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INTRODUCTION: Inguinal hernia repair lacks a standard repair technique, with laparo-endoscopic and open preperitoneal methods showing similar outcomes. Despite higher costs, the popularity of robotic surgery is on the rise, driven by technological advantages. Controversies persist in comparing open repair techniques with the robotic approach, given contradictory results. The objective of this study was to compare postoperative outcomes, including complications, chronic pain, and recurrence, between open and robotic-assisted preperitoneal inguinal hernia repair. METHODS: This single-center retrospective study encompassed patients undergoing elective inguinal hernia repair in a specialized unit, employing both open preperitoneal and robotic-assisted laparoscopic approaches from September 2018 to May 2023. Comparative analysis of short- and long-term outcomes between these techniques was conducted. Additionally, multivariate logistic regression was employed to explore predictors of postoperative complications. RESULTS: A total of 308 patients met the inclusion criteria. 198 (64%) patients underwent surgery using an open preperitoneal approach and 110 (36%) using robot-assisted laparoscopy. Patients in the robot-assisted group were younger (P = 0.006) and had fewer comorbidities (P < 0.001). There were no differences between the groups in terms of postoperative complications (P = 0.133), chronic pain (P = 0.463) or recurrence (P = 0.192). Multivariate analysis identified ASA ≥ III (OR, 1.763; 95%CI, 1.068-3.994; P = 0.027) and inguinoscrotal hernias (OR, 2.371, 95%CI, 1.407-3.944; P = 0.001) as risk factors of postoperative complications. CONCLUSIONS: Both open preperitoneal and robotic-assisted laparoscopic approaches show similar outcomes for complications, chronic pain, and recurrence when performed by experienced surgeons. The open preperitoneal approach, with its quicker operative time, may be advantageous for high-comorbidity cases. Treatment choice should consider patient factors, surgeon experience, and healthcare resources.
Subject(s)
Hernia, Inguinal , Herniorrhaphy , Recurrence , Robotic Surgical Procedures , Humans , Hernia, Inguinal/surgery , Robotic Surgical Procedures/methods , Robotic Surgical Procedures/adverse effects , Male , Retrospective Studies , Middle Aged , Female , Herniorrhaphy/methods , Herniorrhaphy/adverse effects , Aged , Laparoscopy/methods , Laparoscopy/adverse effects , Postoperative Complications/epidemiology , Postoperative Complications/etiology , Adult , Surgical Mesh , Treatment OutcomeABSTRACT
BACKGROUND: The aim of this study is to analyze the data collected on lateral incisional hernias (LIH) in the National Incisional Hernia Registry (EVEREG) and assess current practices and outcomes in LIH repair. METHODS: A retrospective cohort study was conducted using LIH data recorded over 10 years (2012-2022). Comorbidities, hernia characteristics, short-term complications, and recurrences were studied, along with their association with the type of approach used, either open or laparoscopic, in elective surgery. RESULTS: 1742 LIH cases were studied. According to the EHS classification, these included L1 409 (23.5%), L2 388 (22.3%), L3 565 (32.4%), L4 150 (8.6%) and combined 230 (13.2%). An open approach was performed in 1528 (87.7%) cases and laparoscopic in 214 (12.3%). The median age was 66 ± 12.45 years, with a majority of males, 934 (53.6%). The median body mass index was 29 ± 5.18 kg/m2. The most observed comorbidity was arterial hypertension (957 patients, 55%). A specialist was present in 638 interventions (56%). The 24-month follow-up was 17.9%, and recurrence in those cases was 27.2%, with a higher incidence when there was no specialist present during the intervention, onlay mesh position, and larger defect size. CONCLUSIONS: Surgery for LIH is common, although laparoscopic approach remains infrequent. Furthermore, it is associated with a high percentage of recurrences that increases when there is no specialist in abdominal wall surgery present.
ABSTRACT
Streptococcus pneumoniae is an important human pathogen. Currently used conjugate vaccines are effective against invasive disease, but protection is restricted to serotypes included in the formulation, leading to serotype replacement. Furthermore, protection against non-invasive disease is reported to be considerably lower. The development of a serotype-independent vaccine is thus important and Pneumococcal surface protein A (PspA) is a promising vaccine candidate. PspA shows some diversity and can be classified in 6 clades and 3 families, with families 1 and 2 being the most frequent in clinical isolates. The ideal vaccine should thus induce protection against the two most common families of PspA. The aim of this work was to develop a liposome-based vaccine containing PspAs from family 1 and 2 and to characterize its immune response. Liposomes (LP) composed of dipalmitoylphosphatidylcholine (DPPC) and 3ß-[N-(N',N'-dimethylaminoethane)-carbamoyl]cholesterol (DC-Chol) with or without α-galactosylceramide (α-GalCer) were produced by microfluidics, encapsulating PspA from clade 1 (PspA1, family 1) and/or clade 4 (PspA4Pro, family 2) followed by spray-drying with trehalose to form nanocomposite microparticles carriers (NCMP). LP/NCMPs showed good stability and preservation of protein activity. LP/NCMPs containing PspA1 and/or PspA4Pro were used for immunization of mice targeting the lungs. High serum IgG antibody titers against both PspA1 and PspA4Pro were detected in animals immunized with LP/NCMPs containing α-GalCer, with a balance of IgG1 and IgG2a titers. IgG in sera from immunized mice bound to pneumococcal strains from different serotypes and expressing different PspA clades, indicating broad recognition. Mucosal IgG and IgA were also detected. Importantly, immunization with LP/NCMPs induced full protection against strains expressing PspAs from family 1 and 2. Furthermore, CD4+ resident memory T cells were detected in the lungs of the immunized animals that survived the challenge.
Subject(s)
Galactosylceramides , Pneumococcal Infections , Streptococcus pneumoniae , Humans , Animals , Mice , Liposomes , Powders , Pneumococcal Infections/prevention & control , Bacterial Proteins , Immunization , Pneumococcal Vaccines , Immunoglobulin G , Lung , Antibodies, Bacterial , Mice, Inbred BALB CABSTRACT
BACKGROUND: The aim of this study was to analyze the impact of an abdominal wall surgery unit on postoperative complications (within 90 days postoperatively), hernia recurrence and chronic postoperative inguinal pain after elective recurrent inguinal hernia repair. METHODS: We conducted a retrospective cohort study of all adult patients who underwent elective recurrent inguinal hernia repair between January 2010 and October 2021. Short- and long-term outcomes were compared between the group of patients operated on in the abdominal wall surgery unit and the group of patients operated on by other units not specialized in abdominal wall surgery. A logistic regression model was performed for hernia recurrence. RESULTS: A total of 250 patients underwent elective surgery for recurrent inguinal hernia during the study period. The patients in the abdominal wall surgery group were younger (P ≤ 0.001) and had fewer comorbidities (P ≤ 0.001). There were no differences between the groups in terms of complications. The patients in the abdominal wall surgery group presented fewer recurrences (15% vs. 3%; P = 0.001). Surgery performed by the abdominal wall surgery unit was related to fewer recurrences in the multivariate analysis (HR = 0.123; 95% CI = 0.21-0.725; P = 0.021). CONCLUSIONS: Specialization in abdominal wall surgery seems to have a positive impact in terms of recurrence in recurrent inguinal hernia repair. The influence of comorbidities or type of surgery (i.e., outpatient surgery) require further study.
Subject(s)
Abdominal Wall , Hernia, Inguinal , Adult , Humans , Hernia, Inguinal/surgery , Abdominal Wall/surgery , Retrospective Studies , Postoperative Complications/epidemiology , Pain, Postoperative , RecurrenceABSTRACT
PURPOSE: Short-term results have been reported regarding parastomal hernia (PH) prevention with a permanent mesh. Long-term results are scarce. The objective was to assess the long-term PH occurrence after a prophylactic synthetic non-absorbable mesh. METHODS: Long-term data of three randomized controlled trials (RCTs) were collected. The primary outcome was the detection of PH based exclusively on a radiological diagnosis by computed tomography (CT) performed during the long-term follow-up. The Kaplan-Meier method was used for the comparison of time to diagnosis of PH according to the presence of mesh vs. no-mesh and the technique of mesh insertion: open retromuscular, laparoscopic keyhole, and laparoscopic modified Sugarbaker. RESULTS: We studied 121 patients (87 men, median age 70 years), 82 (67.8%) of which developed a PH. The median overall length of follow-up was 48.5 months [interquartile range (IQR) 14.4-104.9], with a median time until PH diagnosis of 17.7 months (IQR 9.3-49.0). The survival analysis did not show significant differences in the time to development of a PH according to the presence or absence of a prophylactic mesh neither in the overall study population (log-rank, P = 0.094) nor in the groups of each technique of mesh insertion, although according to the surgical technique, a higher reduction in the appearance of PH for the open retromuscular technique was found (log-rank, P = 0.001). CONCLUSION: In the long-term follow-up placement of a non-absorbable synthetic prophylactic mesh in the context of an elective end colostomy does not seem effective for preventing PH.
Subject(s)
Hernia, Ventral , Incisional Hernia , Surgical Stomas , Aged , Humans , Male , Colostomy/adverse effects , Colostomy/methods , Follow-Up Studies , Hernia, Ventral/etiology , Hernia, Ventral/prevention & control , Hernia, Ventral/diagnosis , Herniorrhaphy , Incisional Hernia/etiology , Incisional Hernia/prevention & control , Incisional Hernia/surgery , Randomized Controlled Trials as Topic , Surgical Mesh , FemaleABSTRACT
PURPOSE: The aim of this study is to compare the postoperative results, in terms of complications and recurrence, between the anterior and open preperitoneal approaches in emergency femoral hernia. METHODS: This bi-centric retrospective cohort study included patients who underwent emergency femoral hernia repair between January 2010 and December 2018. Postoperative complications and recurrence were analyzed comparing anterior and open preperitoneal approaches. The predictors of complications, mortality and recurrence were investigated using multivariate logistic regression. RESULTS: A total of 204 patients met the inclusion criteria. Open anterior approach was performed in 128 (62.7%) patients and open preperitoneal was performed in 76 (37.3%). Open preperitoneal approach was associated with lower rates of recurrence (P = 0.033) and associated midline laparotomies (P = 0.006). Multivariable analysis identified patients with chronic nephropathy (OR, 3.801; 95%CI, 1.034-13.974; P = 0.044), preoperative bowel obstruction (OR, 2.376; 95%CI, 1.118-5.047; P = 0.024) and required midline laparotomy (OR, 12.467; 95%CI, 11.392-102.372; P = 0.030) as risk factors for complications and ASA ≥ III (OR, 7.820; 95%CI, 1.279-47.804; P = 0.026), COPD (OR, 5.064; 95%CI, 1.188-21.585; P = 0.028), necrotic contents (OR, 36.968; 95%CI, 4.640-294.543; P = 0.001), and required midline laparotomy (OR, 11.047; 95%CI, 1.943-62.809; P = 0.007). as risk factors for 90-day mortality. Male gender (OR, 4.718; 95%CI, 1.668-13.347; P = 0.003) and anterior approach (OR, 5.292; 95%CI, 1.114-25.149; P = 0.036) were risk factors for recurrence. CONCLUSION: Open preperitoneal approach may be superior to anterior approach in the emergency setting because it can avoid the morbidity of associated midline laparotomies, with a lower long-term recurrence rate.
Subject(s)
Hernia, Femoral , Hernia, Inguinal , Humans , Male , Retrospective Studies , Hernia, Femoral/surgery , Hernia, Femoral/complications , Surgical Mesh , Herniorrhaphy/adverse effects , Herniorrhaphy/methods , Postoperative Complications/epidemiology , Postoperative Complications/etiology , Postoperative Complications/surgery , Hernia, Inguinal/surgery , RecurrenceABSTRACT
Events of catastrophic fish mortality in the lakes of the Amazonian floodplains are not uncommon. They are generally associated with thermal inversion of the water column, which is provoked by cold air masses that originate from the south of the continent. These events occur in the period of high water when the lakes are stratified. This paper reports an event of fish mortality that occurred during the low water season in a large floodplain system on the right-hand margin of the Amazon River. Information from seasoned fishers, who live in the same area where the event happened, and hydrological and satellite image analysis was used to identify the potential cause of fish mortality events. The amplitude of the flood pulse and the duration of extreme ebb showed to be the key factors responsible for the occurrence of events of fish mortality. These factors determine connectivity patterns between the floodplain lakes and the river channel, which are essential for maintaining water quality and the biota in the systems.
Subject(s)
Fishes , Lakes , Animals , Rivers , Biota , PerceptionABSTRACT
Background: Ventral hernia repair is one of the most commonly performed surgical procedures worldwide. To reduce the risk of complications, pre- and intra-operative strategies have received increasing focus in recent years. To assess possible preventive surgical strategies, this European Hernia Society endorsed project was launched. The aim of this review was to evaluate the current literature focusing on pre- and intra-operative strategies for surgical site occurrences (SSO) and specifically surgical site infection (SSI) in ventral hernia repair. Methods: A systematic review was conducted and reported in line with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses statement. Databases used were Pubmed and Web of Science. Original retrospective or prospective human adult studies describing at least one intra-operative intervention to reduce SSO after ventral hernia repair were considered eligible. Results: From a total of 4775 results, a total of 18 papers were considered suitable after full text reading. Prehospital chlorhexidine gluconate (CHG) scrub appears to increase the risk of SSO in patients undergoing ventral hernia repair, while there is no association between any type of surgical hat worn and the incidence of postoperative wound events. Intraoperative measures as prophylactic negative pressure therapy, surgical drain placement and the use of quilt sutures seem beneficial for decreasing the incidence of SSO and/or SSI. No positive effect has been shown for antibiotic soaking of a synthetic mesh, nor for the use of fibrin sealants. Conclusion: This review identified a limited amount of literature describing specific preventive measures and techniques during ventral hernia repair. An advantage of prophylactic negative pressure therapy in prevention of SSI was observed, but different tools to decrease SSIs and SSOs continuously further need our full attention to improve patient outcomes and to lower overall costs.
ABSTRACT
Effective and safe therapies to counteract persistent inflammation are necessary. We developed erythrocyte-derived liposomes (EDLs) with intrinsic anti-inflammatory activity. The EDLs were prepared using lipids extracted from erythrocyte membranes, which are rich in omega-3 fatty acids with several health benefits. Diclofenac, a widely used anti-inflammatory drug, was incorporated into EDLs in relevant therapeutic concentrations. The EDLs were also functionalised with folic acid to allow their active targeting of M1 macrophages, which are key players in inflammatory processes. In the presence of lipopolysaccharide (LPS)-stimulated macrophages, empty EDLs and EDLs incorporating diclofenac were able to reduce the levels of important pro-inflammatory cytokines, namely interleukin-6 (IL-6; ≈85% and 77%, respectively) and tumour necrosis factor-alpha (TNF-α; ≈64% and 72%, respectively). Strikingly, cytocompatible concentrations of EDLs presented similar effects to dexamethasone, a potent anti-inflammatory drug, in reducing IL-6 and TNF-α concentrations, demonstrating the EDLs potential to be used as bioactive carriers in the treatment of inflammatory diseases.
Subject(s)
Liposomes , Tumor Necrosis Factor-alpha , Anti-Inflammatory Agents/pharmacology , Anti-Inflammatory Agents/therapeutic use , Cytokines , Diclofenac/pharmacology , Diclofenac/therapeutic use , Erythrocytes , Humans , Inflammation/drug therapy , Interleukin-6Subject(s)
Hernia, Ventral , Incisional Hernia , Hernia, Ventral/surgery , Herniorrhaphy , Humans , Incisional Hernia/surgery , Registries , Surgical MeshABSTRACT
PURPOSE: The objective of this study was to gather information on patient-reported knowledge (PRK) in the field of hernia surgery. METHODS: A prospective quantitative study was designed to explore different aspects of PRK and opinions regarding hernia surgery. Patients referred for the first time to a surgical service with a presumed diagnosis of hernia and eventual hernia repair were eligible, and those who gave consent completed a simple self-assessment questionnaire before the clinical visit. RESULTS: The study population included 449 patients (72.8% men, mean age 61.5). Twenty (4.5%) patients did not have hernia on physical examination. The patient's perceived health status was "neither bad nor good" or "good" in 56.6% of cases. Also, more patients considered that hernia repair would be an easy procedure (35.1%) rather than a difficult one (9.8%). Although patients were referred by their family physicians, 32 (7.1%) answered negatively to the question of coming to the visit to assess the presence of a hernia. The most important reason of the medical visit was to receive medical advice (77.7%), to be operated on as soon as possible (40.1%) or to be included in the surgical waiting list (35.9%). Also, 46.1% of the patients considered that they should undergo a hernia repair and 56.8% that surgery will be a definitive solution. CONCLUSION: PRK of patients referred for the first time to an abdominal wall surgery unit with a presumed diagnosis of hernia was quite limited and there is still a long way towards improving knowledge of hernia surgery.
Subject(s)
Herniorrhaphy , Patient Reported Outcome Measures , Female , Hernia , Herniorrhaphy/adverse effects , Herniorrhaphy/methods , Humans , Male , Middle Aged , Prospective Studies , Surveys and QuestionnairesABSTRACT
Introduction: The current literature has not yet provided a definitive conclusion on the best emergency groin hernia repair. The aim of this study was first to compare the short and long-term outcomes between open preperitoneal and anterior approach in emergency groin hernia repair and second to identify risk factors for postoperative complications, mortality, and recurrence. Materials and Methods: This retrospective cohort study included patients who underwent emergency groin hernia repair between January 2010 and December 2018. Short and long-term outcomes were analyzed comparing approach and repair techniques. The predictors of complications and mortality were investigated using multivariate logistic regression. Cox regression multivariate analysis were used to explore risk factors of recurrence. Results: A total of 316 patients met the inclusion criteria. The most widely used surgical techniques were open preperitoneal mesh repair (34%) and mesh plug (34%), followed by Lichtenstein (19%), plug and patch (7%) and tissue repair (6%). Open preperitoneal mesh repair was associated with lower rates of recurrence (p = 0.02) and associated laparotomies (p < 0.001). Complication and 90-day mortality rate was similar between the techniques. Multivariable analysis identified patients aged 75 years or older (OR, 2.08; 95% CI, 1.14-3.80; p = 0.016) and preoperative bowel obstruction (OR, 2.11; 95% CI, 1.20-3.70; p = 0.010) as risk factors for complications and Comprehensive Complication Index ≥26.2 as risk factor for 90-day mortality (OR, 44.76; 95% CI, 4.51-444.59; p = 0.01). Female gender was the only risk factor for recurrence. Conclusion: Open preperitoneal mesh repair may be superior to other techniques in the emergency setting, because it can avoid the morbidity of associated laparotomies, with a lower long-term recurrence rate.
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AIM: To analyze the outcomes of component separation techniques (CST) to treat incisional hernias (IH) in a large multicenter cohort of patients. METHODS: All IH repair using CST, registered in EVEREG from July 2012 to December 2019, were included. Data on the pre-operative patient characteristics and comorbidities, IH characteristics, surgical technique, complications, and recurrence were collected. Outcomes between anterior (ACS) and posterior component separation (PCS) techniques were compared. Risk factors for complications and recurrences were analyzed. RESULTS: During the study period, 1536 patients underwent CST (45.5% females) with a median age of 64.0 years and median body mass index (BMI) of 29.7 kg/m2. ACS was the most common technique (77.7%). Overall complications were frequent in both ACS and PCS techniques (36.5%), with a higher frequency of wound infection (10.6% vs. 7.0%; P = 0.05) and skin necrosis (4.4% vs. 0.1%; P < 0.0001) with the ACS technique. Main factors leading to major complications were mesh explant (OR 1.72; P = 0.001), previous repair (OR 0.75; P = 0.038), morbid obesity (OR 0.67; P = 0.015), ASA grade (OR 0.62; P < 0.0001), COPD (OR 0.52; P < 0.0001), and longitudinal diameter larger than 10 cm (OR 0.58; P = 0.001). After a minimum follow-up of 6 months (median 15 months; N = 590), 59 (10.0%) recurrences were diagnosed. Operations performed in a non-specialized unit were significantly associated with recurrences (HR 4.903, CI 1.64-14.65; P = 0.004). CONCLUSION: CST is a complex procedure with a high rate of complications. Both ACS and PCS techniques have similar complication and recurrence rates. Operations performed in a specialized unit have better outcomes.
Subject(s)
Abdominal Wall , Hernia, Ventral , Incisional Hernia , Abdominal Muscles/surgery , Abdominal Wall/surgery , Female , Hernia, Ventral/etiology , Herniorrhaphy/methods , Humans , Incisional Hernia/etiology , Incisional Hernia/surgery , Male , Middle Aged , Recurrence , Registries , Retrospective Studies , Surgical Mesh , Treatment OutcomeABSTRACT
PURPOSE: Long delays in waiting lists have a negative impact on the principles of equity and providing timely access to care. This study aimed to assess waiting lists for abdominal wall hernia repair (incisional ventral vs. inguinal hernia) to define explicit prioritization criteria. METHODS: A cross-sectional single-center study was designed. Patients in the waiting list for incisional/ventral hernia (n = 42) and inguinal hernia (n = 50) repair were interviewed by phone and completed health-related quality of life (HRQoL) questionnaires (EQ-5D, COMI-hernia, HerQLes) as a measure of severity. Priority was measured as hernia complexity, patient frailty using the modified frailty index (mFI-11), and the consumption of analgesics for hernia. RESULTS: The mean (SD) time on the waiting list was 5.5 (3.2) months (range 1-14). Complex hernia was present in 34.8% of the patients. HRQoL was moderately poor in patients with incisional/ventral hernia (mean HerQL score 66.1), whereas it was moderately good in patients with inguinal hernia (mean COMI-hernia score 3.40). The use of analgesics was higher in patients with incisional/ventral hernia as compared with those with inguinal hernia (1.48 [0.54] vs. 1.31 [0.51], P = 0.021). Worst values of mFI were associated with inguinal hernia as compared with incisional/ventral hernia (0.21 [0.14] vs. 0.12 [0.11]; P = 0.010). CONCLUSION: Explicit criteria for prioritization in the waiting lists may be the consumption of analgesics for patients with incisional/ventral hernia and frailty for patients with inguinal hernia. A reasonable approach seems to establish separate waiting lists for incisional/ventral hernia and inguinal hernia repair.
Subject(s)
Abdominal Wall , Frailty , Hernia, Inguinal , Hernia, Ventral , Incisional Hernia , Abdominal Wall/surgery , Cross-Sectional Studies , Hernia, Inguinal/surgery , Hernia, Ventral/surgery , Herniorrhaphy , Humans , Incisional Hernia/surgery , Quality of Life , Waiting ListsABSTRACT
AIM: The aim of the present study was to examine the vaccine immune response in ewes supplemented with Bacillus toyonensis BCT-7112T during a period of 5-day supplementation before vaccination against a recombinant Clostridium perfringens epsilon toxin (rETX). METHODS AND RESULTS: Ewes were vaccinated with 200 µg of rETX adjuvanted with 10% aluminium hydroxide. The treat group was orally supplemented with B. toyonensis BCT-7112T (3 × 108 viable spores) for 5 days prior to the first and second vaccination. Ewes supplemented with B. toyonensis BCT-7112T showed higher neutralizing antibody titres than the non-supplemented ewes (P < 0·05), with an increase in serum levels for total IgG anti-rETX by 3·2-fold (P < 0·0001), and for both IgG isotypes IgG1 and IgG2 by 2·1-fold and 2·3-fold (P < 0·01), respectively, compared with the control group. The peripheral blood mononuclear cells of ewes in the supplemented group had a higher (P < 0·05) cytokine mRNA transcription levels for IL-2 (6·4-fold increase), IFN-γ (2·9-fold increase) and transcription factor Bcl6 (2·3-fold increase) compared with the control group. CONCLUSION: We conclude that a 5 days of supplementation with B. toyonensis BCT-7112T prior vaccination is sufficient to significantly improve the humoral immune response of ewes against C. perfringens recombinant ETX vaccine. SIGNIFICANCE AND IMPACT OF THE STUDY: These findings open a new perspective in the utilization of B. toyonensis BCT-7112T as an immunomodulator since a 5 days period of probiotic supplementation is sufficient to improve the vaccine immune response.
Subject(s)
Bacillus , Bacterial Toxins/immunology , Bacterial Vaccines/immunology , Probiotics , Sheep/immunology , Animals , Bacterial Toxins/genetics , Bacterial Vaccines/genetics , Female , Immunity, Humoral , Immunoglobulin G/blood , Immunomodulation , Leukocytes, Mononuclear/immunology , Vaccination , Vaccines, Synthetic/immunologyABSTRACT
O herpesvírus equídeo 1 (EHV-1) apresenta distribuição mundial e causa graves prejuízos à equideocultura. É agente de surtos de doença respiratória, reprodutiva e neurológica, em equídeos jovens e adultos. A glicoproteína D (gD) do envelope viral é essencial para ligação e penetração em células permissivas e direcionamento do sistema imunológico do hospedeiro, induz respostas imunes humorais e celulares, sendo um antígeno apropriado para ser utilizado em vacinas e imunodiagnóstico. O objetivo deste trabalho foi expressar e caracterizar a gD do EHV-1 em Pichia pastoris para posterior utilização como antígeno em técnicas de imunodiagnóstico e formulação de vacinas recombinantes. Uma sequência de DNA que codifica uma forma truncada da gDEHV-1 foi clonada no vetor pPICZαA de expressão em P. pastoris. Obteve-se uma proteína de ~41 kDa, como esperado. A proteína apresentou glicosilação entre 4 kDa e 16 kDa, demonstrada por deglicosilação enzimática. A proteína recombinante foi caracterizada antigenicamente e imunogenicamente por Western blot, utilizando-se anticorpos policlonais equinos anti-EHV-1, e por ELISA indireto em modelo murino, demonstrando que a gD recombinante manteve epítopos similares aos da proteína nativa. Esses resultados sugerem que a gDEHV-1 é um antígeno promissor para uso como imunobiológico no controle do EHV-1.(AU)
Equine herpesvirus 1 (EHV-1) has a worldwide distribution and causes serious damage to horse breeding. It is an agent of respiratory, reproductive and neurological disease outbreaks in young and adult equids. Viral envelope glycoprotein D (gD) is essential for binding and penetration into permissive cells and targeting the host immune system, inducing humoral and cellular immune responses, and is an appropriate antigen for use in vaccines and immunodiagnostics. The objective of this work was to express in Pichia pastoris and to characterize EHV-1 gD for later use as an antigen in immunodiagnostic techniques and formulation of recombinant vaccines. A DNA sequence encoding a truncated form of gDEHV-1 has been cloned into the P. pastoris expression vector pPICZαA. A protein of ~41 kDa was obtained as expected. The protein presented glycosylation between 4 kDa and 16 kDa, demonstrated by enzymatic deglycosylation. The recombinant protein was antigenically and immunogenically characterized by Western blot using equine polyclonal anti-EHV-1 antibodies, and by indirect ELISA in a murine model, demonstrating that the recombinant gD maintained epitopes similar to those of the native protein. These results suggest that gDEHV-1 is a promising antigen for use as an immunobiological in the control of EHV-1.(AU)