Mixed Functional Siloxane Monolayers Prepared via Chemical Vapor Deposition.

Functionalizing surfaces with self-assembled monolayers (SAMs) allows to efficiently bind bioreceptors, for instance, by bio-orthogonal click reactions, which is useful in biosensor fabrication. Control of the bioreceptor concentration on the surface can be achieved by coating an SAM mixture consisting of a functional SAM, which binds the bioreceptor, and a nonfunctional SAM for dilution. In this work, a novel vapor-based coating approach for the preparation of mixed SAM coatings is presented. Sequential evaporation of the SAM precursors, i.e., fluoroalkyl and azidoalkyl silanes, by heating under reduced pressure leads to the formation of a two-dimensional siloxane monolayer network on silicon oxide. The presence of both SAMs in the mixed coatings is confirmed by Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy. As verified by atomic force microscopy, the morphologies of the coatings and the uncoated silicon oxide substrates are similar, indicating a conformal coating. Functionality of the SAM mixture is demonstrated by a reaction with a fluorescent dye, illustrating its potential application in biosensors.

An IR Spectroscopy Study of the Degradation of Surface Bound Azido-Groups in High Vacuum.

Controlled surface functionalization with azides to perform on surface "click chemistry" is desired for a large range of fields such as material engineering and biosensors. In this work, the stability of an azido-containing self-assembled monolayer in high vacuum is investigated using in situ Fourier transform infrared spectroscopy. The intensity of the antisymmetric azide stretching vibration is found to decrease over time, suggesting the degradation of the azido-group in high vacuum. The degradation is further investigated at three different temperatures and at seven different nitrogen pressures ranging from 1 × 10-6 mbar to 5 × 10-3 mbar. The degradation is found to increase at higher temperatures and at lower nitrogen pressures. The latter supporting the theory that the degradation reaction involves the decomposition into molecular nitrogen. For the condition with the highest degradation detected, only 63% of azides is found to remain at the surface after 8 h in vacuum. The findings show a significant loss in control of the surface functionalization. The instability of azides in high vacuum should therefore always be considered when depositing or postprocessing azido-containing layers.

Study of the surface species during thermal and plasma-enhanced atomic layer deposition of titanium oxide films using in situ IR-spectroscopy and in vacuo X-ray photoelectron spectroscopy.

The thermal and plasma-enhanced atomic layer deposition (ALD) growth of titanium oxide using an alkylamine precursor - tetrakis(dimethylamino)titanium (TDMAT) - was investigated. The surface species present during both the precursor and co-reactant pulse were studied with in situ reflection mid-IR spectroscopy (FTIR) and in vacuo X-ray photoelectron spectroscopy (XPS). The thermal process using H2O vapor proceeds through a typical ligand exchange reaction mechanism. The plasma-enhanced ALD processes using H2O-plasma or O2-plasma exhibit an additional decomposition and combustion reaction mechanism. After the plasma exposure, imine (N[double bond, length as m-dash]C) and isocyanate (N[double bond, length as m-dash]C[double bond, length as m-dash]O) surface species were observed by in situ FTIR. In addition, nitrites (NOx) were detected using in vacuo XPS during the O2-plasma process. This study presents the importance of the use of in situ FTIR and in vacuo XPS as complementary techniques to learn more about the ALD reaction mechanism. While in situ FTIR is very sensitive to changes of chemical bonds at the surface, exact identification and quantification could only be done with the aid of in vacuo XPS.

Supercritical Angle Fluorescence Characterization Using Spatially Resolved Fourier Plane Spectroscopy.

Most fluorescent immunoassays require a wash step prior to read-out due to the otherwise overwhelming signal of the large number of unbound (bulk) fluorescent molecules that dominate over the signal from the molecules of interest, usually bound to a substrate. Supercritical angle fluorescence (SAF) sensing is one of the most promising alternatives to total internal reflection fluorescence for fluorescence imaging and sensing. However, detailed experimental investigation of the influence of collection angle on the SAF surface sensitivity, i.e., signal to background ratio (SBR), is still lacking. In this Letter, we present a novel technique that allows to discriminate the emission patterns of free and bound fluorophores simultaneously by collecting both angular and spectral information. The spectrum was probed at multiple positions in the back focal plane using a multimode fiber connected to a spectrometer and the difference in intensity between two fluorophores was used to calculate the SBR. Our study clearly reveals that increasing the angle of SAF collection enhances the surface sensitivity, albeit at the cost of decreased signal intensity. Furthermore, our findings are fully supported by full-field 3D simulations.

Chemical Vapor Deposition of Azidoalkylsilane Monolayer Films.

N3-functionalized monolayers on silicon wafer substrates are prepared via the controlled vapor-phase deposition of 11-azidoundecyltrimethoxysilanes at reduced pressure and elevated temperature. The quality of the layer is assessed using contact angle, attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR), and ellipsometry measurements. At 60 °C, longer deposition times are needed to achieve monolayers with similar N3 density compared to depositions at 145 °C. The monolayers formed via the vapor phase are denser compared to those formed via a solvent-based deposition process. ATR-FTIR measurements confirm the incorporation of azido-alkyl chains in the monolayer and the formation of siloxane bridges with the underlying oxide at both deposition temperatures. X-ray photon spectroscopy shows that the N3 group is oriented upward in the grafted layer. Finally, the density was determined using total reflection X-ray fluorescence after a click reaction with chlorohexyne and amounts to 2.5 × 1014 N3 groups/cm2. In summary, our results demonstrate the formation of a uniform and reproducible N3-containing monolayer on silicon wafers, hereby providing a functional coating that enables click reactions at the substrate.

NPC1 defect results in abnormal platelet formation and function: studies in Niemann-Pick disease type C1 patients and zebrafish.

Niemann-Pick type C is a lysosomal storage disease associated with mutations in NPC1 or NPC2, resulting in an accumulation of cholesterol in the endosomal-lysosomal system. Niemann-Pick type C has a clinical spectrum that ranges from a neonatal rapidly fatal disorder to an adult-onset chronic neurodegenerative disease combined with remarkably, in some cases, hematological defects such as thrombocytopenia, anemia and petechial rash. A role of NPC1 in hematopoiesis was never shown. Here, we describe platelet function abnormalities in three unrelated patients with a proven genetic and biochemical NPC1 defect. Their platelets have reduced aggregations, P-selectin expression and ATP secretions that are compatible with the observed abnormal alpha and reduced dense granules as studied by electron microscopy and CD63 staining after platelet spreading. Their blood counts were normal. NPC1 expression was shown in platelets and megakaryocytes (MKs). In vitro differentiated MKs from NPC1 patients exhibit hyperproliferation of immature MKs with different CD63(+) granules and abnormal cellular accumulation of cholesterol as shown by filipin stainings. The role of NPC1 in megakaryopoiesis was further studied using zebrafish with GFP-labeled thrombocytes or DsRed-labeled erythrocytes. NPC1 depletion in zebrafish resulted in increased cell death in the brain and abnormal cellular accumulation of filipin. NPC1-depleted embryos presented with thrombocytopenia and mild anemia as studied by flow cytometry and real-time QPCR for specific blood cell markers. In conclusion, this is the first report, showing a role of NPC1 in platelet function and formation but further studies are needed to define how cholesterol storage interferes with these processes.

Deletion of NEMO/IKKgamma in liver parenchymal cells causes steatohepatitis and hepatocellular carcinoma.

The IkappaB kinase (IKK) subunit NEMO/IKKgamma is essential for activation of the transcription factor NF-kappaB, which regulates cellular responses to inflammation. The function of NEMO in the adult liver remains elusive. Here we show that ablation of NEMO in liver parenchymal cells caused the spontaneous development of hepatocellular carcinoma in mice. Tumor development was preceded by chronic liver disease resembling human nonalcoholic steatohepatitis (NASH). Antioxidant treatment and genetic ablation of FADD demonstrated that death receptor-mediated and oxidative stress-dependent death of NEMO-deficient hepatocytes triggered disease pathogenesis in this model. These results reveal that NEMO-mediated NF-kappaB activation in hepatocytes has an essential physiological function to prevent the spontaneous development of steatohepatitis and hepatocellular carcinoma, identifying NEMO as a tumor suppressor in the liver.

A multidimensional model of psychobiological interactions in functional dyspepsia: a structural equation modelling approach.

OBJECTIVE: Gastric sensorimotor function, abuse history, 'trait' and 'state' psychological factors and 'somatisation' all play a role in functional dyspepsia (FD) and its associated impaired quality of life (QoL), but their interplay remains poorly understood. We aimed to test a comprehensive, a priori hypothesised model of interactions between these dimensions in FD. DESIGN: In 259 FD patients, we studied gastric sensitivity with a barostat. We measured abuse history (sexual/physical, childhood/adulthood), 'trait' (alexithymia, trait anxiety) and 'state' (positive/negative affect, depression, panic disorder) psychological factors, somatic symptom reporting (somatic symptom count, dyspepsia, irritable bowel syndrome and fatigue symptoms) and QoL (physical, mental) using validated questionnaires. Confirmatory factor analysis (CFA) was used to assess whether four a priori hypothesised latent variables ('abuse', 'trait affectivity', 'state affect' and 'somatic symptom reporting') were adequately supported by the data. Structural equation modelling (SEM) was used to test the a priori hypothesised relationships between these latent variables and the observed variables gastric sensitivity and QoL. RESULTS: Both the CFA and SEM models fitted the data adequately. Abuse exerted its effect directly on 'somatic symptom reporting', rather than indirectly through psychological factors. A reciprocal relationship between 'somatic symptom reporting' and 'state affect' was found. Gastric sensitivity influences 'somatic symptom reporting' but not vice versa. 'Somatic symptom reporting' and 'trait affectivity' are the main determinants of physical and mental QoL, respectively. CONCLUSIONS: We present the first comprehensive model elucidating the complex interactions between multiple dimensions (gastric sensitivity, abuse history, 'state' and 'trait' psychological factors, somatic symptom reporting and QoL) in FD.

Fetal rat hearts do not display acute cardiotoxicity in response to maternal Doxorubicin treatment.

Anthracyclines are used to treat cancers during the second and third trimester of pregnancy. The chemotherapeutic effect of anthracyclines is associated with a dose- and time-dependent cardiotoxicity that is well described for infants and adults. However, data regarding fetal anthracycline-related cardiotoxicity after administration of chemotherapeutics during pregnancy are limited. In this study, we analyzed the acute effect of doxorubicin, an anthracycline derivative, on fetal and maternal rat myocardium. We injected 10 or 20 mg/kg i.v. doxorubicin to pregnant Wistar rats at day 18 of pregnancy; age-matched pregnant rats injected with physiologic saline served as controls. Maternal echocardiography and fetal Doppler scanning were performed before the injection and before sacrifice. Cesarean operation was performed at day 19 or 20, and maternal and fetal blood samples and heart biopsies were collected to measure apoptosis, the impact on cell proliferation, and structural cardiac damage. Acute maternal cardiotoxicity is associated with loss of body weight, moderately deteriorated left ventricular function, induction of apoptosis, and a decrease in cell turnover. Despite a 30% lower fetal body weight and elevated plasma B-type natriuretic peptide concentrations after doxorubicin administration, the fetal hearts had intact microstructure, an unaltered number of apoptotic cells, and preserved cell proliferation compared with controls. Our study suggests that acute treatment using anthracyclines during pregnancy impairs maternal cardiac function, whereas fetal hearts are protected.

Homozygosity for aquaporin 7 G264V in three unrelated children with hyperglyceroluria and a mild platelet secretion defect.

PURPOSE: Aquaporin 7 (AQP7) belongs to the aquaglyceroporin family, which transports glycerol and water. AQP7-deficient mice develop obesity, insulin resistance, and hyperglyceroluria. However, AQP7's pathophysiologic role in humans is not yet known. METHODS: Three children with psychomotor retardation and hyperglyceroluria were screened for AQP7 mutations. The children were from unrelated families. Urine and plasma glycerol levels were measured using a three-step enzymatic approach. Platelet morphology and function were studied using electron microscopy, aggregations, and adenosine triphosphate (ATP) secretion tests. RESULTS: The index patients were homozygous for AQP7 G264V, which has previously been shown to inhibit transport of glycerol in Xenopus oocytes. We also detected a subclinical platelet secretion defect with reduced ATP secretion, and the absence of a secondary aggregation wave after epinephrine stimulation. Electron microscopy revealed round platelets with centrally located granules. Immunostaining showed AQP7 colocalization, with dense granules that seemed to be released after strong platelet activation. Healthy relatives of these patients, who were homozygous (not heterozygous) for G264V, also had hyperglyceroluria and platelet granule abnormalities. CONCLUSION: The discovery of an association between urine glycerol loss and a platelet secretion defect is a novel one, and our findings imply the involvement of AQPs in platelet secretion. Additional studies are needed to define whether AQP7 G264V is also a risk factor for mental disability.

HNF1B deficiency causes ciliary defects in human cholangiocytes.

Heterozygous deletion or mutation in hepatocyte nuclear factor 1 homeobox B/transcription factor 2 (HNF1B/TCF2) causes renal cyst and diabetes syndrome (OMIM #137920). Mice with homozygous liver-specific deletion of Hnf1ß revealed that a complete lack of this factor leads to ductopenia and bile duct dysplasia, in addition to mild hepatocyte defects. However, little is known about the hepatic consequences of deficient HNF1B function in humans. Three patients with heterozygous HNF1B deficiency were found to have normal bile duct formation on radiology and routine liver pathology. Electron microscopy revealed a paucity or absence of normal primary cilia. Therefore, heterozygous HNF1B deficiency is associated with ciliary anomalies in cholangiocytes, and this may cause cholestasis.

Identification of telocytes in the upper lamina propria of the human urinary tract.

The upper lamina propria (ULP) area of interstitial cells (IC) has been studied extensively in bladder, but is rather unexplored in the rest of the urinary tract. This cell layer is intriguing because of the localization directly underneath the urothelium, the intercellular contacts and the close relationship with nerve endings and capillaries. In this study, we examine the ULP layer of IC in human renal pelvis, ureter and urethra, and we make a comparison with ULP IC in bladder. Tissue was obtained from normal areas in nephrectomy, cystectomy and prostatectomy specimens, and processed for morphology, immunohistochemistry and electron microscopy. A morphological and immunohistochemical phenotype for the ULP IC was assessed and region-dependent differences were looked for. The ULP IC in renal pelvis, ureter and urethra had a similar ultrastructural phenotype, which differed somehow from that of bladder IC, that is, thinner and longer cytoplasmic processes, no peripheral actin filaments and presence of dense core granules and microtubules. Together with their immunohistochemical profile, these features are most compatible with the phenotype of telocytes, a recently discovered group of stromal cells. Based on their global ultrastructural and immunohistochemical phenotype, ULP IC in human bladder should also be classified as telocytes. The most striking immunohistochemical finding was the variable expression of oestrogen receptor (ER) and progesterone receptor (PR). The functional relevance of ULP telocytes in the urinary tract remains to be elucidated, and ER and PR might therefore be promising pharmacological research targets.

SCAMP5, NBEA and AMISYN: three candidate genes for autism involved in secretion of large dense-core vesicles.

Autism is a neurodevelopmental disorder characterized by impaired social reciprocity, impaired communication and stereotypical behaviors. Despite strong evidence for a genetic basis, few susceptibility genes have been identified. Here, we describe the positional cloning of SCAMP5, CLIC4 and PPCDC as candidate genes for autism, starting from a person with idiopathic, sporadic autism carrying a de novo chromosomal translocation. One of these genes, SCAMP5 is silenced on the derivative chromosome, and encodes a brain-enriched protein involved in membrane trafficking, similar to the previously identified candidate genes NBEA and AMISYN. Gene silencing of Nbea, Amisyn and Scamp5 in mouse beta-TC3 cells resulted in a 2-fold increase in stimulated secretion of large dense-core vesicles (LDCVs), while overexpression suppressed secretion. Moreover, ultrastructural analysis of blood platelets from the patients with haploinsufficieny of one of the three candidate genes, showed morphological abnormalities of dense-core granules, which closely resemble LDCVs. Taken together, this study shows that in three independent patients with autism three different negative regulators of LDCV secretion are affected, respectively, suggesting that in at least a subgroup of patients the regulation of neuronal vesicle trafficking may be involved in the pathogenesis of autism.

Preserving the morphology and evaluating the quality of liver grafts by hypothermic machine perfusion: a proof-of-concept study using discarded human livers.

The wider use of livers from expanded criteria donors and donation after circulatory death donors may help to improve access to liver transplantation. A prerequisite for safely using these higher risk livers is the development of objective criteria for assessing their condition before transplantation. Compared to simple cold storage, hypothermic machine perfusion (HMP) provides a unique window for evaluating liver grafts between procurement and transplantation. In this proof-of-concept study, we tested basic parameters during HMP that may reflect the condition of human liver grafts, and we assessed their morphology after prolonged HMP. Seventeen discarded human livers were machine-perfused. Eleven livers were nontransplantable (major absolute contraindications and severe macrovesicular steatosis in the majority of the cases). Six livers were found in retrospect to be transplantable but could not be allocated and served as controls. Metabolic parameters (pH, lactate, partial pressure of oxygen, and partial pressure of carbon dioxide), enzyme release in the perfusate [aspartate aminotransferase (AST) and lactate dehydrogenase (LDH)], and arterial/portal resistances were monitored during HMP. Nontransplantable livers released more AST and LDH than transplantable livers. In contrast, arterial/portal vascular resistances and metabolic profiles did not differ between the 2 groups. Morphologically, transplantable livers remained well preserved after 24 hours of HMP. In conclusion, HMP preserves the morphology of human livers for prolonged periods. A biochemical analysis of the perfusate provides information reflecting the extent of the injury endured.

Characterization of upper lamina propria interstitial cells in bladders from patients with neurogenic detrusor overactivity and bladder pain syndrome.

The upper lamina propria (ULP) area of interstitial cells (IC) in bladder has been studied for more than a decade in several species including human beings. Nevertheless there is still lack of uniformity in terminology of this cell layer. The aim of the present study was to add new data to the morphological and immunohistochemical phenotype of these cells and to find out whether this phenotype is changed in bladders from patients with neurogenic detrusor overactivity (NDO) and bladder pain syndrome (BPS). Bladder tissue was obtained from a control group and from patients with NDO and BPS. Samples were processed for morphology, electron microscopy and immunohistochemistry. A morphological and immunohistochemical phenotype for the ULP IC was assessed and changes in this phenotype were looked for in samples from patients with NDO and BPS. The ULP IC were characterized ultrastructurally by the presence of actin filaments with densifications, many caveolae and abundant rough endoplasmic reticulum (RER); on immunohistochemistry ULP IC were immunoreactive for α-sma, vimentin, CD10 and podoplanin and categorized as interstitial Cajal-like cells (ICLC). In NDO and BPS bladders we found a phenotypical shift towards a fibroblastic phenotype which was even more pronounced in the NDO group. In both groups there was also an increased presence in ULP lymphocytes. The ULP area in the human bladder contains a population of ICLC with distinct ultrastructural morphology and immunohistochemical phenotype. Their unique α-sma(+) /desmin(-) /CD34(-) phenotype allows studying this population in various bladder disorders. In bladders form patients with BPS and NDO, we observed these ULP ICLC to shift towards a fibroblast phenotype.

Influence of acute tryptophan depletion on gastric sensorimotor function in humans.

Peripheral serotonin (5-hydrodytryptamine; 5-HT) is involved in the regulation of gastrointestinal motility and sensation, whereas centrally it plays a role in mood regulation. A dysfunctional serotonergic system may provide a plausible link between functional dyspepsia symptoms and its high psychosocial comorbidity such as anxiety and depression. The aim of this study was to evaluate the effect of decreased 5-HT synthesis by acute tryptophan depletion (ATD) on gastric sensorimotor function and nutrient tolerance, anxiety scores, and gastrointestinal mucosal 5-HT concentrations in healthy volunteers. All subjects were studied under a control condition and during ATD. Gastric sensorimotor function and nutrient tolerance were assessed using a barostat (n = 16, mean age 28.8 ± 1.4 yr) and a satiety drinking test (n = 13, mean age 27.3 ± 1.4 yr). Anxiety during the barostat was evaluated using State-Trait Anxiety Inventory (STAI) questionnaire. 5-HT concentrations were measured in fundic and duodenal mucosal biopsies by means of ELISA and immunohistochemistry. ATD significantly decreased plasma tryptophan levels compared with control in every experiment. ATD did not affect gastric sensitivity and compliance but decreased the sensation of nausea during balloon distension (AUC: 17.4 ± 4.3 vs. 11.4 ± 3.4 mm·mmHg, P = 0.030). ATD enhanced the postprandial volume increase (ANOVA, P < 0.05), but this was not accompanied by augmented nutrient tolerance (848 ± 110 vs. 837 ± 99 ml, nonsignificant). ATD had no effect on STAI state anxiety scores. No evidence was found for an effect on the number of enterochromaffin cells, but ATD reduced 5-HT levels in the duodenal mucosa. ATD alters gastric postprandial motor function and distension-induced nausea. These findings confirm involvement of 5-HT in the control of gastric accommodation and sensitivity.

Regional brain activity in functional dyspepsia: a H(2)(15)O-PET study on the role of gastric sensitivity and abuse history.

BACKGROUND & AIMS: Differences in brain activity between health and functional dyspepsia (FD) have been reported; it is unclear whether this is influenced by gastric hypersensitivity or abuse history. Therefore, we aimed to determine the influence of gastric sensitivity and abuse history on gastric sensation scores and brain activity in homeostatic-afferent, emotional-arousal, and cortical-modulatory brain regions in FD. METHODS: Abuse history was assessed using a validated self-report questionnaire. H(2)(15)O positron emission tomography was performed in 25 FD patients (13 hypersensitive and 8 abused) during 3 conditions, that is, no distension, gastric distension at discomfort threshold, and sham distension. Data were analyzed in SPM2. Region of interest analysis was used to confirm differences in prehypothesized regions. RESULTS: No association between hypersensitivity and abuse history was found. Gastric hypersensitivity was associated with significantly higher gastric sensation scores during baseline and sham. A condition-independent difference in ventral posterior cingulate activity was found between groups, as well as distension and sham-specific differences in brainstem and cingulate areas. Abuse history was associated with higher gastric sensation scores in all conditions and with differences in insular, prefrontal, and hippocampus/amygdala activity. CONCLUSIONS: Gastric sensitivity and abuse history independently influence gastric sensation as well as brain activity in FD.

Reproducibility and agreement of pharyngeal automated impedance manometry with videofluoroscopy.

BACKGROUND & AIMS: Automated impedance manometry analysis (AIM) measures swallow function variables that define bolus timing, intrabolus pressure, contractile vigor, and bolus presence; these are combined to derive a swallow risk index (SRI) that is correlated with pharyngeal dysfunction and aspiration. We assessed intra-rater and inter-rater reproducibility of AIM analysis-derived variables; the diagnostic accuracy of AIM-based criteria for detecting aspiration was determined by using expertly scored videofluoroscopy as the standard. METHODS: Data on 50 bolus swallows of 10 mL each were randomly selected from a database of swallows that were simultaneously recorded with impedance, manometry, and videofluoroscopy. Data were divided into 5 subgroups of 10 swallows for analysis: 10 dysphagic liquid, 10 dysphagic liquid with aspiration, 10 dysphagic semisolid, 10 control liquid, and 10 control semisolid. Repeat analyses were performed by 10 observers with varying levels of expertise in manometry by using purpose-designed software (AIMplot). Swallow videos were scored by 4 experts by using the penetration-aspiration scale (PAS) score. Reproducibility of calculation of swallow function variables and the SRI and PAS was assessed by using intraclass correlation coefficient (ICC). The majority consensus of expert PAS scores was used to dichotomously define aspiration (consensus PAS >3). Observer analyses were compared by Cohen κ statistical analysis. RESULTS: The intra-rater and inter-rater reproducibility of swallow function variables was high (SRI mean intra-rater ICC, 0.97 and mean inter-rater ICC, 0.91). SRI >15-20 was optimal for detecting the presence of aspiration during liquid bolus swallows with an almost perfect agreement with expert scoring of videofluoroscopy (κ > 0.8). CONCLUSIONS: AIM analysis has high intra-rater and inter-rater reproducibility, and among observers of varying expertise, SRI predicts the presence of aspiration.

Deficiency of gelatinase B/MMP-9 aggravates lpr-induced lymphoproliferation and lupus-like systemic autoimmune disease.

Gelatinase B/matrix metalloproteinase-9 (MMP-9) is a key enzyme involved in inflammatory, hematological, vascular and neoplastic diseases. In previous studies, we explored the intracellular substrate set or 'degradome' of MMP-9 and found many systemic autoantigens as novel intracellular gelatinase B substrates. Little is known, however, about the functional role of MMP-9 in the development of systemic autoimmunity in vivo. B6(lpr/lpr) mice with defective Fas-mediated apoptosis were used to investigate the functions of MMP-9 in lymphocyte proliferation and in the development of systemic autoimmunity. Combined Fas and gelatinase B deficiency resulted in extreme lymphoproliferative disease with enhanced lymphadenopathy and splenomegaly, and significantly reduced survival compared with single Fas deficiency. At the cellular level, this was corroborated by increased lymph node accumulation of 'double negative' T cells, B cells and myeloid cells. In addition, higher autoantibody titers and more pronounced autoimmune tissue injury were found in the absence of MMP-9, culminating in chronically enhanced systemic lupus erythematosus (SLE)-like autoimmunity. After cleavage by MMP-9 the SLE autoantigens U1snRNP A and ribosomal protein P0 were hardly recognized by plasma samples of both B6(lpr/lpr).MMP-9⁻/⁻ and B6(lpr/lpr).MMP-9+/+ mice, pointing to a destruction of B cell epitopes by MMP-9-mediated proteolysis. In addition, the same loss of immunodominant epitopes was observed with plasma samples from SLE patients, suggesting that MMP-9 suppresses systemic antibody-mediated autoimmunity by clearance of autoepitopes in immunogenic substrates. Thus, new protective functions for MMP-9 were revealed in the suppression of lymphoproliferation and dampening of systemic autoimmunity, cautioning against the long-term use of MMP inhibitors in autoimmune lymphoproliferative syndrome (ALPS) and SLE.

Abuse history, depression, and somatization are associated with gastric sensitivity and gastric emptying in functional dyspepsia.

OBJECTIVE: Gastric sensitivity testing relies on subjective reporting and may therefore be influenced by psychosocial factors and somatization. Furthermore, psychological processes may affect gastric motor function (accommodation to a meal emptying) through efferent brain-gut pathways. This study sought to determine the association of abuse history, psychiatric comorbidity, and somatization with gastric sensorimotor function. METHODS: In 201 patients with functional dyspepsia, gastric sensitivity and accommodation were studied with a barostat. Gastric emptying of solids was studied using a breath test. Sexual and physical abuse history, psychiatric comorbidity (depression and panic disorder), and somatization were assessed using validated questionnaires. Multiple linear regression models were used to identify patient characteristics independently associated with gastric sensitivity and emptying. RESULTS: Age (p = .02), sexual abuse history (p < .001), physical abuse history (p = .004), and somatization (p < .001) were independently associated with gastric discomfort threshold (R(2) = 0.30); a significant depression-by-sexual abuse interaction effect was also found (p = .003). None of the factors studied were associated with gastric accommodation to a meal. Physical abuse history (p = .003) and somatization (p = .048) were independently associated with gastric emptying (R(2) = 0.19). CONCLUSIONS: These results demonstrate the complex relationship among abuse history, psychiatric comorbidity, somatization, and gastric sensorimotor (dys)function. Although the psychobiological mechanisms underlying these relationships remain to be determined, the autonomic nervous, stress hormone, and immune systems may be involved.