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1.
Gut ; 2024 May 16.
Artigo em Inglês | MEDLINE | ID: mdl-38754953

RESUMO

OBJECTIVE: Pancreatic ductal adenocarcinoma (PDAC) has limited therapeutic options, particularly with immune checkpoint inhibitors. Highly chemoresistant 'stem-like' cells, known as cancer stem cells (CSCs), are implicated in PDAC aggressiveness. Thus, comprehending how this subset of cells evades the immune system is crucial for advancing novel therapies. DESIGN: We used the KPC mouse model (LSL-KrasG12D/+; LSL-Trp53R172H/+; Pdx-1-Cre) and primary tumour cell lines to investigate putative CSC populations. Transcriptomic analyses were conducted to pinpoint new genes involved in immune evasion. Overexpressing and knockout cell lines were established with lentiviral vectors. Subsequent in vitro coculture assays, in vivo mouse and zebrafish tumorigenesis studies, and in silico database approaches were performed. RESULTS: Using the KPC mouse model, we functionally confirmed a population of cells marked by EpCAM, Sca-1 and CD133 as authentic CSCs and investigated their transcriptional profile. Immune evasion signatures/genes, notably the gene peptidoglycan recognition protein 1 (PGLYRP1), were significantly overexpressed in these CSCs. Modulating PGLYRP1 impacted CSC immune evasion, affecting their resistance to macrophage-mediated and T-cell-mediated killing and their tumourigenesis in immunocompetent mice. Mechanistically, tumour necrosis factor alpha (TNFα)-regulated PGLYRP1 expression interferes with the immune tumour microenvironment (TME) landscape, promoting myeloid cell-derived immunosuppression and activated T-cell death. Importantly, these findings were not only replicated in human models, but clinically, secreted PGLYRP1 levels were significantly elevated in patients with PDAC. CONCLUSIONS: This study establishes PGLYRP1 as a novel CSC-associated marker crucial for immune evasion, particularly against macrophage phagocytosis and T-cell killing, presenting it as a promising target for PDAC immunotherapy.

2.
Am J Physiol Regul Integr Comp Physiol ; 326(2): R147-R159, 2024 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-38047315

RESUMO

This single-blind, crossover study aimed to measure and evaluate the short-term metabolic responses to continuous and intermittent hypoxic patterns in individuals with obesity. Indirect calorimetry was used to quantify changes in resting metabolic rate (RMR), carbohydrate (CHOox, %CHO), and fat oxidation (FATox, %FAT) in nine individuals with obesity pre and post: 1) breathing normoxic air [normoxic sham control (NS-control)], 2) breathing continuous hypoxia (CH), or 3) breathing intermittent hypoxia (IH). A mean peripheral oxygen saturation ([Formula: see text]) of 80-85% was achieved over a total of 45 min of hypoxia. Throughout each intervention, pulmonary gas exchanges, oxygen consumption (V̇o2) carbon dioxide production (V̇co2), and deoxyhemoglobin concentration (Δ[HHb]) in the vastus lateralis were measured. Both RMR and CHOox measured pre- and postinterventions were unchanged following each treatment: NS-control, CH, or IH (all P > 0.05). Conversely, a significant increase in FATox was evident between pre- and post-IH (+44%, P = 0.048). Although the mean Δ[HHb] values significantly increased during both IH and CH (P < 0.05), the greatest zenith of Δ[HHb] was achieved in IH compared with CH (P = 0.002). Furthermore, there was a positive correlation between Δ[HHb] and the shift in FATox measured pre- and postintervention. It is suggested that during IH, the increased bouts of muscle hypoxia, revealed by elevated Δ[HHb], coupled with cyclic periods of excess posthypoxia oxygen consumption (EPHOC, inherent to the intermittent pattern) played a significant role in driving the increase in FATox post-IH.


Assuntos
Hipóxia , Respiração , Humanos , Método Simples-Cego , Estudos Cross-Over , Hipóxia/metabolismo , Obesidade , Músculo Quadríceps/metabolismo , Oxigênio
3.
FASEB J ; 36(1): e22091, 2022 01.
Artigo em Inglês | MEDLINE | ID: mdl-34919756

RESUMO

Hepatoencephalopathy due to combined oxidative phosphorylation deficiency type 1 (COXPD1) is a recessive mitochondrial translation disorder caused by mutations in GFM1, a nuclear gene encoding mitochondrial elongation factor G1 (EFG1). Patients with COXPD1 typically present hepatoencephalopathy early after birth with rapid disease progression, and usually die within the first few weeks or years of life. We have generated two different mouse models: a Gfm1 knock-in (KI) harboring the p.R671C missense mutation, found in at least 10 patients who survived more than 1 year, and a Gfm1 knock-out (KO) model. Homozygous KO mice (Gfm1-/- ) were embryonically lethal, whereas homozygous KI (Gfm1R671C/R671C ) mice were viable and showed normal growth. R671C mutation in Gfm1 caused drastic reductions in the mitochondrial EFG1 protein content in different organs. Six- to eight-week-old Gfm1R671C/R671C mice showed partial reductions of in organello mitochondrial translation and respiratory complex IV enzyme activity in the liver. Compound heterozygous Gfm1R671C/- showed a more pronounced decrease of EFG1 protein in liver and brain mitochondria, as compared with Gfm1R671C/R671C mice. At 8 weeks of age, their mitochondrial translation rates were significantly reduced in both tissues. Additionally, Gfm1R671C/- mice showed combined oxidative phosphorylation deficiency (reduced complex I and IV enzyme activities in liver and brain), and blue native polyacrylamide gel electrophoresis analysis revealed lower amounts of both affected complexes. We conclude that the compound heterozygous Gfm1R671C/- mouse presents a clear dysfunctional molecular phenotype, showing impaired mitochondrial translation and combined respiratory chain dysfunction, making it a suitable animal model for the study of COXPD1.


Assuntos
Encefalopatia Hepática/metabolismo , Erros Inatos do Metabolismo/metabolismo , Mitocôndrias Hepáticas/metabolismo , Proteínas Mitocondriais/metabolismo , Mutação de Sentido Incorreto , Fosforilação Oxidativa , Fator G para Elongação de Peptídeos/metabolismo , Biossíntese de Proteínas , Substituição de Aminoácidos , Animais , Modelos Animais de Doenças , Complexo IV da Cadeia de Transporte de Elétrons/genética , Complexo IV da Cadeia de Transporte de Elétrons/metabolismo , Encefalopatia Hepática/genética , Erros Inatos do Metabolismo/genética , Camundongos , Camundongos Knockout , Mitocôndrias Hepáticas/genética , Proteínas Mitocondriais/genética , Fator G para Elongação de Peptídeos/genética
4.
Int J Mol Sci ; 24(19)2023 Sep 27.
Artigo em Inglês | MEDLINE | ID: mdl-37834089

RESUMO

Trisomy X is the most frequent sex chromosome anomaly in women, but it is often underdiagnosed postnatally because most patients do not show any clinical manifestation. It is estimated that only 10% of patients with trisomy X are diagnosed by clinical findings. Thus, it has been proposed that the clinical spectrum is not yet fully delimited, and additional uncommon or atypical clinical manifestations could be related to this entity. The present report describes a female carrying trisomy X but presenting atypical manifestations, including severe intellectual disability, short stature, thymus hypoplasia, and congenital hypothyroidism (CH). These clinical findings were initially attributed to trisomy X. However, chromosome microarray analysis (CMA) subsequently revealed that the patient also bears a heterozygous 304-kb deletion at 16p11.2. This pathogenic copy-number variant (CNV) encompasses 13 genes, including TUFM. Some authors recommend that when a phenotype differs from that described for an identified microdeletion, the presence of pathogenic variants in the non-deleted allele should be considered to assess for an autosomal recessive disorder; thus, we used a panel of 697 genes to rule out a pathogenic variant in the non-deleted TUFM allele. We discuss the possible phenotypic modifications that might be related to an additional CNV in individuals with sex chromosome aneuploidy (SCA), as seen in our patient. The presence of karyotype-demonstrated trisomy X and CMA-identified 16p11.2 deletion highlights the importance of always correlating a patient's clinical phenotype with the results of genetic studies. When the phenotype includes unusual manifestations and/or exhibits discrepancies with that described in the literature, as exemplified by our patient, a more extensive analysis should be undertaken to enable a correct diagnosis that will support proper management, genetic counseling, and medical follow-up.


Assuntos
Aberrações dos Cromossomos Sexuais , Trissomia , Humanos , Feminino , Trissomia/diagnóstico , Trissomia/genética , Deleção Cromossômica , Fenótipo , Cariótipo
5.
J Perianesth Nurs ; 38(5): 782-786, 2023 10.
Artigo em Inglês | MEDLINE | ID: mdl-37269281

RESUMO

PURPOSE: To evaluate the effectiveness of the American Society of Perianesthesia Nurses (ASPAN) pediatric competency-based orientation (PCBO) education program on knowledge, confidence, and early recognition of nursing expertise among perianesthesia nurses in an acute care setting. DESIGN: A quasi-experimental pre/post survey-intervention design. METHODS: Sixty perianesthesia nurses with experience ranging from less than 5 years to more than 20 years were included. A chapter review survey was completed to assess knowledge before reviewing the chapters and again after reviewing the ASPAN PCBO materials. A presurvey assessing confidence levels, decision making abilities, and early detection of knowledge regarding pediatric patients' expertise were obtained at the beginning of the study. At the end of the study, a post-study survey was completed to evaluate the effectiveness of the intervention. Random codes were assigned to each participant to ensure that the participants' information was blinded. FINDINGS: Perianesthesia nurses' knowledge increased from pre to post intervention with one of the three sets of chapters (Set 2) statistically significant. Perianesthesia nurses' confidence and recognition of nursing expertise scores increased statistically significant from pre to post intervention. Both confidence (with 33 items [P value ≤ .001]) and recognition of nursing expertise (with 16 items [P value ≤ 0.001]) were statistically significant. CONCLUSIONS: The ASPAN PCBO was shown to be statistically effective at increasing knowledge, building expertise, promoting confidence, and improving decision making skills. The plan is for the ASPAN PCBO to be incorporated into the new-hire perianesthesia orientation didactic and the competency plan.


Assuntos
Competência Clínica , Enfermeiras e Enfermeiros , Humanos , Estados Unidos , Criança , Conhecimento
6.
Molecules ; 27(16)2022 Aug 13.
Artigo em Inglês | MEDLINE | ID: mdl-36014410

RESUMO

Marine organisms are affected by the ubiquitous occurrence of microplastics (MPs) in the environment. Several protocols have been described to extract and quantify MPs in seafood, although their complex matrices, with high level of fat, can compromise the efficiency of MPs extraction. To solve this issue, the present study aimed to develop a detailed methodology suitable to process seafood samples with different levels of fat, namely fish and molluscs, from fresh and canned sources, including the immersive liquids from the cans. Sample digestion was tested using different solutions (10% KOH, 30% H2O2), temperatures (40 °C, 65 °C) and incubation times (24, 48, 72 h). For fat removal, three detergents (two laboratory surfactants and a commercial dish detergent) and 96% ethanol were tested, as well as the manual separation of fat. The methodology optimized in this study combined a digestion with 30% H2O2 at 65 °C, during 24 to 48 h, with a manual separation of the fat remaining after the digestion. All steps from the present methodology were tested in six types of polymers (PE-LD, PET, PE, AC, PS, and lycra), to investigate if these procedures altered the integrity of MPs. Results showed that the optimized methodology will allow for the efficient processing of complex seafood samples with different fat levels, without compromising MPs integrity (recoveries rate higher than 89% for all the polymers tested).


Assuntos
Microplásticos , Poluentes Químicos da Água , Animais , Monitoramento Ambiental/métodos , Peróxido de Hidrogênio , Plásticos , Polímeros , Alimentos Marinhos/análise , Poluentes Químicos da Água/análise
7.
Int J Mol Sci ; 20(21)2019 Oct 23.
Artigo em Inglês | MEDLINE | ID: mdl-31652755

RESUMO

Variation in the location of the 15p region D15Z1 is recognized as a polymorphism in several human populations. We used high-stringency Fluorescence In Situ Hybridization (FISH) to detect D15Z1 in a Mexican cohort. Here, we report the presence of extra D15Z1 sequences on the p-arm of acrocentric chromosomes other than 15 in two groups of Mexican couples, one with healthy offspring (n = 75) and the other with aneuploid offspring (n = 87), mainly trisomy 21. The additional D15Z1 polymorphism was significantly increased in individuals with aneuploid offspring (26.4%), in comparison to individuals with healthy offspring (14%). The most frequent acceptor chromosome of D15Z1 was chromosome 13p, followed by 14p, and finally, 21p. Our results show an overall frequency of 21.6% of this polymorphism in the Mexican population and suggest that its presence might be associated with the mis-segregation of other acrocentric chromosomes and aneuploid offspring. The high frequency of the polymorphism of the D15Z1 sequence on acrocentric chromosomes other than 15 suggests a sequence homogenization of the acrocentric p arms, related to the important function of the centromere and the nucleolar organization region, which flank satellite III DNA.


Assuntos
Aneuploidia , Cromossomos Humanos Par 15/genética , Polimorfismo Genético , Adulto , Feminino , Humanos , Masculino , México , Linhagem
8.
Am J Med Genet A ; 176(1): 161-166, 2018 01.
Artigo em Inglês | MEDLINE | ID: mdl-29150894

RESUMO

Hypochondroplasia (HCH) is a skeletal dysplasia caused by an abnormal function of the fibroblast growth factor receptor 3. Although believed to be relatively common, its prevalence and phenotype are not well established owing to its clinical, radiological, and genetic heterogeneity. Here we report on a molecularly proven HCH family with an affected father and two children. The siblings (male and female) with HCH also had craniosynostosis and cleft palate, respectively. The present report supports the conclusion that the full clinical spectrum of HCH is not completely delineated. It also suggests that secondary, as yet unknown, modifying factors can influence the final phenotype.


Assuntos
Osso e Ossos/anormalidades , Fissura Palatina/diagnóstico , Fissura Palatina/genética , Craniossinostoses/diagnóstico , Craniossinostoses/genética , Nanismo/diagnóstico , Nanismo/genética , Estudos de Associação Genética , Deformidades Congênitas dos Membros/diagnóstico , Deformidades Congênitas dos Membros/genética , Lordose/diagnóstico , Lordose/genética , Adulto , Criança , Fácies , Feminino , Genótipo , Humanos , Cariótipo , Masculino , México , Mutação , Fenótipo , Receptor Tipo 3 de Fator de Crescimento de Fibroblastos/genética , Análise de Sequência de DNA , Síndrome , Tomografia Computadorizada por Raios X
9.
Artigo em Inglês | MEDLINE | ID: mdl-30018007

RESUMO

Consecutive treatments with recombinant follicle-stimulating and luteinizing hormones (rFsh and rLh, respectively) stimulate spermatogenesis and potentiate sperm production in pubescent specimens of the oligospermic Senegalese sole (Solea senegalensis). However, sperm production in response to the hormones is highly variable, and the steroidogenic potential of the testis may be diminished due to sustained hormone supply. Here, we compared the effectiveness of low (9 µg/kg) and high (18 µg/kg) doses of rFsh and rLh to improve sperm production in adult sole during late winter-early spring (onset of the natural spawning period), and in autumn under a controlled temperature of 12 °C (period of testicular recrudescence). Treatment with rFsh over six weeks during spring, followed by a single rLh injection, did not enhance sperm production, possibly because of an advanced stage of sexual maturation of the males, as reflected by high Lh plasma levels (~17 ng/ml) before rFsh treatment. In contrast, in autumn, when the Lh circulating levels were much lower (~3 ng/ml), the low doses of rFsh and rLh generated a four-times increase in sperm production, whereas the high doses of the hormones were ineffective. However, treatment with rLh, regardless of the effect of rFsh, improved the motility of spermatozoa during both spring and autumn. These data confirm that consecutive rFsh and rLh treatments increase sperm production and quality in adult sole males, although they seem to be highly sensitive to the rFsh dose. The efficiency of recombinant gonadotropins also appears to be season-dependent despite the asynchronous nature of the sole testis.


Assuntos
Linguados/fisiologia , Gonadotropinas/farmacologia , Estações do Ano , Espermatozoides/citologia , Animais , Relação Dose-Resposta a Droga , Masculino , Proteínas Recombinantes/farmacologia
10.
Asian Pac J Allergy Immunol ; 34(4): 300-305, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-27001659

RESUMO

BACKGROUND: Chronic Fatigue Syndrome/Myalgic Encephalomyelitis (CFS/ME), and Multiple Sclerosis (MS) may share some similarities in relation to reduced NK cell activity. It is likely that other cells such as regulatory T (Tregs), invariant Natural Killer T (iNKT) and gamma delta T (γδ T) cells may also be dysregulated in CFS/ME and MS. OBJECTIVE: To evaluate and compare specific immune regulatory cells of patients with CFS/ME, patients with MS and healthy controls. METHOD: Sixty three volunteers were included in this study: 24 were CFS/ME patients, 11 were MS patients and 27 were healthy controls. Blood samples were obtained from all participants for flow cytometry analysis of iNKT cells, Tregs and γδ T cell phenotypes. RESULTS: We observed a significant increase in Tregs in the CFS/ME group (p≤0.05) compared to the healthy control group. Total γδ and γδ2 T cells were significantly reduced in MS patients in comparison with the healthy control group. Conversely, CD4+iNKT percentage of iNKT, was significantly increased in the CFS/ME group compared with healthy controls and the double-negative iNKT percentage of iNKT significantly decreased compared with the healthy control group. CONCLUSIONS: This study has not identified any immunological disturbances that are common in both MS and CFS/ME patients. However, the differential expression of cell types between the conditions investigated suggests different pathways of disease. These differences need to be explored in further studies.


Assuntos
Síndrome de Fadiga Crônica/imunologia , Esclerose Múltipla/imunologia , Células T Matadoras Naturais/imunologia , Subpopulações de Linfócitos T/imunologia , Linfócitos T Reguladores/imunologia , Adulto , Separação Celular , Síndrome de Fadiga Crônica/sangue , Feminino , Citometria de Fluxo , Humanos , Imunofenotipagem , Masculino , Pessoa de Meia-Idade , Esclerose Múltipla/sangue , Receptores de Antígenos de Linfócitos T gama-delta/sangue , Receptores de Antígenos de Linfócitos T gama-delta/imunologia
11.
BMC Immunol ; 16: 35, 2015 Jun 02.
Artigo em Inglês | MEDLINE | ID: mdl-26032326

RESUMO

BACKGROUND: Abnormal immune function is often an underlying component of illness pathophysiology and symptom presentation. Functional and phenotypic immune-related alterations may play a role in the obscure pathomechanism of Chronic Fatigue Syndrome/Myalgic Encephalomyelitis (CFS/ME). The objective of this study was to investigate the functional ability of innate and adaptive immune cells in moderate and severe CFS/ME patients. The 1994 Fukuda criteria for CFS/ME were used to define CFS/ME patients. CFS/ME participants were grouped based on illness severity with 15 moderately affected (moderate) and 12 severely affected (severe) CFS/ME patients who were age and sex matched with 18 healthy controls. Flow cytometric protocols were used for immunological analysis of dendritic cells, monocytes and neutrophil function as well as measures of lytic proteins and T, natural killer (NK) and B cell receptors. RESULTS: CFS/ME patients exhibited alterations in NK receptors and adhesion markers and receptors on CD4(+)T and CD8(+)T cells. Moderate CFS/ME patients had increased CD8(+) CD45RA effector memory T cells, SLAM expression on NK cells, KIR2DL5(+) on CD4(+)T cells and BTLA4(+) on CD4(+)T central memory cells. Moderate CFS/ME patients also had reduced CD8(+)T central memory LFA-1, total CD8(+)T KLRG1, naïve CD4(+)T KLRG1 and CD56(dim)CD16(-) NK cell CD2(+) and CD18(+)CD2(+). Severe CFS/ME patients had increased CD18(+)CD11c(-) in the CD56(dim)CD16(-) NK cell phenotype and reduced NKp46 in CD56(bright)CD16(dim) NK cells. CONCLUSIONS: This research accentuated the presence of immunological abnormalities in CFS/ME and highlighted the importance of assessing functional parameters of both innate and adaptive immune systems in the illness.


Assuntos
Síndrome de Fadiga Crônica/imunologia , Síndrome de Fadiga Crônica/patologia , Receptores Imunológicos/metabolismo , Adulto , Linfócitos B Reguladores/imunologia , Contagem de Células Sanguíneas , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Adesão Celular , Separação Celular , Citotoxicidade Imunológica , Células Dendríticas/imunologia , Síndrome de Fadiga Crônica/sangue , Feminino , Citometria de Fluxo , Humanos , Células Matadoras Naturais/imunologia , Masculino , Pessoa de Meia-Idade , Monócitos/imunologia , Neutrófilos/imunologia , Fenótipo , Receptores de Antígenos de Linfócitos B/metabolismo , Receptores KIR2DL5/metabolismo
12.
J Transl Med ; 13: 299, 2015 Sep 14.
Artigo em Inglês | MEDLINE | ID: mdl-26370228

RESUMO

BACKGROUND: Research has identified immunological abnormalities in Chronic Fatigue Syndrome/Myalgic Encephalomyelitis (CFS/ME), a heterogeneous illness with an unknown cause and absence of diagnostic test. There have been no CFS/ME studies examining innate and adaptive immune cells longitudinally in patients with varying severities. This is the first study to investigate immune cells over 6 months while also examining CFS/ME patients of varying symptom severity. METHODS: Participants were grouped into 18 healthy controls, 12 moderate and 12 severe CFS/ME patients and flow cytometry was used to examine cell parameters at 0 and 6 months. RESULTS: Over time, iNKT CD62L expression significantly increased in moderate CFS/ME patients and CD56(bright) NK receptors differed in severe CFS/ME. Naïve CD8(+)T cells, CD8(-)CD4(-) and CD56(-)CD16(-) iNKT phenotypes, γδ2T cells and effector memory subsets were significantly increased in severe CFS/ME patients at 6 months. Severe CFS/ME patients were significantly reduced in CD56(bright)CD16(dim) NKG2D, CD56(dim)CD16(-) KIR2DL2/DL3, CD94(-)CD11a(-) γδ1T cells and CD62L(+)CD11a(-) γδ1T cells at 6 months. CONCLUSIONS: Severe CFS/ME patients differed from controls and moderate CFS/ME patients over time and expressed significant alterations in iNKT cell phenotypes, CD8(+)T cell markers, NK cell receptors and γδT cells at 6 months. This highlights the importance of further assessing these potential immune biomarkers longitudinally in both moderate and severe CFS/ME patients.


Assuntos
Síndrome de Fadiga Crônica/imunologia , Imunidade Adaptativa/imunologia , Adulto , Idoso , Antígeno CD56/metabolismo , Linfócitos T CD8-Positivos/imunologia , Feminino , Citometria de Fluxo , Humanos , Selectina L/metabolismo , Estudos Longitudinais , Masculino , Pessoa de Meia-Idade , Células T Matadoras Naturais/imunologia , Fenótipo , Receptores de Antígenos de Linfócitos T gama-delta/imunologia , Adulto Jovem
13.
Int Immunol ; 26(4): 233-42, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24343819

RESUMO

Perturbations in immune processes are a hallmark of a number of autoimmune and inflammatory disorders. Chronic fatigue syndrome/myalgic encephalomyelitis (CFS/ME) is an inflammatory disorder with possible autoimmune correlates, characterized by reduced NK cell activity, elevations in regulatory T cells (Tregs) and dysregulation in cytokine levels. The purpose of this article is to examine innate and adaptive immune cell phenotypes and functional characteristics that have not been previously examined in CFS/ME patients. Thirty patients with CFS/ME and 25 non-fatigued controls were recruited for this study. Whole blood samples were collected from all participants for the assessment of cell phenotypes, functional properties, receptors, adhesion molecules, antigens and intracellular proteins using flow cytometric protocols. The cells investigated included NK cells, dendritic cells, neutrophils, B cells, T cells, γδT cells and Tregs. Significant changes were observed in B-cell subsets, Tregs, CD4(+)CD73(+)CD39(+) T cells, cytotoxic activity, granzyme B, neutrophil antigens, TNF-α and IFN-γ in the CFS/ME patients in comparison with the non-fatigued controls. Alterations in B cells, Tregs, NK cells and neutrophils suggest significant impairments in immune regulation in CFS/ME and these may have similarities to a number of autoimmune disorders.


Assuntos
Linfócitos B/imunologia , Síndrome de Fadiga Crônica/imunologia , Células Matadoras Naturais/imunologia , Neutrófilos/imunologia , Linfócitos T Reguladores/imunologia , Imunidade Adaptativa , Linfócitos B/virologia , Células Cultivadas , Citotoxicidade Imunológica , Feminino , Regulação da Expressão Gênica/imunologia , Humanos , Imunidade Inata , Imunofenotipagem , Interferon gama/genética , Interferon gama/metabolismo , Células Matadoras Naturais/virologia , Subpopulações de Linfócitos/imunologia , Subpopulações de Linfócitos/virologia , Masculino , Pessoa de Meia-Idade , Linfócitos T Reguladores/virologia , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismo
14.
Int J Med Sci ; 12(10): 764-72, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26516304

RESUMO

Immunological dysregulation is present in Chronic Fatigue Syndrome/Myalgic Encephalomyelitis (CFS/ME), with recent studies also highlighting the importance of examining symptom severity. This research addressed this relationship between CFS/ME severity subgroups, assessing serum immunoglobulins and serum cytokines in severe and moderate CFS/ME patients. Participants included healthy controls (n= 22), moderately (n = 22) and severely (n=19) affected CFS/ME patients. The 1994 Fukuda Criteria defined CFS/ME and severity scales confirmed mobile and housebound CFS/ME patients as moderate and severe respectively. IL-1ß was significantly reduced in severe compared with moderate CFS/ME patients. IL-6 was significantly decreased in moderate CFS/ME patients compared with healthy controls and severe CFS/ME patients. RANTES was significantly increased in moderate CFS/ME patients compared to severe CFS/ME patients. Serum IL-7 and IL-8 were significantly higher in the severe CFS/ME group compared with healthy controls and moderate CFS/ME patients. IFN-γ was significantly increased in severe CFS/ME patients compared with moderately affected patients. This was the first study to show cytokine variation in moderate and severe CFS/ME patients, with significant differences shown between CFS/ME symptom severity groups. This research suggests that distinguishing severity subgroups in CFS/ME research settings may allow for a more stringent analysis of the heterogeneous and otherwise inconsistent illness.


Assuntos
Citocinas/sangue , Síndrome de Fadiga Crônica/imunologia , Imunoglobulinas/sangue , Adulto , Idoso , Quimiocinas/sangue , Feminino , Humanos , Masculino , Pessoa de Meia-Idade
15.
Parasitol Res ; 114(4): 1313-26, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25566774

RESUMO

Chagas disease, caused by the protozoan Trypanosoma cruzi, is a major parasitic disease that affects millions of people in America. However, despite the high impact of this disease on human health, no effective and safe treatment has been found that eliminates the infecting parasite from human patients. Among the possible chemotherapeutic targets that could be considered for study in T. cruzi are the DNA polymerases, in particular DNA polymerase beta (polß), which previous studies have shown to be involved in kinetoplast DNA replication and repair. In this paper, we describe the expression, purification, and biochemical characterization of the Miranda clone polß, corresponding to lineage T. cruzi I (TcI). The recombinant enzyme purified to homogeneity displayed specific activity in the range described for a highly purified mammalian polß. However, the trypanosome enzyme exhibited important differences in biochemical properties compared to the mammalian enzymes, specifically an almost absolute dependency on KCl, high sensitivity to N-ethylmaleimide (NEM), and low sensitivity to ddTTP. Immuno-affinity purification of T. cruzi polymerase beta (Tcpolß) from epimastigote extracts showed that the native enzyme was phosphorylated. In addition, it was demonstrated that Tcpolß interacts with some proteins in a group of about 15 proteins which are required to repair 1-6 bases of gaps of a double strand damaged DNA. It is possible that these proteins form part of a DNA repair complex, analogous to that described in mammals and some trypanosomatids.


Assuntos
Doença de Chagas/parasitologia , DNA Polimerase beta/genética , Regulação Enzimológica da Expressão Gênica , Trypanosoma cruzi/enzimologia , DNA Polimerase beta/efeitos dos fármacos , DNA Polimerase beta/isolamento & purificação , DNA Polimerase beta/metabolismo , DNA de Cinetoplasto/química , DNA de Cinetoplasto/genética , Didesoxinucleotídeos/farmacologia , Inibidores Enzimáticos/farmacologia , Escherichia coli/genética , Escherichia coli/metabolismo , Etilmaleimida/farmacologia , Humanos , Fosforilação , Filogenia , Análise de Sequência de DNA , Nucleotídeos de Timina/farmacologia , Trypanosoma cruzi/genética
16.
Methods Mol Biol ; 2825: 247-262, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38913314

RESUMO

Hodgkin lymphoma (HL) is one of the most common lymphomas, with an incidence of 3 per 100,000 persons. Current treatment uses a cocktail of genotoxic agents, including adriamycin, bleomycin, vinblastine, and dacarbazine (ABVD), along with or without radiotherapy. This treatment regimen has proved to be efficient in killing cancer cells, resulting in HL patients having a survival rate of >90% cancer-free survival at five years. However, this therapy does not have a specific cell target, and it can induce damage in the genome of non-cancerous cells. Previous studies have shown that HL survivors often exhibit karyotypes characterized by complex chromosomal abnormalities that are difficult to analyze by conventional banding. Multicolor fluorescence in situ hybridization (M-FISH) is a powerful tool to analyze complex karyotypes; we used M-FISH to investigate the presence of chromosomal damage in peripheral blood lymphocytes from five healthy individuals and five HL patients before, during, and one year after anti-cancer treatment. Our results show that this anti-cancer treatment-induced genomic chaos that persists in the hematopoietic stem cells from HL patients one year after finishing therapy. This chromosomal instability may play a role in the occurrence of second primary cancers that are observed in 10% of HL survivors. This chapter will describe a protocol for utilizing M-FISH to study treatment-induced genome chaos in Hodgkin's lymphoma (HL) patients, following a brief discussion.


Assuntos
Doença de Hodgkin , Hibridização in Situ Fluorescente , Doença de Hodgkin/genética , Doença de Hodgkin/terapia , Humanos , Hibridização in Situ Fluorescente/métodos , Aberrações Cromossômicas/efeitos da radiação , Doxorrubicina/uso terapêutico , Genoma Humano , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Protocolos de Quimioterapia Combinada Antineoplásica/efeitos adversos , Instabilidade Cromossômica , Linfócitos/efeitos da radiação , Linfócitos/efeitos dos fármacos , Linfócitos/metabolismo , Bleomicina/uso terapêutico
17.
Curr Opin Psychiatry ; 37(3): 225-236, 2024 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-38415715

RESUMO

PURPOSE OF REVIEW: Economic development and urbanisation have prompted many Chinese parents to move from rural to urban regions for better job opportunities. Their children, who remain behind in rural regions, become left-behind children (LBC). With absent parents, children and adolescents are unable to maintain the secure attachment required for healthy social and emotional development, increasing the risk of mental illness. This study aimed to compare risk of self-harm and suicidal ideation in LBC and non-LBC in China. RECENT FINDINGS: Greater risks for poor mental health outcomes including worse depression, loneliness and anxiety have been identified in LBC in cross-sectional studies. Previous studies have also identified higher prevalence of bullying victimization, poorer school performance and worse school attendance amongst LBC. SUMMARY: Findings indicate that prolonged separation from parents put LBC at greater risks of poor mental health. Policy changes to allow children to migrate with their parents and policies to reduce inequalities in job opportunities between urban and rural regions are needed.

18.
Sci Total Environ ; 923: 171443, 2024 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-38447727

RESUMO

Saltmarshes play a crucial role in carbon sequestration and storage, although they are increasingly threatened by climate change-induced sea level rise (SLR). This study assessed the potential variation in Blue Carbon stocks across regional and local scales, and estimated their economic value and potential habitat loss due to SLR based on the IPCC AR6 scenarios for 2050 and 2100 in three estuarine saltmarshes in northern Portugal, the saltmarshes of the Minho, Lima and Cávado estuaries. The combined carbon stock of these saltmarshes was 38,798 ± 2880 t of organic carbon, valued at 3.96 ± 0.38 M€. Local and regional differences in carbon stocks were observed between common species, with the cordgrass Spartina patens and the reed Phragmites australis consistently showing higher values in the Lima saltmarsh in some of the parameters. Overall, the Lima saltmarsh had the highest total carbon per species cover, with S. patens showing the highest values among common species. Bolboschoenus maritimus had the highest values in the Minho saltmarsh, while the other species presented a similar carbon storage capacity. Potential habitat loss due to SLR was most evident in the Cávado saltmarsh over shorter timescales, with a significant risk of inundation even for median values of SLR, while the Lima saltmarsh was shown to be more resistant and resilient. If habitat loss directly equates to carbon loss within these saltmarshes, projected CO2 emissions may range from 22,000 to 43,449 t by 2050 and 33,000 to 130,000 t by 2100 (under the IPCC SSP5-8.5 scenario). The study shows the importance of Blue Carbon site-specific estimates, acknowledging the potential future repercussions from habitat loss due to SLR. It emphasizes the need to consider local and regional variability in Blue Carbon stocks assessments and highlights the critical importance of preserving and rehabilitating these ecosystems to ensure their continued efficacy as vital carbon sinks, thereby contributing to climate change mitigation efforts.

19.
Food Chem ; 448: 139049, 2024 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-38518443

RESUMO

There is limited research on the occurrence of microplastics (MPs) in canned seafood. All types of canned seafood investigated in the present study were contaminated. After sample digestion in 30 % hydrogen peroxide, a total of 40 MPs were recovered. Fibers were the most common type, blue was the dominant colour, and Fourier Transform Infrared Spectroscopy (FTIR) identified polyester as the most common polymer. Considering all samples, an average of 3.5 ± 5.2 MPs/can was obtained, with octopus in tomato sauce and tuna in olive oil presenting the highest contamination (5.2 ± 7.5 MPs/can and 5.2 ± 5.1 MPs/can, respectively). Also, significant differences between the number of MPs in the seafood tissues and immersion liquids were verified. The present study demonstrates MPs occurrence in canned seafood, a potential contamination pathway for humans. More research on the different stages of the canning processing is vital for understanding MPs contamination in cans.


Assuntos
Contaminação de Alimentos , Microplásticos , Alimentos Marinhos , Alimentos Marinhos/análise , Contaminação de Alimentos/análise , Microplásticos/análise , Animais , Alimentos em Conserva/análise
20.
Front Microbiol ; 15: 1345324, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38404599

RESUMO

Introduction: Beer is one of the most consumed alcoholic drinks in the world, and this industry is a growing market that demands different properties to satisfy new consumers. The yeasts are used in different fermented beverages to contribute to new flavors. However, yeast strains used in the beer industry are limited so far, thus the diversity of flavors is very restricted. Therefore, the use of native yeast strains has been taking more importance with the purpose of conferring differentiated organoleptic properties to the product. Based on this observation the potentiality of native Saccharomyces cerevisiae strains obtained from different localities in Chile was researched. Methods: In this work was selected those strains that produced the highest ethanol concentration (nearly 6% v/v), consumed the highest amounts of sugars, and produced the lowest amounts of organic acids in the resulting beers. Finally, we did a beer tasting to select those strains that added different flavors to the final beer compared with a commercial strain used. Results and discussion: In this study, two native strains that produced fruity descriptors are described, which could be used in the future in brewing, craft or industrial production.

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