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1.
Am J Hematol ; 99(4): 745-750, 2024 04.
Artigo em Inglês | MEDLINE | ID: mdl-38264829

RESUMO

Profound immune dysregulation and impaired response to the SARS-CoV-2 vaccine put patients with chronic lymphocytic leukemia (CLL) at risk of severe COVID-19. We compared humoral memory and T-cell responses after booster dose vaccination or breakthrough infection. (Green) Quantitative determination of anti-Spike specific antibodies. Booster doses increased seroconversion rate and antibody titers in all patient categories, ultimately generating humoral responses similar to those observed in the postinfection cohort. In detail, humoral response with overscale median antibody titers arose in >80% of patients in watch and wait, off-therapy in remission, or under treatment with venetoclax single-agent. Anti-CD20 antibodies and active treatment with BTK inhibitors (BTKi) represent limiting factors of humoral response, still memory mounted in ~40% of cases following booster doses or infection. (Blue) Evaluation of SARS-CoV-2-specific T-cell responses. Number of T-cell functional activation markers documented in each patient. The vast majority of patients, including those seronegative, developed T-cell responses, qualitatively similar between treatment groups or between vaccination alone and infection cases. These data highlight the efficacy of booster doses in eliciting T-cell immunity independently of treatment status and support the use of additional vaccination boosters to stimulate humoral immunity in patients on active CLL-directed treatments.


Assuntos
COVID-19 , Leucemia Linfocítica Crônica de Células B , Humanos , SARS-CoV-2 , Leucemia Linfocítica Crônica de Células B/tratamento farmacológico , Vacinas contra COVID-19 , Anticorpos , Subunidade alfa de Receptor de Interleucina-2 , Imunidade Celular , Anticorpos Antivirais , Vacinação
2.
BMC Vet Res ; 17(1): 197, 2021 May 25.
Artigo em Inglês | MEDLINE | ID: mdl-34034728

RESUMO

BACKGROUND: The molecular-based classification of canine mammary carcinomas (CMCs) has been the focus of much current research. Both in canines and humans, the triple-negative (TN) molecular subtype of mammary cancer is defined by a lack of expression of progesterone receptor (PR), oestrogen receptor (ER) and HER2. It has a poor prognosis; no effective targeted therapy is available. Vitamin D displays anticarcinogenic properties, and the expression of its receptor (VDR) has been found in different molecular subtypes, being about 30-40 % of TN breast cancer (TNBC) positive to it. We assessed the VDR expression in the different molecular subtypes of 58 CMCs from 45 female dogs using an immunohistochemical panel for the molecular classification of included: PR, ER, HER2, cytokeratin (CK) 5, CK14, and Ki67. In addition, we studied the relationship among the molecular subtypes of CMCs and clinicopathologic parameters. RESULTS: Investigation showed VDR positivity in 45.0 % of the triple-negative CMCs (TNCMCs), 27.3 % of luminal B and 19.0 % of luminal A. Luminal A was the most molecular subtype represented of the total tumours (36.2 %), followed of TNCMCs (34.5 %), luminal B (20.7 %) and HER2-overexpression (10.3 %). Both HER2-overexpression and TNCMC subtypes were positively related to lymphatic invasion (P = 0.028), simple histologic subtype (P = 0.007), a higher histological grade (P = 0.045) and a trend to higher proliferation index (P = 0.09). CONCLUSIONS: The highest VDR expression was observed in TNCMC, being almost half of them (45 %) positive to this receptor. VDR expression was absent in HER2-overexpression tumours and low in luminal A and B molecular subtypes.


Assuntos
Doenças do Cão/metabolismo , Neoplasias Mamárias Animais/metabolismo , Receptores de Calcitriol/biossíntese , Animais , Doenças do Cão/classificação , Doenças do Cão/genética , Cães , Feminino , Imunofenotipagem , Neoplasias Mamárias Animais/classificação , Neoplasias Mamárias Animais/genética
3.
Ann Rheum Dis ; 77(7): e46, 2018 07.
Artigo em Inglês | MEDLINE | ID: mdl-24675098

RESUMO

This article has been retracted.

4.
Proc Biol Sci ; 284(1855)2017 May 31.
Artigo em Inglês | MEDLINE | ID: mdl-28566485

RESUMO

To survive, web-building spiders rely on their capture threads to restrain prey. Many species use special adhesives for this task, and again the majority of those species cover their threads with viscoelastic glue droplets. Cribellate spiders, by contrast, use a wool of nanofibres as adhesive. Previous studies hypothesized that prey is restrained by van der Waals' forces and entrapment in the nanofibres. A large discrepancy when comparing the adhesive force on artificial surfaces versus prey implied that the real mechanism was still elusive. We observed that insect prey's epicuticular waxes infiltrate the wool of nanofibres, probably induced by capillary forces. The fibre-reinforced composite thus formed led to an adhesion between prey and thread eight times stronger than that between thread and wax-free surfaces. Thus, cribellate spiders employ the originally protective coating of their insect prey as a fatal component of their adhesive and the insect promotes its own capture. We suggest an evolutionary arms race with prey changing the properties of their cuticular waxes to escape the cribellate capture threads that eventually favoured spider threads with viscous glue.


Assuntos
Insetos/química , Seda/química , Aranhas , Ceras/química , Adesividade , Animais
5.
J Exp Biol ; 220(Pt 17): 3142-3153, 2017 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-28659308

RESUMO

During active electrolocation, the weakly electric fish Gnathonemus petersii judges the distance and impedance of nearby objects. Capacitive objects, which modulate local amplitude and waveform of the fish's electric probing signals, cast amplitude and waveform images onto the fish's electroreceptive skin. For an unambiguous estimation of the impedance and distance of an object, the animal has to deal with multiple dependencies of object and image parameters. Based on experimentally recorded amplitude and waveform images, we investigated possible strategies of the fish to unequivocally determine both the distance and the impedance of capacitive objects. We show that the relative slope in amplitude images, but not in waveform images, is independent of object impedance and is a measure of object distance. Distance-invariant impedance estimators were obtained by two different analytical strategies. The peak modulations of both image types were 'calibrated' with the relative slope of the amplitude image. Impedance estimators were obtained whenever these pairs of image features (peak and relative slope) were related dynamically over two consecutive distances. A static impedance estimator termed 'electric colour' is postulated to arise from the relationship of an amplitude and waveform image. Our results confirm that electric colour is indeed unaffected by object distance. For electric colour estimation we suggest a minimalistic approach of just relating the peak modulations of both image types to the basal amplitude and waveform condition. Our results are discussed with regard to the anatomical and physiological organization of the fish's electrosensory neuronal pathways and behavioural strategies of electrolocating fish.


Assuntos
Peixe Elétrico/fisiologia , Impedância Elétrica , Comportamento Espacial , Animais
6.
Reprod Domest Anim ; 52 Suppl 2: 177-186, 2017 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-28402059

RESUMO

Spermatogonial stem cells (SSCs) are an important tool for fertility preservation and species conservation. The ability to expand SSCs by in vitro culture is a crucial premise for their use in assisted reproduction. Because SSCs represent a small proportion of the germ cells in the adult testis, culture success is aided by pre-enrichment through sorting techniques based on cell surface-specific markers. Given the importance of the domestic cat as a model for conservation of endangered wild felids, herein we sought to examine culture conditions as well as molecular markers for cat SSCs. Using a cell culture medium for mouse SSCs supplemented with glial cell-derived neurotrophic factor (GDNF), germ cells from prepuberal cat testes remained viable in culture for up to 43 days. Immunohistochemistry for promyelocytic leukaemia zinc finger (PLZF) protein on foetal, prepuberal and adult testis sections revealed a pattern of expression consistent with the labelling of undifferentiated spermatogonia. Fluorescence-activated cell sorting (FACS) with an antibody against epithelial cell adhesion molecule (EPCAM) was used to sort live cells. Then, the gene expression profile of EPCAM-sorted cells was investigated through RT-qPCR. Notably, EPCAM (+) cells expressed relatively high levels of CKIT (CD117), a surface protein typically expressed in differentiating germ cells but not SSCs. Conversely, EPCAM (-) cells expressed relatively high levels of POU domain class 5 transcription factor 1 (POU1F5 or OCT4), clearly a germ line stem cell marker. These results suggest that cat SSCs would probably be found within the population of EPCAM (-) cells. Future studies should identify additional surface markers that alone or in combination can be used to further enrich SSCs from cat germ cells.


Assuntos
Células-Tronco Germinativas Adultas/química , Biomarcadores/análise , Gatos , Animais , Separação Celular/métodos , Separação Celular/veterinária , Células Cultivadas , Conservação dos Recursos Naturais , Espécies em Perigo de Extinção , Molécula de Adesão da Célula Epitelial , Citometria de Fluxo/veterinária , Imuno-Histoquímica/veterinária , Fatores de Transcrição Kruppel-Like/análise , Masculino , Modelos Animais , Maturidade Sexual , Espermatogônias/química , Testículo/citologia , Transcriptoma
7.
Soc Stud Sci ; 47(4): 556-579, 2017 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-28466752

RESUMO

Rhetorical moves that construct humanoid robots as social agents disclose tensions at the intersection of science and technology studies (STS) and social robotics. The discourse of robotics often constructs robots that are like us (and therefore unlike dumb artefacts). In the discourse of STS, descriptions of how people assimilate robots into their activities are presented directly or indirectly against the backdrop of actor-network theory, which prompts attributing agency to mundane artefacts. In contradistinction to both social robotics and STS, it is suggested here that to view a capacity to partake in dialogical action (to have a 'voice') is necessary for regarding an artefact as authentically social. The theme is explored partly through a critical reinterpretation of an episode that Morana Alac reported and analysed towards demonstrating her bodies-in-interaction concept. This paper turns to 'body' with particular reference to Gibsonian affordances theory so as to identify the level of analysis at which dialogicality enters social interactions.


Assuntos
Relações Interpessoais , Robótica , Comportamento Social , Inteligência Artificial
8.
Langenbecks Arch Surg ; 401(7): 937-942, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27080995

RESUMO

PURPOSE: Focused parathyroidectomy has been proven to be a safe technique for the treatment of single-gland primary hyperparathyroidism (PHPT). The CaPTHUS scoring model has been reported to be an accurate preoperative diagnostic tool for distinguishing single-gland (SGD) from multiglandular disease (MGD), including preoperative serum calcium and PTH values plus ultrasound and Sestamibi scanning. The purpose of the present study was to validate the CaPTHUS model for the population in southern Europe, since the North American and the European populations show different clinicopathological profiles in PHPT. METHODS: This is a retrospective review of a prospectively maintained database of patients diagnosed with PHPT who underwent surgical treatment in a single referral center. Differences between SGD and MGD groups were analyzed using chi-square and Fisher's exact tests for categorical variables and Student's t test for continuous variables. Overall diagnostic accuracy of the scoring model was assessed by the area under the receiver operating characteristic (ROC) curve (AUC). A p < 0.05 level was accepted as significant. RESULTS: From January 2001 to November 2014, 241 patients were included in the study, of whom 92.1 % had SGD and 71.8 % had a CaPTHUS score ≥3. SGD was distinguished from MGD (p < 0.001) using the dichotomous scoring model based on an AUC value of 0.762. Scores ≥3 had a sensitivity of 76.5 % and a positive predictive value of 96 % for SGD. CONCLUSIONS: Despite good test performance, a CaPTHUS score ≥3 does not discard MGD definitely. Intraoperative adjuncts are still needed to further reduce the risk of missing MGD during selective parathyroidectomy.


Assuntos
Hiperparatireoidismo Primário/diagnóstico , Idoso , Cálcio/sangue , Feminino , Humanos , Hiperparatireoidismo Primário/sangue , Hiperparatireoidismo Primário/cirurgia , Masculino , Pessoa de Meia-Idade , Hormônio Paratireóideo/sangue , Paratireoidectomia , Valor Preditivo dos Testes , Curva ROC , Cintilografia , Estudos Retrospectivos , Espanha , Ultrassonografia
9.
Stem Cells ; 32(2): 436-46, 2014 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-24420904

RESUMO

Fanconi anemia (FA) is a complex genetic disease associated with a defective DNA repair pathway known as the FA pathway. In contrast to many other FA proteins, BRCA2 participates downstream in this pathway and has a critical role in homology-directed recombination (HDR). In our current studies, we have observed an extremely low reprogramming efficiency in cells with a hypomorphic mutation in Brca2 (Brca2(Δ) (27/) (Δ27)), that was associated with increased apoptosis and defective generation of nuclear RAD51 foci during the reprogramming process. Gene complementation facilitated the generation of Brca2(Δ) (27/) (Δ27) induced pluripotent stem cells (iPSCs) with a disease-free FA phenotype. Karyotype analyses and comparative genome hybridization arrays of complemented Brca2(Δ) (27/) (Δ27) iPSCs showed, however, the presence of different genetic alterations in these cells, most of which were not evident in their parental Brca2(Δ) (27/) (Δ27) mouse embryonic fibroblasts. Gene-corrected Brca2(Δ) (27/) (Δ27) iPSCs could be differentiated in vitro toward the hematopoietic lineage, although with a more limited efficacy than WT iPSCs or mouse embryonic stem cells, and did not engraft in irradiated Brca2(Δ) (27/) (Δ27) recipients. Our results are consistent with previous studies proposing that HDR is critical for cell reprogramming and demonstrate that reprogramming defects characteristic of Brca2 mutant cells can be efficiently overcome by gene complementation. Finally, based on analysis of the phenotype, genetic stability, and hematopoietic differentiation potential of gene-corrected Brca2(Δ) (27/) (Δ) (27) iPSCs, achievements and limitations in the application of current reprogramming approaches in hematopoietic stem cell therapy are also discussed.


Assuntos
Proteína BRCA2/genética , Anemia de Fanconi/genética , Terapia Genética , Células-Tronco Hematopoéticas , Células-Tronco Pluripotentes Induzidas/citologia , Animais , Proteína BRCA2/biossíntese , Diferenciação Celular/genética , Células Cultivadas , Reprogramação Celular , Dano ao DNA/genética , Anemia de Fanconi/patologia , Anemia de Fanconi/terapia , Fibroblastos/metabolismo , Células-Tronco Hematopoéticas/metabolismo , Células-Tronco Pluripotentes Induzidas/metabolismo , Camundongos
10.
Haemophilia ; 21(2): 249-257, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-25652415

RESUMO

Although 8% of reported FVIII gene (F8) mutations responsible for haemophilia A (HA) affect mRNA processing, very few have been fully characterized at the mRNA level and/or systematically predicted their biological consequences by in silico analysis. This study is aimed to elucidate the effect of potential splice site mutations (PSSM) on the F8 mRNA processing, investigate its correlation with disease severity, and assess their concordance with in silico predictions. We studied the F8 mRNA from 10 HA patient's leucocytes with PSSM by RT-PCR and compared the experimental results with those predicted in silico. The mRNA analysis could explain all the phenotypes observed and demonstrated exon skipping in six cases (c.222G>A, c.601+1delG, c.602-11T>G, c.671-3C>G, c.6115+9C>G and c.6116-1G>A) and activation of cryptic splicing sites, both donor (c.1009+1G>A and c.1009+3A>C) and acceptor sites (c.266-3delC and c.5587-1G>A). In contrast, the in silico analysis was able to predict the score variation of most of the affected splice site, but the precise mechanism could only be correctly determined in two of the 10 mutations analysed. In addition, we have detected aberrant F8 transcripts, even in healthy controls, so this must be taken into account as they could mask the actual contribution of some PSSM. We conclude that F8 mRNA analysis using leucocytes still constitutes an excellent approach to investigate the transcriptional effects of the PSSM in HA, whereas prediction in silico is not always reliable for diagnostic decision-making.


Assuntos
Fator VIII/genética , Hemofilia A/diagnóstico , Hemofilia A/genética , Leucócitos/metabolismo , Mutação , Splicing de RNA , RNA Mensageiro , Biologia Computacional/métodos , Análise Mutacional de DNA , Éxons , Humanos , Íntrons , Fenótipo , Sítios de Splice de RNA
11.
Vet Pathol ; 52(2): 291-4, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24842487

RESUMO

Thymic epithelial cells could play an important role in lymphoid depletion during bovine viral diarrhea virus (BVDV) infection. To evaluate this hypothesis, we examined proliferation of lymphocytes, expression of cytokeratins by thymic epithelial cells, and ultrastructural features at sequential time points after experimental infection of colostrum-deprived calves with the noncytopathogenic BVDV1 strain 7443. Ten clinically healthy Friesian calves were used. Eight were inoculated with the virus, and 2 were used as uninfected controls. Calves were sedated and euthanized in batches between 3 and 14 days postinoculation. At necropsy, thymus samples were collected for structural, immunohistochemical, and ultrastructural study. Thymic lymphoid depletion was accompanied by a decrease in lymphocyte proliferation and immunohistochemical and ultrastructural changes in thymic epithelial cells. Immunohistochemical and ultrastructural results reflect a disturbance of the thymic epithelial cell network, which may explain the decrease in lymphocyte proliferation by defective thymocyte-epithelial cell interactions.


Assuntos
Doença das Mucosas por Vírus da Diarreia Viral Bovina/patologia , Vírus da Diarreia Viral Bovina Tipo 1/patogenicidade , Animais , Doença das Mucosas por Vírus da Diarreia Viral Bovina/virologia , Bovinos , Vírus da Diarreia Viral Bovina Tipo 1/imunologia , Células Epiteliais/patologia , Linfócitos/patologia , Timo/patologia
12.
Am J Physiol Renal Physiol ; 306(4): F422-9, 2014 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-24370590

RESUMO

This study aimed to determine the extent of extraskeletal calcification in uremic Zucker rats, by comparing obese and lean phenotypes, and to evaluate the influence of vitamin E (VitE) on the development of calcifications in both uremic rats and human vascular smooth muscle cells (HVSMCs) cultured in vitro. Zucker rats of lean and obese phenotypes with normal renal function [control (C); C-lean and C-obese groups] and with uremia [5/6 nephrectomy (Nx); Nx-lean and Nx-obese groups] and uremic rats treated with VitE (Nx-lean + VitE and Nx-obese + VitE groups) were studied. Uremic groups were subjected to Nx, fed a 0.9% phosphorus diet, and treated with calcitriol (80 ng/kg ip). The aortic calcium concentration was significantly higher (P < 0.05) in Nx-obese rats (10.0 ± 2.1 mg/g tissue) than in Nx-lean rats (3.6 ± 1.3 mg/g tissue). A decrease in plasma glutathione peroxidase activity was observed in Nx-obese rats compared with Nx-lean rats (217.2 ± 18.2 vs. 382.3 ± 15.5 nmol·min(-1)·ml(-1), P < 0.05). Treatment with VitE restored glutathione peroxidase activity and reduced the aortic calcium concentration to 4.6 ± 1.3 mg/g tissue. The differences in mineral deposition between Nx-lean, Nx-obese, Nx-lean + VitE, and Nx-obese + VitE rats were also evidenced in other soft tissues. In HVSMCs incubated with high phosphate, VitE also prevented oxidative stress and reduced calcium content, bone alkaline phosphatase, and gene expression of core-binding factor-α1. In conclusion, uremic obese rats develop more severe calcifications than uremic lean rats and VitE reduces oxidative stress and vascular calcifications in both rats and cultures of HVSMCs.


Assuntos
Obesidade/patologia , Uremia/patologia , Calcificação Vascular/prevenção & controle , Vitamina E/uso terapêutico , Animais , Osso e Ossos/efeitos dos fármacos , Osso e Ossos/metabolismo , Osso e Ossos/patologia , Cálcio/metabolismo , Glutationa Peroxidase/metabolismo , Miócitos de Músculo Liso/efeitos dos fármacos , Miócitos de Músculo Liso/metabolismo , Miócitos de Músculo Liso/patologia , Obesidade/complicações , Estresse Oxidativo/fisiologia , Ratos , Ratos Zucker , Uremia/complicações , Calcificação Vascular/complicações , Calcificação Vascular/tratamento farmacológico , Vitamina E/farmacologia
13.
Endocr Pract ; 25(8): 862, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30657364
14.
Stem Cell Res Ther ; 14(1): 366, 2023 12 13.
Artigo em Inglês | MEDLINE | ID: mdl-38093328

RESUMO

BACKGROUND: Induced pluripotent stem cell (iPSC)-derived cell therapies are an interesting new area in the field of regenerative medicine. One of the approaches to decrease the costs of iPSC-derived therapies is the use of allogenic homozygous human leukocyte antigen (HLA)-matched donors to generate iPSC lines and to build a clinical-grade iPSC bank covering a high percentage of the Spanish population. METHODS: The Spanish Stem Cell Transplantation Registry was screened for cord blood units (CBUs) homozygous for the most common HLA-A, HLA-B and HLA-DRB1 haplotypes. Seven donors were selected with haplotypes covering 21.37% of the haplotypes of the Spanish population. CD34-positive hematopoietic progenitors were isolated from the mononuclear cell fraction of frozen cord blood units from each donor by density gradient centrifugation and further by immune magnetic labeling and separation using purification columns. Purified CD34 + cells were reprogrammed to iPSCs by transduction with the CTS CytoTune-iPS 2.1 Sendai Reprogramming Kit. RESULTS: The iPSCs generated from the 7 donors were expanded, characterized, banked and registered. Master cell banks (MCBs) and working cell banks (WCBs) from the iPSCs of each donor were produced under GMP conditions in qualified clean rooms. CONCLUSIONS: Here, we present the first clinical-grade, iPSC haplobank in Spain made from CD34 + cells from seven cord blood units homozygous for the most common HLA-A, HLA-B and HLA-DRB1 haplotypes within the Spanish population. We describe their generation by transduction with Sendai viral vectors and their GMP-compliant expansion and banking. These haplolines will constitute starting materials for advanced therapy medicinal product development (ATMP).


Assuntos
Células-Tronco Pluripotentes Induzidas , Humanos , Células-Tronco Pluripotentes Induzidas/metabolismo , Cadeias HLA-DRB1/metabolismo , Antígenos HLA/metabolismo , Antígenos HLA-B/metabolismo , Antígenos HLA-A/metabolismo
15.
Vet Pathol ; 49(5): 811-8, 2012 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-21768605

RESUMO

Thymic depletion, presence of viral antigen, and changes in distribution and cytokine production of thymic macrophages were investigated in calves experimentally infected with a noncytopathogenic bovine viral diarrhea virus type (BVDV) 1 strain. Ten clinically healthy colostrum-deprived calves were used. Eight calves were inoculated with the virus and two were used as uninfected controls. Calves were sedated and euthanized in batches between 3 and 14 days postinoculation. At necropsy, thymus samples were collected for structural, immunohistochemical, and ultrastructural study and TUNEL (terminal deoxynucleotidyl transferase dUTP nick end labeling). From 6 days postinoculation, the thymic cortex was multifocally depleted with increased frequency of pyknosis and karyorrhexis, suggestive of apoptosis and confirmed by the TUNEL technique. Although the onset of lymphoid depletion was coincident with the detection of viral antigen by immunohistochemistry, the number of infected lymphocytes was very low through the experiment. There was an increase in number of macrophages in cortex and medulla, accompanied by ultrastructural changes indicative of phagocyte activation, and a decrease in cells expressing tumor necrosis factor-alpha (TNF-α) and IL-1α. These results suggest that the increase in number of these cells could be related to phagocytosis of cell debris and apoptotic lymphocytes. Furthermore, the results imply that, in contrast to the situation with classical swine fever virus, the lymphocyte apoptosis resulting from bovine viral diarrhea virus infection is not mediated by TNF-α or interleukin-1 alpha (IL-1α) production by virus-infected macrophages. This is the first study that describes this decrease in the number of thymic cells expressing TNF-α and IL-1α in cattle experimentally infected with bovine viral diarrhea virus type 1.


Assuntos
Antígenos Virais/imunologia , Doença das Mucosas por Vírus da Diarreia Viral Bovina/imunologia , Vírus da Diarreia Viral Bovina Tipo 1/imunologia , Macrófagos/imunologia , Timo/imunologia , Animais , Apoptose , Doença das Mucosas por Vírus da Diarreia Viral Bovina/patologia , Doença das Mucosas por Vírus da Diarreia Viral Bovina/virologia , Bovinos , Imuno-Histoquímica/veterinária , Marcação In Situ das Extremidades Cortadas/veterinária , Interleucina-1alfa/metabolismo , Linfócitos/imunologia , Masculino , Fagocitose/imunologia , Timo/patologia , Fator de Necrose Tumoral alfa/metabolismo
16.
Open Life Sci ; 17(1): 917-937, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36045713

RESUMO

Mucormycosis (MCM) is a rare fungal disorder that has recently been increased in parallel with novel COVID-19 infection. MCM with COVID-19 is extremely lethal, particularly in immunocompromised individuals. The collection of available scientific information helps in the management of this co-infection, but still, the main question on COVID-19, whether it is occasional, participatory, concurrent, or coincidental needs to be addressed. Several case reports of these co-infections have been explained as causal associations, but the direct contribution in immunocompromised individuals remains to be explored completely. This review aims to provide an update that serves as a guide for the diagnosis and treatment of MCM patients' co-infection with COVID-19. The initial report has suggested that COVID-19 patients might be susceptible to developing invasive fungal infections by different species, including MCM as a co-infection. In spite of this, co-infection has been explored only in severe cases with common triangles: diabetes, diabetes ketoacidosis, and corticosteroids. Pathogenic mechanisms in the aggressiveness of MCM infection involves the reduction of phagocytic activity, attainable quantities of ferritin attributed with transferrin in diabetic ketoacidosis, and fungal heme oxygenase, which enhances iron absorption for its metabolism. Therefore, severe COVID-19 cases are associated with increased risk factors of invasive fungal co-infections. In addition, COVID-19 infection leads to reduction in cluster of differentiation, especially CD4+ and CD8+ T cell counts, which may be highly implicated in fungal co-infections. Thus, the progress in MCM management is dependent on a different strategy, including reduction or stopping of implicit predisposing factors, early intake of active antifungal drugs at appropriate doses, and complete elimination via surgical debridement of infected tissues.

17.
Stem Cell Res ; 53: 102333, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33862537

RESUMO

Limb-girdle muscular dystrophy recessive 1 (LGMDR1) represents one of the most common types of LGMD in the population, where patients develop a progressive muscle degeneration. The disease is caused by mutations in calpain 3 gene, with over 500 mutations reported to date. However, the molecular events that lead to muscle wasting are not clear, nor the reasons for the great clinical variability among patients, and this has so far hindered the development of effective therapies. Here we generate human induced pluripotent stem cells (iPSCs) from skin fibroblasts of 2 healthy controls and 4 LGMDR1 patients with different mutations. The generated lines were able to differentiate into myogenic progenitors and myotubes in vitro and in vivo, upon a transient PAX7 overexpressing protocol. Thus, we have generated myogenic cellular models of LGMDR1 that harbor different CAPN3 mutations within a human genetic background, and which do not derive from muscular biopsies. These models will allow us to investigate disease mechanisms and test therapies. Despite the variability found among iPSC lines that was unrelated to CAPN3 mutations, we found that patient-derived myogenic progenitors and myotubes express lower levels of DMD, which codes a key protein in satellite cell regulation and myotube maturation.


Assuntos
Células-Tronco Pluripotentes Induzidas , Distrofia Muscular do Cíngulo dos Membros , Humanos , Fibras Musculares Esqueléticas , Distrofia Muscular do Cíngulo dos Membros/genética , Mutação
18.
NPJ Parkinsons Dis ; 7(1): 55, 2021 Jul 02.
Artigo em Inglês | MEDLINE | ID: mdl-34215735

RESUMO

A deeper understanding of early disease mechanisms occurring in Parkinson's disease (PD) is needed to reveal restorative targets. Here we report that human induced pluripotent stem cell (iPSC)-derived dopaminergic neurons (DAn) obtained from healthy individuals or patients harboring LRRK2 PD-causing mutation can create highly complex networks with evident signs of functional maturation over time. Compared to control neuronal networks, LRRK2 PD patients' networks displayed an elevated bursting behavior, in the absence of neurodegeneration. By combining functional calcium imaging, biophysical modeling, and DAn-lineage tracing, we found a decrease in DAn neurite density that triggered overall functional alterations in PD neuronal networks. Our data implicate early dysfunction as a prime focus that may contribute to the initiation of downstream degenerative pathways preceding DAn loss in PD, highlighting a potential window of opportunity for pre-symptomatic assessment of chronic degenerative diseases.

19.
Hum Reprod ; 25(1): 158-67, 2010 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-19815622

RESUMO

BACKGROUND: Given the significant drawbacks of using human embryonic stem (hES) cells for regenerative medicine, the search for alternative sources of multipotent cells is ongoing. Studies in mice have shown that multipotent ES-like cells can be derived from neonatal and adult testis. Here we report the derivation of ES-like cells from adult human testis. METHODS: Testis material was donated for research by four men undergoing bilateral castration as part of prostate cancer treatment. Testicular cells were cultured using StemPro medium. Colonies that appeared sharp edged and compact were collected and subcultured under hES-specific conditions. Molecular characterization of these colonies was performed using RT-PCR and immunohistochemistry. (Epi)genetic stability was tested using bisulphite sequencing and karyotype analysis. Directed differentiation protocols in vitro were performed to investigate the potency of these cells and the cells were injected into immunocompromised mice to investigate their tumorigenicity. RESULTS: In testicular cell cultures from all four men, sharp-edged and compact colonies appeared between 3 and 8 weeks. Subcultured cells from these colonies showed alkaline phosphatase activity and expressed hES cell-specific genes (Pou5f1, Sox2, Cripto1, Dnmt3b), proteins and carbohydrate antigens (POU5F1, NANOG, SOX2 and TRA-1-60, TRA-1-81, SSEA4). These ES-like cells were able to differentiate in vitro into derivatives of all three germ layers including neural, epithelial, osteogenic, myogenic, adipocyte and pancreatic lineages. The pancreatic beta cells were able to produce insulin in response to glucose and osteogenic-differentiated cells showed deposition of phosphate and calcium, demonstrating their functional capacity. Although we observed small areas with differentiated cell types of human origin, we never observed extensive teratomas upon injection of testis-derived ES-like cells into immunocompromised mice. CONCLUSIONS: Multipotent cells can be established from adult human testis. Their easy accessibility and ethical acceptability as well as their non-tumorigenic and autogenic nature make these cells an attractive alternative to human ES cells for future stem cell therapies.


Assuntos
Técnicas de Cultura de Células , Células-Tronco Embrionárias/citologia , Células-Tronco Multipotentes/citologia , Testículo/citologia , Idoso , Idoso de 80 Anos ou mais , Animais , Diferenciação Celular , Proliferação de Células , Células-Tronco Embrionárias/metabolismo , Humanos , Cariotipagem , Masculino , Camundongos , Pessoa de Meia-Idade , Células-Tronco Multipotentes/metabolismo
20.
Transplant Proc ; 52(2): 534-536, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-32081355

RESUMO

OBJECTIVE: To analyze predictors of survival involved in liver retransplantation (LRT), including the Rosen Model (RM). MATERIALS AND METHODS: This was a descriptive, observational, and unicentric study based on predictors of survival including patients who underwent LRT in a tertiary medical center between April 2002 and December 2018. Recipient, donor, and transplant data were collected, and RM score was calculated for every patient. Fisher exact test and Student t test were used for qualitative and quantitative variables, respectively. The Shapiro-Wilks test was applied to verify the normality of the sample. Survival differences between subgroups were checked using the log-rank test. Statistical significance was stated at P < .05. RESULTS: Among 32 retransplanted patients in this period, 17 (53.1%) survived more than 12 months after LRT. The results of statistical associations between prognostic factors and overall survival highlighted that an older recipient age was significantly correlated with a lower overall survival. The 3-month overall survival was 84.3%. Nineteen patients had a low risk according to RM, with a 3-month survival rate of 78.9%. Eight had a RM intermediate risk, with a survival rate of 21%. Despite the aforementioned data, the log-rank test did not find statistical differences in survival (P = .488). CONCLUSION: We should consider older recipient age as a negative prognostic factor of overall survival. Also, we should contemplate intermediate risk according to RM as an adverse predictor regarding survival in LRT. Both data are of interest regarding the indication or not of LRT and prioritization on the waiting list.


Assuntos
Transplante de Fígado/mortalidade , Seleção de Pacientes , Reoperação/mortalidade , Listas de Espera/mortalidade , Adulto , Feminino , Humanos , Transplante de Fígado/métodos , Masculino , Pessoa de Meia-Idade , Reoperação/métodos , Taxa de Sobrevida
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