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OBJECTIVES: Infertility is a disease of the male or female reproductive systems. Male reproductive workup is based on routine semen analysis, although of limited value. The 2021 WHO Manual incorporated Sperm DNA Fragmentation (SDF) assessment, and highlighted the need for individual laboratories to define suitable thresholds. This study aimed to present an alternative to address this issue, determine an SDF cut-off value with fertile donors, and characterize SDF in a patient cohort and their relationship with semen parameters. STUDY DESIGN: A service unit was established to remotely perform TUNEL assay in a 2 step-process. Semen samples were received at andrology laboratories, subjected to routine semen analysis (WHO, 2010), partially processed and transported to the service unit for SDF evaluation. Using this setting, studies were done in fertile donors (n = 15) to define the cut-off value, and in men undergoing infertility workup (n = 318). RESULTS: A cut-off value of 9.17 % was determined with the fertile donor cohort. With this cut-off, a 64.46 % abnormal SDF incidence was determined in the patient cohort. SDF negatively correlated with sperm number, vitality and motility, and positively with abnormal morphology and male age (P < 0.05). TUNEL-positive cases depicted lower sperm quality and higher male age (P < 0.05). A similar abnormal SDF incidence was determined among patients with semen abnormalities. Asthenozoospermic and ≥40 years patient samples depicted higher (P < 0.05) SDF than those of the general population. SDF incidence was also high in normozoospermic patients. CONCLUSIONS: Using a 2-step remote approach with a standardized procedure and an SDF cut-off value established with fertile donors, high SDF incidence in semen samples depicting normal and abnormal quality were identified in men consulting for infertility, highlighting the relevance of its evaluation as part of the male fertility workup.
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Fragmentación del ADN , Etiquetado Corte-Fin in Situ , Infertilidad Masculina , Análisis de Semen , Espermatozoides , Humanos , Masculino , Adulto , Análisis de Semen/métodos , Infertilidad Masculina/diagnóstico , Persona de Mediana Edad , Motilidad EspermáticaRESUMEN
Background: Children with special health care needs including Down Syndrome, Autism Spectrum Disorder and Down Syndrome experience difficulties in receiving dental treatment. Silver Diamine Fluoride (SDF) and Silver Fluoride (SF) are a minimally invasive treatments options to arrest dental caries without sedation; local or general anaesthesia (GA). Aim: Evaluation of Brazilian's parents' acceptance of the use of SF in CSHCN. Methods: After receiving education on SF, 100 Parents of CSHCN completed a questionnaire concerning their acceptance of SF, in different dental situation. Result: Majority of parents (74,5%) agreed to the use of SF for their children. SF was more acceptable on posterior teeth (74,5%) when compared to its use on anterior teeth (43,1%). Parents accepted to use SF in order: to reduce infection and pain (82,4%); to avoid dental injection (72,5%) and treatment under GA (84,3%). The Majority of parents accepted the properties of SF (82,4%) and Silver (80,4%). Conclusion: Silver Fluoride was accepted as a treatment option for caries, by Brazilian parents of CSHCN. SF should be considered as a treatment option for caries limited to dentine for CSHCN, taking into consideration the individual needs and opinions with regard to aesthetics and exposure to fluoride and silver.
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Introduction: Providing conventional, restorative dental care to children with special healthcare needs (CSHCN) often requires sedation using general anesthesia. Saliva consistency, diet, and oral hygiene practice are different for CSHCN, and limited evidence is available on the efficacy of silver fluoride (SF) for the management of carious lesions for this vulnerable population. Methods: Parents of CSHCN were educated about silver fluoride as a treatment option for caries. In total, 550 carious lesions from 100 participants were identified and scored according to the Nyvad Caries criteria. A total of 100 lesions with Nyvad scores 1, 2, and 3 were treated with a single application of silver fluoride and observed postoperatively at 1, 3, and 6 weeks. Result: The results indicate statistically significant (p < 0.05) differences in lesion remineralization over the 6-week follow-up period. At the 6-week follow-up, more than 85% of all lesions were remineralized across all children, regardless of condition or original Nyvad score of 1, 2, or 3. Conclusion: A single application of silver fluoride has demonstrated effectiveness in remineralization and inactivation of carious lesions over 6 weeks among Brazilian CSHCN. Silver fluoride should be considered an option for the management of carious lesions among CSHCN. Further studies are recommended, including larger sample sizes, longer follow-up times, a second application of SF, and different special needs conditions.
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CCR5Δ32 and SDF1-3'A polymorphisms were investigated in a cohort of viremia controllers, without the use of therapy, along with their influence on CD4+ T lymphocytes (TLs), CD8+ TLs, and plasma viral load (VL). The samples were analyzed from 32 HIV-1-infected individuals classified as viremia controllers 1 and 2 and viremia non-controllers, from both sexes, mostly heterosexuals, paired with 300 individuals from a control group. CCR5∆32 polymorphism was identified by PCR amplification of a fragment of 189 bp for the wild-type allele and 157 bp for the allele with the ∆32 deletion. SDF1-3'A polymorphism was identified by PCR, followed by enzymatic digestion (restriction fragment length polymorphism) with the Msp I enzyme. The relative quantification of gene expression was performed by real-time PCR. The distribution of allele and genotype frequencies did not show significant differences between the groups. The gene expression of CCR5 and SDF1 was not different between the profiles of AIDS progression. There was no significant correlation between the progression markers (CD4+ TL/CD8+ TL and VL) and the CCR5∆32 polymorphism carrier status. The 3'A allele variant was associated with a marked loss of CD4+ TLs and a higher plasma VL. Neither CCR5∆32 nor SDF1-3'A was associated with viremia control or the controlling phenotype.
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Síndrome de Inmunodeficiencia Adquirida , Quimiocina CXCL12 , Infecciones por VIH , Receptores CCR5 , Femenino , Humanos , Masculino , Síndrome de Inmunodeficiencia Adquirida/genética , Biomarcadores , Brasil , Quimiocina CXCL12/genética , Progresión de la Enfermedad , Frecuencia de los Genes , VIH-1 , Receptores CCR5/genética , ViremiaRESUMEN
BACKGROUND: Cancer-associated fibroblasts (CAFs), one of the main members of stromal cells in tumor microenvironment are proposed to play a central role in promoting tumor metastasis. It is unclear whether and how CAFs mediates tumor metastasis or chemoresistance in human ovarian cancer. METHODS: CAFs were extracted from human ovarian cancer tissues (OCs) of patients with different kinds of histological types. RESULTS: We found that CAFs showed more aggressive potency than those tumor cells, both of which were isolated from the same ovarian cancer specimen. Moreover, when co-cultured with CAFs, cell migration abilities of ovarian cancer cells (SKOV3, OVCAR3 and HEY) were significantly increased. Next, we preliminarily detected a higher CAFs density in sections of metastatic lesions than those in primary tumor site of primary OCs clinically. However, no significant difference of stromal derived factors-1α (SDF-1α) production from CAFs was found between primary and metastatic lesions. Additionally, in contrast with tumor cells, CAFs exhibited obvious apoptosis resistance when treated with cisplatin. Furthermore, we found that cisplatin-induced cytotoxicity and apoptosis were significantly inhibited by co-cultured with recombinant human SDF-1α in SKOV3 in a time and dose-dependent manner, and this effect was suppressed by the CXCR4 antagonist AMD3100. CONCLUSIONS: CAFs might be involved in the malignant metastasis in human ovarian cancer through promoting cell migration in tumor cells. And their resistance to cytotoxic agents might be mediated by paracrine SDF-1α/CXCR4 signaling in ovarian cancer.
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Fibroblastos Asociados al Cáncer , Neoplasias Ováricas , Humanos , Femenino , Neoplasias Ováricas/tratamiento farmacológico , Neoplasias Ováricas/patología , Fibroblastos Asociados al Cáncer/patología , Quimiocina CXCL12 , Cisplatino/farmacología , Apoptosis , Línea Celular Tumoral , Movimiento Celular , Fibroblastos , Proliferación Celular , Microambiente TumoralRESUMEN
PURPOSE: The aim of this study was to investigate the involvement of the SDF-1/CXCR4 axis in the process of BMMSC homing in prostate cancer (PCa) in vivo and in vitro. METHODS: After verification of BMMSCs, we fixed the concentration gradient of SDF-1 for BMMSC cultivation to analyze CXCR4 expression by qRT-PCR and flow cytometric analysis. Furthermore, we developed a non-contact co-culture system and explored the participation of the SDF-1/CXCR4 axis in PCa using qRT-PCR, flow cytometry, and ELISA. In addition, A green fluorescent protein (GFP)-transplanted methylnitrosourea (MNU)-induced PCa mouse model was established to investigate the CXCR4 expression in vivo. RESULTS: The CXCR4 expression was up-regulated with the increase in SDF-1 concentrations, and elevated SDF-1 had a significant promoting effect on cell proliferation and migration in BMMSCs. Moreover, the CXCR4 expression of BMMSCs was significantly increased in the non-contact co-culture model with vascular endothelial cells (VECs), and analysis of this model also showed that the proliferation and migration of BMMSCs were promoted in the presence of VECs. The ELISA assay showed that the SDF-1 levels in the co-culture model at 48 h were significantly increased. Twenty of the GFP-transplanted mice were divided into a PCa group and a control group, and four GFP-transplanted mice were observed to have prostate tumorigenesis. It also showed that CXCR4 was obviously increased in the prostate tissue of PCa mice. CONCLUSION: Our findings suggest that BMMSCs could home and promote the proliferation and migration of PCa through the SDF-1/CXCR4 axis in vivo and in vitro.
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Células Madre Mesenquimatosas , Neoplasias de la Próstata , Animales , Células de la Médula Ósea , Movimiento Celular , Quimiocina CXCL12/metabolismo , Células Endoteliales/metabolismo , Humanos , Masculino , Ratones , Neoplasias de la Próstata/metabolismo , Receptores CXCR4/metabolismo , Transducción de SeñalRESUMEN
This study evaluated the effects of land-use change (L-UCH) on dung beetle community structure (Scarabaeinae) in a disturbed dry ecosystem in southern Ecuador. Five different L-UCH classes were analyzed by capturing the dung beetle species at each site using 120 pitfall traps in total. To determine dung beetle abundance and diversity at each L-UCH, a general linear model (GLM) and a redundancy analysis (RDA) were applied, which correlated environmental and edaphic conditions to the community structure. Furthermore, changes in dung-producing vertebrate fauna were examined, which varied significantly between the different L-UCH classes due to the specific anthropogenic use or level of ecosystem disturbance. The results indicated that soil organic matter, pH, potassium, and phosphorus (RDA: component 1), as well as temperature and altitude (RDA: component 2) significantly affect the abundance of beetles (GLM: p value < 0.001), besides the food availability (dung). The highest abundance and diversity (Simpson's index > 0.4, Shannon-Wiener index > 1.10) was found in highly disturbed sites, where soils were generally more compacted, but with a greater food supply due to the introduced farm animals. At highly disturbed sites, the species Canthon balteatus, Dichotomius problematicus, and Onthphagus confuses were found specifically, which makes them useful as bio-indicators for disturbed dry forest ecosystems in southern Ecuador.
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New microcirculatory imaging techniques allowed direct observation of microcirculation at the bedside. This study presents a new device that assists the operator with the unprecedented Hands Free technique. To this end, a replica of Handheld Vital Microscopy was developed to simulate the method of capturing the image in the sublingual area, the most used site to assess microcirculation in critically ill patients. We achieved a reduction in the displacement of micros cope replica with a Hands Free method. The immediate consequence is an increase in the stability of HVM replica by 75 times, or more, over the current 4 seconds, during its contact with the sublingual tissue. The device also offers better control of th e pressure of the tip of the HVM replica over the sublingual area. The results demonstrated that the Hands Free technique, operating in the same sublingual area for 900 seconds, should allow for future research aimed at therapeutic maneuvers in patients with serious illnesses.
Las nuevas técnicas de imagen microcirculatoria permitieron la observación directa de la microcirculación junto a la cama del paciente. Este estudio sin precedentes presenta un nuevo dispositivo que ayuda al operador con la técnica manos libres. Con este fin, se desarrolló una réplica de la Microscopía Vital Portátil para simular el método de captura de la imagen en el área sublingual, el sitio más utilizado para evaluar la microcirculación en pacientes críticos. Logramos una reducción en el desplazamiento de la réplica del microscopio con el método de manos libres. La consecuencia inmediata es un aumento en la estabilidad de la réplica de HVM en 75 veces, o más, durante los 4 segundos actuales, durante su contacto con el tejido sublingual. El dispositivo también ofrece un mejor control de la presión de la punta de la réplica de HVM sobre el área sublingual. Los resultados demostraron que la técnica de manos libres, que opera en la misma área sublingual durante 900 segundos, debería permitir futuras investigaciones destinadas a maniobras terapéuticas en pacientes con enfermedades graves.
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Humanos , Cirugía Asistida por Computador/métodos , Microcirculación/fisiología , Procesamiento de Imagen Asistido por Computador , Enfermedad Crítica , Microscopía por Video , Microscopía Intravital , Suelo de la Boca/irrigación sanguíneaRESUMEN
Varicocele has been extensively described and studied as the most important reversible cause of male infertility. Its impact on semen parameters, pregnancy rates, and assisted reproductive outcomes have been associated with multifactorial aspects, most of them converging to increase of reactive oxygen species (ROS). More recently, sperm DNA fragmentation has gained significant attention and potential clinical use, although the body of evidence still needs further evolution. The associations between sperm DNA damage and a variety of disorders, including varicocele itself, share common pathways to ROS increase. This mini-review discusses different aspects related to the etiology of ROS and its relation to varicocele and potential mechanisms of DNA damage.
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BACKGROUND AND AIM: The endoplasmic reticulum (ER) stress response and the unfolded protein response (UPR) are essential cellular mechanisms to ensure the proper functioning of ER in adverse conditions. However, activation of these pathways has also been associated with insulin resistance and cell death in pathological conditions such as diabetes mellitus. In the present study, we investigated whether stromal cell-derived factor 2 (SDF2)-an ER stress-responsive factor-is related to ER response in placental cells exposed to maternal gestational diabetes mellitus (GDM) or to a hyperglycaemic in vitro condition. OBJECTIVE: The study aimed to investigate the role of SDF2 in BeWo cells , a trophoblast cell line originating from choriocarcinoma , and in placental tissue under hyperglycaemic conditions. METHODS: Protein levels of SDF2 and UPR factors, glucose-related protein 78 (GRP78) and eukaryotic initiation factor 2 alpha (elF2 alpha) were evaluated in the placentae of pregnant women diagnosed with GDM and treated by diet-control (insulin was added when necessary). The mRNA expression of SDF2 and UPR factors CHOP and sXBP1 were assessed in cultured BeWo cells challenged with glucose and treated with or without insulin. RESULTS: SDF2 expression was increased in the placentae of GDM women treated with diet. However, its values were similar to those of normoglycemic controls when the GDM women were treated with insulin and diet. BeWo cells cultured with high glucose and insulin showed decreased SDF2 expression, while high glucose increased CHOP and sXBP1 expression, which was then significantly reverted with insulin treatment. CONCLUSION: Our findings extend the understanding of ER stress and SDF2 expression in placentae exposed to hyperglycaemia, highlighting the relevance of insulin in reducing the levels of ER stress factors in placental cells. Understanding the effect of ER stress partners such as SDF2 on signalling pathways involved in gestation, complicated by hyperglycaemia, is pivotal for basic biomedical research and may lead to new therapeutic possibilities.
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Glucemia/metabolismo , Diabetes Gestacional/metabolismo , Estrés del Retículo Endoplásmico , Proteínas/metabolismo , Trofoblastos/metabolismo , Estudios de Casos y Controles , Línea Celular Tumoral , Estudios Transversales , Diabetes Gestacional/sangre , Diabetes Gestacional/patología , Diabetes Gestacional/terapia , Dieta Saludable , Chaperón BiP del Retículo Endoplásmico , Estrés del Retículo Endoplásmico/efectos de los fármacos , Factor 2 Eucariótico de Iniciación/metabolismo , Femenino , Proteínas de Choque Térmico/metabolismo , Humanos , Hipoglucemiantes/uso terapéutico , Insulina/uso terapéutico , Embarazo , Proteínas/genética , Transducción de Señal , Factor de Transcripción CHOP/genética , Factor de Transcripción CHOP/metabolismo , Trofoblastos/efectos de los fármacos , Trofoblastos/patología , Proteína 1 de Unión a la X-Box/genética , Proteína 1 de Unión a la X-Box/metabolismoRESUMEN
Telocytes are cells present in the stroma of various tissues including the prostate. The detection of telocytes is still very much dependent on obtaining ultrastructural data that show the presence of telopodes, which are cytoplasmic projections that alternate between dilated regions, the podoms, and thin segments, the podomers. These structures are the distinctive characteristics of the telocytes. Thus, in vitro assays are important for the study of telocytes, which are more easily identified in culture, which also enables the experimental manipulation of these cells. The isolation of telocytes per se does not allow the analysis of the behavior of these cells in relation to other cell types in a given organ. In this sense, in the prostate, explants could be a useful tool for the study of telocytes. The present study obtained prostatic explants and evaluated the influence of recombinant proteins, scattering factor (SCF) and stromal-derived factor 1 (SDF-1), which could impact on the migration of CD34-positive cells. Telocytes migrate out of explants and SDF-1 stimulates the proliferation and formation of telocyte networks in vitro. Telocytes are not smooth muscle cell progenitors in the prostate; on the contrary, they are CD90- and CD44-negative cells and, hence, have limited progenitor capacity. The present study demonstrated that explants are useful tools to elucidate the nature of telocytes and their functions.
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Quimiocina CXCL12/metabolismo , Factor de Crecimiento de Hepatocito/metabolismo , Telocitos/metabolismo , Animales , Antígenos CD34/metabolismo , Técnicas de Cultivo de Célula/métodos , Gerbillinae , Masculino , Próstata/metabolismo , Receptor alfa de Factor de Crecimiento Derivado de Plaquetas/metabolismo , Telocitos/fisiologíaRESUMEN
BACKGROUND: The lung infections by Staphylococcus aureus are strongly associated with its ability to produce enterotoxins. However, little is known about the mechanisms underlying trafficking of bone marrow (BM) neutrophils during airway inflammation induced by Staphylococcal enterotoxin B (SEB). We therefore aimed to investigate the effects of mouse airways SEB exposure on BM neutrophil counts and its adhesive properties as well as on the release of cytokines/chemokines that orchestrate BM neutrophils trafficking to lung tissue. METHODS: Male BALB/c mice were intranasally exposed to SEB (1⯵g), and at 4, 16 and 24â¯h thereafter, BM, circulating blood, bronchoalveolar lavage (BAL) fluid and lung tissue were collected. BM neutrophils adhesion, MAC-1 and LFA1-α expressions (by flow cytometry) as well as measurement of cytokine and/or chemokines levels were assayed after SEB-airway exposure. RESULTS: Prior exposure to SEB promoted a marked influx of neutrophils to BAL and lung tissue, which was accompanied by increased counts of BM immature neutrophils and blood neutrophilia. BM neutrophil expressions of LFA1-α and MAC-1 were unchanged by SEB exposure whereas a significant enhancement of adhesion properties to VCAM-1 was observed. The early phase of airway SEB exposure was accompanied by high levels of GM-CSF, G-CSF, IFN-γ, TNF-α and KC/CXCL1, while the latter phase by the equilibrated actions of SDF1-α and MIP-2. CONCLUSION: Mouse airways exposure to SEB induces BM cytokines/chemokines release and their integrated actions enhance the adhesion of BM neutrophils leading to acute lung injury.
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Células de la Médula Ósea/inmunología , Citocinas/metabolismo , Neutrófilos/inmunología , Neumonía Estafilocócica/inmunología , Staphylococcus aureus/inmunología , Administración Intranasal , Animales , Médula Ósea/inmunología , Médula Ósea/patología , Células de la Médula Ósea/metabolismo , Líquido del Lavado Bronquioalveolar/citología , Líquido del Lavado Bronquioalveolar/inmunología , Citocinas/inmunología , Enterotoxinas/administración & dosificación , Enterotoxinas/inmunología , Humanos , Recuento de Leucocitos , Pulmón/citología , Pulmón/inmunología , Pulmón/patología , Masculino , Ratones , Infiltración Neutrófila/inmunología , Neutrófilos/metabolismo , Neumonía Estafilocócica/microbiología , Neumonía Estafilocócica/patología , Staphylococcus aureus/metabolismoRESUMEN
Androgens induce rat prostate induction from the urogenital sinus epithelium at embryonic day 17.5. Subsequent morphogenesis, including epithelial cord growth, branching, and canalization, results from concerted paracrine interactions with the stroma. A significant number of paracrine factors bind heparan sulfate (HS). We hypothesized that interfering with overall sulfation could disrupt the signaling mediated by HS-binding factors and that the undersulfated environment would allow investigation of individual exogenous morphogens. First, we investigated whether acinar morphogenesis involved HS-proteoglycan expression and found that syndecans 1 and 3 were upregulated in RWPE1 cells in the transition from two- to three-dimensional (3D) Matrigel, capable of promoting spheroid formation. We then investigated whether sodium chlorate, a general sulfation inhibitor, interfered with spheroid formation by RWPE1 cells and acinar morphogenesis in ex vivo ventral prostate (VP) organ culture. As expected, treatment with sodium chlorate inhibited spheroid formation by RWPE1 cells in 3D culture. Chlorate also inhibited ex vivo VP epithelial branching and canalization, resulting in long branchless epithelial structures. We then investigated whether the HS-binding factors, FGF10, TGFß1, and SDF1, could reverse the effect of sodium chlorate. Although no effect was seen in the FGF10- and TGFß1-treated samples, SDF1 promoted epithelial canalization in the low sulfated environment, highlighting its specific role in lumen formation. Altogether, the results show that sodium chlorate perturbed prostate morphogenesis and allowed investigation of factors involved in branching and/or canalization, implicating SDF1 signaling in epithelial canalization.
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Quimiocina CXCL12/metabolismo , Células Epiteliales/metabolismo , Morfogénesis/fisiología , Próstata/metabolismo , Próstata/fisiología , Animales , Línea Celular , Colágeno/metabolismo , Combinación de Medicamentos , Células Epiteliales/fisiología , Epitelio/metabolismo , Epitelio/fisiología , Factor 10 de Crecimiento de Fibroblastos/metabolismo , Regulación del Desarrollo de la Expresión Génica/fisiología , Proteoglicanos de Heparán Sulfato/metabolismo , Heparitina Sulfato/metabolismo , Humanos , Laminina/metabolismo , Masculino , Técnicas de Cultivo de Órganos/métodos , Organogénesis/fisiología , Proteoglicanos/metabolismo , Ratas , Ratas Wistar , Transducción de Señal/fisiología , Factor de Crecimiento Transformador beta1/metabolismoRESUMEN
SDF-1α (stromal cell-derived factor-1α) is a CXCR4-receptor agonist and DPP4 (dipeptidyl peptidase 4) substrate. SDF-1α, particularly when combined with sitagliptin to block the metabolism of SDF-1α by DPP4, stimulates proliferation of cardiac fibroblasts via the CXCR4 receptor; this effect is greater in cells from spontaneously hypertensive rats versus Wistar-Kyoto normotensive rats. Emerging evidence indicates that ubiquitin(1-76) exists in plasma and is a potent CXCR4-receptor agonist. Therefore, we hypothesized that ubiquitin(1-76), similar to SDF-1α, should increase proliferation of cardiac fibroblasts. Contrary to our working hypothesis, ubiquitin(1-76) did not stimulate cardiac fibroblast proliferation, yet unexpectedly antagonized the proproliferative effects of SDF-1α combined with sitagliptin. In this regard, ubiquitin(1-76) was more potent in spontaneously hypertensive versus Wistar-Kyoto cells. In the presence of 6bk (selective inhibitor of insulin-degrading enzyme [IDE]; an enzyme known to convert ubiquitin(1-76) to ubiquitin(1-74)), ubiquitin(1-76) no longer antagonized the proproliferative effects of SDF-1α/sitagliptin. Ubiquitin(1-74) also antagonized the proproliferative effects of SDF-1α/sitagliptin, and this effect of ubiquitin(1-74) was not blocked by 6bk and was >10-fold more potent compared with ubiquitin(1-76). Neither ubiquitin(1-76) nor ubiquitin(1-74) inhibited the proproliferative effects of the non-CXCR4 receptor agonist neuropeptide Y (activates Y1 receptors). Cardiac fibroblasts expressed IDE mRNA, protein, and activity and converted ubiquitin(1-76) to ubiquitin(1-74). Spontaneously hypertensive fibroblasts expressed greater IDE activity. Extracellular ubiquitin(1-76) blocks the proproliferative effects of SDF-1α/sitagliptin via its conversion by IDE to ubiquitin(1-74), a potent CXCR4 antagonist. Thus, IDE inhibitors, particularly when combined with DPP4 inhibitors or hypertension, could increase the risk of cardiac fibrosis.
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Proliferación Celular , Quimiocina CXCL12/metabolismo , Fibroblastos , Hipertensión/metabolismo , Insulisina , Miocardio/patología , Receptores CXCR4 , Animales , Presión Sanguínea/fisiología , Proliferación Celular/efectos de los fármacos , Proliferación Celular/fisiología , Células Cultivadas , Dipeptidil Peptidasa 4/metabolismo , Inhibidores de la Dipeptidil-Peptidasa IV/farmacología , Fibroblastos/efectos de los fármacos , Fibroblastos/metabolismo , Fibrosis , Insulisina/antagonistas & inhibidores , Insulisina/metabolismo , Neuropéptido Y/metabolismo , Ratas , Ratas Endogámicas SHR , Ratas Endogámicas WKY , Receptores CXCR4/agonistas , Receptores CXCR4/metabolismo , Transducción de Señal , Fosfato de Sitagliptina/farmacología , Ubiquitina/metabolismoRESUMEN
AIM: To examine the interference of ß-blockers with the chemokine stromal cell-derived factor-1 (SDF-1) found in cell homing receptors, C-X-C chemokine receptor type 4 (CXCR-4) and CXCR-7, and regulatory proteins of homing pathways, we administered atenolol, carvedilol, metoprolol, and propranolol for 30 days using an orogastric tube to hypertensive rats. METHOD: We collected blood samples before and after treatment and quantified the levels of SDF-1 with enzyme-linked immunosorbent assay (ELISA). On day 30 of treatment, the spontaneously hypertensive rats (SHR) were euthanized, and heart, liver, lung, and kidney tissues were biopsied. Proteins were isolated for determining the expression of CXCR-4, CXCR-7, GRK-2 (G protein-coupled receptors kinase 2), ß-arrestins (ß1-AR and ß2-AR), and nuclear factor kappa B (NFκB). RESULTS: We found that the study drugs modulated these proteins, and metoprolol and propranolol strongly affected the expression of ß1-AR (P = .0102) and ß2-AR (P = .0034). CONCLUSION: ß-blockers modulated tissue expression of the proteins and their interactions following 30 days of treatment. It evidences that this class of drugs can interfere with proteins of cell homing pathways.
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Antagonistas Adrenérgicos beta/farmacología , Antihipertensivos/farmacología , Movimiento Celular/efectos de los fármacos , Hipertensión/tratamiento farmacológico , Antagonistas de Receptores Adrenérgicos beta 1/farmacología , Animales , Atenolol/farmacología , Carbazoles/farmacología , Carvedilol , Quimiocina CXCL12/sangre , Modelos Animales de Enfermedad , Quinasa 2 del Receptor Acoplado a Proteína-G/metabolismo , Hipertensión/sangre , Hipertensión/fisiopatología , Riñón/efectos de los fármacos , Riñón/metabolismo , Hígado/efectos de los fármacos , Hígado/metabolismo , Pulmón/efectos de los fármacos , Pulmón/metabolismo , Metoprolol/farmacología , Miocardio/metabolismo , FN-kappa B/metabolismo , Propanolaminas/farmacología , Propranolol/farmacología , Ratas Endogámicas SHR , Receptores CXCR/metabolismo , Receptores CXCR4/metabolismo , Transducción de Señal/efectos de los fármacos , beta-Arrestina 1/metabolismo , Arrestina beta 2/metabolismoRESUMEN
PURPOSE: We hypothesized that the chemokine SDF1/CXCR4 system was present in feline cumulus-oocyte complexes (COCs) and that COCs cultured with SDF1 would directly upregulate gene expression in the ovulatory cascade. METHODS: Ovaries (n = 50) were obtained from adult domestic cats during the breeding season and COCs were recovered from antral follicles. Because IVM media triggers cumulus-oocyte expansion, culture conditions needed to be optimized to study periovulatory genes. After optimization, the effects of 25 ng/ml SDF1 and the CXCR4 inhibitor were examined in a COC culture for 3, 12, and 24 h. RESULTS: MEM-hepes with 1% of charcoal stripped-FBS was the optimized culture medium, assessed by the expansion of COCs at 24 h in the gonadotropin (GNT) group but not in the media with serum alone. The mRNA expression of HAS2, TNFAIP6, PTX3, and AREG peaked at 3 h in GNT group as compared to all other groups (p < 0.05). COCs cultured with SDF1 showed increased HAS2 and TNFAIP6 mRNA expression at 3 h compared to negative controls and to the CXCR4 inhibitor group. CXCR4 and SDF1 immunostaining was present in both cumulus cells and the oocyte. CONCLUSIONS: These results demonstrate that GNT stimulation upregulates key periovulatory genes and expansion in feline COCs from antral follicles, and support the use of this culture system to examine molecular processes within the COC. In addition, SDF1 directly promotes key periovulatory genes in feline COCs, suggesting that the SDF1-CXCR4 pathway may extend its function beyond a chemoattractant, and may play a direct role within the COC.
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Quimiocina CXCL12/metabolismo , Células del Cúmulo/fisiología , Regulación de la Expresión Génica , Oocitos/fisiología , Ovulación/genética , Animales , Gatos , Técnicas de Cultivo de Célula/métodos , Quimiocina CXCL12/farmacología , Medios de Cultivo/química , Medios de Cultivo/farmacología , Células del Cúmulo/citología , Células del Cúmulo/efectos de los fármacos , Femenino , Regulación de la Expresión Génica/efectos de los fármacos , Gonadotropinas/farmacología , Oocitos/citología , Oocitos/efectos de los fármacos , Receptores CXCR4/metabolismoRESUMEN
This study assessed the effects of diet supplementation with industrial processing by-products of acerola (Malpighia emarginata D.C.), cashew (Anacardium occidentale L.) and guava (Psidium guajava L.) fruit on the intestinal health and lipid metabolism of female Wistar rats with diet-induced dyslipidaemia. Female rats were randomly divided into five groups: healthy control, dyslipidaemic control and dyslipidaemic experimental receiving acerola, cashew or guava processing by-products. Fruit processing by-products were administered (400 mg/kg body weight) via orogastric administration for 28 consecutive days. Acerola, cashew and guava by-products caused body weight reduction (3·42, 3·08 and 5·20 %, respectively) in dyslipidaemic female rats. Dyslipidaemic female rats receiving fruit by-products, especially from acerola, presented decreased faecal pH, visceral fat, liver fat and serum lipid levels, as well as increased faecal moisture, faecal fat excretion, faecal Bifidobacterium spp. and Lactobacillus spp. counts and amounts of organic acids in faeces. Administration of the tested fruit processing by-products protected colon and liver from tissue damage (e.g. destruction of liver and colon cells and increased fat deposition in hepatocytes) induced by dyslipidaemic diet. Dietary fibres and phenolic compounds in tested fruit by-products may be associated with these positive effects. The industrial fruit processing by-products studied, mainly from acerola, exert functional properties that could enable their use to protect the harmful effects on intestinal health and lipid metabolism caused by dyslipidaemic diet.
Asunto(s)
Anacardium/química , Suplementos Dietéticos , Dislipidemias/terapia , Mucosa Intestinal/metabolismo , Metabolismo de los Lípidos , Magnoliopsida/química , Psidium/química , Animales , Peso Corporal , Fibras de la Dieta/análisis , Femenino , Frutas/química , Concentración de Iones de Hidrógeno , Lípidos/química , Hígado/metabolismo , Extractos Vegetales , Ratas , Ratas WistarRESUMEN
PURPOSE: Transcatheter arterial embolization (TAE) has been widely used in treating non-curative hepatocellular carcinoma (HCC). However, it is noticed that TAE may cause invasion of some cancer cells into circulation, resulting in distal metastasis and poor therapeutic outcome. Here, we aimed to reduce the side effects of TAE using the inhibitors for epidermal growth factor receptor (EGFR). METHODS: Transient hepatic artery ligation (HAL) was used as a mouse model for TAE. EGFR inhibitors were applied. Tumor size, presence of tumor cells in circulation, distal tumor formation, and activation of genes associated with tumor cell invasion and metastasis were analyzed. RESULTS: Inhibitors for EGFR significantly reduced the size of primary tumor, presence of tumor cells in circulation, and distal tumor formation after HAL. Further studies showed that EGFR inhibition suppressed several genes associated with tumor cell invasion and metastasis, such as vascular endothelial growth factor-A, stromal cell-derived factor 1, and Slug. CONCLUSION: EGFR inhibitor application may reduce circulating cancer cells during TAE and thus improve the therapy for advanced HCC.
Asunto(s)
Carcinoma Hepatocelular/patología , Embolización Terapéutica/efectos adversos , Receptores ErbB/antagonistas & inhibidores , Neoplasias Hepáticas/patología , Células Neoplásicas Circulantes/efectos de los fármacos , Animales , Antineoplásicos Inmunológicos/farmacología , Cetuximab/farmacología , Células Hep G2 , Humanos , Masculino , Ratones , Ratones Desnudos , Células Neoplásicas Circulantes/patología , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
BACKGROUND: Sperm DNA fragmentation (SDF) is recognized as a leading cause of male infertility because it can impair the paternal genome through distinct pathophysiological mechanisms. Current evidence supports SDF as a major factor in the pathophysiology of several conditions, including varicocele, unexplained infertility, assisted reproductive technology failure, and environmental lifestyle factors, although the mechanisms involved have not been fully described yet. Measurement of the levels of DNA fragmentation in semen provides valuable information on the integrity of paternal chromatin and may guide therapeutic strategies. A recently published clinical practice guideline (CPG) highlighted how to use the information provided by SDF testing in daily practice, which triggered a series of commentaries by leading infertility experts. These commentaries contained an abundance of information and conflicting views about the clinical utility of SDF testing, which underline the complex nature of SDF. METHODS: A search of papers published in response to the CPG entitled "Clinical utility of sperm DNA fragmentation testing: practice recommendations based on clinical scenarios" was performed within the Translational Andrology and Urology (TAU) website (http://tau.amegroups.com/). The start and end dates for the search were May 2017 and August 2017, respectively. Each commentary meeting our inclusion criteria was rated as "supportive without reservation", "supportive with reservation", "not supportive" or "neutral". We recorded whether articles discussed either SDF characteristics as a laboratory test method or clinical scenarios, or both. Subsequently, we extracted the particulars from each commentary and utilized the 'Strengths-Weaknesses-Opportunities-Threats' (SWOT) analysis to understand the perceived advantages and drawbacks of SDF as a specialized sperm function method in clinical practice. RESULTS: Fifty-eight fertility experts from six continents and twenty-two countries contributed commentaries. Overall, participants (87.9%; n=51) were supportive of the recommendations provided by the CPG on the utility of SDF testing based on clinical scenarios. The majority of participants made explicit remarks about both the clinical scenarios and SDF assays' characteristics. Among 'not supportive' and 'supportive with reservation' participants, 75% (n=30/40) and 77.5% (n=31/40) expressed concerns related to technical limitations of SDF testing methods and clinical utility of the test in one or more clinical scenarios discussed in the CPG, respectively. The SWOT analysis revealed that the CPG provides a reasonable evidence-based proposal for integration of SDF testing in the routine daily practice. It also uncovered gaps of knowledge and threats limiting the widespread application of SDF in everyday practice, thus allowing the identification of opportunities to further refine SDF testing and its clinical utility. CONCLUSIONS: The understanding of the role of SDF in male infertility requires an in-depth analysis of the multifactorial pathophysiological processes and the theories involved. The SWOT analysis allowed an objective evaluation of CPG on the clinical utility of SDF testing based on clinical scenarios and its accompanying commentaries written by global experts in all possible angles. Implementation of SDF testing in the clinic may not only increase the outcome of ART but more importantly improve the health of both fathers to be and resulting offspring.
RESUMEN
BACKGROUND: A rare phenotype of clinical non-progressors to AIDS is not well understood and the new protocol for universal treatment, may block the understanding of viral control thus it is crucial to define this controversial group. METHODS: A cohort of 30 persons followed a criteria for viremia control groups 1 (VC1; n = 2) and 2 (VC2; n = 7) and non-viral controllers (NC; n = 21) including number of years of diagnosis, LTCD4+, LTCD8+ counts, plasma viral load and the absence of ART; 241 uninfected control persons were matched to age and sex. Infected persons were regularly examined and submitted to two or three annual laboratory measurements. Polymorphisms and allele frequencies of CCR5Δ32 and SDF1-3'A were detected in the genomic DNA. Plasma levels of cytokines (IL-2, IL-4, IL-5, IL-9, IL-10, IL-13, IL-17 and IFN-y) were measured. RESULTS: The group investigated is originated from a miscigenetic population and demographic and social characteristics were not significantly relevant. LTCD4+ median values were higher among VC than NC, but significantly lower than uninfected controls. Evolution of LTCD4+ and LTCD8+ counts, showed a slight increase of LTCD4+ among VC, but a significant decrease in the NC. The percentage of annual change in LTCD4+ was also significantly different between the groups. LTCD4+/LTCD8+ ratio was inverted but not significant among the VC, thus the ratio may be a useful biomarker for the VC. A clear signature indicated a change from Th1 to Th2 cytokine profiles from VC to NC, respectively. CONCLUSIONS: The knowledge of viral controllers characteristics in different population groups is important to define a strict universal definition for the sake of learning about the pathogenesis of HIV-1. Data on LTCD4+ seems to be stable and repetitive from published data, but the LTCD8+ response and the significance of LTCD4+/LTCD8+ ratio values are in need to further exploration as biomarkers. The change from Th1 to Th2 cytokine profile may help to design and adjust specific treatment protocols for the group.