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1.
Nat Immunol ; 19(6): 617-624, 2018 06.
Artículo en Inglés | MEDLINE | ID: mdl-29760533

RESUMEN

Humoral immune responses to microbial polysaccharide surface antigens can prevent bacterial infection but are typically strain specific and fail to mediate broad protection against different serotypes. Here we describe a panel of affinity-matured monoclonal human antibodies from peripheral blood immunoglobulin M-positive (IgM+) and IgA+ memory B cells and clonally related intestinal plasmablasts, directed against the lipopolysaccharide (LPS) O-antigen of Klebsiella pneumoniae, an opportunistic pathogen and major cause of antibiotic-resistant nosocomial infections. The antibodies showed distinct patterns of in vivo cross-specificity and protection against different clinically relevant K. pneumoniae serotypes. However, cross-specificity was not limited to K. pneumoniae, as K. pneumoniae-specific antibodies recognized diverse intestinal microbes and neutralized not only K. pneumoniae LPS but also non-K. pneumoniae LPS. Our data suggest that the recognition of minimal glycan epitopes abundantly expressed on microbial surfaces might serve as an efficient humoral immunological mechanism to control invading pathogens and the large diversity of the human microbiota with a limited set of cross-specific antibodies.


Asunto(s)
Anticuerpos Antibacterianos/inmunología , Especificidad de Anticuerpos/inmunología , Klebsiella pneumoniae/inmunología , Antígenos O/inmunología , Anticuerpos Monoclonales/inmunología , Reacciones Cruzadas/inmunología , Humanos
2.
J Biol Chem ; 300(5): 107259, 2024 May.
Artículo en Inglés | MEDLINE | ID: mdl-38582453

RESUMEN

Selenoprotein I (SELENOI) catalyzes the final reaction of the CDP-ethanolamine branch of the Kennedy pathway, generating the phospholipids phosphatidylethanolamine (PE) and plasmenyl-PE. Plasmenyl-PE is a key component of myelin and is characterized by a vinyl ether bond that preferentially reacts with oxidants, thus serves as a sacrificial antioxidant. In humans, multiple loss-of-function mutations in genes affecting plasmenyl-PE metabolism have been implicated in hereditary spastic paraplegia, including SELENOI. Herein, we developed a mouse model of nervous system-restricted SELENOI deficiency that circumvents embryonic lethality caused by constitutive deletion and recapitulates phenotypic features of hereditary spastic paraplegia. Resulting mice exhibited pronounced alterations in brain lipid composition, which coincided with motor deficits and neuropathology including hypomyelination, elevated reactive gliosis, and microcephaly. Further studies revealed increased lipid peroxidation in oligodendrocyte lineage cells and disrupted oligodendrocyte maturation both in vivo and in vitro. Altogether, these findings detail a critical role for SELENOI-derived plasmenyl-PE in myelination that is of paramount importance for neurodevelopment.


Asunto(s)
Homeostasis , Metabolismo de los Lípidos , Vaina de Mielina , Oligodendroglía , Selenoproteínas , Animales , Humanos , Ratones , Encéfalo/metabolismo , Encéfalo/patología , Peroxidación de Lípido , Ratones Noqueados , Vaina de Mielina/metabolismo , Oligodendroglía/metabolismo , Oligodendroglía/patología , Fosfatidiletanolaminas/metabolismo , Éteres Fosfolípidos/metabolismo , Plasmalógenos/metabolismo , Selenoproteínas/metabolismo , Selenoproteínas/genética , Paraplejía Espástica Hereditaria/metabolismo , Paraplejía Espástica Hereditaria/genética , Paraplejía Espástica Hereditaria/patología
3.
Mol Cell Proteomics ; 22(9): 100627, 2023 09.
Artículo en Inglés | MEDLINE | ID: mdl-37532177

RESUMEN

As the first in-person Asia Oceania Human Proteomics Organization (AOHUPO) congress since 2018, the 11th AOHUPO congress was an opportune time for the research community to reconnect and to renew friendships after the long period of restricted travel due to the global pandemic. Moreover, this congress was a great opportunity for the many AO regional proteomics and mass spectrometry scientists to meet in Singapore to exchange ideas and to present their latest findings. Cohosted by the Singapore Society for Mass Spectrometry and the Malaysian Proteomics Society and held in conjunction with the seventh Asia Oceania Agricultural Proteomics Organization Congress and Singapore Society for Mass Spectrometry 2023, the meeting featured both human and agricultural proteomics. Over five hundred scientists from the AO region converged on the MAX Atria @ Singapore EXPO, Changi, Singapore from May 8 to 10 for the main congress. The diverse program was made up of 64 invited speakers and panellists for seven plenary lectures, 27 concurrent symposia, precongress and postcongress workshops, and 174 poster presentations. The AOHUPO society were able to celebrate not only their 20th anniversary but also the outstanding academic research from biological and agricultural proteomics and related 'omics fields being conducted across the Asia-Oceania region.


Asunto(s)
Proteoma , Proteómica , Humanos , Asia , Proteómica/métodos , Espectrometría de Masas , Oceanía
4.
Rapid Commun Mass Spectrom ; 38(9): e9721, 2024 May 15.
Artículo en Inglés | MEDLINE | ID: mdl-38525810

RESUMEN

RATIONALE: The application of matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) to murine lungs is challenging due to the spongy nature of the tissue. Lungs consist of interconnected air sacs (alveoli) lined by a single layer of flattened epithelial cells, which requires inflation to maintain its natural structure. Therefore, a protocol that is compatible with both lung instillation and high spatial resolution is essential to enable multi-omic studies on murine lung disease models using MALDI-MSI. METHODS AND RESULTS: To maintain the structural integrity of the tissue, murine lungs were inflated with 8% (w/v) gelatin for lipid MSI of fresh frozen tissues or 4% (v/v) paraformaldehyde neutral buffer for N-glycan and peptide MSI of FFPE tissues. Tissues were sectioned and prepared for enzymatic digestion and/or matrix deposition. Glycerol-free PNGase F was applied for N-glycan MSI, while Trypsin Gold was applied for peptide MSI using the iMatrixSpray and ImagePrep Station, respectively. For lipid, N-glycan and peptide MSI, α-cyano-4-hydroxycinnamic acid matrix was deposited using the iMatrixSpray. MS data were acquired with 20 µm spatial resolution using a timsTOF fleX MS instrument followed by MS fragmentation of lipids, N-glycans and peptides. For lipid MSI, trapped ion mobility spectrometry was used to separate isomeric/isobaric lipid species. SCiLS™ Lab was used to visualize all MSI data. For analyte identification, MetaboScape®, GlycoMod and Mascot were used to annotate MS fragmentation spectra of lipids, N-glycans and tryptic peptides, respectively. CONCLUSIONS: Our protocol provides instructions on sample preparation for high spatial resolution MALDI-MSI, MS/MS data acquisition and lipid, N-glycan and peptide annotation and identification from murine lungs. This protocol will allow non-biased analyses of diseased lungs from preclinical murine models and provide further insight into disease models.


Asunto(s)
Péptidos , Espectrometría de Masas en Tándem , Animales , Ratones , Péptidos/análisis , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/métodos , Polisacáridos/análisis , Pulmón/química , Lípidos
5.
Graefes Arch Clin Exp Ophthalmol ; 262(2): 505-517, 2024 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-37530850

RESUMEN

BACKGROUND: This study uses bootstrapping to evaluate the technical variability (in terms of model parameter variation) of Zernike corneal surface fit parameters based on Casia2 biometric data. METHODS: Using a dataset containing N = 6953 Casia2 biometric measurements from a cataractous population, a Fringe Zernike polynomial surface of radial degree 10 (36 components) was fitted to the height data. The fit error (height - reconstruction) was bootstrapped 100 times after normalisation. After reversal of normalisation, the bootstrapped fit errors were added to the reconstructed height, and characteristic surface parameters (flat/steep axis, radii, and asphericities in both axes) extracted. The median parameters refer to a robust surface representation for later estimates of elevation, whereas the SD of the 100 bootstraps refers to the variability of the surface fit. RESULTS: Bootstrapping gave median radius and asphericity values of 7.74/7.68 mm and -0.20/-0.24 for the corneal front surface in the flat/steep meridian and 6.52/6.37 mm and -0.22/-0.31 for the corneal back surface. The respective SD values for the 100 bootstraps were 0.0032/0.0028 mm and 0.0093/0.0082 for the front and 0.0126/0.0115 mm and 0.0366/0.0312 for the back surface. The uncertainties for the back surface are systematically larger as compared to the uncertainties of the front surface. CONCLUSION: As measured with the Casia2 tomographer, the fit parameters for the corneal back surface exhibit a larger degree of variability compared with those for the front surface. Further studies are needed to show whether these uncertainties are representative for the situation where actual repeat measurements are possible.


Asunto(s)
Córnea , Tomografía de Coherencia Óptica , Humanos , Topografía de la Córnea , Biometría
6.
Graefes Arch Clin Exp Ophthalmol ; 262(3): 835-846, 2024 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-37658183

RESUMEN

BACKGROUND: Intraocular lenses (IOLs) require proper positioning in the eye to provide good imaging performance. This is especially important for premium IOLs. The purpose of this study was to develop prediction models for estimating IOL decentration, tilt and the axial IOL equator position (IOLEQ) based on preoperative biometric and tomographic measures. METHODS: Based on a dataset (N = 250) containing preoperative IOLMaster 700 and pre-/postoperative Casia2 measurements from a cataractous population, we implemented shallow feedforward neural networks and multilinear regression models to predict the IOL decentration, tilt and IOLEQ from the preoperative biometric and tomography measures. After identifying the relevant predictors using a stepwise linear regression approach and training of the models (150 training and 50 validation data points), the performance was evaluated using an N = 50 subset of test data. RESULTS: In general, all models performed well. Prediction of IOL decentration shows the lowest performance, whereas prediction of IOL tilt and especially IOLEQ showed superior performance. According to the 95% confidence intervals, decentration/tilt/IOLEQ could be predicted within 0.3 mm/1.5°/0.3 mm. The neural network performed slightly better compared to the regression, but without significance for decentration and tilt. CONCLUSION: Neural network or linear regression-based prediction models for IOL decentration, tilt and axial lens position could be used for modern IOL power calculation schemes dealing with 'real' IOL positions and for indications for premium lenses, for which misplacement is known to induce photic effects and image distortion.


Asunto(s)
Cristalino , Lentes Intraoculares , Humanos , Tomografía de Coherencia Óptica , Biometría , Ojo Artificial
7.
Graefes Arch Clin Exp Ophthalmol ; 262(5): 1553-1565, 2024 May.
Artículo en Inglés | MEDLINE | ID: mdl-38150030

RESUMEN

BACKGROUND: Phakic lenses (PIOLs, the most common and only disclosed type being the implantable collamer lens, ICL) are used in patients with large or excessive ametropia in cases where laser refractive surgery is contraindicated. The purpose of this study was to present a strategy based on anterior segment OCT data for calculating the refraction correction (REF) and the change in lateral magnification (ΔM) with ICL implantation. METHODS: Based on a dataset (N = 3659) containing Casia 2 measurements, we developed a vergence-based calculation scheme to derive the REF and gain or loss in ΔM on implantation of a PIOL having power PIOLP. The calculation concept is based on either a thick or thin lens model for the cornea and the PIOL. In a Monte-Carlo simulation considering, all PIOL steps listed in the US patent 5,913,898, nonlinear regression models for REF and ΔM were defined for each PIOL datapoint. RESULTS: The calculation shows that simplifying the PIOL to a thin lens could cause some inaccuracies in REF (up to ½ dpt) and ΔM for PIOLs with high positive power. The full range of listed ICL powers (- 17 to 17 dpt) could correct REF in a range from - 17 to 12 dpt with a change in ΔM from 17 to - 25%. The linear regression considering anterior segment biometric data and the PIOLP was not capable of properly characterizing REF and ΔM, whereas the nonlinear model with a quadratic term for the PIOLP showed a good performance for both REF and ΔM prediction. CONCLUSION: Where PIOL design data are available, the calculation concept should consider the PIOL as thick lens model. For daily use, a nonlinear regression model can properly predict REF and ΔM for the entire range of PIOL steps if a vergence calculation is unavailable.


Asunto(s)
Cristalino , Lentes Intraoculares Fáquicas , Humanos , Implantación de Lentes Intraoculares , Tomografía de Coherencia Óptica , Cristalino/cirugía , Refracción Ocular
8.
Graefes Arch Clin Exp Ophthalmol ; 262(8): 2533-2539, 2024 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-38456928

RESUMEN

PURPOSE: This study aimed to estimate the corneal keratometric index in the eyes of cataract surgery patients who received zero-power intraocular lenses (IOLs). METHODOLOGY: This retrospective study analyzed postoperative equivalent spherical refraction and axial length, mean anterior curvature radius and aqueous humor refractive index to calculate the theoretical corneal keratometric index value (nk). Data was collected from 2 centers located in France and Germany. RESULTS: Thirty-six eyes were analyzed. The results revealed a mean corneal keratometric index of 1.329 ± 0.005 for traditional axial length (AL) and 1.331 ± 0.005 for Cooke modified axial length (CMAL). Results ranged from minimum values of 1.318/1.320 to maximum values of 1.340/1.340. CONCLUSION: The corneal keratometric index is a crucial parameter for ophthalmic procedures and calculations, particularly for IOL power calculation. Notably, the estimated corneal keratometric index value of 1.329/1.331 in this study is lower than the commonly used 1.3375 index. These findings align with recent research demonstrating that the theoretical corneal keratometric index should be approximately 1.329 using traditional AL and 1.331 using CMAL, based on the ratio between the mean anterior and posterior corneal curvature radii (1.22).


Asunto(s)
Extracción de Catarata , Córnea , Lentes Intraoculares , Refracción Ocular , Humanos , Estudios Retrospectivos , Femenino , Masculino , Refracción Ocular/fisiología , Anciano , Persona de Mediana Edad , Longitud Axial del Ojo/patología , Agudeza Visual/fisiología , Anciano de 80 o más Años , Topografía de la Córnea/métodos , Implantación de Lentes Intraoculares , Diseño de Prótesis , Estudios de Seguimiento , Biometría/métodos
9.
Biofouling ; 40(7): 402-414, 2024 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-38991845

RESUMEN

Microbial fouling involves the physicochemical interactions between microorganisms and solid surfaces. An electromagnetic field (EMF) may change the diffusion rates of microbial cells and the electrical double layer around the cells and contacting surfaces. In the current study, polycardanol exhibiting antibiofouling activity was modified with ferromagnetic iron oxide (IO) to investigate the EMF effects on bacterial adhesion. When there was a flow of electrolyte that contained bacterial cells, flow-induced EMF was generated according to Faraday's principle. It was observed that the IO-ionic solution (IS)-modified surfaces, with an induced current of 44, 53, 66 nA, showed decreases in the adhesion of bacteria cells more than the unmodified (polycardanol) and IO-nanoparticles-modified ones. In addition to the EMF effects, the nano-scale uniform roughness of the modified surfaces appeared to play an important role in the reduction of cell adhesion. The results demonstrated that the IOIS-modified surface (3.2 × 10-6 mM IO) had the highest antibiofouling activity.


Asunto(s)
Adhesión Bacteriana , Incrustaciones Biológicas , Campos Electromagnéticos , Fenoles , Propiedades de Superficie , Incrustaciones Biológicas/prevención & control , Adhesión Bacteriana/efectos de los fármacos , Fenoles/química , Fenoles/farmacología , Compuestos Férricos/química , Biopelículas/efectos de los fármacos
10.
Proc Natl Acad Sci U S A ; 118(17)2021 04 27.
Artículo en Inglés | MEDLINE | ID: mdl-33875601

RESUMEN

Leukocyte homing driven by the chemokine CCL21 is pivotal for adaptive immunity because it controls dendritic cell (DC) and T cell migration through CCR7. ACKR4 scavenges CCL21 and has been shown to play an essential role in DC trafficking at the steady state and during immune responses to tumors and cutaneous inflammation. However, the mechanism by which ACKR4 regulates peripheral DC migration is unknown, and the extent to which it regulates CCL21 in steady-state skin and lymph nodes (LNs) is contested. Specifically, our previous findings that CCL21 levels are increased in LNs of ACKR4-deficient mice [I. Comerford et al., Blood 116, 4130-4140 (2010)] were refuted [M. H. Ulvmar et al., Nat. Immunol. 15, 623-630 (2014)], and no differences in CCL21 levels in steady-state skin of ACKR4-deficient mice were reported despite compromised CCR7-dependent DC egress in these animals [S. A. Bryce et al., J. Immunol. 196, 3341-3353 (2016)]. Here, we resolve these issues and reveal that two forms of CCL21, full-length immobilized and cleaved soluble CCL21, exist in steady-state barrier tissues, and both are regulated by ACKR4. Without ACKR4, extracellular CCL21 gradients in barrier sites are saturated and nonfunctional, DCs cannot home directly to lymphatic vessels, and excess soluble CCL21 from peripheral tissues pollutes downstream LNs. The results identify the mechanism by which ACKR4 controls DC migration in barrier tissues and reveal a complex mode of CCL21 regulation in vivo, which enhances understanding of functional chemokine gradient formation.


Asunto(s)
Movimiento Celular , Quimiocina CCL21/metabolismo , Células Dendríticas/fisiología , Ganglios Linfáticos/metabolismo , Receptores CCR/metabolismo , Animales , Ratones Endogámicos C57BL
11.
Clin Exp Ophthalmol ; 2024 May 13.
Artículo en Inglés | MEDLINE | ID: mdl-38741026

RESUMEN

BACKGROUND: To compare results from different corneal astigmatism measurement instruments; to reconstruct corneal astigmatism from the postimplantation spectacle refraction and toric intraocular lens (IOL) power; and to derive models for mapping measured corneal astigmatism to reconstructed corneal astigmatism. METHODS: Retrospective single centre study involving 150 eyes treated with a toric IOL (Alcon SN6AT, DFT or TFNT). Measurements included IOLMaster 700 keratometry (IOLMK) and total keratometry (IOLMTK), Pentacam keratometry (PK) and total corneal refractive power in 3 and 4 mm zones (PTCRP3 and PTCRP4), and Aladdin keratometry (AK). Regression-based models mapping the measured C0 and C45 components (Alpin's method) to reconstructed corneal astigmatism were derived. RESULTS: Mean C0 components were 0.50/0.59/0.51 dioptres (D) for IOLMK/PK/AK; 0.2/0.26/0.31 D for IOLMTK/PTCRP3/PTCRP4; and 0.26 D for reconstructed corneal astigmatism. All corresponding C45 components ranged around 0. The prediction models had main diagonal elements lower than 1 with some crosstalk between C0 and C45 (nonzero off-diagonal elements). Root-mean-squared residuals were 0.44/0.45/0.48/0.51/0.50/0.47 D for IOLMK/IOLMTK/PK/PTCRP3/PTCRP4/AK. CONCLUSIONS: Results from the different modalities are not consistent. On average IOLMTK/PTCRP3/PTCRP4 match reconstructed corneal astigmatism, whereas IOLMK/PK/AK show systematic C0 offsets of around 0.25 D. IOLMTK/PTCRP3/PTCRP4. Prediction models can reduce but not fully eliminate residual astigmatism after toric IOL implantation.

12.
Semin Cell Dev Biol ; 115: 54-61, 2021 07.
Artículo en Inglés | MEDLINE | ID: mdl-33214077

RESUMEN

Selenium (Se) is an essential micronutrient that plays a key role in regulating the immune system. T cells are of particular interest due to their important role in promoting adaptive immunity against pathogens and cancer as well as regulating tolerance, all of which are influenced by dietary Se levels. The biological effects of Se are mainly exerted through the actions of the proteins into which it is inserted, i.e. selenoproteins. Thus, the roles that selenoproteins play in regulating T cell biology and molecular mechanisms involved have emerged as important areas of research for understanding how selenium affects immunity. Members of this diverse family of proteins exhibit a wide variety of functions within T cells that include regulating calcium flux induced by T cell receptor (TCR) engagement, shaping the redox tone of T cells before, during, and after activation, and linking TCR-induced activation to metabolic reprogramming required for T cell proliferation and differentiation. This review summarizes recent insights into the roles that selenoproteins play in these processes and their implications in understanding how Se may influence immunity.


Asunto(s)
Metabolismo/inmunología , Selenio/metabolismo , Selenoproteínas/metabolismo , Linfocitos T/metabolismo , Animales , Diferenciación Celular , Proliferación Celular , Modelos Animales de Enfermedad , Humanos , Ratones
13.
J Proteome Res ; 22(8): 2694-2702, 2023 08 04.
Artículo en Inglés | MEDLINE | ID: mdl-37417588

RESUMEN

Abnormal N-glycosylation has been shown to play an important role in the pathogenesis of multiple diseases. However, little is known about the relationship between N-glycosylation and knee osteoarthritis (KOA) progression at the tissue level. Thus, the aim of this study was to quantify the cartilage histomorphometric changes in formalin-fixed paraffin-embedded (FFPE) tissue collected from the lateral and medial compartments of the tibial plateau KOA patients (n = 8). Subsequently, N-glycans were analyzed by matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) followed by in situ MS/MS fragmentation. Overall, the Osteoarthritis Research Society International (OARSI) histological grade and cartilage surface fibrillation index were significantly higher, and chondrocyte size in the superficial zone was much larger, for the medial high-loaded cartilage compared to the lateral less-loaded cartilage. Among 92 putative N-glycans observed by MALDI-MSI, 3 complex-type N-glycans, (Hex)4(HexNAc)3, (Hex)4(HexNAc)4, and (Hex)5(HexNAc)4, and 1 oligomannose-type N-glycan, (Hex)9(HexNAc)2, were significantly higher in intensity in the medial cartilage compared to the lateral cartilage, whereas 2 tetra-antennary fucosylated-type N-glycans, (Hex)3(HexNAc)6(Fuc)2 and (Hex)3(HexNAc)6(Fuc)3, were significantly higher in intensity in the lateral cartilage than the medial cartilage. Our findings indicate that complex-type N-glycans are associated with higher severity of cartilage degeneration and may influence the cellular processes of KOA.


Asunto(s)
Osteoartritis de la Rodilla , Humanos , Osteoartritis de la Rodilla/patología , Espectrometría de Masas en Tándem , Cartílago/química , Cartílago/patología , Polisacáridos/química , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/métodos
14.
J Biol Chem ; 298(3): 101634, 2022 03.
Artículo en Inglés | MEDLINE | ID: mdl-35085550

RESUMEN

While miRs have been extensively studied in the context of malignancy and tumor progression, their functions in regulating T-cell activation are less clear. In initial studies, we found reduced levels of miR-15a/16 at 3 to 18 h post-T-cell receptor (TCR) stimulation, suggesting a role for decreased levels of this miR pair in shaping T-cell activation. To further explore this, we developed an inducible miR15a/16 transgenic mouse model to determine how elevating miR-15a/16 levels during early stages of activation would affect T-cell proliferation and to identify TCR signaling pathways regulated by this miR pair. Doxycycline (DOX)-induced expression of miR-15a/16 from 0 to 18 h post-TCR stimulation decreased ex vivo T-cell proliferation as well as in vivo antigen-specific T-cell proliferation. We also combined bioinformatics and proteomics approaches to identify the mitogen-activated protein kinase kinase 1 (MEK1) (Map2k1) as a target of miR-15a/16. MEK1 targeting by miR-15a/16 was confirmed using miR mimics that decreased Map2k1 mRNA containing the 3'-UTR target nucleotide sequence (UGCUGCUA) but did not decrease Map2k1 containing a mutated control sequence (AAAAAAAA). Phosphorylation of downstream signaling molecules, extracellular signal-regulated protein kinase 1/2 (ERK1/2) and Elk1, was also decreased by DOX-induced miR-15a/16 expression. In addition to MEK1, ERK1 was subsequently found to be targeted by miR-15a/16, with DOX-induced miR-15a/16 reducing total ERK1 levels in T cells. These findings show that TCR stimulation reduces miR-15a/16 levels at early stages of T-cell activation to facilitate increased MEK1 and ERK1, which promotes the sustained MEK1-ERK1/2-Elk1 signaling required for optimal proliferation.


Asunto(s)
Sistema de Señalización de MAP Quinasas , MicroARNs , Linfocitos T , Regiones no Traducidas 3' , Animales , Activación de Linfocitos , MAP Quinasa Quinasa 1/genética , MAP Quinasa Quinasa 1/inmunología , MAP Quinasa Quinasa 1/metabolismo , Sistema de Señalización de MAP Quinasas/inmunología , Ratones , MicroARNs/genética , MicroARNs/inmunología , MicroARNs/metabolismo , Receptores de Antígenos de Linfocitos T/genética , Receptores de Antígenos de Linfocitos T/inmunología , Receptores de Antígenos de Linfocitos T/metabolismo , Linfocitos T/inmunología , Linfocitos T/metabolismo , Proteína Elk-1 con Dominio ets/inmunología , Proteína Elk-1 con Dominio ets/metabolismo
15.
Curr Issues Mol Biol ; 45(4): 3603-3627, 2023 Apr 20.
Artículo en Inglés | MEDLINE | ID: mdl-37185759

RESUMEN

Nearly 90% of cervical cancers are linked to human papillomavirus (HPV). Uncovering the protein signatures in each histological phase of cervical oncogenesis provides a path to biomarker discovery. The proteomes extracted from formalin-fixed paraffin-embedded tissues of the normal cervix, HPV16/18-associated squamous intraepithelial lesion (SIL), and squamous cell carcinoma (SCC) were compared using liquid chromatography-mass spectrometry (LC-MS). A total of 3597 proteins were identified, with 589, 550, and 1570 proteins unique to the normal cervix, SIL, and SCC groups, respectively, while 332 proteins overlapped between the three groups. In the transition from normal cervix to SIL, all 39 differentially expressed proteins were downregulated, while all 51 proteins discovered were upregulated in SIL to SCC. The binding process was the top molecular function, while chromatin silencing in the SIL vs. normal group, and nucleosome assembly in SCC vs. SIL groups was the top biological process. The PI3 kinase pathway appears crucial in initiating neoplastic transformation, while viral carcinogenesis and necroptosis are important for cell proliferation, migration, and metastasis in cervical cancer development. Annexin A2 and cornulin were selected for validation based on LC-MS results. The former was downregulated in the SIL vs. normal cervix and upregulated in the progression from SIL to SCC. In contrast, cornulin exhibited the highest expression in the normal cervix and lowest in SCC. Although other proteins, such as histones, collagen, and vimentin, were differentially expressed, their ubiquitous expression in most cells precluded further analysis. Immunohistochemical analysis of tissue microarrays found no significant difference in Annexin A2 expression between the groups. Conversely, cornulin exhibited the strongest expression in the normal cervix and lowest in SCC, supporting its role as a tumor suppressor and potential biomarker for disease progression.

16.
Anal Chem ; 95(34): 12640-12647, 2023 08 29.
Artículo en Inglés | MEDLINE | ID: mdl-37583288

RESUMEN

N-glycan alterations contribute to the progression of several joint diseases, including knee osteoarthritis (KOA). However, molecular changes in KOA subchondral trabecular bone, when exposed to different joint loading forces, are still unknown. The aim of this study was, therefore, to demonstrate the feasibility to differentiate N-glycan changes in subchondral trabecular bone from four different joint loading forces of the tibial plateau regions (i.e., Lateral Anterior (L-A), Lateral Posterior (L-P), Medial Anterior (M-A), and Medial Posterior (M-P)) in KOA patients (n = 10) using matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) at 20 µm spatial resolution. The degree of cartilage degeneration was evaluated histologically, and the subchondral bone tissue microarrays (TMAs) were subsequently manually constructed from formalin-fixed paraffin-embedded (FFPE) KOA osteochondral (i.e., cartilage-subchondral bone) tissues. Overall, the Osteoarthritis Research Society International (OARSI) histological grade was significantly higher and the size of chondrocytes in the superficial zone was much larger for both M-A and M-P compared to L-A and L-P of cartilage (p = 0.006, p = 0.030, p = 0.028, and p = 0.010; respectively). Among the 65 putative N-glycans observed by MALDI-MSI, 2 core fucosylated bi-antennary N-glycans, m/z 1809.64; (Hex)5(HexNAc)4(Fuc)1 and 2100.73; (NeuAc)1(Hex)5(HexNAc)4(Fuc)1, were significantly higher in intensity in M-A compared to L-A of the trabecular bone (p = 0.027, and p = 0.038, respectively). These N-glycans were then further structurally characterized by in situ MS/MS fragmentation post-MALDI-MSI. Our results demonstrate, for the first time, N-glycan alterations can occur at different joint loading forces in the KOA tibial plateau and the feasibility of subchondral bone TMA construction for N-glycan MALDI-MSI.


Asunto(s)
Osteoartritis de la Rodilla , Humanos , Osteoartritis de la Rodilla/diagnóstico por imagen , Osteoartritis de la Rodilla/patología , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/métodos , Espectrometría de Masas en Tándem , Polisacáridos/química , Tibia/patología
17.
Yeast ; 40(5-6): 171-181, 2023 May.
Artículo en Inglés | MEDLINE | ID: mdl-37070695

RESUMEN

Beer refermentation in bottles is an industrial process utilized by breweries where yeast and fermentable extract are added to green beer. The beer is refermented for a minimum of 2 weeks before distribution, with the physiological state of the yeast a critical factor for successful refermentation. Ideally, fresh yeast that is propagated from a dedicated propagation plant should be used for refermentation in bottles. Here, we explored the applicability of the fluorescent and redox-sensitive dye, resazurin, to assess cellular metabolism in yeast and its ability to differentiate between growth stages. We applied this assay, with other markers of yeast physiology, to evaluate yeast quality during a full-scale industrial propagation. Resazurin allowed the discrimination between the different growth phases in yeast and afforded a more in-depth understanding of yeast metabolism during propagation. This assay can be used to optimize the yeast propagation process and cropping time to improve beer quality.


Asunto(s)
Cerveza , Saccharomyces cerevisiae , Saccharomyces cerevisiae/metabolismo , Fermentación , Cerveza/análisis , Oxidación-Reducción
18.
Graefes Arch Clin Exp Ophthalmol ; 261(4): 999-1010, 2023 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-36307658

RESUMEN

PURPOSE: To compare biometric measures from 2 modern swept-source OCT biometers (IOLMaster700 (Z, Carl-Zeiss-Meditec) and Anterion (H, Heidelberg Engineering)) and evaluate the effect of measurement differences on the resulting lens power (IOLP). METHODS: Biometric measurements were made on a large study population with both instruments. We compared axial length (AL), central corneal thickness (CCT), anterior chamber depth (ACD), lens thickness (LT) and corneal front and back surface curvature measurements. Corneal curvature was converted to power vectors and total power derived using the Gullstrand formula. A paraxial lens power calculation formula and a prediction for the IOL axial position according to the Castrop formula were used to estimate differences in IOLP targeting for emmetropia. RESULTS: There were no systematic differences between measurements of AL (- 0.0146 ± 0.0286 mm) and LT (0.0383 ± 0.0595 mm), whereas CCT yielded lower (7.8 ± 6.6 µm) and ACD higher (0.1200 ± 0.0531 mm) values with H. With H, CCT was lower for thicker corneas. The mean corneal front surface radius did not differ (- 0.4 ± 41.6 µm), but the corneal back surface yielded a steeper radius (- 397.0 ± 74.6 µm) with H, giving lower mean total power (- 0.3469 ± 0.2689 dpt). The astigmatic vector components in 0°/90° and 45°/135° were the same between both instruments for the front/back surface or total power. CONCLUSION: The biometric measures used in standard formulae (AL, corneal front surface curvature/power) are consistent between instruments. However, modern formulae involving ACD, CCT or corneal back surface curvature may yield differences in IOLP, and therefore, formula constant optimisation customised to the biometer type is required.


Asunto(s)
Cristalino , Lentes Intraoculares , Humanos , Longitud Axial del Ojo/anatomía & histología , Tomografía de Coherencia Óptica/métodos , Córnea , Biometría/métodos , Reproducibilidad de los Resultados , Cámara Anterior
19.
Clin Exp Ophthalmol ; 51(6): 559-565, 2023 08.
Artículo en Inglés | MEDLINE | ID: mdl-37264533

RESUMEN

BACKGROUND: We evaluated whether the best-fit intraocular lens (IOL) power formula for the first operated eye (BF1) was also the most accurate formula for the second eye. METHODS: This was a retrospective study of 152 patients who underwent uncomplicated delayed bilateral cataract surgery with a minimum delay of 3 weeks using only one 1-piece IOL (HOYA, Vivinex) at the Medical University of Vienna, Austria. Seven different formulae (Barrett Universal II, Castrop, Haigis, Hoffer Q, Holladay 1, Kane, and SRK/T) were investigated to test the formula selection approach with regard to the calculated mean and median absolute prediction errors (MAE/MedAE). RESULTS: The mean intraindividual difference in axial length was 0.2 mm (±0.3 mm). BF1 coincided with the best-fit formula for the second eye (BF2) in 56% of patients (p < 0.05). Using BF1 for the second eye led to a lower MedAE (0.22 dioptre, D) than using a formula at random (0.33 D) and was less accurate than using the best-fit formula for each eye separately (0.1 D). The MedAEs of all formulae were generally low, ranging from 0.28 to 0.35 D. CONCLUSION: Using BF1 for the second eye led to a lower MedAE than the random selection of a formula. Therefore, BF1 can be used for the second eye if the surgeon is unsure of the choice of formula.


Asunto(s)
Catarata , Lentes Intraoculares , Facoemulsificación , Humanos , Implantación de Lentes Intraoculares , Refracción Ocular , Estudios Retrospectivos , Biometría , Óptica y Fotónica , Longitud Axial del Ojo , Catarata/complicaciones
20.
Chaos ; 33(4)2023 Apr 01.
Artículo en Inglés | MEDLINE | ID: mdl-37097944

RESUMEN

This paper introduces the Graphics Processing Unit (GPU)-based tool Geo-Temporal eXplorer (GTX), integrating a set of highly interactive techniques for visual analytics of large geo-referenced complex networks from the climate research domain. The visual exploration of these networks faces a multitude of challenges related to the geo-reference and the size of these networks with up to several million edges and the manifold types of such networks. In this paper, solutions for the interactive visual analysis for several distinct types of large complex networks will be discussed, in particular, time-dependent, multi-scale, and multi-layered ensemble networks. Custom-tailored for climate researchers, the GTX tool supports heterogeneous tasks based on interactive, GPU-based solutions for on-the-fly large network data processing, analysis, and visualization. These solutions are illustrated for two use cases: multi-scale climatic process and climate infection risk networks. This tool helps one to reduce the complexity of the highly interrelated climate information and unveils hidden and temporal links in the climate system, not available using standard and linear tools (such as empirical orthogonal function analysis).

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