of ATPeq. X (10(11) platelets)-1. There was no correlation between energy consumption and optical aggregation. In contrast, the rate of single platelet disappearance, which is a measure for the early formation of small aggregates, correlated closely with the rate of energy consumption. Compared with secretion, however, the energy requirement of single platelet disappearance was minor, since 2mM-EDTA completely prevented this response but decreased the energy consumption only slightly. An increase of 0.5-1.0 mumol of ATPeq. X (10(11) platelets)-1 was seen before single platelet disappearance and the three secretion responses were initiated, indicating an increase in energy consuming processes that preceded these responses.(ABSTRACT TRUNCATED AT 400 WORDS)
Asunto(s)
Plaquetas/metabolismo , Metabolismo Energético/efectos de los fármacos , Trombina/farmacología , Acetilglucosaminidasa/sangre , Adenosina Trifosfato/sangre , Plaquetas/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Humanos , Técnicas In Vitro , Agregación Plaquetaria/efectos de los fármacos , Serotonina/sangre , Termodinámica , Factores de Tiempo , beta-Tromboglobulina/metabolismoRESUMEN
Despite the use of preservatives, platelets are severely damaged during cryopreservation and, following freezing, function poorly in a number of in vitro tests. We report here that cryopreserved platelets show diminished aggregation in response to collagen. This may be a consequence of a secretion defect as evidenced by a 20 to 30 percent loss of dense- and alpha-granule content (p less than 0.05) and an impaired secretion mechanism. Analysis of adenine nucleotides confirmed the defect in dense granule adenosine triphosphate (ATP) and adenosine diphosphate (ADP) content (storage pool), but in addition revealed a 50 percent fall in cytosolic ATP (metabolic pool). In contrast, the adenylate energy charge, (ATP + 1/2 ADP)/(ATP + ADP + adenosine monophosphate), was normal. We concluded that platelet cryopreservation leads to a secretion defect, probably as a result of activation during freezing and thawing procedures.
Asunto(s)
Plaquetas/efectos de los fármacos , Gránulos Citoplasmáticos/efectos de los fármacos , Dimetilsulfóxido/efectos adversos , Adenosina Difosfato/sangre , Adenosina Monofosfato/sangre , Adenosina Trifosfato/sangre , Plaquetas/metabolismo , Plaquetas/fisiología , Colágeno/farmacología , Gránulos Citoplasmáticos/metabolismo , Gránulos Citoplasmáticos/fisiología , Congelación/métodos , Humanos , Inosina Monofosfato/sangre , Agregación PlaquetariaRESUMEN
The degradation of platelet-activating factor (PAF) in plasma is catalyzed by PAF-acetylhydrolase resulting in lyso-PAF which is biologically inactive. Normally, most of the PAF-degrading activity is associated with low-density lipoproteins (LDL). The enzyme activity was measured in the plasma of a patient with abetalipoproteinemia, a disorder characterized by the absence of apolipoprotein-B-containing lipoproteins (chylomicrons, VLDL and LDL). Here we report that the plasma of the patient has a normal activity of PAF-acetylhydrolase. The enzyme activity is bound to high-density lipoproteins (HDL) and shows the kinetic properties of the LDL-associated enzyme of healthy subjects. Following administration of artificial triglyceride-rich particles (ATRP), part of the enzyme activity is found associated with ATRP, indicating that PAF-acetylhydrolase can transfer from HDL to triglyceride-containing lipid complexes in vivo.
Asunto(s)
Abetalipoproteinemia/enzimología , Lipoproteínas HDL/química , Lipoproteínas LDL/química , Fosfolipasas A/sangre , 1-Alquil-2-acetilglicerofosfocolina Esterasa , Adulto , Femenino , Humanos , Infusiones Intravenosas , Cinética , Valores de Referencia , Triglicéridos/farmacologíaRESUMEN
Among the different platelet responses, secretion requires the greatest amount of metabolic energy. The velocities of dense, alpha- and acid hydrolase granule secretion vary in parallel with the increase in energy consumption seen in thrombin-stimulated cells. This covariance is preceded by a phase in which energy consumption is increased without the extracellular appearance of secretion markers. By treating the platelets with thrombin and hirudin we have stimulated the platelets for short intervals and succeeded in separating shape change, single platelet disappearance and secretion to a great extent. In this report we show that the early increase in energy consumption reflects the energy requirement of aggregation but not of shape change. The cost of 100% of single platelet disappearance is 2.8 mumol of ATPeq. X (10(11) platelets)-1. Concurrent analysis of phosphorylation of Mr 20 000 and 47 000 proteins and of 32P-labelled phosphatidylinositol metabolites led to the following observations. Firstly, shape change is neither accompanied by an increase in protein phosphorylation nor by changes in the steady state levels of 32P-labelled phosphatidylinositol metabolites. Secondly, when aggregation occurs both proteins are phosphorylated, but the phosphatidylinositol metabolites do not change. Thirdly, when secretion follows, more phosphorylation of the Mr 47 000 protein occurs and initially only phosphatidic acid accumulates. At a later stage of the secretion responses, more protein phosphorylation and phosphatidic acid accumulation become evident, and are now accompanied by alterations in the steady state levels of 32P-labelled (poly)phosphoinositides. Hence, the early increase in energy consumption coincides with protein phosphorylation and, at a later stage, with alterations in (poly)phosphoinositides metabolites. This demonstrates that metabolic energy is directly involved in stimulus-response coupling in aggregating platelets.