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1.
Nat Commun ; 12(1): 7151, 2021 12 09.
Article in English | MEDLINE | ID: mdl-34887420

ABSTRACT

The heterozygous Phospholamban p.Arg14del mutation is found in patients with dilated or arrhythmogenic cardiomyopathy. This mutation triggers cardiac contractile dysfunction and arrhythmogenesis by affecting intracellular Ca2+ dynamics. Little is known about the physiological processes preceding induced cardiomyopathy, which is characterized by sub-epicardial accumulation of fibrofatty tissue, and a specific drug treatment is currently lacking. Here, we address these issues using a knock-in Phospholamban p.Arg14del zebrafish model. Hearts from adult zebrafish with this mutation display age-related remodeling with sub-epicardial inflammation and fibrosis. Echocardiography reveals contractile variations before overt structural changes occur, which correlates at the cellular level with action potential duration alternans. These functional alterations are preceded by diminished Ca2+ transient amplitudes in embryonic hearts as well as an increase in diastolic Ca2+ level, slower Ca2+ transient decay and longer Ca2+ transients in cells of adult hearts. We find that istaroxime treatment ameliorates the in vivo Ca2+ dysregulation, rescues the cellular action potential duration alternans, while it improves cardiac relaxation. Thus, we present insight into the pathophysiology of Phospholamban p.Arg14del cardiomyopathy.


Subject(s)
Calcium-Binding Proteins/genetics , Calcium/metabolism , Cardiomyopathy, Dilated/genetics , Etiocholanolone/analogs & derivatives , Zebrafish/metabolism , Animals , Calcium-Binding Proteins/metabolism , Cardiomyopathy, Dilated/metabolism , Cardiomyopathy, Dilated/physiopathology , Disease Models, Animal , Echocardiography , Etiocholanolone/administration & dosage , Female , Gene Knock-In Techniques , Humans , Male , Myocardial Contraction , Myocardium/metabolism , Sequence Deletion , Zebrafish/genetics
2.
Clin Transl Sci ; 9(6): 321-327, 2016 12.
Article in English | MEDLINE | ID: mdl-27743499

ABSTRACT

Therapeutics promoting myelin synthesis may enhance recovery in demyelinating diseases, such as multiple sclerosis. However, no suitable method exists to quantify myelination. The turnover of galactosylceramide (myelin component) is indicative of myelination in mice, but its turnover has not been determined in humans. Here, six healthy subjects consumed 120 mL 70% D2 O daily for 70 days to label galactosylceramide. We then used mass spectrometry and compartmental modeling to quantify the turnover rate of galactosylceramide in cerebrospinal fluid. Maximum deuterium enrichment of body water ranged from 1.5-3.9%, whereas that of galactosylceramide was much lower: 0.05-0.14%. This suggests a slow turnover rate, which was confirmed by the model-estimated galactosylceramide turnover rate of 0.00168 day-1 , which corresponds to a half-life of 413 days. Additional studies in patients with multiple sclerosis are needed to investigate whether galactosylceramide turnover could be used as an outcome measure in clinical trials with remyelination therapies.


Subject(s)
Ceramides/cerebrospinal fluid , Deuterium/metabolism , Healthy Volunteers , Isotope Labeling , Monosaccharides/cerebrospinal fluid , Adult , Aged , Body Water , Female , Humans , Kinetics , Male , Middle Aged , Models, Biological , Young Adult
3.
Expert Rev Cardiovasc Ther ; 14(2): 245-53, 2016.
Article in English | MEDLINE | ID: mdl-26560188

ABSTRACT

Drug-induced Torsade de Pointes arrhythmia is a life-threatening adverse effect feared by pharmaceutical companies. For the last decade, the cardiac safety guidelines have imposed human ether-a-go-go-related gene channel blockade and prolongation of QT interval as surrogates for proarrhythmic risk propensity of a new chemical entity. Suffering from a lack of specificity, this assessment strategy led to a great amount of false positive outcomes. Therefore, this review will discuss new pharmaceutical strategies: the cardiac safety proposal that recently emerged, the Comprehensive in vitro Proarrhythmia Assay, combining in vitro assays that integrate effects on main cardiac ion channels, with computational models of human ventricular action potential as well as assays using human stem cell-derived cardiomyocytes for an improved prediction of drug's proarrhythmic liability, alternative pharmacological perspectives as well as the current treatment of drug-induced long QT syndrome.


Subject(s)
Arrhythmias, Cardiac/prevention & control , Long QT Syndrome/prevention & control , Torsades de Pointes/prevention & control , Action Potentials/drug effects , Animals , Arrhythmias, Cardiac/chemically induced , Computer Simulation , Electrocardiography , Heart Ventricles/physiopathology , Humans , Ion Channels/metabolism , Long QT Syndrome/chemically induced , Myocytes, Cardiac/metabolism , Torsades de Pointes/chemically induced
4.
Lab Anim ; 47(2): 79-88, 2013 Apr.
Article in English | MEDLINE | ID: mdl-23492513

ABSTRACT

The aetiology of insulin resistance is still an enigma. Mouse models are frequently employed to study the underlying pathology. The most commonly used methods to monitor insulin resistance are the HOMA-IR, glucose or insulin tolerance tests and the hyperinsulinemic euglycaemic clamp (HIEC). Unfortunately, these tests disturb steady state glucose metabolism. Here we describe a method in which blood glucose kinetics can be determined in fasted mice without noticeably perturbing glucose homeostasis. The method involves an intraperitoneal injection of a trace amount of [6,6-(2)H2]glucose and can be performed repeatedly in individual mice. The validity and performance of this novel method was tested in mice fed on chow or high-fat diet for a period of five weeks. After administering the mice with [6,6-(2)H2]glucose, decay of the glucose label was followed in small volumes of blood collected by tail tip bleeding during a 90-minute period. The total amount of blood collected was less than 120 µL. This novel approach confirmed in detail the well-known increase in insulin resistance induced by a high-fat diet. The mice showed reduced glucose clearance rate, and reduced hepatic and peripheral insulin sensitivity. To compensate for this insulin resistance, ß-cell function was slightly increased. We conclude that this refinement of existing methods enables detailed information of glucose homeostasis in mice. Insulin resistance can be accurately determined while mechanistic insight is obtained in underlying pathology. In addition, this novel approach reduces the number of mice needed for longitudinal studies of insulin sensitivity and glucose metabolism.


Subject(s)
Blood Glucose/analysis , Glucose Tolerance Test/methods , Insulin Resistance , Mice/metabolism , Models, Animal , Animals , Diet, High-Fat/adverse effects , Enzyme-Linked Immunosorbent Assay , Gas Chromatography-Mass Spectrometry , Injections, Intraperitoneal , Insulin/blood , Kinetics , Longitudinal Studies , Male , Mice, Inbred C57BL , Models, Biological
5.
Osteoarthritis Cartilage ; 20(8): 846-53, 2012 Aug.
Article in English | MEDLINE | ID: mdl-22595228

ABSTRACT

OBJECTIVE: Adipose tissue is an endocrine tissue releasing adipokines suggested to be involved in the pathogenesis of osteoarthritis (OA). Nevertheless, their relative contribution and exact mechanisms are still ambiguous. The aim of this study is to compare serum adipokine levels between end-stage knee OA patients and controls and to relate these serum levels to local parameters of cartilage damage and synovial inflammation. METHODS: Serum was collected from 172 severe knee OA patients, shortly before total knee replacement (TKR) surgery and from 132 controls without radiographic knee OA [Kellgren & Lawrence (K&L) = 0]. Serum adiponectin, leptin, and resistin levels were measured by enzyme-linked immunosorbent assay (ELISA). Cartilage and synovial tissue were collected at TKR surgery and assessed for cartilage degeneration and synovial inflammation by histochemistry and biochemical analyses. RESULTS: The adipokine levels were all distinctly higher in OA patients as compared to controls. Especially adiponectin and leptin were associated with female gender (stand beta = 0.239 and 0.467, respectively, P < 0.001) and body mass index (BMI) (stand beta = -0.189 and 0.396, respectively, P < 0.001). No associations between serum levels of adipokines and cartilage damage (histochemistry, proteoglycan content) were found whereas weak but positive associations with synovial inflammation were found [adiponectin and interleukin-1ß (IL-1ß), stand beta = 0.172, P = 0.02; resistin and histology, stand beta = 0.183, P = 0.034, adjusted for demographics]. CONCLUSION: This study suggests an important involvement of adipokines in OA patients considering their high serum levels compared to controls. Associations of systemic adipokines with local synovial tissue inflammation were found, although not represented by similar relations with cartilage damage, suggesting that adipokines are of relevance in the inflammatory component of OA.


Subject(s)
Adipokines/blood , Cartilage, Articular/pathology , Osteoarthritis, Knee/blood , Adiponectin/blood , Aged , Aged, 80 and over , Arthroplasty, Replacement, Knee , Body Mass Index , Case-Control Studies , Enzyme-Linked Immunosorbent Assay , Female , Humans , Interleukin-1beta/blood , Leptin/blood , Male , Middle Aged , Proteoglycans/blood , Resistin/blood , Synovial Fluid/chemistry
6.
Osteoarthritis Cartilage ; 20(3): 233-40, 2012 Mar.
Article in English | MEDLINE | ID: mdl-22227209

ABSTRACT

OBJECTIVES: Age is the most prominent predisposition for development of osteoarthritis (OA). Age-related changes of articular cartilage are likely to play a role. Advanced glycation endproducts (AGEs) accumulate in cartilage matrix with increasing age and adversely affect the biomechanical properties of the cartilage matrix and influence chondrocyte activity. In clinical studies AGEing of cartilage and its relation to actual cartilage damage can only be measured by surrogate markers (e.g., serum, skin or urine AGE levels and imaging or biochemical markers of cartilage damage). In this study actual cartilage AGE levels were directly related to actual cartilage damage by use of cartilage obtained at joint replacement surgery. METHODS: Cartilage and urine samples were obtained from 69 patients undergoing total knee replacement. Samples were analyzed for pentosidine as marker of AGE. Cartilage damage was evaluated macroscopically, histologically, and biochemically. RESULTS: Cartilage and urine pentosidine both increased with increasing age. The higher the macroscopic, histological, and biochemical cartilage damage the lower the cartilage pentosidine levels were. In multiple regression analysis age is not found to be a confounder. CONCLUSION: There is an inverse relation between cartilage AGEs and actual cartilage damage in end-stage OA. This is likely due to ongoing (ineffective) increased turnover of cartilage matrix proteins even in end stage disease.


Subject(s)
Arginine/analogs & derivatives , Cartilage, Articular/metabolism , Lysine/analogs & derivatives , Osteoarthritis, Knee/metabolism , Aged , Aging/metabolism , Arginine/metabolism , Arginine/urine , Arthroplasty, Replacement, Knee , Biomarkers/metabolism , Biomarkers/urine , Cartilage, Articular/pathology , Collagen/metabolism , Female , Glycation End Products, Advanced/metabolism , Glycation End Products, Advanced/urine , Humans , Knee Joint/metabolism , Knee Joint/pathology , Lysine/metabolism , Lysine/urine , Male , Middle Aged , Osteoarthritis, Knee/pathology , Osteoarthritis, Knee/surgery , Severity of Illness Index
7.
Br J Pharmacol ; 165(2): 467-78, 2012 Jan.
Article in English | MEDLINE | ID: mdl-21718297

ABSTRACT

BACKGROUND AND PURPOSE: Drug development requires the testing of new chemical entities for adverse effects. For cardiac safety screening, improved assays are urgently needed. Isolated adult cardiomyocytes (CM) and human embryonic stem cell-derived cardiomyocytes (hESC-CM) could be used to identify pro-arrhythmic compounds. In the present study, five assays were employed to investigate their sensitivity and specificity for evaluating the pro-arrhythmic properties of I(Kr) blockers, using moxifloxacin (safe compound) and dofetilide or E-4031 (unsafe compounds). EXPERIMENTAL APPROACH: Assays included the anaesthetized remodelled chronic complete AV block (CAVB) dog, the anaesthetized methoxamine-sensitized unremodelled rabbit, multi-cellular hESC-CM clusters, isolated CM obtained from CAVB dogs and isolated CM obtained from the normal rabbit. Arrhythmic outcome was defined as Torsade de Pointes (TdP) in the animal models and early afterdepolarizations (EADs) in the cell models. KEY RESULTS: At clinically relevant concentrations (5-12 µM), moxifloxacin was free of pro-arrhythmic properties in all assays with the exception of the isolated CM, in which 10 µM induced EADs in 35% of the CAVB CM and in 23% of the rabbit CM. At supra-therapeutic concentrations (≥100 µM), moxifloxacin was pro-arrhythmic in the isolated rabbit CM (33%), in the hESC-CM clusters (18%), and in the methoxamine rabbit (17%). Dofetilide and E-4031 induced EADs or TdP in all assays (50-83%), and the induction correlated with a significant increase in beat-to-beat variability of repolarization. CONCLUSION AND IMPLICATIONS: Isolated cardiomyocytes lack specificity to discriminate between TdP liability of the I(Kr) blocking drugs moxifloxacin and dofetilide or E4031.


Subject(s)
Anti-Arrhythmia Agents/pharmacology , Aza Compounds/pharmacology , Myocytes, Cardiac/drug effects , Phenethylamines/pharmacology , Piperidines/pharmacology , Potassium Channel Blockers/pharmacology , Pyridines/pharmacology , Quinolines/pharmacology , Sulfonamides/pharmacology , Torsades de Pointes/chemically induced , Action Potentials/drug effects , Animals , Cell Line , Disease Models, Animal , Dogs , Embryonic Stem Cells/cytology , Female , Fluoroquinolones , Heart/drug effects , Heart/physiopathology , Heart Block/physiopathology , Humans , Methoxamine , Moxifloxacin , Myocytes, Cardiac/physiology , Rabbits , Torsades de Pointes/physiopathology , Ventricular Remodeling/drug effects
8.
Br J Cancer ; 105(12): 1856-63, 2011 Dec 06.
Article in English | MEDLINE | ID: mdl-22045186

ABSTRACT

BACKGROUND: High vascular endothelial growth factor (VEGFA) levels at the time of diagnosis confer a worse prognosis to multiple malignancies. Our aim was to investigate the role of VEGFA in promoting tumour growth through interaction with its environment. METHODS: HL-60 cells were transduced with VEGFA165 or control vector using retroviral constructs. Control cells (n=7) or VEGFA165 cells (n=7) were subcutaneously injected into NOD/SCID mice. Immunohistochemistry of markers for angiogenesis (CD31) and cell proliferation (Ki67) and gene expression profiling of tumours were performed. Paracrine effects were investigated by mouse-specific cytokine arrays. RESULTS: In vivo we observed a twofold increase in tumour weight when VEGFA165 was overexpressed (P=0.001), combined with increased angiogenesis (P=0.002) and enhanced tumour cell proliferation (P=0.001). Gene expression profiling revealed human genes involved in TGF-ß signalling differentially expressed between both tumour groups, that is, TGFBR2 and SMAD5 were lower expressed whereas the inhibitory SMAD7 was higher expressed with VEGFA165. An increased expression of mouse-derived cytokines IFNG and interleukin 7 was found in VEGFA165 tumours, both described to induce SMAD7 expression. CONCLUSION: These results suggest a role for VEGFA-driven tumour growth by TGF-ß signalling inhibition via paracrine mechanisms in vivo, and underscore the importance of stromal interaction in the VEGFA-induced phenotype.


Subject(s)
Neoplasms, Experimental/pathology , Signal Transduction , Stromal Cells/pathology , Transforming Growth Factor beta/metabolism , Vascular Endothelial Growth Factor A/physiology , Animals , Cell Line, Tumor , Enzyme-Linked Immunosorbent Assay , Flow Cytometry , Gene Expression Profiling , Immunohistochemistry , Mice , Mice, Inbred NOD , Mice, SCID , Neoplasms, Experimental/metabolism , Real-Time Polymerase Chain Reaction
9.
Eur J Trauma Emerg Surg ; 37(4): 373-7, 2011 Aug.
Article in English | MEDLINE | ID: mdl-21949543

ABSTRACT

PURPOSE OF THE STUDY: To evaluate the value of a pelvic X-ray compared to clinical examination in diagnosing pelvic ring fractures, using computed tomography (CT) as the gold standard, in alert [Glasgow Coma Scale (GCS) ≥ 13] adult blunt trauma patients in the emergency room. METHODS: A systematic literature search was performed in PubMed and Embase. The results were screened on their titles and abstracts using in- and exclusion criteria. Subsequently, the selected articles were critically appraised for their relevance and validity. RESULTS: Two studies investigating the diagnostic value of clinical examination and pelvic X-ray compared to CT were identified. Both studies demonstrate higher negative predictive values for clinical examination [0.99 (95% confidence interval [CI] 0.98-1.0) and 1.0 (95% CI 0.99-1.0)] compared to the negative predictive values of pelvic X-ray [0.98 (95% CI 0.93-0.99) and 0.99 (95% CI 0.99-1.0)]. The positive predictive values for clinical examination were low [0.18 (95% CI 0.16-0.23) and 0.35 (95% CI 0.30-0.42)] compared to pelvic X-ray [0.97 (95% CI 0.96-0.98) and 0.97 (95% CI 0.90-0.99)]. CONCLUSIONS: In alert blunt trauma patients, pelvic X-ray only has additional diagnostic value for the detection of pelvic ring fractures if the clinical examination is positive. Pelvic X-ray should not be performed if the clinical examination is negative. In this manner, the expenditure of time, costs, and radiation are optimized.

10.
Eur J Obstet Gynecol Reprod Biol ; 158(2): 343-9, 2011 Oct.
Article in English | MEDLINE | ID: mdl-21664028

ABSTRACT

OBJECTIVE: To estimate the prevalence and identify the factors associated with previous pelvic organ prolapse (POP) and/or incontinence surgery. STUDY DESIGN: In a cross-sectional study, all women who were aged 45-85 years and registered in eight general practices were invited to participate. They completed standardised questionnaires (the urinary distress inventory (UDI) and the defaecatory distress inventory (DDI)) and answered questions on previous pelvic floor surgery. RESULTS: Out of 2979 women eligible for this study, 1380 women were included. Previous surgery had been performed in 119 women. The prevalence of surgery increased with age, with a prevalence of 20.3% in the age group 76-85 years. Pelvic floor symptoms were more prevalent in women who had undergone previous surgery, with higher UDI and DDI scores. Factors associated with previous surgery were age, higher BMI, POP symptoms during pregnancy and previous hernia surgery. CONCLUSION: In The Netherlands, approximately one in five women will undergo surgery for POP and/or incontinence during her lifetime. The women who underwent surgery were more likely to have symptoms of pelvic floor dysfunction than those who did not undergo surgery.


Subject(s)
Pelvic Organ Prolapse/surgery , Urinary Incontinence/surgery , Age Factors , Aged , Aged, 80 and over , Cross-Sectional Studies , Female , Humans , Middle Aged , Netherlands , Pelvic Floor/surgery , Pelvic Organ Prolapse/epidemiology , Prevalence , Urinary Incontinence/epidemiology
11.
J Biomech Eng ; 132(7): 071012, 2010 Jul.
Article in English | MEDLINE | ID: mdl-20590290

ABSTRACT

We use simple walking models, based on mechanical principles, to study the preferred strategy selection in human stumble recovery. Humans typically apply an elevating strategy in response to a stumble in early swing and midswing, for which the perturbed step is lengthened in a continuation of the original step. A lowering strategy is executed for stumbles occurring at midswing or late swing, for which the perturbed swing foot is immediately placed on the ground and the recovery is executed in the subsequent step. There is no clear understanding of why either strategy is preferred over the other. We hypothesize that the human strategy preference is the result of an attempt to minimize the cost of successful recovery. We evaluate five hypothesized measures for recovery cost, focusing on the energetic cost of active recovery limb placement. We determine all hypothesized cost measures as a function of the chosen recovery strategy and the timing of the stumble during gait. Minimization of the cost measures based on the required torque, impulse, power and torque/time results in a humanlike strategy preference. The cost measure based on swing work does not predict a favorable strategy as a function of the gait phase.


Subject(s)
Gait/physiology , Walking/physiology , Computer Simulation , Foot , Humans , Models, Biological , Physical Therapy Modalities , Physiological Phenomena , Research , Torque
12.
Acta Physiol (Oxf) ; 199(3): 243-56, 2010 Jul 01.
Article in English | MEDLINE | ID: mdl-20331539

ABSTRACT

Inward rectifier currents based on K(IR)2.x subunits are regarded as essential components for establishing a stable and negative resting membrane potential in many excitable cell types. Pharmacological inhibition, null mutation in mice and dominant positive and negative mutations in patients reveal some of the important functions of these channels in their native tissues. Here we review the complex mammalian expression pattern of K(IR)2.x subunits and relate these to the outcomes of functional inhibition of the resultant channels. Correlations between expression and function in muscle and bone tissue are observed, while we recognize a discrepancy between neuronal expression and function.


Subject(s)
Potassium Channels, Inwardly Rectifying/genetics , Potassium Channels, Inwardly Rectifying/physiology , Animals , Humans , Mice , Mice, Knockout , Mutation/physiology , Phenotype , Potassium Channel Blockers/pharmacology , Potassium Channels, Inwardly Rectifying/biosynthesis , Potassium Channels, Inwardly Rectifying/drug effects
13.
Infection ; 38(2): 128-30, 2010 Apr.
Article in English | MEDLINE | ID: mdl-20213287

ABSTRACT

Mendelian susceptibility to mycobacterial disease (MSMD) is a rare disorder with predisposition to severe, sometimes lethal, disease caused by otherwise poorly virulent, non-tuberculous environmental mycobacteria and poorly virulent salmonellae. In patients with MSMD, mutations have been identified in five genes that encode for the proteins IL-12/IL-23p40, IL-12/ IL-23Rbeta1, IFN-R1, IFN-gammaR2 and STAT1. These proteins play important roles in the type-1 cytokine pathway, which is crucial for human host defence against intracellular pathogens such as mycobacteria and salmonellae. We report a girl with mild Mycobacterium bovis Bacille Calmette-Guérin (BCG) disease and Salmonella enteritidis cervical lymphadenitis. Despite treatment, she has remained a fecal carrier of S. enteritidis for the past 14 years. She was found to have complete IL-12/IL-23Rbeta1 deficiency. A homozygous r.518G>C IL12RB1 mutation was identified, leading to a non-functional R173P substitution in the IL-12/IL-23Rbeta1 protein. This mutation abrogated IL-12/IL-23Rbeta1 cell-surface expression and resulted in complete lack of T cell responsiveness to both IL-12 and IL-23.


Subject(s)
Interleukin-12 Receptor beta 1 Subunit/deficiency , Lymphadenitis/microbiology , Mycobacterium bovis/isolation & purification , Receptors, Interleukin/deficiency , Salmonella Infections/microbiology , Salmonella enteritidis/isolation & purification , Tuberculosis/microbiology , Adult , Female , Humans , Interleukin-12 Receptor beta 1 Subunit/genetics , Point Mutation , Receptors, Interleukin/genetics , Salmonella Infections/immunology , Tuberculosis/immunology
14.
Br J Pharmacol ; 159(7): 1532-41, 2010 Apr.
Article in English | MEDLINE | ID: mdl-20180941

ABSTRACT

BACKGROUND AND PURPOSE: Pentamidine is a drug used in treatment of protozoal infections. Pentamidine treatment may cause sudden cardiac death by provoking cardiac arrhythmias associated with QTc prolongation and U-wave alterations. This proarrhythmic effect was linked to inhibition of hERG trafficking, but not to acute block of ion channels contributing to the action potential. Because the U-wave has been linked to the cardiac inward rectifier current (I(K1)), we examined the action and mechanism of pentamidine-mediated I(K1) block. EXPERIMENTAL APPROACH: Patch clamp measurements of I(K1) were made on cultured adult canine ventricular cardiomyocytes, K(IR)2.1-HEK293 cells and K(IR)2.x inside-out patches. Pentamidine binding to cytoplasmic amino acid residues of K(IR)2.1 channels was studied by molecular modelling. KEY RESULTS: Pentamidine application (24 h) decreased I(K1) in cultured canine cardiomyocytes and K(IR)2.1-HEK293 cells under whole cell clamp conditions. Pentamidine inhibited I(K1) in K(IR)2.1-HEK293 cells 10 min after application. When applied to the cytoplasmic side under inside-out patch clamp conditions, pentamidine block of I(K1) was acute (IC(50)= 0.17 microM). Molecular modelling predicted pentamidine-channel interactions in the cytoplasmic pore region of K(IR)2.1 at amino acids E224, D259 and E299. Mutation of these conserved residues to alanine reduced pentamidine block of I(K1). Block was independent of the presence of spermine. K(IR)2.2, and K(IR)2.3 based I(K1) was also sensitive to pentamidine blockade. CONCLUSIONS AND IMPLICATIONS: Pentamidine inhibits cardiac I(K1) by interacting with three negatively charged amino acids in the cytoplasmic pore region. Our findings may provide new insights for development of specific I(K1) blocking compounds.


Subject(s)
Antiprotozoal Agents/pharmacology , Cytoplasm/drug effects , Pentamidine/pharmacology , Potassium Channels, Inwardly Rectifying/antagonists & inhibitors , Animals , Blotting, Western , Cell Line , Cytoplasm/metabolism , Dogs , Humans , Mutation , Patch-Clamp Techniques , Potassium Channels, Inwardly Rectifying/genetics
15.
Eur J Obstet Gynecol Reprod Biol ; 149(1): 112-6, 2010 Mar.
Article in English | MEDLINE | ID: mdl-20045242

ABSTRACT

OBJECTIVE: To evaluate factors influencing pelvic organ prolapse (POP) recurrence after repair surgery with a porcine dermal collagen implant (Pelvicol). STUDY DESIGN: From December 2003 to August 2005, 78 patients with mainly stage 3 pelvic organ prolapse by the Pelvic Organ Prolapse Quantification (POP-Q) system underwent vaginal repair using porcine dermal implant. Assessments were made preoperatively and at 6 weeks and one year follow-up. These included physical examination with POP-Q, Incontinence Impact Questionnaire, Urogenital Distress Inventory and the Defaecatory Distress Inventory and questions from the Patient Global Impression of Improvement. Data were recorded on changes in functional status, mesh erosion and adverse events. Procedural success was defined as POP-Q stage 0/1. RESULTS: 71 patients returned for one year follow-up. The overall cure rate was 74.6%. The chance of a successful operation in the younger group was significantly higher than in the older group (OR: 7.5; 95% CI 1.38, 40.69), but this effect is lower and no longer significant after adjusting for compartment (post, anterior, post+anterior) (adjusted OR: 5.5; 95% CI 0.92, 32.6). CONCLUSIONS: A low complication rate and satisfactory overall results were observed in a group of women after POP repair surgery with Pelvicol. Whether these results are better or worse than with conventional surgery or non-resorbable mesh implantation is unclear and can only be determined in a randomized controlled trial. There was a strong tendency towards better results in the younger women than in the older women, but the reason for this phenomenon is unclear.


Subject(s)
Pelvic Organ Prolapse/surgery , Skin Transplantation , Suburethral Slings , Surgical Mesh , Adult , Age Factors , Aged , Aged, 80 and over , Female , Humans , Middle Aged , Patient Satisfaction , Prosthesis Implantation , Quality of Life , Recurrence , Severity of Illness Index , Surveys and Questionnaires , Treatment Outcome
16.
Neurourol Urodyn ; 29(1): 30-9, 2010.
Article in English | MEDLINE | ID: mdl-20025017

ABSTRACT

AIMS: In this review we try to shed light on the following questions: *How frequently are symptoms of overactive bladder (OAB) and is detrusor overactivity (DO) present in patients with pelvic organ prolapse (POP) and is there a difference from women without POP? *Does the presence of OAB symptoms depend on the prolapsed compartment and/or stage of the prolapse? *What is the possible pathophysiology of OAB in POP? *Do OAB symptoms and DO change after conservative or surgical treatment of POP? METHODS: We searched on Medline and Embase for relevant studies. We only included studies in which actual data about OAB symptoms were available. All data for prolapse surgery were without the results of concomitant stress urinary incontinence (SUI) surgery. RESULTS: Community- and hospital-based studies showed that the prevalence of OAB symptoms was greater in patients with POP than without POP. No evidence was found for a relationship between the compartment or stage of the prolapse and the presence of OAB symptoms. All treatments for POP (surgery, pessaries) resulted in an improvement in OAB symptoms. It is unclear what predicts whether OAB symptoms disappear or not. When there is concomitant DO and POP, following POP surgery DO disappear in a proportion of the patients. Bladder outlet obstruction is likely to be the most important mechanism by which POP induces OAB symptoms and DO signs. However, several other mechanisms might also play a role. CONCLUSIONS: There are strong indications that there is a causal relationship between OAB and POP.


Subject(s)
Pelvic Floor/physiopathology , Pelvic Organ Prolapse/complications , Urinary Bladder Neck Obstruction/etiology , Urinary Bladder, Overactive/etiology , Urinary Bladder/physiopathology , Female , Humans , Pelvic Organ Prolapse/epidemiology , Pelvic Organ Prolapse/physiopathology , Pelvic Organ Prolapse/therapy , Pessaries , Prevalence , Risk Factors , Severity of Illness Index , Treatment Outcome , Urinary Bladder Neck Obstruction/epidemiology , Urinary Bladder Neck Obstruction/physiopathology , Urinary Bladder Neck Obstruction/therapy , Urinary Bladder, Overactive/epidemiology , Urinary Bladder, Overactive/physiopathology , Urinary Bladder, Overactive/therapy , Urogenital Surgical Procedures
17.
Br J Pharmacol ; 158(2): 532-40, 2009 Sep.
Article in English | MEDLINE | ID: mdl-19732063

ABSTRACT

BACKGROUND AND PURPOSE: (13)C-urea may be a suitable marker to assess the in vivo fate of colon-targeted dosage forms given by mouth. We postulated that release in the colon (urease-rich segment) of (13)C-urea from colon-targeted capsules would lead to fermentation of (13)C-urea by bacterial ureases into (13)CO(2). Subsequent absorption into the blood and circulation would lead to detectable (13)C (as (13)CO(2)) in breath. If, however, release of (13)C-urea occurred in the small intestine (urease-poor segment), we expected detectable (13)C (as (13)C-urea) in blood but no breath (13)C (as (13)CO(2)). The differential kinetics of (13)C-urea could thus potentially describe both release kinetics and indicate the gastrointestinal segment of release. EXPERIMENTAL APPROACH: The in vivo study consisted of three experiments, during which the same group of four volunteers participated. KEY RESULTS: The kinetic model was internally valid. The appearance of (13)C-in breath CO(2) (F(fermented)) and the appearance of (13)C in blood as (13)C-urea (F(not fermented)) show a high inverse correlation (Pearson's r=-0.981, P= 0.06). The total recovery of (13)C (F(fermented)+F(not fermented)) averaged 99%, indicating complete recovery of the administered (13)C via breath and blood. (13)CO(2) exhalation was observed in all subjects. This indicates that (13)C-urea was available in urease-rich segments, such as the caecum or colon. CONCLUSIONS AND IMPLICATIONS: In this proof-of-concept study, (13)C-urea was able to provide information on both the release kinetics of a colon-targeted oral dosage form and the gastrointestinal segment where it was released.


Subject(s)
Colon/metabolism , Drug Delivery Systems , Urea/pharmacokinetics , Administration, Oral , Adolescent , Adult , Aged , Breath Tests , Capsules , Carbon Dioxide/metabolism , Carbon Isotopes , Gastrointestinal Tract/metabolism , Humans , Middle Aged , Models, Biological , Urease/metabolism , Young Adult
18.
Osteoarthritis Cartilage ; 17(4): 482-8, 2009 Apr.
Article in English | MEDLINE | ID: mdl-18926729

ABSTRACT

OBJECTIVE: Recent in vitro studies showed that celecoxib, a selective cyclooxygenase (COX)-2 inhibitor, protects human osteoarthritic cartilage tissue from degeneration. The objective was to substantiate these beneficial effects in an in vivo (clinical) study with celecoxib treatment of patients with severe knee osteoarthritis (OA) and subsequent evaluation of cartilage tissue ex vivo. METHODS: Patients with knee OA were treated 4 weeks prior to total knee replacement surgery with either celecoxib 200mg b.d., indomethacin 50mg b.d., or received no treatment. During surgery cartilage and synovium were collected and analyzed in detail ex vivo. RESULTS: When compared to non-treated patients, patients treated with celecoxib showed significant beneficial effects on proteoglycan synthesis, -release, and -content, confirming the in vitro data. In the indomethacin group, no significant differences were found compared to the control group. On the contrary, a tendency towards a lower content and lower synthesis rate was found. In the treated groups prostaglandin-E(2) levels were lower than in the control group, indicating COX-2 inhibition. Ex vivo release of interleukin-1 beta (IL-1 beta) and tumour necrosis factor-alpha by synovial tissue was decreased by treatment with celecoxib, whereas in the indomethacin group only IL-1 beta release was decreased. CONCLUSION: Using this novel approach we were able to demonstrate an in vivo generated chondrobeneficial effect of celecoxib in patients with end stage knee OA.


Subject(s)
Cartilage, Articular/drug effects , Cyclooxygenase 2 Inhibitors/therapeutic use , Osteoarthritis, Knee/drug therapy , Aged , Anti-Inflammatory Agents, Non-Steroidal/therapeutic use , Arthroplasty, Replacement, Knee , Cartilage, Articular/metabolism , Cartilage, Articular/pathology , Celecoxib , Dinoprostone/biosynthesis , Female , Humans , Indomethacin/therapeutic use , Interleukin-1beta/metabolism , Male , Matrix Metalloproteinases/metabolism , Middle Aged , Nitric Oxide/biosynthesis , Osteoarthritis, Knee/metabolism , Osteoarthritis, Knee/pathology , Osteoarthritis, Knee/surgery , Proteoglycans/metabolism , Pyrazoles/therapeutic use , Sulfonamides/therapeutic use , Synovial Membrane/drug effects , Synovial Membrane/metabolism , Tumor Necrosis Factor-alpha/metabolism
19.
J Chem Theory Comput ; 5(4): 1068-78, 2009 Apr 14.
Article in English | MEDLINE | ID: mdl-26609616

ABSTRACT

Finite temperature Born-Oppenheimer DFT-based molecular dynamics simulations are presented for the vibrational spectroscopy of the prototype gas-phase Ala2H(+) and Ala3H(+) protonated peptides. The dynamics and the vibrational signatures are used to interpret IR-MPD spectra recorded in the NH/OH stretch region. Molecular dynamics simulations are one way to go beyond the harmonic approximations commonly applied for the calculations of infrared spectra, naturally including all anharmonicities, i.e. mode couplings, vibrational and dipole anharmonicities. The dynamics of the peptides allows understanding of the evolution of the shape and width of the N-H bands when increasing the size of the peptide, as demonstrated here with the two small prototypes Ala2H(+) and Ala3H(+). Hence, the conformational dynamics of Ala2(+) at room temperature participates to the broadening of the IR active bands. The complex N-H broadband of Ala3H(+) is shown to result from the dynamics of the N-H groups in the different peptide families, with a special role from breaking/reforming of hydrogen bonds involving the N-H groups. Taking this dynamics into account is thus mandatory for the understanding of this band in the 300-400 K experimental spectrum.

20.
Eur J Clin Invest ; 38(11): 863-8, 2008 Nov.
Article in English | MEDLINE | ID: mdl-19021705

ABSTRACT

BACKGROUND: Information about the extent of carbohydrate digestion and fermentation is critical to our ability to explore the metabolic effects of carbohydrate fermentation in vivo. We used cooked (13)C-labelled barley kernels, which are rich in indigestible carbohydrates, to develop a method which makes it possible to distinguish between and to assess carbohydrate digestion and fermentation. MATERIALS AND METHODS: Seventeen volunteers ingested 86 g (dry weight) of cooked naturally (13)C enriched barley kernels after an overnight fast. (13)CO(2) and H(2) in breath samples were measured every half hour for 12 h. The data of (13)CO(2) in breath before the start of the fermentation were used to fit the curve representing the digestion phase. The difference between the area under curve (AUC) of the fitted digestion curve and the AUC of the observed curve was regarded to represent the fermentation part. Different approaches were applied to determine the proportion of the (13)C-dose available for digestion and fermentation. RESULTS: Four hours after intake of barley, H(2)-excretion in breath started to rise. Within 12 h, 24-48% of the (13)C-dose was recovered as (13)CO(2), of which 18-19% was derived from colonic fermentation and the rest from digestion. By extrapolating the curve to baseline, it was estimated that eventually 24-25% of the total available (13)C in barley would be derived from colon fermentation. CONCLUSION: Curve fitting, using (13)CO(2)- and H(2)-breath data, is a feasible and non-invasive method to assess carbohydrate digestion and fermentation after consumption of (13)C enriched starchy food.


Subject(s)
Carbon Dioxide/analysis , Dietary Carbohydrates/metabolism , Hydrogen/analysis , Starch/metabolism , Adult , Area Under Curve , Breath Tests , Carbon Isotopes , Dietary Carbohydrates/administration & dosage , Digestion , Female , Fermentation , Hordeum , Humans , Male
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