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Introduction: Post-thoracotomy air leaks remain a significant challenge in thoracic surgery. Aim: This randomized controlled trial assessed the efficacy of autologous fibrin glue in reducing air leaks following thoracotomy procedures. Material and methods: Conducted as a single-center, single-blind, randomized clinical trial, the study enrolled adult patients undergoing lung resection or decortication at a thoracic surgery clinic. Participants were randomly assigned to either the intervention group, receiving autologous fibrin glue application during surgery, or the control group, undergoing standard surgical procedures without glue application. Key inclusion criteria were adult patients undergoing elective thoracotomy for lung resection or decortication, while exclusion criteria included patients with severe comorbidities or contraindications to fibrin glue. Results: A total of 40 patients were enrolled and randomized equally to the two groups. The group treated with autologous fibrin glue demonstrated a significant reduction in the duration of air leakage and chest tube drainage, along with a shorter hospital stay, compared to the control group. There were no statistically significant differences in postoperative complications between the groups. Conclusions: The application of autologous fibrin glue during thoracotomy procedures significantly reduces postoperative air leaks and hospitalization duration without increasing complication rates. This finding suggests a beneficial role of fibrin glue in thoracic procedures requiring lung resection or decortication.
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Background: Fat graft surgery is one of the most effective procedures in plastic surgery, and since some patients request multiple surgeries and these cases sometimes take hours, it endangers the viability of the fat graft. In this study, we intend to evaluate the viability of adipose tissue aspirated with a syringe at refrigerator (4°C) and freezer (-20 °C) temperatures. Methods: This was a cross-sectional study. After receiving the ethics committee's approval (IR.MUMS.MEDICAL.REC.1401.423), 17 volunteers entered the study. The harvested fat tissue sample was divided into 3 parts, and each of them was transferred to 3 separate sterile tubes. The first tube was sent to the laboratory for preliminary examination of fresh fat, and the second tube was transferred to a 4°C refrigerator for 72 hours. The sample from the third tube was first passed through a strainer and after drying, it was transferred to a -20°C freezer for 72 hours. After treatment with trypsin, we placed the sample inside the centrifuge using the Coleman method. Finally, 3 layers were formed, and the white middle layer was extracted as a fat cell suspension. Tissue samples were stained with trypan blue, and the percentage of viable cells was calculated using an optical microscope. Results: There was a significant difference between the mean number and percentage of viable cells in all 3 groups. Samples in the 4°C refrigerator had significantly more cellular viability than those in the -20°C freezer (mean difference, 72.842%; P < 0.001). Conclusion: Our findings showed that after 72 hours at 4°C, adipose tissue has significantly higher survival than at -20°C (98.93% vs 75.31%). Since the survival of fat cells is one of the direct determinants of fat retention, it can affect the results after surgery. The present study recommends fresh adipose tissue for immediate transplantation unless there is an urgent need for cold storage.
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Introduction: Hypocalcemia is a common complication of total thyroidectomy (TT). This study was designed to investigate the effect of preoperative vitamin.D (Vit.D) status on the occurrence of post-total thyroidectomy hypocalcemia. Materials and Methods: Patients who underwent TT without parathyroidectomy were divided into three groups based on their preoperative Serum Vit.D levels (<20 ng/ml, 20-30 ng/ml, and ≥30 ng/ml were considered deficient, insufficient, and normal Vit.D levels, respectively). Serum levels of calcium and phosphorus were measured before and 24 hours after surgery in all patients. The patients were examined for clinical symptoms and signs of hypocalcemia postoperatively. In cases with positive clinical symptoms and signs of hypocalcemia and/or calcium levels <8 mg/dl, PTH level was measured before starting calcium infusion, while serum calcium and phosphorus levels were also measured 24 hours later. Results: Among 100 patients enrolled in this study, 81% were females. The mean age was 36.60±8.32 years. Before surgery, the mean Vit.D level was 26.9±16.89 ng/ml, while 47% of cases had normal Vit.D level, 32% had insufficient vitamin levels, and 21% had Vit.D deficiency. Twenty-four hours after surgery, the calcium (P=0.356) and phosphorus (P=0.743) levels were not significantly different between the three Vit.D groups. A comparison of postoperative PTH levels between the three Vit.D groups showed no significant difference (P=0.596). Conclusions: Based on our findings, preoperative serum Vit.D levels did not affect postoperative serum calcium levels.
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Introduction: The rate of olfactory loss related to COVID-19 was reported between 4-89 percent. There is no approved treatment for patients who experience anosmia after the mentioned infection. This systematic review aimed to assess the therapeutic effects of corticosteroids on anosmia in COVID-19 patients. Materials and Methods: Databases including PubMed, ISI Web of Sciences, Scopus, and Cochrane Library. Databases were searched up to September 2022 to find out randomized controlled trials that assessed the effect of corticosteroids on post-COVID anosmia/hyposmia. Only studies published in the English language were entered in this review. Results: Among the six relevant trials with a total population of 712, one study administered the combination therapy of both systemic and nasal corticosteroids, while others used intranasal corticosteroids. No significant difference was observed between the intervention (IG) and control (CG) groups in terms of duration of improvement from anosmia (mean difference:-1.799). The pooled effect of self-rating olfactory scores was assessed at 2 weeks and at the end point of the studies which revealed no significant effect in favor of the IG (pooled effect in 2 weeks: 0.739; in the endpoint: 1.32). The objective evaluation with different tools indicated that IG obtained higher scores at the endpoint of treatment. The pooled results showed that the number of patients who recovered from anosmia is higher in IG compared to CG (Odds Ratio: 1.719). Conclusion: It appears that the duration of corticosteroid therapy more than two weeks may be a considerable effect on the recovery of smell dysfunction in COVID-19 patients.
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Introduction: Allergic rhinitis (AR) is a prevalent chronic disease affecting a significant portion of the global population. The substantial economic burden associated with treating AR necessitates the exploration of alternative therapies. Probiotics have gained attention due to their availability, minimal adverse effects, and cost-effectiveness. The present study aims to investigate the role of synbiotics as adjunctive agents in the treatment of AR when added to standard treatment. Method: Thirty patients with persistent allergic rhinitis (PAR) were randomly assigned to receive routine diet therapy plus synbiotics or routine diet therapy plus placebo per day for 4 months. The data analysis was conducted using SPSS Version 20. Result: This study revealed a notable difference in immunoglobulin (Ig)E levels between the placebo and synbiotics groups (p = 0.035) following the intervention. Although a statistically significant difference (p = 0.039) was observed in the changes before and after the intervention (synbiotics and placebo) in the SNOT22 questionnaire, this finding was not observed for the MiniRQLQ questionnaire. For the MiniRQLQ questionnaire, the within-group analysis showed significant changes in activity variables (p = 0.023), ocular symptoms (p = 0.036), and practical problems (p = 0.043) exclusively in the synbiotics group. Additionally, changes in nasal symptoms were observed in both synbiotics (p = 0.006) and placebo (p = 0.007) groups. Conclusion: This study suggests that synbiotics supplementation for 4 months can impact IgE levels compared with placebo in individuals with PAR, while also exhibiting positive effects on symptomology.
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BACKGROUND: The therapeutic potential of oral colchicine administration may help combat COVID-19 infection due to reduced disease severity and mortality risk. OBJECTIVE: This randomized trial aimed to assess the effect of colchicine treatment on the inflammatory and hematologic markers as well as clinical features in non-hospitalized patients with mild-to-moderate COVID-19 disease. METHODS: In the present placebo-controlled randomized trial, 80 non-hospitalized COVID-19 patients were enrolled and followed for 14 days. Subjects randomly received oral colchicine or placebo tablets once a day for two weeks. The fever and cough clinical signs, as well as C-reactive protein (CRP) and lymphopenia, were evaluated through the follow-up. RESULTS: No significant between-group differences were observed in terms of the duration of clinical symptoms, CRP, and lymphopenia at 0, 7, and 14 days of intervention. Although the proportion of participants with fever, cough, positive CRP, and lymphopenia was higher reduced in the colchicine group than the placebo during treatment, no significant differences were found between groups. Due to no adverse effects detected in this trial, colchicine therapy was well-tolerated and safe. CONCLUSION: Our findings revealed that colchicine adjuvant therapy had no beneficial effect on clinical and para-clinical parameters in non-hospitalized COVID-19 patients during 14 days of intervention. The present trial does not support colchicine as a potential treatment against COVID-19 disease.
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Therapeutic approaches with immune-modulatory effects such as probiotics and prebiotics adjuvant therapy may be essential to combat against COVID-19 pandemic. The present trial aimed to reveal the efficacy of synbiotic supplementation on clinical and paraclinical outcomes of hospitalized COVID-19 patients. The current randomized placebo-controlled trial enrolled 78 hospitalized patients with confirmed COVID-19 infection. Participants were randomly allocated to intervention and control groups that received synbiotic or placebo capsules twice daily for 2 weeks, respectively. The synbiotic capsule contains multi-strain probiotics such as Lactobacillus (L.) rhamnosus, L. helveticus, L. casei, Bifidobacterium (B.) lactis, L. acidophilus, B. breve, L. bulgaricus, B. longum, L. plantarum, B. bifidum, L. gasseri, and Streptococcus (S.) thermophilus (109 CFU), as well as fructooligosaccharides prebiotic agent. Besides COVID-19 clinical features, levels of proinflammatory interleukin-6 (IL-6), C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), liver and renal function markers, as well as hematological parameters, were assessed during follow-up. The serum level of IL-6 was significantly decreased in the intervention group compared to the placebo after 2 weeks of intervention (p = 0.002). A significant difference was found regarding the count of white blood cells (WBC) within the synbiotic group from pre to post-treatment (p = 0.004). The levels of ESR (p = 0.935) and CRP (p = 0.952) had a higher reduction trend in the synbiotic group relative to the placebo, with no significant between-group differences. Other findings had no statistical differences between groups. Our results provide the support that synbiotic adjuvant therapy for 2 weeks can be effective to modulate inflammatory responses against COVID-19 infection.
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COVID-19 , Probióticos , Simbióticos , Humanos , Interleucina-6 , Pandemias , Proteína C-Reativa/metabolismo , Método Duplo-CegoRESUMO
BACKGROUND: Polycystic Ovary Syndrome (PCOS) is one of the most prevalent endocrine illnesses among women of reproductive age. PCOS is linked to several issues, including hypothyroidism and metabolic disorders. Hypothyroidism seems to be associated with insulin resistance and other metabolic factors. OBJECTIVE: The present study aimed to evaluate the incidence of hypothyroidism in PCOS patients and compare it with healthy controls. Moreover, the impact of hypothyroidism on metabolic parameters, particularly insulin resistance, in PCOS patients was also examined. METHODS: This study was conducted on 41 women with PCOS and 41 healthy women as controls from March to November, 2018. Participants' demographic information was recorded. Thyroid function tests were compared between the case and control groups. Metabolic parameters were examined between hypothyroid and euthyroid PCOS individuals. RESULTS: Patients with PCOS displayed a greater incidence of hypothyroidism and a higher level of anti-thyroid peroxidase antibodies compared to the control group. High-density lipoprotein (HDL) cholesterol was substantially higher in hypothyroid PCOS patients than in non-hypothyroid individuals, although no significant changes were observed in other metabolic markers. Hypothyroid PCOS patients and those without hypothyroidism did not differ in insulin resistance. Autoimmunity was not found to be linked to a higher risk of metabolic problems. CONCLUSION: As evidenced by the results of this study, PCOS patients had a higher prevalence of subclinical hypothyroidism than healthy subjects. Metabolic indicators, except for HDL, were not different between PCOS patients with and without hypothyroidism.
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Hipotireoidismo , Resistência à Insulina , Síndrome Metabólica , Síndrome do Ovário Policístico , Feminino , Humanos , Síndrome do Ovário Policístico/complicações , Síndrome do Ovário Policístico/epidemiologia , Tireotropina , Hipotireoidismo/complicações , Hipotireoidismo/epidemiologia , Síndrome Metabólica/complicações , Índice de Massa CorporalRESUMO
Background: Colorectal cancer (CRC) is the most prevalent cancer with high mortality worldwide. We aimed to evaluate the incidence of CRC based on the positive fecal immunochemical test (FIT) result in the Iranian population. Methods: The present study was conducted on the health assessment data recorded in the SINA (Integrated Health Information System) in 2018 and 2019 from individuals who had participated in the national program, including asymptomatic people aged 50-69 years or had risk factors of CRC such as family or past personal history of CRC as well as symptomatic individuals, for the early detection and prevention of CRC in Mashhad, Iran. Results: The study participants included 140,463 eligible individuals, of whom 8258 (5.88%) and 145 (2.21%) were positive for FIT and diagnosed with colon cancer, respectively. Unfortunately, only 654 people had undergone colonoscopy. Our results indicated that age, fast food intake (≥two units per day), family history of CRC in first or second-degree relatives, some gastrointestinal diseases such as inflammatory bowel disease (IBD) and CRC, as well as bleeding per anus, constipation, abdominal cramp, and losing body weight were associated with increased risk of positive FIT. However, some other factors, including having a hard job, physical activity, and Iranian nationality (compared to non-Iranians), were associated with a low risk of positive FIT screening tests for CRC. Conclusion: A high number of high-risk persons in Mashhad were positive for the FIT test in 2018-2019, and many of them were diagnosed with CRC, according to the colonoscopy results. Therefore, screening is highly recommended as the first step in the early detection of CRC.
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BACKGROUND: The coronavirus disease 2019 seems to change antibiotic resistance pattern. Certain conditions in the Covid-19 era may be contributing to the rise of antimicrobial resistance (AMR). Due to the limited information on the impact of Covid-19 on antimicrobial resistance (AMR), the purpose of this research was to investigate the trend in antimicrobial resistance changes of E. coli, P. aeruginosa, K. pneumoniae, and A. baumannii in Hasheminezhad hospital. This hospital was a Corona center in Mashhad at the onset of this epidemic. METHODS: 1672 clinical samples were collected between January 21, 2020 and January 30, 2022from patients hospitalized at Hasheminezhad Hospital in Mashhad, Conventional microbiological procedures for identifying gram-negative bacteria and antibiotic susceptibility testing were used, according to the clinical and laboratory standards institute (CLSI) 2021. The two years of the pandemic, from the initial stage of the outbreak until the 6th peak, (January 2020 to and January 2022) were divided into 9 periods according to the seasons. RESULTS: Highest resistance rates were seen in E. coli (615 samples), K. pneumoniae (351 samples), P. aeruginosa (362 samples) and A. baumannii (344 samples) to Ampicillin (89.6%), Ampicillin (98%), Imipenem (91.8%), and Ceftazidime (94.6%), respectively. The largest change in antibiotic resistance was seen between Summer 2020 and Summer 2021 for K. pneumoniae with about a 30% rise in antibiotic resistance to Ceftriaxone. CONCLUSIONS: All 4 species evaluated in this study, have shown rising AMR rates during the first year of the pandemic in the northeast of Iran. This study revealed that E. coli, P. aeruginosa, K. pneumoniae, and A. baumannii strains in Northern Iran have a higher level of antibiotic resistance than what was measured in similar studies conducted before the pandemic. This will further restrict treatment choices and jeopardize global public health.
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Acinetobacter baumannii , COVID-19 , Ampicilina/farmacologia , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , COVID-19/epidemiologia , Ceftazidima/farmacologia , Ceftriaxona/farmacologia , Farmacorresistência Bacteriana , Escherichia coli , Humanos , Imipenem/farmacologia , Irã (Geográfico)/epidemiologia , Klebsiella pneumoniae , Testes de Sensibilidade Microbiana , Pandemias , Pseudomonas aeruginosaRESUMO
BACKGROUND AND AIMS: Olfactory dysfunction is a recognized manifestation in patients infected with Coronavirus Disease 2019 (COVID-19). This investigation aimed to assess the effect of mometasone furoate intranasal spray on the improvement of smell dysfunction in post-COVID-19 patients. MATERIALS AND METHODS: This randomized placebo-controlled trial included 80 non-hospitalized adult patients who had persistent anosmia or severe microsmia for more than 4 weeks due to COVID-19 infection. The participants were randomly allocated to the intervention or placebo group to receive mometasone furoate nasal spray or sodium chloride intranasal spray during 4 weeks of follow-up, respectively. The patients' olfactory dysfunction was assessed in terms of visual analog scale (VAS), and smell test score according to the modified version of the University of Pennsylvania smell identification test for the Iranian population. RESULTS: A total of 70 participants completed the follow-up period and were analyzed in this study. By comparing the olfactory scores including smell test and VAS scores, no significant differences were found between case and control groups at baseline, 2, and 4 weeks intervals. However, the change of both olfactory scores at pre to post-treatment intervals and 2-4 weeks was significantly higher in the mometasone group relative to the placebo group. At post-treatment, the frequency of anosmia was 22.9% reduced in the case group compared to the control group. CONCLUSION: Overall, there was no significant difference in olfactory dysfunction between the two groups during follow-up. However, based on the significant between-group difference in terms of olfactory scores changes, it seems that the nasal corticosteroids may be a positive effect on the recovery process of patients who received more than 2 weeks. LEVEL OF EVIDENCE: 2 Laryngoscope, 132:2209-2216, 2022.
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COVID-19 , Transtornos do Olfato , Corticosteroides , Adulto , Anosmia/tratamento farmacológico , Anosmia/etiologia , COVID-19/complicações , Humanos , Irã (Geográfico) , Furoato de Mometasona , Sprays Nasais , Transtornos do Olfato/tratamento farmacológico , Transtornos do Olfato/etiologia , Olfato , Cloreto de SódioRESUMO
INTRODUCTION: The previous reports on clusterin (CLU) levels in various types of cancer have been controversial and heterogeneous. The present meta-analysis has aimed to evaluate the association between soluble CLU levels and the risk of different human cancers based on observational studies. METHODS: A systematic literature review was conducted to determine the relevant eligible studies in English language from health-related electronic databases up to January 2021. Random effects models were used to calculate the summary standard mean difference (SMD) with 95% confidence intervals (CIs) to identify the correlation between CLU levels and cancer risk. The meta-regression, sensitivity, Galbraith, and subgroup analyses were performed to explore the source of between-study heterogeneity. Furthermore, the funnel plot and Egger's linear regression tests were carried out to evaluate the risk of publication bias. RESULTS: According to 16 eligible articles, 3331 patients and 839 healthy controls were included in our meta-analysis. Overall, the CLU levels were significantly higher in various cancer cases compared to the healthy groups (SMD = 1.50, 95% CI = 0.47-2.53). Moreover, subgroup analysis based on types of cancer showed a significant correlation between CLU levels and the risk of digestive system cancers (SMD = 1.54, 95% CI = 0.91-2.18, P <0.001), especially in HCC (SMD = 1.89, 95% CI = 0.76-3.03, P = 0.001), and CRC (SMD = 1.63, 95% CI = 0.0-3.23, P = 0.048). CONCLUSION: The present meta-analysis indicates a significant association of CLU levels with the risk of digestive system cancers such as hepatocellular carcinoma and colorectal cancer. Therefore, CLU can be monitored as a novel molecular biomarker for the prognosis and diagnosis of various types of cancers particularly in the digestive system.
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Carcinoma Hepatocelular , Neoplasias Hepáticas , Clusterina , Humanos , Prognóstico , RiscoRESUMO
The crucial challenge in tuberculosis (TB) as a chronic infectious disease is to present a novel vaccine candidate that improves current vaccination and provides efficient protection in individuals. The present study aimed to evaluate the immune efficacy of multi-subunit vaccines containing chitosan (CHT)- or trimethyl chitosan (TMC)-coated PLGA nanospheres to stimulate cell-mediated and mucosal responses against Mycobacterium Tuberculosis (Mtb) in an animal model. The surface-modified PLGA nanoparticles (NPs) containing tri-fusion protein from three Mtb antigens were produced by the double emulsion technique. The subcutaneously or nasally administered PLGA vaccines in the absence or presence of BCG were assessed to compare the levels of mucosal IgA, IgG1, and IgG2a production as well as secretion of IFN-γ, IL-17, IL-4, and TGF-ß cytokines. According to the release profile, the tri-fusion encapsulated in modified PLGA NPs demonstrated a biphasic release profile including initial burst release on the first day and sustained release within 18 days. All designed PLGA vaccines induced a shift of Th1/Th2 balance toward Th1-dominant response. Although immunized mice through subcutaneous injection elicited higher cell-mediated responses relative to the nasal vaccination, the intranasally administered groups stimulated robust mucosal IgA immunity. The modified PLGA NPs using TMC cationic polymer were more efficient to elevate Th1 and mucosal responses in comparison with the CHT-coated PLGA nanospheres. Our findings highlighted that the tri-fusion loaded in TMC-PLGA NPs may represent an efficient prophylactic vaccine and can be considered as a novel candidate against TB.
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Quitosana , Nanosferas , Tuberculose , Administração Intranasal , Animais , Camundongos , Ácido Poliglicólico , Copolímero de Ácido Poliláctico e Ácido Poliglicólico , Tuberculose/prevenção & controle , Vacinas de Subunidades AntigênicasRESUMO
BACKGROUND: Primary hyperparathyroidism (PHPT) can lead to renal and skeletal disorders, as well as insulin resistance and impaired glucose metabolism. The current study aimed to assess the effects of parathyroidectomy on insulin resistance in patients with PHPT. MATERIALS AND METHODS: The present study was conducted on 65 patients with PHPT and indications for parathyroidectomy who were referred to the endocrinology clinics of Mashhad University of Medical Sciences. Thereafter, the demographic characteristics of the patients were recorded. Blood tests, including haemoglobin A1c (HbA1c), fasting blood glucose (FBG) and insulin levels, were assessed one week before and three months after the surgery. The insulin resistance score (HOMA-IR) was calculated and compared using the relevant formula. RESULTS: A total of 65 participants with a mean age of 45.44 ± 9.59 years were included in the current study. In one-month postoperative tests, mean scores of FBG (p < .05), insulin level (p < .05) and HbA1c (p < .05) were significantly reduced. Moreover, the HOMA-IR index decreased in 51 patients after the surgery. CONCLUSION: According to our findings, parathyroidectomy can be effective in the reduction of insulin resistance and corresponding complications in patients with PHPT in the present short-term study. However, it has yet to be confirmed as a treatment method for insulin resistance in these patients. Future long-term studies are required to be done to investigate the effect of parathyroidectomy on insulin resistance.
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Hiperparatireoidismo Primário , Resistência à Insulina , Adulto , Hemoglobinas Glicadas , Humanos , Hiperparatireoidismo Primário/complicações , Hiperparatireoidismo Primário/cirurgia , Pessoa de Meia-Idade , Paratireoidectomia , Período Pós-OperatórioRESUMO
OBJECTIVES: The present study aimed to determine the immunoadjuvant efficacy of mixed cross-linked dextran microspheres (CDM) and tetanus toxoid (TT)-loaded trimethyl chitosan (TMC) nanospheres in dry powder form. MATERIALS AND METHODS: The TMC nanoparticles (NPs) containing TT were produced using the ionic gelation method. Co-administration of TT-loaded TMC NPs and CDM as an absorption enhancer was performed to improve immunity against the antigen. Dry powder formulations were delivered via the nasal route in a rabbit model. RESULTS: Among immunization groups, mixing of CDM with TT encapsulated in TMC NPs could elicit the highest titer of systemic IgG antibody. Furthermore, the addition of CDM to TT-loaded TMC enhanced the sIgA response relative to the TT solution. CONCLUSION: The TMC NPs had a considerable effect on mucosal and systemic immunity against the TT antigen. Therefore, the CDM excipient can be utilized for nasal immunization to elevate systemic and mucosal responses.
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BACKGROUND: The engineered chimeric peptides including functional multi-epitope structures fused by various peptide linkers are widely applied in biotechnological research to improve the expression level and biological activity of chimera. OBJECTIVE: The aim of our study was to evaluate the effect of helical and flexible linkers on solubility, expression level and folding of multi-epitope chimera containing four epitopes of Human T Lymphotropic Virus Type 1 (HTLV-1). METHODS: For this purpose, the chimera sequences connected by the helical or flexible linker were inserted into different plasmid vectors and expressed in E. coli strains. The expressed products were analyzed using SDS-PAGE and Western blot techniques. Additionally, the molecular modeling study of the chimera with helical or flexible linker was performed using iterative threading assembly refinement (I-TASSER) to attain their three-dimensional structures. RESULTS: Comparison of the chimera expression indicated that the insertion of a flexible (GGGGS)3 linker among chimera epitopes could significantly enhance the level of expression, whereas, the low-level of chimera expression was observed for chimera containing the contiguous helical (EAAAK)5 linker. According to the results of sequence alignment and plasmid stability test, the structure and function of a consecutive helical linker among chimera epitopes were similar to porins as the outer-membrane pore-forming proteins. The molecular modeling results confirmed our experimental study. CONCLUSION: This investigation illustrated the key role of linker design in determining the expression level of multi-epitope chimera and conformational folding.
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Epitopos , Vírus Linfotrópico T Tipo 1 Humano/genética , Engenharia de Proteínas , Proteínas Recombinantes de Fusão , Proteínas Virais , Epitopos/biossíntese , Epitopos/química , Epitopos/genética , Humanos , Estrutura Secundária de Proteína , Proteínas Recombinantes de Fusão/biossíntese , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/genética , Proteínas Virais/biossíntese , Proteínas Virais/química , Proteínas Virais/genéticaRESUMO
In this investigation, the immunogenicity of HTLV-1 fusion epitope-loaded PLGA nanoparticles (NPs) was assessed in the absence or presence of co-encapsulated CpG ODN adjuvant, in a mice model. For this purpose, the multi-epitope chimera including Tax, env, and gag immunodominant HTLV-1 epitopes was encapsulated in biodegradable PLGA NPs with or without CpG adjuvant. PLGA nanospheres produced by a double emulsion method had a size of <200â¯nm, and encapsulation efficiency of chimera antigen was 85%. The release profile of radiolabeled chimera indicated that only 17.4% and 20.1% of chimera were released from PLGA NPs without or with co-encapsulated CPG ODN during one month, respectively. The PLGA formulations significantly elevated titers of IgG1, IgG2a, and sIgA antibodies, as well as IL-10, and IFN-γ cytokines and also reduced the amount of TGF-ß1 production relative to the other vaccines. Additionally, co-delivery of chimera and CpG ODN in PLGA NPs significantly promoted cellular and mucosal responses compared to the incorporation of CpG and chimera antigen. In summary, these results revealed that the sustained release of chimera from PLGA as an efficient polymeric system elicited potent cell-mediated and mucosal immunity without inflammatory responses against HTLV-1. Therefore, the proper design of vaccine formulation and immunization strategy are crucial factors to construct an efficient vaccine.
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Infecções por HTLV-I/imunologia , Vírus Linfotrópico T Tipo 1 Humano/imunologia , Imunidade nas Mucosas/imunologia , Nanopartículas/química , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/química , Vacinas/imunologia , Adjuvantes Imunológicos/administração & dosagem , Adjuvantes Imunológicos/química , Animais , Antígenos/imunologia , Epitopos/química , Epitopos/imunologia , Imunização/métodos , Imunoglobulina A Secretora/imunologia , Imunoglobulina G/imunologia , Interferon gama/imunologia , Interleucina-10/imunologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Nanosferas/química , Oligodesoxirribonucleotídeos/química , Fator de Crescimento Transformador beta1/imunologia , Vacinação/métodos , Vacinas/químicaRESUMO
This study reports on the immunogenicity assessment of a novel chimeric peptide vaccine including Tax, gp21, gp46, and gag immunodominant epitopes of human T-cell lymphotropic virus type 1 (HTLV-1) to induce immunity against HTLV-1 after subcutaneous (SC) or intranasal administration in a mice model. Additionally, to elevate the efficacy of the HTLV-1 vaccine, the chimera was physically mixed with monophosphoryl lipid A (MPLA) or ISCOMATRIX (IMX) adjuvants. For this purpose, the ISCOMATRIX with a size range of 40-60â¯nm were prepared using lipid film hydration method. Our investigation revealed that the mixture of IMX and chimera could significantly increase antibody titers containing IgG2a, and mucosal IgA, as well as IFN-γ and IL-10 cytokines and decrease the level of TGF-ß1, compared to other vaccine formulations. The intranasal delivery of chimera vaccine in the absence or presence adjuvants stimulated potent mucosal sIgA titer relative to subcutaneous immunization. Furthermore, the SC or nasal delivery of various vaccine formulations could shift the immunity toward cell-mediated responses, as evident by higher IgG2a and IFN-γ, as well as suppressed TGF-ß1 level. Our findings suggest that proper design, construction, and immunization of multi-epitope vaccine are essential for developing an effective HTLV-1 vaccine.