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PURPOSE: The implantation of non-absorbable meshes is the gold standard technique for ventral hernia (VH) repairs. However, emergency surgeries are often related to contaminated/infected fields, where the implantation of prosthetic materials may not be recommendable. Our aim was to evaluate the results of polyvinylidene fluoride (PVDF) meshes used for contaminated and/or complicated VH repairs in the acute setting. METHODS: We conducted a retrospective analysis of patients with VH who underwent emergency surgery involving PVDF meshes, in a tertiary hospital (from November 2013 to September 2019). We analyzed postoperative complications and 1-year recurrence rates. We evaluated the relationships between contamination grade, mesh placement, infectious complications, and recurrences. RESULTS: We gathered data on 123 patients; their mean age was 62.3 years, their mean BMI was 31.1 kg/m2, and their mean CeDAR index was 51.6. 96.4% of patients had a grade 2-3 ventral hernia according to the Rosen index. The mean defect width was 8 cm (IQR 2-18). 93 cases (75.6%) were described as contaminated or dirty surgeries. A PVDF mesh was placed using an IPOM technique in 56.3% of cases, and via interposition location in 39.9%. The one-month recurrence rate was 5.7% and recurrence after one year was 19.1%. The overall mortality rate was 27.6%. Risk of recurrence was related to patients with a Rosen score over 2 (p < 0.001), as well as with postoperative SSI (p = 0.045). Higher recurrence rates were not related to PVDF mesh placement. CONCLUSION: The use of PVDF meshes for emergency VH repairs in contaminated surgeries seems safe and useful, with reasonable recurrence rates, and acceptable infectious complication rates, similar to those published in the literature.
Assuntos
Hérnia Ventral , Herniorrafia , Hérnia Ventral/cirurgia , Herniorrafia/efeitos adversos , Humanos , Pessoa de Meia-Idade , Polivinil , Recidiva , Estudos Retrospectivos , Telas Cirúrgicas , Resultado do TratamentoRESUMO
Minimally invasive glaucoma surgery for chronic open-angle glaucoma treatment tries to reduce the amount of chronic topical drug treatment, delaying invasive glaucoma surgery and diminishing the number of surgical complications. XEN45 uses subconjunctival drainage in order to reduce intraocular pressure. The report is presented of a patient following phacoemulsification, intraocular lens implantation and XEN45 implant. After four days acute malignant glaucoma was observed, with stage 3 athalamia requiring pars plana vitrectomy and secondary Ahmed valve implantation. The lower rate of complications due to minimally invasive glaucoma surgery has increased its use. Nevertheless, it is not free of severe adverse events, even requiring a multidisciplinary approach as in our malignant glaucoma report.
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Implantes para Drenagem de Glaucoma/efeitos adversos , Glaucoma/etiologia , Complicações Pós-Operatórias/etiologia , Idoso , Feminino , HumanosRESUMO
BACKGROUND: Osteomyelitis is a difficult-to-cure infection with a high relapse rate despite combined medical and surgical therapies. Some severity factors, duration of antimicrobial therapy and type of surgical procedure might influence osteomyelitis relapse. METHODS: 116 patients with osteomyelitis were followed for ≥1 year after hospital discharge. Demographic, microbiological and clinical data, eight severity factors and treatment (surgical and antibiotic) were analyzed. RESULTS: Mean age was 53 years and 74.1% were men. Tibia (62.1%) and S. aureus (58.5%) were the most commonly involved bone and bacteria, respectively. Mean follow-up was 67.1 months. Forty-six patients underwent bone debridement, 61 debridement plus flap coverage and 9 antimicrobial therapy only. Twenty-six patients (22.4%) relapsed, at a mean of 11.2 months since hospital discharge. Duration > 3 months (p = 0.025), number of severity factors (P = 0.02) and absence of surgery (P = 0.004) were associated with osteomyelitis relapse in the univariate analysis. In the Cox regression analysis, osteomyelitis duration > 3 months (P = 0.012), bone exposure (P = 0.0003) and type of surgery (P < 0.0001) were associated with relapse. Regarding the surgical modalities, bone debridement with muscle flap was associated with better osteomyelitis outcomes, as compared with no surgery (P < 0.0001) and debridement only (P = 0.004). CONCLUSIONS: Osteomyelitis extending for > 3 months, bone exposure and treatment other than surgical debridement with muscular flap are risk factors for osteomyelitis relapse.
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Infecções Bacterianas/diagnóstico , Infecções Bacterianas/epidemiologia , Osteomielite/diagnóstico , Osteomielite/epidemiologia , Adulto , Idoso , Antibacterianos/uso terapêutico , Infecções Bacterianas/terapia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Osteomielite/microbiologia , Osteomielite/terapia , Prognóstico , Recidiva , Fatores de Risco , Staphylococcus aureus/isolamento & purificaçãoRESUMO
OBJECTIVE: Osteomyelitis is a difficult-to-cure infection, with high relapse rate despite adequate therapy. Large published osteomyelitis series in adults are rare. METHODS: A total of 344 adult osteomyelitis patients were studied and followed > 12 months after hospital discharge. Demographic, microbiological, clinical, therapeutic and outcome data were analyzed. RESULTS: Mean age was 52.5 ± 18.3 years and 233 (67.7%) were male. Main osteomyelitis types were post-surgical (31.1%), post-traumatic (26.2%) and hematogenous (23%). Tibia (24.1%) and femur (21.8%), and methicillin-susceptible S. aureus (29.6%) were the most commonly involved bone and bacteria, respectively. Median follow-up was 12.0 (IQR 0-48) months. Inflammatory markers were increased in 73.6%. Overall, patients were treated by IV and oral routes with one (IV: 44.5%, oral: 26.7%), two (IV: 30.1%, oral: 21.8%) or ≥ 2 (IV: 15.2%, oral: 6.1%) antibiotics. Median duration on IV/oral antimicrobials was 28.0 (IQR 24-28) and 19.5 (IQR 4-56) days, respectively. Anti-staphylococcal ß-lactams cloxacillin/cefazolin (19.2%) and ciprofloxacin (5.5%) were the most frequently used IV and orally, respectively. Overall 234 (68.0%) underwent surgery, 113 (32.8%) debridement, 97 (27.4%) debridement + muscle flap and 24 (7%) amputation. At the end of follow-up 208 patients (60.6%) did not have relapsed. Operated patients had significantly less relapses (p<0.0001). A total of 23 (6.7%) died, 11 (3.2%) by infectious complications and 48 (14%) were lost in the follow-up. CONCLUSIONS: Osteomyelitis is due to different causes complicating its therapy. Risk factors or causal microorganism could influence its treatment and outcome. Aggressive surgery along with adequate antimicrobial therapy are mandatory for cure.
Assuntos
Infecções Bacterianas/tratamento farmacológico , Infecções Bacterianas/microbiologia , Osteomielite/tratamento farmacológico , Osteomielite/microbiologia , Administração Oral , Adulto , Idoso , Idoso de 80 Anos ou mais , Antibacterianos/administração & dosagem , Antibacterianos/uso terapêutico , Infecções Bacterianas/cirurgia , Osso e Ossos/microbiologia , Desbridamento , Feminino , Seguimentos , Humanos , Mediadores da Inflamação/sangue , Injeções Intravenosas , Masculino , Staphylococcus aureus Resistente à Meticilina , Pessoa de Meia-Idade , Osteomielite/cirurgia , Recidiva , Estudos Retrospectivos , Resultado do TratamentoRESUMO
In teleosts, spermatogenesis is regulated by pituitary gonadotropins and sex steroids. 5α-dihydrotestosterone (DHT), derived from testosterone (T) through the action of 5α-reductase, has recently been suggested to play a physiologically important role in some fish species. In this study, gilthead seabream, Sparus aurata L., males received an implant of 1µgT/g body mass (bm) or vehicle alone and, 7days later, 1mg finasteride (FIN, an inhibitor of 5α-reductase)/kg bm or vehicle. Serum levels of T, 11-ketotestosterone (11KT), DHT and 17ß-estradiol (E2), and the mRNA levels of the main enzymes involved in their synthesis, were analysed. T promoted a transient increase in the serum levels of T, 11KT and E2 but a decrease in those of DHT at day 15 following T injection, in accordance with the up-regulation of mRNA levels of the enzymes involved in T transformation to 11KT (coding genes: cyp11b1 and hsd11b) and the down-regulation of mRNA levels of the enzyme responsible for T transformation to DHT (coding gene: srd5a). Interestingly, a similar effect was observed when FIN was injected. However, when fish were injected with T and FIN successively (T+FIN), control levels were not recovered at the end of the experimental period (28days). DHT seems to regulate E2 serum levels via the down-regulation of mRNA levels of aromatase (coding gene: cyp19a1a), which is needed for the transformation of T into E2. The testis histology, together with the proliferative rates recorded upon T, FIN or T+FIN treatment, suggests that DHT is involved in the onset of the meiotic phase of spermatogenesis.
Assuntos
3-Oxo-5-alfa-Esteroide 4-Desidrogenase/genética , Inibidores de 5-alfa Redutase/farmacologia , Androgênios/farmacologia , Di-Hidrotestosterona/sangue , Finasterida/farmacologia , Testosterona/farmacologia , 11-beta-Hidroxiesteroide Desidrogenase Tipo 1/genética , Androgênios/sangue , Animais , Aromatase/genética , Estradiol/sangue , Proteínas de Peixes/genética , Masculino , Dourada , Esteroide 11-beta-Hidroxilase/genética , Testículo/efeitos dos fármacos , Testículo/crescimento & desenvolvimento , Testosterona/análogos & derivados , Testosterona/sangueAssuntos
Adenocarcinoma Mucinoso/cirurgia , Carcinoma de Células em Anel de Sinete/cirurgia , Neoplasias do Íleo/cirurgia , Ileostomia/efeitos adversos , Complicações Pós-Operatórias/cirurgia , Reoperação/métodos , Adenocarcinoma Mucinoso/etiologia , Idoso , Carcinoma de Células em Anel de Sinete/etiologia , Humanos , Neoplasias do Íleo/etiologia , Masculino , Complicações Pós-Operatórias/etiologiaRESUMO
BACKGROUND: Survival after orthotopic liver transplantation (LT) for hepatocellular carcinoma (HCC) is influenced by tumor recurrence. This study examines the survival of patients who underwent LT for HCC and developed recurrence of tumor after transplantation. METHODS: A retrospective analysis was performed of the 200 patients who underwent LT secondary to HCC from 1990 to 2014. We excluded 19 patients from the study owing to early postoperative deaths in the 1st month. We divided our sample into 2 groups according to the presence of recurrence. We performed a univariate analysis to identify variables that are significantly associated with the risk of recurrence. Afterward we use multivariate analysis regression analysis to find independent significance. RESULTS: Univariate analysis shows significant relationship between high Edmondson-Steiner grades (G3-G4) and the development of tumor recurrence. Tumor size, vascular invasion, and capsular invasion were found to be independent risk factors of tumor recurrence in the multivariate analysis. CONCLUSIONS: Tumor recurrence defines survival of patients who underwent LT for HCC. In this study we discuss which histologic factor are associated with higher risk of tumor recurrence, and therefore a negative the impact on patient's survival.
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Carcinoma Hepatocelular/cirurgia , Neoplasias Hepáticas/cirurgia , Transplante de Fígado , Recidiva Local de Neoplasia/etiologia , Adulto , Idoso , Carcinoma Hepatocelular/mortalidade , Carcinoma Hepatocelular/patologia , Feminino , Humanos , Neoplasias Hepáticas/mortalidade , Neoplasias Hepáticas/patologia , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Recidiva Local de Neoplasia/cirurgia , Estudos Retrospectivos , Fatores de RiscoRESUMO
17α-Ethynylestradiol (EE2), which is used in oral contraceptives and hormone replacement therapy, is a well documented estrogenic endocrine disruptor and an aquatic contaminant. In the present study, adult male specimens of the marine hermaphrodite teleost gilthead (Sparus aurata L.) were fed a diet containing tamoxifen (Tmx), an estrogen receptor ligand used in cancer therapy, alone or combined with EE2, for 25 days and then fed a commercial diet for a further 25 days (recovery period). The effects of short (5days) and long (25 days) treatments on several reproductive and gonad immune parameters and the reversibility of the disruptive effects after the recovery period were examined. Our data showed that Tmx acted as an estrogenic endocrine disruptor as revealed by the increase in the hepatic transcription of the vitellogenin gene in males, the serum levels of 17ß-estradiol and the gonad expression levels of the estrogen receptor α and G protein-coupled estrogen receptor genes, and the recruitment of leukocytes into the gonad, a well known estrogenic-dependent process in gilthead seabream males. On the other hand, Tmx also increased sperm concentration and motility as well as the serum levels of androgens and the expression levels of genes that codify for androgenic enzymes, while decreasing the expression levels of the gene that code for gonadal aromatase. When applied simultaneously, Tmx and EE2 could act in synergy or counteract, each other, depending on the parameter measured. The disruptive effect of EE2 and/or Tmx was not reversible after a 25 day recovery period.
Assuntos
Disruptores Endócrinos/toxicidade , Antagonistas de Estrogênios/toxicidade , Etinilestradiol/toxicidade , Reprodução/efeitos dos fármacos , Espermatogênese/efeitos dos fármacos , Tamoxifeno/toxicidade , Animais , Disruptores Endócrinos/sangue , Antagonistas de Estrogênios/sangue , Etinilestradiol/sangue , Masculino , Reprodução/fisiologia , Dourada , Espermatogênese/fisiologia , Tamoxifeno/sangue , Testículo/efeitos dos fármacos , Testículo/fisiologia , Testículo/ultraestruturaRESUMO
BACKGROUND: Familial adenomatous polyposis (FAP) is an autosomal dominant cancer predisposition syndrome characterised by the progressive development of multiple colorectal adenomatous polyps and an increased incidence of colorectal carcinoma. It is often accompanied by other benign or malignant extracolonic manifestations, including gastric and duodenal tumours, osteomas, desmoid tumours, retinal pigmentation, and thyroid and adrenocortical tumours METHODS AND RESULTS: We report the case of a 42-year-old white female with FAP who was referred to our Endocrine Surgery Unit for surgery because of a palpable mass in the left side of the neck. An ultrasound-guided fine needle aspiration biopsy showed a cribriform-morular variant (CMV) of papillary thyroid carcinoma (PTC). The incidence, clinical presentation, histology and treatment options for this rare histological subtype are discussed. CONCLUSIONS: The diagnosis of CMV of PTC is very strongly related to the FAP syndrome and must be suspected when a thyroid node appears in FAP patients. Likewise, any patient without known FAP who presents this histology in a surgically biopsied or resected thyroid node should undergo total colonoscopy for screening of colonic polyposis and genetic study of the APC gene sequence.
RESUMO
We report a case of hookworm parasitosis in a Spanish patient who before imprisonment had lived in Brazil. The diagnosis was established from a progressive manifestation of asthenia, together with significant weight loss. Laboratory tests showed hypochromic microcytic anemia and eosinophilia. Consequently, the patient was admitted to hospital in order to complete the study, where several hookworm eggs were later found in feces. The patient was subsequently treated with Albendazole and iron,achieving clinical cure, normalization of biochemical parameters and eventual eradication of the parasite. We believe it is important for the prison doctor to bear this and other parasitosis in mind when facing the case of inmates who are originally from tropical countries, where these parasites are endemic. In addition, it might be appropriate to implement parasite screening programs in the immigrant population headed by the Prison Health Service, even when said population is asymptomatic.
Assuntos
Anemia Ferropriva/parasitologia , Infecções por Uncinaria/complicações , Prisioneiros , Adulto , Humanos , MasculinoRESUMO
INTRODUCTION: Selenium is an essential micronutrient for human health, being a cofactor for enzymes with antioxidant activity that protect the organism from oxidative damage. An inadequate intake of this mineral has been associated with the onset and progression of chronic diseases such as hypertension, diabetes, coronary diseases, asthma, and cancer. For this reason, knowledge of the plasma and erythrocyte selenium levels of a population makes a relevant contribution to assessment of its nutritional status. OBJECTIVE: The objective of the present study was to determine the nutritional status of selenium and risk of selenium deficiency in a healthy adult population in Spain by examining food and nutrient intake and analyzing biochemical parameters related to selenium metabolism, including plasma and erythrocyte levels and selenium-dependent glutathione peroxidase (GPx) enzymatic activity. MATERIAL AND METHODS: We studied 84 healthy adults (31 males and 53 females) from the province of Granada, determining their plasma and erythrocyte selenium concentrations and the association of these levels with the enzymatic activity of glutathione peroxidase (GPx) and with life style factors. We also gathered data on their food and nutrient intake and the results of biochemical analyses. Correlations were studied among all of these variables. RESULTS: The mean plasma selenium concentration was 76.6 ± 17.3 µg/L (87.3 ± 17.4 µg/L in males, 67.3 ± 10.7 µg/L in females), whereas the mean erythrocyte selenium concentration was 104.6 µg/L (107.9 ± 26.1 µg/L in males and 101.7 ± 21.7 µg/L in females). The nutritional status of selenium was defined by the plasma concentration required to reach maximum GPx activity, establishing 90 µg/L as reference value. According to this criterion, 50% of the men and 53% of the women were selenium deficient. CONCLUSIONS: Selenium is subjected to multiple regulation mechanisms. Erythrocyte selenium is a good marker of longer term selenium status, while plasma selenium appears to be a marker of short-term nutritional status. The present findings indicate a positive correlation between plasma selenium concentration and the practice of physical activity. Bioavailability studies are required to establish appropriate reference levels of this mineral for the Spanish population.
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Estado Nutricional , Selênio/deficiência , Adulto , Interpretação Estatística de Dados , Ingestão de Alimentos , Eritrócitos/química , Feminino , Glutationa Peroxidase/sangue , Humanos , Masculino , Espectrometria de Massas , Pessoa de Meia-Idade , Atividade Motora , Risco , Medição de Risco , Espanha/epidemiologia , Adulto JovemRESUMO
INTRODUCTION: Few studies have evaluated the efficacy and reliability of weight loss-focussed prepared food dishes in obese post-menopausal women. OBJECTIVE: To compare the efficacy of a weight loss programme based on a balanced hypocaloric diet using prepared dishes* with that of a similar programme based on standard commercially available foods and with a non-intervened control group. A further aim was to evaluate the subjectivity of participants in the preparation of the diet-adjusted dishes based on usually consumed products. SUBJECTS: Obese post-menopausal women aged between 55 and 65 years. DESIGN: Controlled longitudinal interventional study. METHOD: The sample of 75 female volunteers were divided into three groups of 25 women: a control group, who continued to consume their usual non-dietary adjusted meals (CG), an intervened group, treated with a diet adjusted to their individual requirements and based on standard commercially available food (SG), and another intervened group, treated with a similarly adjusted diet but based on prepared dishes (PG). Data were gathered on anthropometric variables, consumption habits and physical activity levels, and clinical-nutritional controls were conducted at the start and every two weeks to the end of the 8-week study in order to evaluate biochemical changes. RESULTS: The weight loss was slightly higher in the prepared-dishes group (PG) than in the standard food diet group (SG), but the difference was not statistically significant, whereas it was considerably higher in both groups than in the non-dietary adjusted control group (CG) and this difference was highly significant (losses of 7.60 kg in PG and 7.01 kg in SG versus 2.10 kg in CG (p < 0.01). However, the PG showed a significantly higher (p < 0.01) loss of fatty mass and abdominal circumference versus the SG women. CONCLUSION: More weight was lost by the two groups treated with a diet based on prepared dishes or usual food items in comparison to untreated controls, but the diet based on prepared dishes obtained more reliable and higher quality outcomes, achieving a positive change at fatty compartment level and in the abdominal circumference.
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Estado Nutricional , Obesidade/dietoterapia , Pós-Menopausa/fisiologia , Redução de Peso , Idoso , Antropometria , Restrição Calórica/métodos , Dieta , Dieta Redutora , Ingestão de Energia , Comportamento Alimentar , Feminino , Humanos , Estudos Longitudinais , Pessoa de Meia-Idade , Resultado do Tratamento , Circunferência da Cintura/fisiologiaRESUMO
Sex steroids are known to deeply alter processes other than fish reproduction, including fish growth, intermediary metabolism, osmoregulation and immunity. We have previously reported that 17ß-estradiol (E(2)), the main fish estrogen, promotes the mobilization of acidophilic granulocytes from the head kidney, the bone marrow equivalent in fish, to the gonad in the bony fish gilthead seabream (Sparus aurata L.). The aim of this study was to investigate the effects of E(2) and 17α-ethinylestradiol (EE(2)), an endocrine disruptor with strong estrogenic effects commonly found in the aquatic environment, on the ability of gilthead seabream endothelial cells (ECs) to promote leukocyte infiltration. E(2) and EE(2) were seen to affect ECs in different ways. Thus, E(2) was able to increase the production of nitric oxide (NO) and up-regulate the expression of the key activation markers, interleukin-1ß, CC chemokine ligand 4, interleukin-8, E-selectin and matrix metalloproteinase 9, when used alone or combined with bacterial DNA. In contrast, EE(2) failed to affect NO release and reduced the up-regulation of the above genes promoted by bacterial DNA. Moreover, we found that leukocyte adhesion to ECs was enhanced by E(2) treatment. Collectively, these results suggest that estrogens modulate fish leukocyte trafficking during an inflammatory process by activating ECs.
Assuntos
Quimiotaxia de Leucócito/imunologia , Células Endoteliais/imunologia , Estrogênios/imunologia , Inflamação/imunologia , Dourada/imunologia , Animais , Adesão Celular/efeitos dos fármacos , Quimiotaxia de Leucócito/efeitos dos fármacos , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/metabolismo , Estradiol/imunologia , Estradiol/metabolismo , Estradiol/farmacologia , Estrogênios/metabolismo , Estrogênios/farmacologia , Etinilestradiol/imunologia , Etinilestradiol/metabolismo , Etinilestradiol/farmacologia , Perfilação da Expressão Gênica , Inflamação/metabolismo , Óxido Nítrico/biossíntese , Receptores de Estrogênio/biossíntese , Receptores de Estrogênio/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Dourada/genética , Dourada/metabolismoRESUMO
The role of sex steroids in the modulation of fish immune responses has received little attention. Previous studies have demonstrated that 17ß-estradiol (E(2)) is able to alter the response of gilthead seabream leukocytes to infectious agents. We have used suppression subtractive hybridization to identify genes upregulated by E(2) (50 ng/ml) in macrophage cultures from gilthead seabream. We isolated 393 up-regulated cDNA fragments that led to the identification of 162 candidate estrogen-responsive genes. Functional analyses revealed the presence of several enriched immune processes and molecular pathways. The E(2) up-regulation of some immune-relevant genes was further confirmed by real time RT-PCR. Bioinformatics analysis revealed the ability of E(2) to orchestrate profound alterations in the macrophage expression profile, especially immune-related processes and pathways. This is the first report on E(2)-dependent modifications of fish macrophage transcriptome and lends weight to a suggested role for estrogen in the immune system, the possible significance of which is discussed.
Assuntos
Estrogênios/farmacologia , Perfilação da Expressão Gênica , Macrófagos/efeitos dos fármacos , Dourada/genética , Actinas/genética , Actinas/metabolismo , Animais , Células Cultivadas , Simulação por Computador , Etiquetas de Sequências Expressas , Proteínas de Peixes/genética , Proteínas de Peixes/metabolismo , Expressão Gênica/efeitos dos fármacos , Genes MHC Classe I , Peptídeos e Proteínas de Sinalização Intercelular/genética , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Metaloproteinase 13 da Matriz/genética , Metaloproteinase 13 da Matriz/metabolismo , Receptores Imunológicos/genética , Receptores Imunológicos/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de DNA , Transcrição Gênica/efeitos dos fármacosRESUMO
In mammals, estrogens regulate the immune system, either directly or indirectly via several leukocyte types through autocrine/paracrine mechanisms. In the gilthead seabream (Sparus aurata L.) gonad, an intensive remodeling process accompanied by the massive infiltration of acidophilic granulocytes (AG) is partially triggered by 17ß-estradiol (E(2)). Once AG infiltrated the gonad, show impaired activities. In this study we first demonstrate that neither testicular nor head-kidney AG express any of the three estrogen receptor (ER) genes (ERa, ERb1 and ERb2) described in the gilthead seabream, while head-kidney macrophages (Mc) and lymphocytes (Ly) constitutively express ERa gene. Moreover, Mc are important in the immune-modulatory role of E(2), as suggested by its ability to induce ERb2 gene expression and up-regulate the expression of genes coding for ERa, ERb1, pro-inflammatory cytokines, chemokines and tissue remodeling molecules. Furthermore, the soluble factors produced by E(2)-treated Mc decreased in head-kidney phagocytes, their phagocytic ability and capacity, while no effects were observed on their reactive oxygen intermediate (ROI) production or their migratory capabilities. However, the role of Ly in the regulation of AG migration and the modulation of phagocytic and ROI production activities triggered by E(2) can not be ruled out, so that further studies are necessary to clarify these issues.
Assuntos
Estradiol/imunologia , Regulação da Expressão Gênica , Ativação de Macrófagos/imunologia , Macrófagos/imunologia , Fagócitos/imunologia , Dourada/imunologia , Animais , Células Cultivadas , Receptores de Estrogênio/genética , Receptores de Estrogênio/imunologia , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Although there are some commercial vaccines available against infectious pancreatic necrosis virus (IPNV), the disease still continues to be a major problem for aquaculture development worldwide. In the current work, we constructed a DNA vaccine against IPNV (pIPNV-PP) by cloning the long open reading frame of the polyprotein encoded by the viral RNA segment A. In vitro, the vaccine is properly translated giving the functional IPNV polyprotein since preVP2, VP2 and VP3 proteins were detected because of the VP4-protease cleavage. EPC cells transfected with the vaccine plasmid expressed the viral proteins and induced the expression of type I interferon (IFN)-induced Mx genes. Furthermore, IPNV synthesized proteins seemed to assemble in virus-like particles as evidenced by electron microscopy. In vivo, rainbow trout specimens were intramuscularly injected with the vaccine and expression of immune-relevant genes, the presence of neutralizing antibodies and effect on viral load was determined. The pIPNV-PP vaccine was expressed at the injection site and up-regulated MHC Ialpha, MHC IIalpha, type-I interferon (IFN), Mx, CD4 and CD8alpha gene expression in the muscle, head kidney or spleen, although to a much lower extent than the up-regulations observed in response to an effective DNA vaccine against viral hemorrhagic septicaemia virus (VHSV). However, the IPNV vaccine was also very effective in terms of acquired immunity since it elicited neutralizing antibodies (in 6 out of 8 trout fingerlings) and decreased 665-fold the viral load after IPNV infection. The effectiveness of this new IPNV DNA vaccine and its possible mechanism of action are discussed and compared to other viral vaccines.
Assuntos
Infecções por Birnaviridae/prevenção & controle , Doenças dos Peixes/prevenção & controle , Vírus da Necrose Pancreática Infecciosa/imunologia , Vacinas de DNA/imunologia , Vacinas Virais/imunologia , Estruturas Animais/imunologia , Animais , Anticorpos Neutralizantes/sangue , Anticorpos Antivirais/sangue , Infecções por Birnaviridae/imunologia , Antígenos CD4/biossíntese , Antígenos CD8/biossíntese , Linhagem Celular , Doenças dos Peixes/imunologia , Antígenos de Histocompatibilidade Classe I/biossíntese , Antígenos de Histocompatibilidade Classe II/biossíntese , Vírus da Necrose Pancreática Infecciosa/genética , Injeções Intramusculares , Interferon Tipo I/biossíntese , Oncorhynchus mykiss , Poliproteínas/biossíntese , Rhabdoviridae/genética , Rhabdoviridae/imunologia , Vacinas de DNA/administração & dosagem , Vacinas de DNA/genética , Proteínas Virais/biossíntese , Vacinas Virais/administração & dosagem , Vacinas Virais/genéticaRESUMO
A new tool for DNA transfer to fish cell lines such as epithelioma papulosum cyprini (EPC) and rainbow trout gonad (RTG2), has been optimized by testing commercially available polyethylenimine (PEI) polymers as transfectant reagents. Deacylated 25 kDa PEI polymers were selected amongst the most active and then low toxicity deacylated PEIs fractions around 15 kDa were obtained by gel filtration chromatography to increase 3-4-fold their initial in vitro transfection efficiency. The EPC and plasmids coding for reporter genes were first used to optimize variable values for best expression by transfection with deacylated low toxicity PEI while both EPC/RTG2 and a plasmid coding for the glycoprotein G gene of the fish pathogen, viral haemorrhagic septicemia virus (VHSV) were then used to demonstrate some of their practical applications. Due to its relatively low price, defined chemical composition and availability, low toxicity deacylated PEI might be used for numerous applications for all those studying fish cell immunology in vitro as well as in vivo.
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Carpas/fisiologia , Oncorhynchus mykiss/fisiologia , Polietilenoimina , Transfecção/métodos , Animais , Linhagem Celular Tumoral , Regulação da Expressão Gênica , Peso Molecular , Polietilenoimina/químicaRESUMO
The gilthead seabream is a protandrous hermaphrodite seasonal breeding teleost with a bisexual gonad that offers an interesting model for studying the two first reproductive cycles (RCs), in which the specimens develops as males. During the first RC (RC1), 11-ketotestosterone (11-KT) and testosterone (T), the main androgens in fish, play different and specific roles in the testicular physiology as they peak at different stages of RC1. Moreover, the profiles of T serum levels during the second RC (RC2) demonstrated that this androgen is not essential in the testicular regression process that occurs during this cycle. However, changes in serum levels suggest that 17beta-estradiol (E2) orchestrates this process during both RCs. Moreover, the E2 serum levels recorded during RC2 indicate that there is a threshold level that determines the initiation of ovarian development during this cycle without promoting complete feminization. We analysed triglyceride, protein, glucose and lactate serum levels in order to establish a relationship between the mobilization and transfer of nutrients, the hormonal changes that take place during the RC and body composition, finding that in vivo serum levels of metabolites change significantly throughout the first two RCs, although the physiological relevance of such changes is still unknown. Triglyceride levels seem to be affected by the beginning of ovary development during RC2 but not by testicular recrudescence during RC1. Moreover, glucose and lactate might be important sources of energy during the resting and testicular involution stages.