RESUMO
The objective of the study was to investigate the effects of a dacitic tuff breccia (DTB) on Eimeria-infected broilers. A total of 600 one-day-old Cobb 500 male chickens were randomly assigned to 5 treatments with 10 replicates of 12 birds. Treatments were: an unchallenged control (UC), a challenged (CC) control (0% DTB), and 3 challenged groups with 0.125, 0.25, or 0.5% DTB. At d 14, birds in the CC and DTB groups were orally gavaged with mixed Eimeria spp., while the UC received water. Growth performance was evaluated during prechallenge, challenge, and postchallenge periods (0-14 d; 14-20 d; and 20-26 d, respectively). Gastrointestinal permeability was measured at 5 days postinfection (dpi). Intestinal histology and nutrient digestibility of dry matter (DM), crude protein (CP), and ileal digestible energy (IDE) were measured at 6 dpi. Liver activity of glutathione peroxidase (GSH-Px) was determined at 6 dpi, and concentrations of reduced (GSH) and oxidized glutathione (GSSG) were analyzed at 6 and 12 dpi. Data were analyzed using a linear mixed model analysis and Tukey's test (P ≤ 0.05). From 0 to 14 d, similar average daily gain (ADG) and average daily feed intake (ADFI, P > 0.05) were observed. Gain:feed ratio (GF) was higher in 0.125, 0.25, and 0.5% of DTB than the CC and UC (P < 0.001). From 14 to 20 d, the UC had the highest ADG, ADFI, and GF (P < 0.001). At 5 dpi, intestinal permeability was higher in the challenged groups than the UC. Additionally, the UC showed the highest apparent ileal digestibility of CP, whereas 0.125% DTB had higher CP digestibility than the CC and 0.5% DTB (P < 0.001). At 6 dpi, 0.125% DTB increased GSH-Px activity compared to the CC, 0.5% DTB, and UC (P < 0.001). At 12 dpi, 0.125% DTB showed increased GSH concentration compared to the CC, 0.25% DTB, and 0.5% DTB (P < 0.01). The mild coccidia infection negatively impacted growth performance, apparent ileal nutrient digestibility, intestinal histology, and gastrointestinal integrity in broilers. The use of 0.125% DTB exhibited potential in improving antioxidant responses, apparent ileal digestibility of CP, and growth performance.
Assuntos
Eimeria , Animais , Masculino , Eimeria/fisiologia , Antioxidantes/farmacologia , Dieta/veterinária , Galinhas/fisiologia , Intestinos , Ração Animal/análise , Suplementos Nutricionais/análiseRESUMO
BACKGROUND: Intestinal ultrasound (IUS) is an increasingly used non-invasive tool to evaluate Crohn's disease (CD) activity. Recently, two IUS scores that evaluate inflammatory activity have emerged: the Simple Ultrasound Activity Score for CD (SUS-CD) and the International Bowel Ultrasound Segmental Activity Score (IBUS-SAS). We aimed to compare the accuracy of SUS-CD, IBUS-SAS and contrast-enhanced ultrasound (CEUS) in predicting inflammatory activity in the terminal ileum in ileocolonoscopy in CD patients. METHODS: Retrospective study including all consecutive CD patients submitted to IUS with CEUS directed to the terminal ileum performed by a single operator between April 2016 and March 2020. Segmental SUS-CD and IBUS-SAS were calculated. A time-intensity curve of the contrast bowel wall enhancement was created with measurement of peak intensity using CEUS. The CD endoscopic activity in ileocolonoscopy was graded by Simple Endoscopic Score for CD (SES-CD) as inactive (SES-CD < 7) or active (SES-CD ≥ 7). RESULTS: Fifty patients were included, 54.0% were female, with mean age of 34 ± 12 years, and most had isolated ileal disease (60.0%), and a nonstricturing, nonpenetrating behaviour (44.0%). Most of the patients (60.0%) had active endoscopic disease (SES-CD ≥ 7). SUS-CD and IBUS-SAS were not different between patients with active or inactive endoscopic disease (p = 0.15; 0.57, respectively), having a poor accuracy to correlate endoscopic activity (area under de curve (AUC) 0.62; 0.55, respectively). Peak intensity in CEUS was significantly different in patients with active or inactive endoscopic disease (p = 0.004), having a good accuracy to correlate endoscopic activity (AUC 0.80). CONCLUSION: Unlike CEUS, SUS-CD and IBUS-SAS were not able to accurately correlate endoscopic activity in terminal ileum in CD. Therefore, CEUS is a non-invasive emerging method that should be increasingly integrated in the ultrasonographic evaluation of CD patients.
Assuntos
Doença de Crohn , Doenças do Íleo , Adulto , Doença de Crohn/diagnóstico por imagem , Feminino , Humanos , Íleo/diagnóstico por imagem , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Índice de Gravidade de Doença , Ultrassonografia , Adulto JovemRESUMO
Phubbing is defined as ignoring other individuals by using a mobile phone during a face-to-face conversation. The Phubbing Scale (PS) was developed to assess this practice. In this study, we analyze the psychometric properties of the 8-item version of the PS (PS-8) in the Portuguese population, providing validity evidence based on internal structure and on relationships with other variables, and examining item properties, reliability, and measurement invariance across gender. Participants were 391 Portuguese adults (132 men, 259 women) who completed a battery of questionnaires. Confirmatory factor analysis yielded satisfactory goodness-of-fit indices for the two-factor structure (communication disturbance and phone obsession), which was invariant across gender. Item homogeneity and reliability of factor scores (McDonald's omega) were satisfactory. Validity evidence based on relationships with other variables was provided by positive associations with time spent on the Internet on weekdays and at the weekend, time spent on social networking sites, number of social networks used, Internet addiction, problematic mobile phone use, Facebook intrusion, fear of missing out, and depression. These associations show the addictive component of phubbing and its relationship with mental health. The PS-8 is a short and easy-to-administer scale with adequate psychometric properties for measuring phubbing in the Portuguese population. (AU)
Assuntos
Humanos , Masculino , Feminino , Adolescente , Adulto , Pessoa de Meia-Idade , Reprodutibilidade dos Testes , Comportamento Aditivo/psicologia , Comunicação , Telefone Celular/estatística & dados numéricos , Portugal , Psicometria , Fatores Sexuais , Inquéritos e Questionários , Internet/estatística & dados numéricosRESUMO
Endometriosis was induced (autotransplant) in Wistar rats. After 21 days, the rats were randomly divided into two groups (16 female rats each). Control group was forced-fed 0.9% sodium chloride solution, and the ginger group was forced-fed 0.5 mg/100 g of Zingiber officinale Roscoe fresh extract, both by gavage, for 14 days, in addition to their normal diet. After that, an anesthetic dose (ketamine/xylazine) was administered until euthanasia. Peritoneal lavage fluid was collected to evaluate tumor necrosis factor (TNF)-α and interleukin (IL)-6, and autotransplant was measured and excised to evaluate histology. The final mean volumes were larger in the control group (120.92 mm3 ± 78.91) than in the ginger group (40.50 mm3 ± 19.57) (P = .01). The endometriosis foci increased in the control group from 45.10 mm3 ± 29.96 at 21 days postimplantation to 120.92 mm3 ± 78.91 on the day of euthanasia (P = .02). In the ginger group, a slight increase was observed from 38.43 mm3 ± 19.96 to 40.50 mm3 ± 19.57, without statistical difference (P = .83). In addition, a greater increase in growth of the endometriosis foci was found when compared with the control (75.81 mm3 ± 58.95) and ginger groups (2.07 mm3 ± 18.87) (P = .004). No difference was found in TNF-α (P = .51) and in IL-6 (P = .12). The degree of lesion atrophy was higher in the ginger group (1 ± 0.92) than in the control group (2.25 ± 1.16) (P = .03). The ginger extract reduced and atrophied autotransplanted endometriosis foci, but did not reduce IL-6 and TNF-α in the peritoneal lavage fluid.
Assuntos
Endometriose , Zingiber officinale , Animais , Endometriose/tratamento farmacológico , Feminino , Humanos , Extratos Vegetais , Ratos , Ratos WistarRESUMO
Mesenchymal stem cells (MSC) are modern tools in regenerative therapies of humans and animals owed to their immunomodulatory properties, which are activated in a pro-inflammatory environment. Different preconditioning strategies had been devised to enhance the immunomodulatory properties of MSC. In this research, we evaluated the immunological attributes of equine adipose MSC (eAMSC) before and after preconditioning in vitro with prostaglandin E2 (PGE2), substance P (SP), their combination and IFNγ. PGE2/SP was the best combination to keep or enhance the mesodermal lineage differentiation of eAMSC. Alongside with this, preconditioning of eMSC with PGE2 and SP did not affect expression of stemness MSC surface phenotype: CD90+, CD44+, MHC class I+, MHC class II- and CD45-, assessed by cytometry. Both naïve and preconditioned eAMSC expressed genes related with immune properties, such as MHC-I, PTGES, IL6, IL1A, TNFα and IL8 assessed by qPCR. Only TNFα was under expressed in treated cells, while the other markers were either overexpressed or not changed. In no cases MHC-II expression was detected. The antiproliferative effect of preconditioned eAMSC exposed to activated peripheral blood mononuclear cells (PBMC) showed that SP treatment significantly inhibited proliferation of LPS stimulated PBMC. When eAMSC were stimulated with Poly I:C, all the treatments significantly inhibited proliferation of stimulated PBMC (p < 0.05). Direct contact (coculture) between the preconditioned eAMSC and PBMC, induced a shift of significantly more (CD4/CD25/FOXP3)+ T-regulatory PBMC than naïve eAMSC. In the experiments of this research, we investigated the secreted proteomic profile of naïve and preconditioned eAMSC, 42 up-regulated and 40 down-regulated proteins were found in the proteomic assay. Our proteomic data revealed profound changes in the secretory pattern of MSC exposed to different treatments, compared to naïve eAMSC as well as among treatments. In overall, compared to naïve cells, the protein profile of preconditioned cells resembled the mesenchymal-epithelial transition (MET). Here we showed that the combined use of PGE2 and SP provoked in overall the highest expression of anti-inflammatory markers as well as lead to an increased acquisition of a T-regulatory phenotype in preconditioned eAMSC without affecting their "stemness".
Assuntos
Dinoprostona/imunologia , Cavalos/imunologia , Células-Tronco Mesenquimais/imunologia , Proteínas/metabolismo , Substância P/imunologia , Animais , Biomarcadores/metabolismo , Diferenciação Celular/imunologia , Proliferação de Células , Citometria de Fluxo/veterinária , Interferon gama/imunologia , Masculino , Células-Tronco Mesenquimais/metabolismo , Mesoderma/citologia , Proteoma , Via Secretória/imunologiaRESUMO
Tectona grandis L.f. es una especie forestal de madera dura que, a pesar de no ser nativa de América, su plasticidad de adaptación ha permitido que en Ecuador se establezcan plantaciones intensivas con fines de exportación. Una compleja enfermedad con características de marchitez vascular está matando miles de árboles de diferentes edades en el país. Se planteó conocer el agente fitopatógeno causante de la patogénesis y muerte de los árboles. Se aislaron los hongos Fusarium sp. y Ceratocystis fimbriata Ellis & Halst. desde árboles enfermos. A nivel de invernadero se plantearon los postulados de Koch, para el efecto 30 plantas de T. grandis de cuatro meses de edad por cada tratamiento, se inocularon con los microrganismos como se describe a continuación: T1= C. fimbriata, T2 = Fusarium sp., T3 = C. fimbriata + Fusarium sp., T4 = agar-agar (control). Se empleó un diseño completo al azar (DCA) y las plantas se evaluaron a los 45 días después de inoculadas. Los tratamientos C. fimbriata, y C. fimbriata + Fusarium sp., mostraron mayores volúmenes aparentes de necrosis, con 1.52 cm3 y 1.93 cm3, y plantas muertas por la infección durante el tiempo de evaluación, mientras que Fusarium sp. mostró baja o nula patogenicidad y comportamiento similar al control, con 0.27 cm3 y 0.16 cm3 respectivamente. Estos resultados sugieren que el agente causal de la enfermedad de marchitez vascular en T. grandis es C. fimbriata y sería el primer reporte del fitopatógeno atacando teca en Ecuador. (AU)
Tectona grandisL.f. it is a hardwood forest species, which despite not being native to America, its adaptive plasticity has allowed intensive plantations to be established for export purposes in Ecuador. A complex disease with characteristics of vascular wilt is killing thousands of trees of different ages in the country. It was proposed to know the phytopathogenic agent causing the pathogenesis and death of the trees. Fusarium sp. andCeratocystis fimbriata Ellis & Halst. were isolated from diseased trees. At the greenhouselevel, Koch's postulates were proposed, for the effect 30 four month old T. grandisplants for each treatment were inoculated with the microorganisms as described below: T1 = C. fimbriata, T2 = Fusariumsp ., T3 = C. fimbriata+ Fusariumsp., T4 = agar-agar (control). A complete randomized design (DCA) was used and the plants were evaluated 45 days after inoculation. The treatments C. fimbriata, and C. fimbriata+ Fusariumsp. showed higher apparent volumes of necrosis, with 1.52 cm3and 1.93 cm3, and plants killed by the infection during the evaluation time, while Fusariumsp. showed low or no pathogenicity and behavior similar to the control, with 0.27 cm3and 0.16 cm3respectively. These results suggest that the causative agent of vascular wilt disease in T. grandisis C. fimbriataand it would be the first report of phytopathogen attacking teak in Ecuador. (AU)
Assuntos
Árvores/microbiologia , Ceratocystis/patogenicidade , Florestas , Equador , Fusarium/patogenicidadeRESUMO
AIM: To test the diagnostic performance of a deep learning-based system for the detection of clinically significant pulmonary nodules/masses on chest radiographs. MATERIALS AND METHODS: Using a retrospective study of 100 patients (47 with clinically significant pulmonary nodules/masses and 53 control subjects without pulmonary nodules), two radiologists verified clinically significantly pulmonary nodules/masses according to chest computed tomography (CT) findings. A computer-aided diagnosis (CAD) software using a deep-learning approach was used to detect pulmonary nodules/masses to determine the diagnostic performance in four algorithms (heat map, abnormal probability, nodule probability, and mass probability). RESULTS: A total of 100 cases were included in the analysis. Among the four algorithms, mass algorithm could achieve a 76.6% sensitivity (36/47, 11 false negative) and 88.68% specificity (47/53, six false-positive) in the detection of pulmonary nodules/masses at the optimal probability score cut-off of 0.2884. Compared to the other three algorithms, mass probability algorithm had best predictive ability for pulmonary nodule/mass detection at the optimal probability score cut-off of 0.2884 (AUCMass: 0.916 versus AUCHeat map: 0.682, p<0.001; AUCMass: 0.916 versus AUCAbnormal: 0.810, p=0.002; AUCMass: 0.916 versus AUCNodule: 0.813, p=0.014). CONCLUSION: In conclusion, the deep-learning based computer-aided diagnosis system will likely play a vital role in the early detection and diagnosis of pulmonary nodules/masses on chest radiographs. In future applications, these algorithms could support triage workflow via double reading to improve sensitivity and specificity during the diagnostic process.
Assuntos
Aprendizado Profundo , Neoplasias Pulmonares/diagnóstico por imagem , Nódulos Pulmonares Múltiplos/diagnóstico por imagem , Radiografia Torácica/métodos , Nódulo Pulmonar Solitário/diagnóstico por imagem , Tomografia Computadorizada por Raios X/métodos , Algoritmos , Diagnóstico por Computador/métodos , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Melhoria de Qualidade , Interpretação de Imagem Radiográfica Assistida por Computador/métodos , Estudos Retrospectivos , Sensibilidade e Especificidade , Software , TaiwanRESUMO
Herpes simplex virus type 1 (HSV-1) is a ubiquitous infectious agent that can establish latency in neurons, and in some cases, viral retrograde transport results in infection of the central nervous system (CNS). Several antivirals have been identified with the ability to inhibit HSV-1 replication in human cells to a greater or lesser degree, most of which are nucleoside analogues that unfortunately exhibit teratogenic potential, embryotoxicity, carcinogenic or antiproliferative activities and resistances in immunocompromised patients, specially. In the present study, we assessed two amidic derivatives of valproic acid (VPA) - valpromide (VPD) and valnoctamide (VCD) - which are already used in clinic treatments, as feasible HSV-1 antivirals in glial cells. Both VPD and VCD have exhibited increased efficacy in bipolar disorders and as anticonvulsant drugs compared to VPA, while being less teratogenic and hepatotoxic. Cytotoxicity assays carried out in our laboratory showed that VPD and VCD were not toxic in a human oligodendroglioma cell line (HOG), at least at the concentrations established for human treatments. Infectivity assays showed a significant inhibition of HSV-1 infection in HOG cells after VPD and VCD treatment, being more pronounced in VPD-treated cells, comparable to the effects obtained with acyclovir. Furthermore, the same antiherpetic effects of VPD were observed in other oligodendrocytic cell lines and rat primary oligodendrocytes (OPCs), confirming the results obtained in HOG cells. Altogether, our results allow us to propose VPD as a potential antiherpetic drug that is able to act directly on oligodendrocytes of the CNS.
Assuntos
Amidas/farmacologia , Antivirais/farmacologia , Herpesvirus Humano 1/efeitos dos fármacos , Oligodendroglia/virologia , Ácido Valproico/análogos & derivados , Amidas/química , Animais , Antivirais/química , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Humanos , Estrutura Molecular , Oligodendroglia/efeitos dos fármacos , Ratos , Ácido Valproico/química , Ácido Valproico/farmacologia , Proteínas Virais/genética , Internalização do Vírus/efeitos dos fármacos , Replicação Viral/efeitos dos fármacosRESUMO
Adult stromal mesenchymal stem cells (MSCs) have been postulated as responsible for cell renewal in highly and continuously regenerative tissues such as the endometrium. MSCs have been identified in the endometrium of many species including humans, rodents, pets and some farm animals, but not in horses. The objective of this work was to isolate such cells from the endometrium of mares and to compare their main biological attributes with horse adipose-derived MSCs. Here we successfully isolated and characterized endometrial MSCs (eMSCs) from mares. Said cells showed fibroblast-like morphology, grew on plastic, had doubling population times of 46.4 ± 3.38 h, underwent tri-lineage (osteo, chondro and adipogenic) differentiation after appropriate inductions, migrated toward the attraction of fetal calf serum and displayed a pattern of surface markers commonly accepted for horse MSCs. All these are properties of MSCs. Some of these attributes were shared with equine adipose-derived MSCs, but the migration pattern of eMSC at 12 and 24 h after stimulation was reduced in comparison with adipose MSCs. Also, expression of CD44, CD90 and MHCI surface markers were dramatically down-regulated in eMSCs. In conclusion, equine-derived endometrial MSC share biological attributes with adipose MSC of this species, but displayed a different surface marker phenotype and an impaired migration ability. Conceivably, this phenotype is distinctive for MSC of this origin.
Assuntos
Tecido Adiposo/citologia , Endométrio/citologia , Cavalos/fisiologia , Proteínas de Membrana/metabolismo , Células-Tronco Mesenquimais/fisiologia , Animais , Biomarcadores , Movimento Celular , Células Cultivadas , Feminino , Regulação da Expressão Gênica , Proteínas de Membrana/genéticaRESUMO
In Poster Sessions, the first-author name was missing from the authorship group originally listed for Abstract PS-01-004 (page S52), entitled "Clinicopathological features of microinvasive breast cancer". The correct authorship group is shown above.
RESUMO
In broiler breeder roosters, the weight of the testes is positively associated with daily sperm production and fertility. In birds, the testes are located in the coelomic cavity, not being accessible to reproductive evaluation as mammalian testes. The reproductive evaluation of roosters is based on phenotypic traits. Any changes on testes will be reflected on fertility levels. The aim of this paper was to evaluate the potential of association of phenotypic traits as body weight (BW), comb scores (COS), cloaca scores (CLS) and feet scores (FS) with testicular morphometric parameters in adult Cobb® Mx roosters in two ages (25 and 45 weeks of age). The low BW roosters had lower testes weight (TW), smaller seminiferous tubule diameter (STD) and lower comb and cloaca scores than the medium and high BW roosters (P≤0.05). Heavy roosters had the highest COS and bigger seminiferous epithelium height (SEH). There was no statistical difference between the BW categories in FS and volumetric proportion of seminiferous tubule (ST) and interstitial tissue (IT). Considering the age effect, roosters with 45 weeks of age had lower TW and ST than 25-week roosters. Positive, moderate and significant correlation was found between testes weight and phenotypic characteristics as BW, COS and CLS in 45week roosters. A positive, moderate and significant correlation was found between body weight and histologic testicular characteristics as STD and SHE in both evaluated ages. In conclusion, there is a regression in the testes weight with age and it is more intense in Cobb® Mx roosters with lower BW. Furthermore, it is concluded that the BW positively influences the testes weight and histological quality of the testes. COS and CLS can be used with moderate potential prediction to identify roosters with low testes weight at 45 weeks of age.(AU)
Em galos de matriz pesada sabe-se que o peso dos testículos pode ser associado positivamente com a produção diária de espermatozoides e que alterações nos testículos irão refletir na fertilidade. Nas aves, os testículos estão localizados dentro da cavidade celomática, e, portanto, não são acessíveis para exame andrológico direto, como em mamíferos. A avaliação reprodutiva de galos se baseia principalmente em características fenotípicas. Buscou-se avaliar o potencial de associação de alguns parâmetros fenotípicos, como peso corporal (PC), escores de crista (ECR), escores de cloaca (ECL) e escores de pés (EP), com parâmetros morfológicos de testículos em galos Cobb® Mx em duas idades (25 e 45 semanas de idade). Galos leves apresentaram menor peso testicular (PT), menor diâmetro de túbulo seminífero (DT), e menores ECR e ECL que galos médios e pesados (P≤0.05). Galos pesados apresentaram os maiores ECR e maiores valores para altura do epitélio seminífero (AE), e DT. Não houve diferença estatisticamente significativa entre as categorias de peso para escores de pé e proporções de túbulo seminífero (TS) e interstício (INT). Considerando-se o efeito da idade, galos com 45 semanas de idade apresentaram menores PT e TS que galos com 25 semanas de idade. Correlação positiva, moderada e significativa foi encontrada entre peso de testículo e características fenotípicas como ECR, ECL e PC em galos de 45 semanas de idade. Foi encontrada correlação positiva, moderada e significativa entre PC e características testiculares como DT e AE nas duas idades avaliadas. Conclui-se que em galos Cobb® Mx ocorre regressão do testículo com o avanço da idade, e ela é mais intensa em galos com menor peso corporal. Conclui-se que o PC tem influência positiva no peso e índices histomorfométricos dos testículos, e que escores de crista e de cloaca podem ser utilizados com potencial moderado de predição para identificar galos com baixo peso de testículos em galos com 45 semanas de idade.(AU)
Assuntos
Animais , Masculino , Galinhas/anatomia & histologia , Testículo/anatomia & histologia , Testículo/crescimento & desenvolvimento , Peso Corporal , Cloaca/anatomia & histologiaRESUMO
A 7-month-old domestic medium hair cat presented with facial asymmetry affecting the bony and soft tissue structures of the right side of the head including the maxilla, nose, eye and pinna of the ear. Additionally, neurological dysfunction of the facial and vestibulocochlear nerves on the affected side was present. A congenital malformation affecting the first and second embryologic pharyngeal arches was suspected. This is the first case of hemifacial microsomia of likely congenital origin reported in a cat.
Assuntos
Doenças do Gato/patologia , Síndrome de Goldenhar/veterinária , Animais , Doenças do Gato/congênito , Doenças do Gato/diagnóstico por imagem , Doenças do Gato/cirurgia , Gatos , Orelha Média/patologia , Orelha Média/cirurgia , Ossos Faciais/anormalidades , Ossos Faciais/diagnóstico por imagem , Ossos Faciais/patologia , Síndrome de Goldenhar/diagnóstico por imagem , Síndrome de Goldenhar/patologia , Síndrome de Goldenhar/cirurgia , Masculino , Pólipos Nasais/patologia , Pólipos Nasais/cirurgia , Pólipos Nasais/veterinária , Tomografia Computadorizada por Raios X/veterináriaRESUMO
This study evaluated the effect of three reversible meiotic inhibitors (MINs) and their interaction with gonadotrophins (Gns) on the meiotic maturation and developmental competence of porcine oocytes. In experiment 1, the oocytes were matured for 22 hr in the presence or absence of dbcAMP (1 mM), cycloheximide (7 µM) or cilostamide (20 µM) with or without Gns, and for an additional 22 hr in the absence of MINs and Gns. At 22 hr of maturation, regardless of the presence of Gns, a higher proportion (p < .001) of oocytes cultured in the presence of MINs were effectively arrested at the germinal vesicle stage compared with the oocytes cultured without MINs. At 44 hr of maturation, the proportion of oocytes that reached MII was higher (p < .05) in groups with Gns compared with groups without Gns. In experiment 2, oocytes that were matured as in experiment 1 were inseminated and cultured for 7 days to evaluate fertilization parameters and blastocyst formation. Only oocytes from the dbcAMP + Gns group had higher (p < .05) efficiency of fertilization compared with the other treatment groups. The presence of dbcAMP during maturation also increased (p < .05) blastocyst formation and efficiency of blastocyst formation in both the presence and absence of Gns. These results indicate that the interaction of Gns with the tested MINs improved meiotic progression. In addition, regardless of supplementation with Gns, the presence of dbcAMP during the first maturation period increased and even doubled the capacity of oocytes to develop to the blastocyst stage.
Assuntos
Desenvolvimento Embrionário/efeitos dos fármacos , Fertilização in vitro/veterinária , Gonadotropinas/farmacologia , Técnicas de Maturação in Vitro de Oócitos/veterinária , Oócitos/efeitos dos fármacos , Animais , Blastocisto/efeitos dos fármacos , Bucladesina/farmacologia , Cicloeximida/farmacologia , Feminino , Fertilização/efeitos dos fármacos , Gonadotropinas/administração & dosagem , Masculino , Quinolonas/farmacologia , SuínosRESUMO
Stem cells have been postulated as responsible for cell regeneration in highly and continuously regenerative tissues such as the endometrium. Few studies in cattle have identified and specified the presence of stem cells in the endometrium during the oestrous cycle. The aim of this study was to investigate the presence of mesenchymal stem cells (MSCs) in the bovine endometrium during the follicular phase (FP) of the oestrous cycle. Uterine tissue was collected in the time-frame comprising day 18 of the cycle and ovulation (day 0). We isolated, cultured and expanded four primary cell lines from endometrium and identified byRT-qPCR the expression of OCT4, SOX2 but not NANOG (undifferentiated/embryonic markers), CD44 (MSCs marker) and c-KIT (stem cell marker) genes; and the encoded Oct4, Sox2 and Cd44 proteins by Western blot or immunostaining of paraffin-embedded tissue in endometrium. We demonstrated that cells isolated from bovine endometrium displayed essentially the same gene expression pattern; however, at the protein level, Oct4 and Cd44 were not detected. Besides, they showed typical functional characteristics of MSCs such as fibroblast-like morphology, plastic adherence, high proliferative capacity, clone formation in vitro and the ability to differentiate into chondrogenic, osteogenic and adipogenic lineages. We obtained for the first time an extensive characterization of undifferentiated cells populations contained in the bovine endometrium during the FP of the oestrous cycle.
Assuntos
Endométrio/citologia , Fase Folicular , Células-Tronco Mesenquimais/citologia , Animais , Bovinos , Diferenciação Celular , Células Cultivadas , Ciclo Estral , Feminino , Expressão GênicaRESUMO
Defective apoptosis might be involved in the pathogenesis of multiple sclerosis (MS). We evaluated apoptosis-related molecules in MS patients before and after autologous haematopoietic stem cell transplantation (AHSCT) using BCNU, Etoposide, AraC and Melphalan (BEAM) or cyclophosphamide (CY)-based conditioning regimens. Patients were followed for clinical and immunological parameters for 2 years after AHSCT. At baseline, MS patients had decreased proapoptotic BAD, BAX and FASL and increased A1 gene expression when compared with healthy counterparts. In the BEAM group, BAK, BIK, BIMEL , FAS, FASL, A1, BCL2, BCLXL , CFLIPL and CIAP2 genes were up-regulated after AHSCT. With the exception of BIK, BIMEL and A1, all genes reached levels similar to controls at day + 720 post-transplantation. Furthermore, in these patients, we observed increased CD8+ Fas+ T cell frequencies after AHSCT when compared to baseline. In the CY group, we observed increased BAX, BCLW, CFLIPL and CIAP1 and decreased BIK and BID gene expressions after transplantation. At day + 720 post-AHSCT, the expression of BAX, FAS, FASL, BCL2, BCLXL and CIAP1 was similar to that of controls. Protein analyses showed increased Bcl-2 expression before transplantation. At 1 year post-AHSCT, expression of Bak, Bim, Bcl-2, Bcl-xL and cFlip-L was decreased when compared to baseline values. In summary, our findings suggest that normalization of apoptosis-related molecules is associated with the early therapeutic effects of AHSCT in MS patients. These mechanisms may be involved in the re-establishment of immune tolerance during the first 2 years post-transplantation.
Assuntos
Apoptose/genética , Proteína 5 Relacionada à Autofagia/genética , Esclerose Múltipla/genética , Adulto , Linfócitos T CD8-Positivos/efeitos dos fármacos , Ciclofosfamida/uso terapêutico , Feminino , Expressão Gênica/genética , Transplante de Células-Tronco Hematopoéticas/métodos , Humanos , Masculino , Pessoa de Meia-Idade , Esclerose Múltipla/tratamento farmacológico , Condicionamento Pré-Transplante/métodos , Transplante Autólogo/métodos , Adulto JovemRESUMO
BACKGROUND AND PURPOSE: Recombinant IFN-ß is one of the first-line treatments in multiple sclerosis (MS), despite its lack of efficacy in some patients. In this context, mesenchymal stem cells (MSCs) represent a promising therapeutic alternative due to their immunomodulatory properties and multipotency. Moreover, by taking advantage of their pathotropism, these cells can be genetically modified to be used as carriers for delivering or secreting therapeutic drugs into injured tissues. Here, we report the therapeutic effect of systemic delivery of adipose-derived MSCs (AdMSCs), transduced with the IFN-ß gene, into mice with experimental autoimmune encephalomyelitis (EAE). EXPERIMENTAL APPROACH: Relapsing-remitting and chronic progressive EAE were induced in mice. Cells were injected i.v. Disease severity, inflammation and tissue damage were assessed clinically, by flow cytometry of spleens and histopathological evaluation of the CNS respectively. KEY RESULTS: Genetic engineering did not modify the biological characteristics of these AdMSCs (morphology, growth rate, immunophenotype and multipotency). Furthermore, the transduction of IFN-ß to AdMSCs maintained and, in some cases, enhanced the functional properties of AdMSCs by ameliorating the symptoms of MS in EAE models and by decreasing indications of peripheral and central neuro-inflammation. CONCLUSION AND IMPLICATIONS: Gene therapy was found to be more effective than cell therapy in ameliorating several clinical parameters in both EAE models, presumably due to the continuous expression of IFN-ß. Furthermore, it has significant advantages over AdMSC therapy, and also over systemic IFN-ß treatment, by providing long-term expression of the cytokine at therapeutic concentrations and reducing the frequency of injections, while minimizing dose-limiting side effects.
Assuntos
Encefalomielite Autoimune Experimental/terapia , Terapia Genética/métodos , Interferon beta/genética , Transplante de Células-Tronco Mesenquimais/métodos , Esclerose Múltipla Recidivante-Remitente/terapia , Tecido Adiposo/citologia , Animais , Modelos Animais de Doenças , Encefalomielite Autoimune Experimental/genética , Encefalomielite Autoimune Experimental/fisiopatologia , Feminino , Citometria de Fluxo , Células-Tronco Mesenquimais/citologia , Camundongos , Camundongos Endogâmicos C57BL , Esclerose Múltipla Crônica Progressiva/genética , Esclerose Múltipla Crônica Progressiva/fisiopatologia , Esclerose Múltipla Crônica Progressiva/terapia , Esclerose Múltipla Recidivante-Remitente/genética , Esclerose Múltipla Recidivante-Remitente/fisiopatologia , Índice de Gravidade de DoençaRESUMO
Despite all the knowledge, the cellular and molecular mechanisms involved in myeloproliferative neoplasm (MPN) pathophysiology remain unclear. Authors have shown galectin-1 (Gal-1) and 3 playing roles in tumour angiogenesis and fibrosis, which were correlated with poor prognosis in patients with MPN. In the present study LGALS1 and LGALS3 were differently expressed between polycythemia vera, essential thrombocythemia (ET) and primary myelofibrosis (PMF) diseases. Increased LGALS3 expression was associated with a negative JAK2 V617F status mutation in leucocytes from PMF but not in patients with ET without this mutation. However, a positive Janus kinase 2 (JAK2) V617F cell line established from patients with ET (SET-2 cells) when treated with JAK inhibitor presented high levels of LGALS3. Additionally, high LGALS1 expression was found in CD34(+) cells but not in leucocytes from patients with PMF, in absence of JAK2 V617F mutation, and also in SET-2 cells treated with JAK inhibitor. Thus, our findings indicate that differential expression of LGALS1 and/or LGALS3 in patients with MPN is linked with JAK2 V617F status mutation in these diseases and state of cell differentiation.
Assuntos
Galectina 1/genética , Galectina 3/genética , Janus Quinase 2/genética , Policitemia Vera/genética , Mielofibrose Primária/genética , Trombocitemia Essencial/genética , Adulto , Substituição de Aminoácidos , Antígenos CD34/genética , Proteínas Sanguíneas , Medula Óssea/patologia , Neoplasias da Medula Óssea/diagnóstico , Neoplasias da Medula Óssea/genética , Neoplasias da Medula Óssea/metabolismo , Linhagem Celular , Galectina 1/metabolismo , Galectina 3/metabolismo , Galectinas , Regulação Neoplásica da Expressão Gênica , Humanos , Janus Quinase 2/metabolismo , Masculino , Pessoa de Meia-Idade , Mutação , Transtornos Mieloproliferativos/diagnóstico , Transtornos Mieloproliferativos/genética , Transtornos Mieloproliferativos/metabolismo , Policitemia Vera/diagnóstico , Policitemia Vera/metabolismo , Mielofibrose Primária/diagnóstico , Mielofibrose Primária/metabolismo , Trombocitemia Essencial/diagnóstico , Trombocitemia Essencial/metabolismoRESUMO
The most common radiological error in examinations of the spine is the failure to diagnose fractures. This is the third most frequent reason for lawsuits brought against radiologists for negligence, after the failure to diagnose breast cancer and the failure to diagnose lung cancer. The thousands of radiological reports of spinal examinations done every year affect not only patients' health, but also their permission to be off work and their compensation. For this reason, it is our responsibility to know why errors are committed and how to detect them in order to avoid their repetition. In this article, we show the spectrum of the most common errors in our experience in double reading spinal examinations, and we try to determine what causes these errors.
Assuntos
Erros de Diagnóstico , Fraturas da Coluna Vertebral/diagnóstico por imagem , HumanosRESUMO
BACKGROUND: Nonmelanoma skin cancer (NMSC) is the most common cancer in Germany, but detailed information on survival is lacking. OBJECTIVES: To provide survival estimates for female and male patients with basal cell carcinoma (BCC), squamous cell carcinoma (SCC), sarcoma, adenocarcinoma and Merkel cell carcinoma. Further subgroup analyses were carried out by age, tumour stage and body site. METHODS: In total 459 640 patients diagnosed with NMSC in 1997-2011 were included from population-based cancer registers, covering a population of 33 million inhabitants. Age-standardized absolute and relative 5-year and 10-year survival were calculated using period analysis. RESULTS: The absolute and relative 5-year survival were 87·1% and 102·9% for BCC, 77·6% and 93·6% for SCC, 82·1% and 96·0% for sarcoma, 71·4% and 85·7% for adenocarcinoma and 60·0% and 70·7% for Merkel cell carcinoma, respectively. Higher age, female sex and advanced stage were associated with lower survival. CONCLUSIONS: A comprehensive overview of NMSC survival in Germany is provided. The differences between the NMSC subtypes require a more differentiated consideration of patient survival. The survival advantage of patients with BCC may be related to health-promoting factors related to the BCC diagnosis, such as changes to a healthier lifestyle.