Improved Immunogenicity of Tetanus Toxoid by Brucella abortus S19 LPS Adjuvant

ABSTRACT Background: Adjuvants are used to increase the immunogenicity of new generation vaccines, especially those based on recombinant proteins. Despite immunostimulatory properties, the use of bacterial lipopolysaccharide (LPS) as an adjuvant has been hampered due to its toxicity and pyrogenicity. Brucella abortus LPS is less toxic and has no pyrogenic properties compared to LPS from other gram negative bacteria. Objectives: To evaluate the adjuvant effect of B. abortus (vaccine strain, S19) LPS for tetanus toxoid antigen (TT) and to investigate the protective effect of different tetanus vaccine preparations. Methods: LPS was extracted and purified from B. abortus S19 and KDO, glycan, phosphate content, and protein contamination were measured. Adipic acid dihydrazide (ADH) was used as a linker for the conjugation of TT to LPS. Different amounts of B. abortus LPS, TT, TT conjugated with LPS, and TT mixed with LPS or complete Freund's adjuvant (CFA) were injected into mice and antibody production against TT was measured. The protective effect of induced antibodies was determined by LD50. Results: Immunization of mice with TT+LPS produced the highest anti-TT antibody titer in comparison to the group immunized with TT without any adjuvant or the groups immunized with TT-LPS or TT+CFA. Tetanus toxid-S19 LPS also produced a 100% protective effect against TT in immunized mice. Conclusion: These data indicate that B. abortus LPS enhances the immune responses to TT and suggest the possible use of B. abortus LPS as an adjuvant in vaccine preparations

Outcomes of Phaco-viscocanalostomy in Primary Open Angle Glaucoma versus Pseudoexfoliation Glaucoma

Purpose: Viscocanalostomy represents an alternative to standard penetrating glaucoma surgery. The aim of this study is to compare the outcomes of combined phacoemulsification and viscocanalostomy in eyes with primary open-angle glaucoma (POAG) versus eyes with pseudoexfoliation glaucoma (PEXG). Methods: In this prospective non-randomized comparative study, eyes with cataract and POAG or PEXG were enrolled. Pre- and postoperative data including best corrected visual acuity (BCVA), intraocular pressure (IOP), and the number of antiglaucoma medications administered were recorded at each visit. All patients underwent phacoviscocanalostomy. Complete success was defined as the IOP of 21 mmHg or less without the administration of medication while a qualified success reported the same IOP parameters either with or without the administration of medication. Results: Fifty-four eyes with POAG and fifty-four with PEXG underwent phacoviscocanalostomy. The mean follow-up time was 23.36 ± 8.8 months (range, 6–40 months). The mean postoperative IOP reduced significantly in both groups, although the mean IOP reduction was significantly greater in PEXG eyes (14.7 ± 8.9 vs 10.1 ± 7.7 mmHg) (P = 0.05). At the final follow-up visit, the mean postoperative IOP was 14.1 ± 2.1 and 16.6 ± 3.5 mmHg in the PEXG and POAG eyes, respectively (P = 0.001). A complete success rate of 88.9% and 75.9% was achieved in PEXG and POAG eyes, respectively (P = 0.07). The qualified success rate was 100% in the PEXG and 85.2% in POAG groups (P = 0.03). Conclusion: Phacoviscocanalostomy achieved significant IOP reduction and visual improvement in both POAG and PEXG patients. Our results indicated that in terms of IOP reduction, this procedure was more effective in treating PEXG

Association of glutathione S-transferase polymorphisms with the severity of mustard lung

Introduction: Glutathione S-transferase (GST) is one of the major detoxifiers in alveoli. Polymorphism in GST genes can influence the ability of individuals to suppress oxidative stress and inflammation. The present study was aimed to explore the hypothesis that the genetic polymorphisms of GST T1, M1 and P1 are associated with the severity of the mustard lung in the sulfur mustard-exposed individuals. Methods: Blood samples were taken from 185 sulfur mustard-exposed and 57 unexposed subjects. According to the stage of the mustard lung, sulfur mustard-exposed patients were categorized in the mild/moderate and severe/very severe groups. A multiplex PCR method was conducted to identify GSTM1 and GSTT1 null genotypes. To determine the polymorphisms of GSTP1 in exon 5 (Ile105Val) and exon 6 (Ala114Val), RFLP-PCR method was performed. Results: The frequency of GSTM1 homozygous deletion was significantly higher in the severe/very severe patients compared with the mild/moderate subjects (66.3% vs. 48%, P = 0.013). The GSTM1 null genotype was associated with the severity of mustard lung (adjusted odds ratio [OR], 2.257; 95% CI, 1.219-4.180). There was no significant association between GSTT1 and GSTP1 polymorphisms with the severity of the mustard lung. Conclusion: The different distribution of GSTM1 null genotype in severe/very severe and mild/moderate groups indicated that the severity of the mustard lung might be associated with the genetic polymorphism(s)

Inhibitory effects of thymol and carvacrol on heme degradation and oxidative products due to tartrazine: In silico and in vitro studies

The pathology of many diseases arises from oxidative stress and cell destruction. Antioxidant application is one of the most important ways for oxidative stress prevention in the cells and its consequent effects. The present study investigated the natural antioxidants inhibitory effects of thymol and carvacrol on human hemoglobin treated with tartrazine. Purified hemoglobin from human blood samples was treated with tartrazine alone or in combination with mentioned natural antioxidants (thymol and carvacrol). Treated samples were picked up at regular time intervals and changes were followed by UV–visible and fluorescence spectroscopic assays, and circular dichroism spectroscopy (CD). The result of fluorescence spectroscopy revealed that thymol and carvacrol prevented the production of heme-degradation products and advanced glycation end products (AGEs) caused by hemoglobin oxidation with tartrazine. The results of UV–visible and fluorescence spectroscopy revealed the positive effect of these antioxidants on preserving Hb folding, heme, and especially the porphyrin ring surrounding the microenvironment. The results of the circular dichroism (CD) assay showed the protection of alpha helix structure in hemoglobin treated with thymol and carvacrol compared to the control sample. The mentioned antioxidants caused hemoglobin resistance against tartrazine's destructive effect by preventing both heme degradation and glycemic toxins formation and thus reducing the rate of oxidative processes. This matter can be important for various pharmaceutical, health, and cosmetic industries

Comparing Serum Zinc and Cortisol Concentrations in Patients with Major Depressive Disorder Treated with Fluoxetine and Cognitive-Behavioral Therapy: A Randomized Clinical Trial

Background and purpose: Major depressive disorder (MDD) is a massive challenge for community mental health. Serum zinc and cortisol are suitable biomarkers for assessing the response to a given treatment. In the present study, serum concentrations of these markers were compared between two groups of patients treated with fluoxetine and cognitive behavioral therapy. Materials and methods: In this randomized clinical trial study, 40 female MDD (DSM-V criteria) patients were divided into two groups: fluoxetine therapy (20 mg/daily/3 months, n=20) and group CBT (90-min/once a week/12 weeks, n=20). Severity of depression was studied by Beck Depression Inventory (BDI-II) and serum cortisol and zinc concentrations were measured before and after treatments. Data were analyzed in Graphpad Prism 9. Results: BDI-II scores significantly reduced in both fluoxetine therapy (from 23.2±1.5 to 11.1± 1.1) and CBT (from 26.8±0.6 to 9.5±0.8) groups after treatments. Serum cortisol level did not change in patients treated with fluoxetine, but it decreased in CBT group (from 453 ± 0.09 to 386.4±0.07). Serum zinc level decreased in the fluoxetine group (from 91.6±4.8 to 72.8±2.8) and increased in the CBT group (from 78.9±2.1 to 86.5±2.6). Post intervention inter-group comparison showed greater decrease in BDI-II score and cortisol concentration in CBT group than the fluoxetine therapy group. Conclusion: The decrease in BDI-II score and cortisol level and increase in serum zinc were higher in CBT group than fluoxetine group. Therefore, CBT seems to be suitable for treatment of MDD. Further studies are needed with larger sample size and longer study periods. (Clinical Trials Registry Number: IRCT20190710044171N1

Human Papillomavirus Type16- L1 VLP Production in Insect Cells

Objective(s):  Infection by high-risk papillomavirus is regarded as the major risk factor in the development of cervical cancer. Recombinant DNA technology allows expression of the L1 major capsid protein of HPV in different expression systems, which has intrinsic capacity to self-assemble into viral-like particles (VLP). VLPS are non-infectious, highly immunogenic and can elicit neutralizing antibodies. VLP-based HPV vaccines can prevent persistent HPV infections and cervical cancer. In this study recombinant HPV-16 L1 protein was produced in Sf9 insect cells and VLP formation was confirmed. Materials and Methods: Complete HPV-16 L1 gene was inserted into pFast HTa plasmid and transformed into DH10BAC Escherichia coli containing bacmid and helper plasmid. The recombinant Bacmid colonies turned to white and non-recombinant colonies harboring L1 gene remained blue in the presence of X-gal and IPTG in colony selection strategy. To confirm the recombinant bacmid production, PCR was applied using specific L1 primers. To produce recombinant baculovirus, the recombinant bacmid DNA was extracted and transfected into Sf9 cells using Cellfectin. The expression of L1 in Sf9 cells was identified through SDS-PAGE and western blot analysis using specific L1 monoclonal antibody. Self-assembled HPV16L-VLPs in Sf9 cells was confirmed by electron microscopy. Results:The recombinant protein L1 was predominantly ~60 KD in SDS-PAGE with distinct immunoreactivity in western blot analysis and formed VLPS as confirmed by electron microscopy. Conclusion:Application of recombinant baculovirus containing HPV-16 L1 gene will certainly prove to be a constructive tool in production of VLPs for prophylactic vaccine development as well as diagnostic tests

Serum cortisol level and adrenal reserve as a predictor of patients’ outcome after successful cardiopulmonary resuscitation

Introduction: It is thought that pituitary-adrenal axis has a fundamental role in outcome of cardiopulmonary arrest (CPA). This study designed to evaluate the correlation between adrenal reserve and post-resuscitation outcome. Methods: In this clinical trial study, 52 consecutive patients with CPA were enrolled in two emergency departments (EDs) over a 3-month period. Plasma cortisol level was measured at the beginning of CPR. Intravenous adrenocorticotropic hormone (ACTH) stimulation test was carried out after successful CPR, and blood samples were taken at 30 and 60 minutes, and 24 hours thereafter. Patients were divided into two groups: in-hospital death or hospital discharge. Results: In patients who died, baseline and post-ACTH serum cortisol after 30 and 60 minutes and 24 hours were higher than patients who discharged from the hospital, but it was not statistically significant except to that of minute 60 (P = 0.49). A model of multivariate logistic regression analysis showed that age and need for vasopressor infusion correlated with mortality. Conclusion: Current study could not show the statistically significant difference in initial and post-ACTH serum cortisol levels between survivor and non-survivor patients with cardiac arrest who had initial successful CPR, except to that of minute 60

Comparison of the study parameters between responders and nonresponders at baseline.

<p>Comparison of the study parameters between responders and nonresponders at baseline.</p

Expression of Recombinant Human Programmed Cell Death Protein-1 and Development of Camel Polyclonal Antibody

Aims Programmed cell death protein-1 (PD-1) is a membrane receptor expressed on the surface of T and B lymphocytes, monocytes, natural killers, and dendritic cells. In cancer, the PD-1/PD-L1 system prevents the proliferation of T lymphocytes and causes the release of cytokines and cytotoxicity, which leads to the apoptosis of tumor-specific T cells, thereby preventing the immune response to cancer cells.  Methods & Materials In this study, the extracellular part of the humanized PD-1 protein was cloned and expressed, and the protein was injected as an antigen into a camel (Camelus dromedarius) to obtain a camel polyclonal antibody against PD-1 protein.  Findings The obtained results indicate the proper expression of the protein in the prokaryotic system. Also, using various tests, such as ELISA and western blot, it was confirmed that the polyclonal antibody obtained from camel can identify PD-1 protein.  Conclusion This study showed that because of the advantages, such as the ability to bind multiple epitopes, camel polyclonal antibodies can be used in antibody-based research for effective and strong molecular applications to detect PD-1 receptors

Change in the study parameters from baseline to three months of metformin therapy in responders and non-responders receiving atorvastatin.

<p>Change in the study parameters from baseline to three months of metformin therapy in responders and non-responders receiving atorvastatin.</p