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1.
Amino Acids ; 54(7): 1001-1011, 2022 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-35386060

RESUMEN

Hypertension is a major risk factor for kidney and cardiovascular disease. The treatment of hypertensive individuals by selected ACE inhibitors and certain di-and tripeptides halts the progression of renal deterioration and extends life-span. Renal reabsorption of these low molecular weight substrates are mediated by the PEPT1 and PEPT2 cotransporters. This study aims to investigate whether hypertension and ageing affects renal PEPT cotransporters at gene, protein expression and distribution as well as function in the superficial cortex and the outer medulla of the kidney. Membrane vesicles from the brush border (BBMV) and outer medulla (OMMV) were isolated from the kidneys of young Wistar Kyoto (Y-WKY), young spontaneously hypertensive (Y-SHR), and middle aged SHR (M-SHR) rats. Transport activity was measured using the substrate, ß-Ala-Lys (AMCA). Gene expression levels of PEPT genes were assessed with qRT-PCR while renal localisation of PEPT cotransporters was examined by immunohistochemistry with Western Blot validation. The Km and Vmax of renal PEPT1 were decreased significantly in SHR compared to WKY BBMV, whilst the Vmax of PEPT2 showed differences between SHR and WKY. By contrast to the reported cortical distribution of PEPT1, PEPT1-staining was detected in the outer medulla, whilst PEPT2 was expressed primarily in the cortex of all SHR; PEPT1 was significantly upregulated in the cortex of Y-SHR. These outcomes are indicative of a redistribution of PEPT1 and PEPT2 in the kidney proximal tubule under hypertensive conditions that has potential repercussions for nutrient handling and the therapeutic use of ACE inhibitors in hypertensive individuals.


Asunto(s)
Hipertensión , Simportadores , Inhibidores de la Enzima Convertidora de Angiotensina , Animales , Hipertensión/genética , Hipertensión/metabolismo , Riñón/metabolismo , Transportador de Péptidos 1/genética , Transportador de Péptidos 1/metabolismo , Péptidos/metabolismo , Ratas , Ratas Endogámicas SHR , Ratas Endogámicas WKY , Roedores/metabolismo , Simportadores/genética , Simportadores/metabolismo
2.
Avian Pathol ; 51(2): 141-145, 2022 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-34881666

RESUMEN

Infectious laryngotracheitis virus (ILTV) DNA has been detected in blood fractions, but the cell phenotype with which the virus is associated is unknown. This study investigated the presence of ILTV antigen in peripheral blood cells of six acutely ILTV-infected chickens (5 or 9 days post ocular inoculation with a virulent isolate) and three sham-inoculated chickens using immunofluorescent staining. Blood fractions were separated by Ficoll-Paque density gradient centrifugation, and smears were prepared from erythrocyte and leukocyte fractions. The smears were stained for ILTV glycoprotein E and the leukocyte markers CD4, CD8, Bu-1 (B cell), KUL01 (monocyte/macrophage), TCRγδ, and TCRαß/Vß2 and examined under a confocal microscope. In samples from infected birds, ILTV gE-specific fluorescence was localized in B cells and all evaluated T cell types, but not in monocytes and erythrocytes. The percentage of CD4, CD8, TCRγδ, TCRαß/Vß1, TCRαß/Vß2 and B cells positive for ILTV antigen ranged from 13.3% to 22.3%. None of the samples from the sham-inoculated chickens exhibited fluorescence for ILTV gE. The results of this pilot study suggest that ILTV has a tropism for peripheral blood T and B cells. Further research is required to investigate whether these cells support ILTV productive replication. RESEARCH HIGHLIGHTSSelective tropism of ILTV for peripheral blood cells was demonstrated in acutely infected birds.The ILTV antigen gE was detected in blood CD4, CD8, TCRγδ, TCRαß and B cells but not in monocytes and erythrocytes.The highest percentage of ILTV antigen was observed in CD4 cells (22.3%) followed by TCRαß/Vß1 (20.6%), CD8 (15.4%), TCRαß/Vß2 or B cells (14.4%) and TCRγδ cells (13.3%).


Asunto(s)
Infecciones por Herpesviridae , Herpesvirus Gallináceo 1 , Enfermedades de las Aves de Corral , Vacunas Virales , Animales , Pollos , Glicoproteínas , Infecciones por Herpesviridae/veterinaria , Linfocitos , Proyectos Piloto
3.
Amino Acids ; 53(2): 183-193, 2021 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-33404911

RESUMEN

Some dipeptides have been implicated in myocardial protection, but little is known about their membrane transporter PEPT2. The aim of this study was to determine whether the expression and activity of the cardiac-type PEPT2 cotransporter could be affected by ageing and/or hypertension. Sarcolemmal vesicles (SV) were isolated from the hearts of all rat groups using a standard procedure to investigate the transport activity and protein abundance by fluorescence spectroscopy and Western blot, respectively. SLC15A2 "PEPT2" gene expression was relatively quantified by RT-qPCR. In the Wistar rat groups, the protein and gene expression of PEPT2 were upregulated with ageing. These changes were accompanied by corresponding increases in the competitive inhibition and the transport rate (Vmax) of ß-Ala-Lys (AMCA) into SV isolated from middle-aged hearts. Although, the transport rate of ß-Ala-Lys (AMCA) into SV isolated from old hearts was significantly the lowest compared to middle-aged and young adult hearts, the inhibition percentage of ß-Ala-Lys (AMCA) transport by Gly-Gln was the highest. In the WKY and SHR rat groups, Y-SHR hypertrophied hearts showed an increase in PEPT2 gene expression accompanied by a significant decrease in protein expression and activity. With advanced age, however, M-SHR hypertrophied hearts revealed significantly lower gene expression, but higher protein expression and activity than Y-SHR hearts. These findings suggest that increased expression of PEPT2 cotransporter in all types of middle-aged hearts could be exploited to facilitate di-and tripeptide transport by PEPT2 in these hearts, which subsequently could result in improved myocardial protection in these populations.


Asunto(s)
Envejecimiento/metabolismo , Cardiomegalia/metabolismo , Hipertensión/metabolismo , Simportadores/metabolismo , Animales , Transporte Biológico , Cardiomegalia/genética , Dipéptidos/metabolismo , Hipertensión/genética , Masculino , Ratas , Ratas Endogámicas SHR , Ratas Endogámicas WKY , Simportadores/genética
4.
Mol Cell Biochem ; 476(5): 2039-2045, 2021 May.
Artículo en Inglés | MEDLINE | ID: mdl-33515384

RESUMEN

Homocysteine (Hcy) is a breakdown product of methionine metabolism. The risk of cardiovascular disease (CVD) correlates with an increase in plasma Hcy levels. The aim of this study was to investigate whether 1% methionine supplementation of adult rats altered intracellular reactive oxygen species (ROS) generation, intracellular Ca2+ content, and contractile activity in freshly isolated cardiomyocytes. This was measured under normal conditions and during oxidative stress in freshly isolated cardiomyocytes. Single rat cardiomyocytes from both sexes were isolated by enzymatic and mechanical dispersion techniques. Fluorescence microscopy was used to measure ROS production and intracellular Ca2+ concentration. Cell contraction was measured using a video camera. During exposure to 200 µM, H2O2 female cardiomyocytes produced significantly fewer ROS and had a higher intracellular Ca2+ concentration compared to male cardiomyocytes in control and methionine-fed conditions. The contractility of cardiomyocytes isolated from male rats was insignificantly decreased after methionine feeding compared to control, while the contractility of cardiomyocytes from female rats insignificantly reduced after methionine feeding and acute exposure to oxidative stress. These findings provide evidence that during exposure to 200 µM H2O2, cardiomyocytes from female rats produce less ROS and have higher intracellular Ca2+ levels. There were no significant effects on contractility in cardiomyocytes from either gender and under any of the different conditions.


Asunto(s)
Calcio/metabolismo , Metionina/farmacología , Contracción Miocárdica/efectos de los fármacos , Miocitos Cardíacos/metabolismo , Estrés Oxidativo/efectos de los fármacos , Alimentación Animal , Animales , Células Cultivadas , Femenino , Masculino , Ratas , Ratas Wistar
5.
Spinal Cord ; 59(10): 1053-1060, 2021 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-33495580

RESUMEN

STUDY DESIGN: Retrospective chart audit. OBJECTIVES: Firstly determining the prevalence of scoliosis in myelomeningocele (MMC) patients of the University Hospitals Leuven. Secondly analyzing whether there are differences concerning distribution of radiological level, ambulatory status, hydrocephalus, tethered cord, and syringomyelia in MMC patients with/without scoliosis. SETTING: University Hospitals Leuven, spina bifida convention. METHODS: The following data were collected: age, gender, radiograph type, age at the time of the radiograph, position during radiograph, presence of fusion, age at the time of fusion, diagnosis of hydrocephalus, tethered cord, or syringomyelia, radiological level of MMC, ambulatory status, main Cobb angle, main curve convexity, and main curve location. Correlation between prevalence of scoliosis and ambulatory status, neurological comorbidities, and radiological level were investigated. RESULTS: There were 116 patients remaining, after excluding patients without MMC or useful images. The scoliosis prevalence in MMC patients was 78.4% (95% CI, 71.0-85.8) for Cobb angle ≥10°; 60.3% (95% CI, 51.4-69.2) for ≥20°, 52.6% (95% CI, 43.5-61.7) for ≥30°, and 36.6% (95% CI, 27.7-45.5) for an angle ≥40°. Wheelchair users had 4 to 8 times more chance of having scoliosis than patients able to walk on all surfaces without aid. Thoracolumbar and lumbar radiological levels had a slightly higher prevalence of scoliosis than sacral levels. CONCLUSIONS: The high prevalence of scoliosis warrants a thorough screening and follow-up for MMC. There was no statistically significant difference between hydrocephalus, tethered cord, or syringomyelia regarding scoliosis. Future studies should focus on the interactions of the neurological comorbidities associated with MMC and scoliosis.


Asunto(s)
Meningomielocele , Escoliosis , Traumatismos de la Médula Espinal , Bélgica , Humanos , Meningomielocele/complicaciones , Meningomielocele/diagnóstico por imagen , Meningomielocele/epidemiología , Prevalencia , Estudios Retrospectivos , Escoliosis/complicaciones , Escoliosis/diagnóstico por imagen , Escoliosis/epidemiología
6.
Acta Orthop Belg ; 87(2): 255-261, 2021 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-34529378

RESUMEN

Scoliosis is an important problem in children with cerebral palsy (CP). However, the choice for a spinal fusion has to be weighed against the risks of major surgery in this vulnerable population. Paediatricians are frequently consulted preoperatively to assess the (respiratory) risk involved, but data on this question specific for CP are rare. Therefore, we investigated the complication rate after scoliosis surgery in children with CP, compared to idiopathic scoliosis (IS), and we searched for risk factors for the most common complications. In a retrospective monocenter study we analyzed the medical records from all children with CP and IS undergoing scoliosis surgery from 2010 until 2014. Duration of hospitalization and postoperative complications were compared within a 6-month follow-up. Univariate and multivariate logistic regression models were used to identify risk factors. The study included 44 patients with CP (mean age 15y0mo) and 78 patients with IS (mean age 14y6mo). Children with CP experience a higher rate of complications : respiratory and cardiovascular, as well as wound infections and decubitus ulcers (p<0.05). Postoperative pneumonia was the most frequent complication in both patient groups (43% and 18% in CP and IS respectively), with epilepsy being a significant risk factor (OR 3.85, p= 0.037) in children with CP. Intensive care unit and total hospital stay were longer in the CP group (p<0.001). These results may add information on perioperative care and surgical decision making.


Asunto(s)
Parálisis Cerebral , Escoliosis , Fusión Vertebral , Adolescente , Parálisis Cerebral/complicaciones , Parálisis Cerebral/epidemiología , Niño , Humanos , Complicaciones Posoperatorias/epidemiología , Estudios Retrospectivos , Escoliosis/complicaciones , Escoliosis/cirugía , Fusión Vertebral/efectos adversos , Resultado del Tratamiento
7.
Acta Orthop Belg ; 87(1): 175-179, 2021 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-34129772

RESUMEN

Obtaining a spine that is well balanced after fusion for scoliotic deformity is primordial for the patients' quality of life. A simple T-shaped instrument combined with standard intraoperative fluoroscopy can be of great help to evaluate the coronal alignment quickly. The aim of this study was to evaluate if a T-shaped device could predict the postoperative coronal balance. Before finalization of the rod fixation, the balance was checked by verifying the relationship between the T-shaped instrument and the upper instrumented vertebra (UIV), and final adjustments were made to correct the coronal balance. A retrospective study was conducted on 48 patients who underwent surgery to correct scoliotic deformity. Intraoperative and postoperative coronal alignment was measured independently by two observers. The mean intraoperative horizontal offset measured between T-shaped instrument and the center of the UIV was 1,69mm to the right with a standard deviation (SD) of 12,43 mm. On postoperative full spine radiographs, the mean offset between the centra sacral vertical line and the center of the UIV was 2,44mm to the left with a SD of 13,10mm. There is no significant difference in coronal balance between both measurements (p=0,12). With this technique we were able to predict the postoperative coronal balance in all but one patient (97,92%). We conclude that the use of a simple T-shaped instrument can provide adequate intraoperative assessment of coronal balance in correcting scoliotic deformity. Level of evidence : IV - case series.


Asunto(s)
Calidad de Vida , Fusión Vertebral , Humanos , Radiografía , Estudios Retrospectivos , Sacro
8.
Mol Cell Biochem ; 452(1-2): 71-82, 2019 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-30019300

RESUMEN

Renal PEPT1 and PEPT2 cotransporters play an important role in the balance of circulating body oligopeptides and selected peptidomimetic drugs. We aim to comprehensively characterise age-related changes of the renal PEPT cotransporters at the gene, protein, and functional level. Brush border membrane vesicles (BBMV) and outer medulla membrane vesicles (OMMV) were isolated from the kidneys of young, middle-aged and old rats. The protein expression of PEPT1 was not only increased in BBMV from old rats, but PEPT1 also appeared in OMMV from middle-aged and old rats. SLC15A1 gene expression in the renal cortex increased in middle-aged group. PEPT2 protein expression was not only increased with ageing, but PEPT2 also was found in BBMV from middle-aged and old groups. SLC15A2 gene expression in the renal outer medulla increased in the old group. These changes in the expressions and localisations of PEPT1 and PEPT2 could explain the changes to transport activity in BBMV and OMMV. These findings provide novel insights that would be useful for maintaining protein nutrition and optimising the delivery of some peptidomimetic drugs in elderly individuals.


Asunto(s)
Envejecimiento/patología , Riñón/patología , Transportador de Péptidos 1/metabolismo , Simportadores/metabolismo , Envejecimiento/metabolismo , Animales , Transporte Biológico , Riñón/metabolismo , Masculino , Microvellosidades/metabolismo , Microvellosidades/patología , Transportador de Péptidos 1/genética , Ratas , Ratas Wistar , Simportadores/genética
9.
Methods ; 140-141: 119-125, 2018 05 01.
Artículo en Inglés | MEDLINE | ID: mdl-29242135

RESUMEN

Image mean square displacement analysis (iMSD) is a method allowing the mapping of diffusion dynamics of molecules in living cells. However, it can also be used to obtain quantitative information on the diffusion processes of fluorescently labelled molecules and how their diffusion dynamics change when the cell environment is modified. In this paper, we describe the use of iMSD to obtain quantitative data of the diffusion dynamics of a small cytoskeletal protein, profilin 1 (pfn1), at the membrane of live cells and how its diffusion is perturbed when the cells are treated with Cytochalasin D and/or the interactions of pfn1 are modified when its actin and polyphosphoinositide binding sites are mutated (pfn1-R88A). Using total internal reflection fluorescence microscopy images, we obtained data on isotropic and confined diffusion coefficients, the proportion of cell areas where isotropic diffusion is the major diffusion mode compared to the confined diffusion mode, the size of the confinement zones and the size of the domains of dynamic partitioning of pfn1. Using these quantitative data, we could demonstrate a decreased isotropic diffusion coefficient for the cells treated with Cytochalasin D and for the pfn1-R88A mutant. We could also see changes in the modes of diffusion between the different conditions and changes in the size of the zones of pfn1 confinements for the pfn1 treated with Cytochalasin D. All of this information was acquired in only a few minutes of imaging per cell and without the need to record thousands of single molecule trajectories.


Asunto(s)
Membrana Celular/metabolismo , Microscopía Intravital/métodos , Profilinas/metabolismo , Imagen Individual de Molécula/métodos , Espectrometría de Fluorescencia/métodos , Línea Celular Tumoral , Membrana Celular/efectos de los fármacos , Citocalasina D/metabolismo , Difusión/efectos de los fármacos , Proteínas Fluorescentes Verdes/química , Proteínas Fluorescentes Verdes/genética , Células HEK293 , Humanos , Microscopía Intravital/instrumentación , Microscopía Fluorescente/instrumentación , Microscopía Fluorescente/métodos , Simulación de Dinámica Molecular , Mutagénesis Sitio-Dirigida , Mutación , Inhibidores de la Síntesis del Ácido Nucleico/farmacología , Profilinas/genética , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Imagen Individual de Molécula/instrumentación , Espectrometría de Fluorescencia/instrumentación
10.
Biochim Biophys Acta Biomembr ; 1860(5): 960-964, 2018 May.
Artículo en Inglés | MEDLINE | ID: mdl-29291378

RESUMEN

Integral membrane proteins PEPT1 and PEPT2 are essential for reabsorbing almost all hydrolysed or filtered di- and tripeptides alongside a wide range of peptidomimetic drugs in the kidney. The aim of this study was to investigate the potential use of the fluorophore-conjugated dipeptide ß-Ala-Lys (AMCA) as a biosensor for measuring peptide transport activity in brush border membrane vesicles isolated from the outer cortex (BBMV-OC) and outer medulla (BBMV-OM) (representing PEPT1 and PEPT2 respectively). The vesicles were isolated using a dual magnesium precipitation and centrifugation technique. Intravesicular fluorescence accumulation was measured after incubating extra-vesicular media at pH6.6 and different concentrations of ß-Ala-Lys (AMCA) with vesicles pre-equilibrated at pH7.4. Both BBMV-OC and BMMV-OM showed accumulation of an intravesicular fluorescence signal after 20min incubation. Changing the extra-vesicular pH to 7.4 caused a significant reduction in the ß-Ala-Lys (AMCA) uptake into BBMV-OC at concentrations >100µM. When different concentrations of dipeptide, Gly-Gln was added, there was a significant inhibition of 100µM ß-Ala-Lys (AMCA) uptake into BBMV-OC and BMMV-OM, reaching 69% and 80%, respectively. Kinetic analysis of ß-Ala-Lys (AMCA) at 20min showed that the Km and Vmax were 783.7±115.7µM and 2191.2±133.9ΔF/min/mg for BBMV-OC, while BMMV-OM showed significantly higher affinity, but lower capacity at Km=93.6±21.9µM and Vmax=935.8±50.2ΔF/min/mg. These findings demonstrate the applicability of ß-Ala-Lys (AMCA) as a biosensor to measure the transport activity of the renal-type PEPT1 and PEPT2 in BBMV-OC and BMMV-OM respectively.


Asunto(s)
Cumarinas/farmacocinética , Corteza Renal/metabolismo , Médula Renal/metabolismo , Microvellosidades/metabolismo , Oligopéptidos/farmacocinética , Transportador de Péptidos 1/metabolismo , Simportadores/metabolismo , Animales , Transporte Biológico , Colorantes Fluorescentes/farmacocinética , Riñón/metabolismo , Masculino , Ratas , Ratas Wistar
11.
Chemistry ; 24(31): 7834-7839, 2018 Jun 04.
Artículo en Inglés | MEDLINE | ID: mdl-29644728

RESUMEN

Antifreeze glycoproteins (AFGPs) are polymeric natural products that have drawn considerable interest in diverse research fields owing to their potent ice recrystallization inhibition (IRI) activity. Self-assembled materials have emerged as a promising class of biomimetic ice growth inhibitor, yet the development of AFGP-based supramolecular materials that emulate the aggregative behavior of AFGPs have not yet been reported. This work reports the first example of the 1D self-assembly and IRI activity of AFGP-functionalized perylene bisimides (AFGP-PBIs). Glycopeptide-functionalized PBIs underwent 1D self-assembly in water and showed modest IRI activity, which could be tuned through substitution of the PBI core. This work presents essential proof-of-principle for the development of novel IRIs as potential supramolecular cryoprotectants and glycoprotein mimics.


Asunto(s)
Proteínas Anticongelantes/química , Glicopéptidos/química , Hielo , Imidas/química , Perileno/análogos & derivados , Agua/química , Cristalización , Perileno/química , Multimerización de Proteína , Termodinámica
12.
J Membr Biol ; 250(6): 641-649, 2017 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-28988287

RESUMEN

Tri- and dipeptides are transported in the kidney by PEPT1 and PEPT2 isoforms. The aim of this study was to investigate differences in transport kinetics between renal brush border (BBMV) and outer medulla (OMMV) membrane vesicles (where PEPT1 and PEPT2 are sequentially available) for a range of di- and tripeptides and peptidomimetic drugs. This was accomplished through the use of the potential-sensitive fluorescent dye 3,3'-dipropylthiacarbocyanine iodide [DiS-C3-(3)]. BBMV and OMMV were prepared from the rat kidney using standard techniques. The presence of PEPT1 in BBMV and PEPT2 in OMMV was confirmed using Western blotting. Fluorescence changes were measured when extravesicular medium at pH 6.6 containing 0-1 mM substrates was added to a cuvette containing vesicles pre-equilibrated at pH 7.4 and 2.71 µM DiS-C3-(3). An increase in fluorescence intensity occurred upon substrate addition reflecting the expected positive change in membrane potential difference. Of the range of substrates studied, OMMV manifested the highest affinity to cefadroxil and valacyclovir (K m 4.3 ± 1.2 and 11.7 ± 3.2 µM, respectively) compared to other substrates, whilst the BBMV showed a higher affinity to Gly-His (K m 15.4 ± 3.1 µM) compared to other substrates. In addition, OMMV showed higher affinity and capacity to Gly-Gln (K m 47.1 ± 9.8 µM, 55.5 ± 2.8 ΔF/s/mg protein) than BBMV (K m 78.1 ± 13.3 µM and 35.5 ± 1.7 ΔF/s/mg protein, respectively). In conclusion, this study successfully separated the expression of PEPT1 and PEPT2 into different vesicle preparations inferring their activity in different regions of the renal proximal tubule.


Asunto(s)
Peptidomiméticos , Animales , Transporte Biológico , Riñón/metabolismo , Cinética , Potenciales de la Membrana/fisiología , Ratas
13.
Int J Mol Sci ; 17(4)2016 Apr 20.
Artículo en Inglés | MEDLINE | ID: mdl-27104521

RESUMEN

Mental health disorders have become worldwide health priorities. It is estimated that in the next 20 years they will account for a 16 trillion United State dollars (US$) loss. Up to now, the underlying pathophysiology of psychiatric disorders remains elusive. Altered cytoskeleton proteins expression that may influence the assembly, organization and maintenance of cytoskeletal integrity has been reported in major depressive disorders, schizophrenia and to some extent bipolar disorders. The use of quantitative proteomics, dynamic microscopy and super-resolution microscopy to investigate disease-specific protein signatures holds great promise to improve our understanding of these disorders. In this review, we present the currently available quantitative proteomic approaches use in neurology, gel-based, stable isotope-labelling and label-free methodologies and evaluate their strengths and limitations. We also reported on enrichment/subfractionation methods that target the cytoskeleton associated proteins and discuss the need of alternative methods for further characterization of the neurocytoskeletal proteome. Finally, we present live cell imaging approaches and emerging dynamic microscopy technology that will provide the tools necessary to investigate protein interactions and their dynamics in the whole cells. While these areas of research are still in their infancy, they offer huge potential towards the understanding of the neuronal network stability and its modification across neuropsychiatric disorders.


Asunto(s)
Proteínas del Citoesqueleto/metabolismo , Citoesqueleto/metabolismo , Trastornos Mentales/metabolismo , Proteómica/métodos , Citoesqueleto/ultraestructura , Humanos , Trastornos Mentales/patología , Microscopía Fluorescente , Modelos Biológicos , Red Nerviosa , Proteoma , Imagen de Lapso de Tiempo
14.
Biophys J ; 108(6): 1448-1458, 2015 Mar 24.
Artículo en Inglés | MEDLINE | ID: mdl-25809257

RESUMEN

The image-mean square displacement technique applies the calculation of the mean square displacement commonly used in single-molecule tracking to images without resolving single particles. The image-mean square displacement plot obtained is similar to the mean square displacement plot obtained using the single-particle tracking technique. This plot is then used to reconstruct the protein diffusion law and to identify whether the labeled molecules are undergoing pure isotropic, restricted, corralled, transiently confined, or directed diffusion. In our study total internal reflection fluorescence microscopy images were taken of Cholera toxin subunit B (CtxB) membrane-labeled NIH 3T3 mouse fibroblasts and MDA 231 MB cells. We found a population of CTxB undergoing purely isotropic diffusion and one displaying restricted diffusion with corral sizes ranging from 150 to ∼1800 nm. We show that the diffusion rate of CTxB bound to GM1 is independent of the size of the confinement, suggesting that the mechanism of confinement is different from the mechanism controlling the diffusion rate of CtxB. We highlight the potential effect of continuous illumination on the diffusion mode of CTxB. We also show that aggregation of CTxB/GM1 in large complexes occurs and that these aggregates tend to have slower diffusion rates.


Asunto(s)
Membrana Celular/metabolismo , Toxina del Cólera/metabolismo , Gangliósido G(M1)/metabolismo , Animales , Línea Celular Tumoral , Simulación por Computador , Difusión , Humanos , Ratones , Microscopía Fluorescente , Modelos Moleculares , Células 3T3 NIH
15.
Pediatr Dermatol ; 31(4): 493-6, 2014.
Artículo en Inglés | MEDLINE | ID: mdl-24915996

RESUMEN

Conradi-Hünermann-Happle syndrome (X-linked dominant chondrodysplasia punctata, CDPX2 [Online Mendelian Inheritance in Man 302960]) is a rare genodermatosis that presents with blaschkolinear ichthyosis, cicatricial alopecia, chondrodysplasia punctata, asymmetric shortening of the bones, and cataracts. In this case report we describe a child presenting with a patterned alopecia in which supplementary signs and clinical examination of the mother led to the suspicion of Conradi-Hünermann-Happle syndrome. Mutation analysis revealed a heterozygous novel missense mutation, c.204G>T (p.W68C), in exon 2.


Asunto(s)
Alopecia/genética , Condrodisplasia Punctata/genética , Mutación Missense , Esteroide Isomerasas/genética , Condrodisplasia Punctata/diagnóstico , Exones/genética , Femenino , Heterocigoto , Humanos , Lactante
16.
J Clin Med ; 13(12)2024 Jun 14.
Artículo en Inglés | MEDLINE | ID: mdl-38929996

RESUMEN

Spinal deformities are considered an important complication of neuromuscular disorders such as spinal muscular atrophy (SMA). SMA patients typically develop progressive early-onset scoliosis, which is associated with increased functional decline, discomfort, and respiratory dysfunction. Over the second decade of the twenty-first century, a lot has changed in terms of the therapeutic options available to people with SMA. Specifically, the use of pharmaceutical agents such as nusinersen (Spinraza), onasemnogene abeparvovec (Zolgensma), and risdiplam (Evrysdi) has dramatically changed the landscape for SMA patients. These medications significantly alter motor- and respiratory functioning, as well as the natural progression of spinal deformities. When evaluating these agents and their impact on the development of scoliosis and motor functioning, it is important to consider the timing of treatment initiation. In patients treated after they had already developed symptoms, a shift of phenotype to a less severe subtype has been observed. This results in a delay in the onset of scoliosis for the less severe SMA types and an increase in early-onset scoliosis for the severe types in patients who would typically not live to develop scoliosis. Patients who receive treatment before they develop symptoms achieve almost normal motor functioning and will likely have a significant decrease in scoliosis prevalence or at least delay its onset.

17.
J Clin Med ; 13(15)2024 Jul 27.
Artículo en Inglés | MEDLINE | ID: mdl-39124669

RESUMEN

Background: The progression of scoliosis has been observed in skeletally mature patients with cerebral palsy (CP). The aims of this systematic review were to determine the incidence of curve progression of untreated scoliosis after skeletal maturity, to estimate the average annual increase and to identify factors that influence the progression. Methods: A systematic literature search was performed in PubMed, Embase and the Cochrane Library for original research articles published between 1968 and May 2024 with a retrospective, prospective or cross-sectional design, investigating CP patients that were followed up beyond the age of 15 years. The search was limited to articles in English, French, German and Dutch. Articles were excluded if the study population concerned neuromuscular diseases other than CP. After an assessment of the methodological quality of each study, estimates of annual curve progression and the effect of the investigated risk factors for progression were recorded systematically and synthetized. Results: Fifteen studies met the inclusion criteria, resulting in a total sample size of 2569 participants. The study populations of the included original research articles were small and heterogeneous in terms of patient age and the type and severity of CP. Curve progression after skeletal maturity occurred in all included studies. A greater curve magnitude at the end of adolescence and a severe motor deficit (an inability to walk or GMFCS IV-V) were identified as significant risk factors for the progression of scoliosis after skeletal maturity. If at least one of these risk factors was present, scoliotic curves progressed after skeletal maturity in up to 74% of patients, with an average annual increase of 1.4 to 3.5 degrees per year. No significant association was found between curve progression and the physiologic type of CP, the type of scoliotic curve, previous hip surgery, positioning and gravity, weight and length, sex, epilepsy, or pelvic obliquity. Findings on the effect of hip instability were inconsistent: a positive correlation was found with the progression of scoliosis overall, but not after skeletal maturity in particular. A significant selection bias should be considered in the calculation of average annual curve progression, as patients that received interventions to halt curve progression were excluded from follow-up. Conclusions: The identification of risk factors in patients with CP and scoliosis can aid in predicting curve progression and managing follow-ups in clinical practice. Based on the findings in this review a radiographic follow-up once every 3 years is recommended for skeletally mature CP patients with at least one risk factor, and once every 5 years if no risk factors are present.

18.
Spine Deform ; 2024 Jun 17.
Artículo en Inglés | MEDLINE | ID: mdl-38886262

RESUMEN

PURPOSE: The insertion of ilio-sacral (IS) screws for distal anchoring in the instrumentation of pediatric neuromuscular scoliosis (NS) presents a significant challenge, often leading to elevated rates of complications. Utilizing computed tomography (CT) navigation and preoperative planning technology is proposed as a potential solution to mitigate these challenges. This study aims to assess the precision of IS screw placement through CT-graphic measurements, both with and without preoperative planning, followed by navigated IS screw insertion, in pediatric neuromuscular scoliosis. METHODS: Thirty-two treated patients were grouped based on surgical procedure: planned (P): 19 patients (n = 38 screws) and non-planned (NP): 13 patients (n = 26 screws). All screw placements (P and NP) took place under CT navigation. IS screw trajectories of P-group were drawn preoperatively on CT images with the cranial trajectory planning program and fused with the intraoperative CT images. There are several important anatomical structures that should be avoided when placing the IS screw (L5 root, spinal canal, L5S1 facet, SI joint, neurovascular structures anteriorly to the sacrum, S1 root in the S1 foramen and the intestines). Each trajectory was evaluated based on seven radiographical parameters whom we have enlisted partially based on the essentials of a good trajectory described by Miladi et al. (1: Ilium; 2: SI joint; 3: Promontorium; 4: Sacral plate; 5: Anterior sacral cortex; 6: S1 foramen; 7: Spinal canal). An independent sample T test was executed to compare both groups. RESULTS: The trajectories in the P group showed a significantly (P < 0.05) higher overall similarity and optimality (12.1 ± 2.1 vs 9.1 ± 2.2 points) compared to the non-planned trajectory. CONCLUSIONS: Preoperative planning and navigated placement of IS screws on fusion images with intraoperative CT, results in a better trajectory of the ilio-sacral screws.

19.
Acta Orthop Belg ; 79(2): 216-21, 2013 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-23821975

RESUMEN

The authors conducted a retrospective study comparing the corrective effect of two sublaminar techniques on scoliosis: the classical one, based on metal wire, and a more recent one, based on polyester tape (thoracic Universal Clamp), known to be safer (less risk of neurological damage, less laminar breakthrough) and compatible with MRI. Lumbar screws were used in both groups. The authors composed two groups of 25 scoliosis patients, matched for gender, age, aetiology, anterior release, number of levels fused, number of infections, major curve and flexibility: there was no significant difference. Only the follow-up period was different: 55 months in the metal wire group, versus 17 months in the polyester tape group (p < 0.001), but this was immaterial because the curves were compared one year after surgery. After one year there was no significant difference between both groups, as to correction in the coronal or in the sagittal plane. This means that the polyester tape technique offers an interesting alternative, given that it yields supplementary advantages, as mentioned above.


Asunto(s)
Procedimientos Ortopédicos/instrumentación , Procedimientos Ortopédicos/métodos , Escoliosis/cirugía , Adolescente , Hilos Ortopédicos , Humanos , Masculino , Metales , Poliésteres , Estudios Retrospectivos , Cinta Quirúrgica
20.
Spine Deform ; 11(4): 927-932, 2023 07.
Artículo en Inglés | MEDLINE | ID: mdl-37000346

RESUMEN

PURPOSE: Although the functional outcome (e.g. the return to daily activities) plays an important role in the evaluation of treatment success for the paediatric patient, clinicians currently cannot make accurate and objective predictions regarding the very early (≤ 6 weeks) functional outcome and its recovery over time. The purpose of the present study is to objectively measure initial postoperative physical activity levels and examine the relationship with patient characteristics, fusion levels and pain. METHODS: Step count (SC) was obtained pre- (Pre-Op) and postoperatively (Post-3W: 3 weeks after surgery; Post-6W: 6 weeks after surgery) using an accelerometer. Patients were grouped based on LIV (thoracic (T-group) and lumbar (L-group)) and fusion length (FL ≤ 10 levels = SF-group and FL ≥ 11 levels = LF-group). Differences in the daily SC between groups (LIV and FL) and the three timepoints was investigated using a two-way ANOVA. RESULTS: The SC was significantly lower at both Post-3W (p < 0.001) and Post-6W (p < 0.001) compared to the preoperative SC, and significantly (p < 0.001) increased from Post-3W to Post-6W (Pre-Op = 13,049 ± 3214 steps/day; Post-3W = 6486 ± 2925 steps/day; Post-6W = 8723 ± 3020 steps/day). At both post-op timepoints the T-group had a higher SC compared to the L-group. CONCLUSION: A fusion surgery with the LIV at L2 or below has a negative impact on the very early postoperative activity levels. The initial functional outcome level of AIS patients was not related to the presently collected patient characteristics. This suggests that objective activity trackers provide novel information and could have an added value in very early rehabilitation programs.


Asunto(s)
Monitores de Ejercicio , Recuperación de la Función , Escoliosis , Fusión Vertebral , Adolescente , Niño , Humanos , Cifosis/cirugía , Radiografía , Escoliosis/cirugía , Vértebras Torácicas/cirugía
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