Reactivity Prediction of Cu-Catalyzed Halogen Atom Transfer Reactions Using Data-Driven Techniques.

In catalysis, linear free energy relationships (LFERs) are commonly used to identify reaction descriptors that enable the prediction of outcomes and the design of more effective catalysts. Herein, LFERs are established for the reductive cleavage of the C(sp3)-X bond in alkyl halides (RX) by Cu complexes. This reaction represents the activation step in atom transfer radical polymerization and atom transfer radical addition/cyclization. The values of the activation rate constant, kact, for 107 Cu complex/RX couples in 5 different solvents spanning over 13 orders of magnitude were effectively interpolated by the equation: log kact = sC(I + C + S), where I, C, and S are, respectively, the initiator, catalyst, and solvent parameters, and sC is the catalyst-specific sensitivity parameter. Furthermore, each of these parameters was correlated to relevant descriptors, which included the bond dissociation free energy of RX and its Tolman cone angle θ, the electron affinity of X, the radical stabilization energy, the standard reduction potential of the Cu complex, the polarizability parameter π* of the solvent, and the distortion energy of the complex in its transition state. This set of descriptors establishes the fundamental properties of Cu complexes and RX that determine their reactivity and that need to be considered when designing novel systems for atom transfer radical reactions. Finally, a multivariate linear regression (MLR) approach was adopted to develop an objective model that surpassed the predictive capability of the LFER equation. Thus, the MLR model was employed to predict kact values for >2000 Cu complex/RX pairs.

Making ATRP More Practical: Oxygen Tolerance.

Atom-transfer radical polymerization (ATRP) is a well-known technique for the controlled polymerization of vinyl monomers under mild conditions. However, as with any other radical polymerization, ATRP typically requires rigorous oxygen exclusion, making it time-consuming and challenging to use by nonexperts. In this Account, we discuss various approaches to achieving oxygen tolerance in ATRP, presenting the overall progress in the field.Copper-mediated ATRP, which we first discovered in the late 1990s, uses a CuI/L activator that reversibly reacts with the dormant C(sp3)-X polymer chain end, forming a X-CuII/L deactivator and a propagating radical. Oxygen interferes with activation and chain propagation by quenching the radicals and oxidizing the activator. At ATRP equilibrium, the activator is present at a much higher concentration than the propagating radicals. Thus, oxidation of the activator is the dominant inhibition pathway. In conventional ATRP, this reaction is irreversible, so oxygen must be strictly excluded to achieve good results.Over the last two decades, our group has developed several ATRP techniques based on the concept of regenerating the activator. When the oxidized activator is continuously converted back to its active reduced form, then the catalytic system itself can act as an oxygen scavenger. Regeneration can be accomplished by reducing agents and photo-, electro-, and mechanochemical stimuli. This family of methods offers a degree of oxygen tolerance, but most of them can tolerate only a limited amount of oxygen and do not allow polymerization in an open vessel.More recently, we discovered that enzymes can be used in auxiliary catalytic systems that directly deoxygenate the reaction medium and protect the polymerization process. We developed a method that uses glucose oxidase (GOx), glucose, and sodium pyruvate to very effectively scavenge oxygen and enable open-vessel ATRP. By adding a second enzyme, horseradish peroxidase (HPR), we managed to extend the role of the auxiliary enzymatic system to generating carbon-based radicals and changed ATRP from an oxygen-sensitive to an oxygen-fueled reaction.While performing control experiments for the enzymatic methods, we noticed that using sodium pyruvate under UV irradiation triggers polymerization without the presence of GOx. This serendipitous discovery allowed us to develop the first oxygen-proof, small-molecule-based, photoinduced ATRP system. It has oxygen tolerance similar to that of the enzymatic methods, exhibits superior compatibility with both aqueous media and organic solvents, and avoids problems associated with purifying polymers from enzymes. The system was able to rapidly polymerize N-isopropylacrylamide, a challenging monomer, with a high degree of control.These contributions have substantially simplified the use of ATRP, making it more practical and accessible to everyone.

The role of geographic information system and global positioning system in dementia care and research: a scoping review.

BACKGROUND: Geographic Information System (GIS) and Global Positioning System (GPS), vital tools for supporting public health research, provide a framework to collect, analyze and visualize the interaction between different levels of the health care system. The extent to which GIS and GPS applications have been used in dementia care and research is not yet investigated. This scoping review aims to elaborate on the role and types of GIS and GPS applications in dementia care and research. METHODS: A scoping review was conducted based on Arksey and O'Malley's framework. All published articles in peer-reviewed journals were searched in PubMed, Scopus, and Web of Science, subject to involving at least one GIS/GPS approach focused on dementia. Eligible studies were reviewed, grouped, and synthesized to identify GIS and GPS applications. The PRISMA standard was used to report the study. RESULTS: Ninety-two studies met our inclusion criteria, and their data were extracted. Six types of GIS/GPS applications had been reported in dementia literature including mapping and surveillance (n = 59), data preparation (n = 26), dementia care provision (n = 18), basic research (n = 18), contextual and risk factor analysis (n = 4), and planning (n = 1). Thematic mapping and GPS were most frequently used techniques in the dementia field. CONCLUSIONS: Even though the applications of GIS/GPS methodologies in dementia care and research are growing, there is limited research on GIS/GPS utilization in dementia care, risk factor analysis, and dementia policy planning. GIS and GPS are space-based systems, so they have a strong capacity for developing innovative research based on spatial analysis in the area of dementia. The existing research has been summarized in this review which could help researchers to know the GIS/GPS capabilities in dementia research.

Spray freeze-drying for inhalation application: process and formulation variables.

High porous particles with specific aerodynamic properties were processed by the spray freeze-drying (SFD) method. Comprehensive knowledge about all aspects of the SFD method is required for particle engineering of various pharmaceutical products with good flow properties. In this review, different types of the SFD method, the most frequently employed excipients, properties of particles prepared by this method, and most recent approaches concerning SFD are summarized. Generally, this technique can prepare spherical-shaped particles with a highly porous interior structure, responsible for the very low density of powders. Increasing the solubility of spray freeze-dried formulations achieves the desired efficacy. Also, due to the high efficiency of SFD, by determining the different features of this method and optimizing the process by model-based studies, desirable results for various inhaled products can be achieved and significant progress can be made in the field of pulmonary drug delivery.

The association between living in socially fragmented neighbourhoods and psychological distress among immigrant and non-immigrant people aged 45 and over in NSW, Australia.

OBJECTIVE: Immigrants' mental health is a growing public health concern. Neighbourhood characteristics in the host society may contribute to the poor mental health observed among immigrants. In this study, we aimed to investigate the association between neighbourhood-level social fragmentation and socioeconomic characteristics with psychological distress among immigrants and non-immigrants living in Australia. METHODOLOGY: We conducted cross-sectional secondary data analysis of 228,039 participants from the Sax Institute's 45 and Up Study, with psychological distress measured with the Kessler 10 (K10) and area-level social fragmentation and economic advantage/disadvantage measured at the statistical area level 1 (areas containing approximately 400 people). Multilevel logistic models were used to examine the extent to which differences across the least and most fragmented and economic advantage/disadvantage neighbourhoods contributed to the prevalence of high psychological distress (K10 score ⩾ 22). RESULTS: Immigrants accounted for about 23% of the sample. Slightly more immigrants (34.8%) compared to non-immigrants (32.9%) lived in fragmented areas. Although immigrants were over represented in areas with socioeconomic advantage (40% vs 33.9%), the prevalence of high psychological distress in neighbourhoods with higher social fragmentation and socioeconomic disadvantage was higher in immigrants than non-immigrants. Immigrants had 17% (95% confidence interval = [12%, 22%]) higher odds of having high psychological distress compared to non-immigrants. There was no evidence of an interaction between social fragmentation or socioeconomic disadvantage and immigrant status. Living in fragmented or socioeconomically disadvantaged areas was associated with higher psychological distress among immigrants and non-immigrants. English as a second language and low annual income were significant predictors of psychological distress in immigrants over and above area-level characteristics. CONCLUSION: Immigrants are vulnerable to mental health issues, but the characteristics of the area they live in are also important. Helping immigrants settle into well-integrated and economically advantaged areas may decrease the possibility of mental health issues.

Fat mass changes during menopause: a metaanalysis.

OBJECTIVE: Data: Fat mass has been shown to increase in aging women; however, the extent to which menopausal status mediates these changes remains unclear. The purpose of this review was to determine (1) how fat mass differs in quantity and distribution between premenopausal and postmenopausal women, (2) whether and how age and/or menopausal status moderates any observed differences, and (3) which type of fat mass measure is best suited to the detection of differences in fat mass between groups. STUDY: This review with metaanalyses is reported according to Metaanalysis of Observational Studies in Epidemiology guidelines. STUDY APPRAISAL AND SYNTHESIS METHODS: Studies (published up to May 2018) were identified via PubMed to provide fat mass measures in premenopausal and postmenopausal women. We included 201 cross-sectional studies in the metaanalysis, which provided a combined sample size of 1,049,919 individuals and consisted of 478,734 premenopausal women and 571,185 postmenopausal women. Eleven longitudinal studies were included in the metaanalyses, which provided a combined sample size of 2472 women who were premenopausal at baseline and postmenopausal at follow up. RESULTS: The main findings of this review were that fat mass significantly increased between premenopausal and postmenopausal women across most measures, which included body mass index (1.14 kg/m2; 95% confidence interval, 0.95-1.32 kg/m2), bodyweight (1 kg; 95% confidence interval, 0.44-1.57 kg), body fat percentage (2.88%; 95% confidence interval, 2.13-3.63%), waist circumference (4.63 cm; 95% confidence interval, 3.90-5.35 cm), hip circumference (2.01 cm; 95% confidence interval, 1.36-2.65 cm), waist-hip ratio (0.04; 95% confidence interval, 0.03-0.05), visceral fat (26.90 cm2; 95% confidence interval, 13.12-40.68), and trunk fat percentage (5.49%; 95% confidence interval, 3.91-7.06 cm2). The exception was total leg fat percentage, which significantly decreased (-3.19%; 95% confidence interval, -5.98 to -0.41%). No interactive effects were observed between menopausal status and age across all fat mass measures. CONCLUSION: The change in fat mass quantity between premenopausal and postmenopausal women was attributable predominantly to increasing age; menopause had no significant additional influence. However, the decrease in total leg fat percentage and increase in measures of central fat are indicative of a possible change in fat mass distribution after menopause. These changes are likely to, at least in part, be due to hormonal shifts that occur during midlife when women have a higher androgen (ie, testosterone) to estradiol ratio after menopause, which has been linked to enhanced central adiposity deposition. Evidently, these findings suggest attention should be paid to the accumulation of central fat after menopause, whereas increases in total fat mass should be monitored consistently across the lifespan.

Expression and hydroxyurea-triggered induction of EGFP upon CRISPR/Cas9-mediated integration into the γ-globin gene of K562 cells.

OBJECTIVE: To knock-in an EGFP cassette into the γ-globin genes of K562 cells via CRISPR/Cas9, and to assess expression and hydroxyurea (HU)-mediated induction of the targeted EGFP transgene. RESULTS: The EGFP cassettes were specifically knocked into the Gγ gene. EGFP expression was detected in the targeted cell population and isolated clones. Furthermore, EGFP transcript and fluorescence levels were significantly induced following HU-treatment. CONCLUSION: This system is readily utilizable for genome scale studies of cis-acting regulatory elements which are implicated in γ-globin expression or HU-mediated induction.

The cerebellum shrinks faster than normal ageing in Alzheimer's disease but not in mild cognitive impairment.

BACKGROUND: While acceleration in age-related cerebral atrophy has been well documented in Alzheimer's disease, the cerebellar contributions to this effect have not been thoroughly investigated. OBJECTIVE: This study investigated cerebellar volume and atrophy rate using magnetic resonance imaging in individuals with normal cognition (CN), mild cognitive impairment (MCI), and Alzheimer's disease (AD). METHODS: Two hundred twenty-nine CN, 398 MCI and 191 AD participants of stage I ADNI database with screening scans were evaluated for cerebellar volume. Of those, 758 individuals with two or more follow-up scans were categorized into stable, converted, and reverted CN, MCI and AD and evaluated for cerebellar atrophy rate. RESULTS: Cerebellar volume was 2.5% larger in CN than in those with AD but there were no differences between CN and MCI and MCI and AD in cross-sectional analysis. Similarly, the atrophy rate was 49% larger in AD and 64% larger in MCI who converted to AD but no difference was detected between CN and MCI. There were no association between education and APOEe4 and cerebellar volume or cerebellar atrophy across the diagnostic groups. CONCLUSION: Cerebellar atrophy contributes to Alzheimer's clinical progression but mostly at the late stage of the disease. However, even in the late stage shrinkage rate is less than the average of the shrinkage in the cerebrum and is not associated with AD moderators. This suggests that cerebellar involvement is secondary to cerebral involvement and can be due to network connection spread regardless of the primary pathology. Hum Brain Mapp, 2017. © 2017 Wiley Periodicals, Inc. Hum Brain Mapp 38:3141-3150, 2017. © 2017 The Authors Human Brain Mapping Published by Wiley Periodicals, Inc. Hum Brain Mapp 38:3141-3150, 2017. © 2017 Wiley Periodicals, Inc.

Numerical solutions of the nonlinear fractional-order brusselator system by Bernstein polynomials.

In this paper we propose the Bernstein polynomials to achieve the numerical solutions of nonlinear fractional-order chaotic system known by fractional-order Brusselator system. We use operational matrices of fractional integration and multiplication of Bernstein polynomials, which turns the nonlinear fractional-order Brusselator system to a system of algebraic equations. Two illustrative examples are given in order to demonstrate the accuracy and simplicity of the proposed techniques.

The Integrated Atlas of Dementia Care in the Australian Capital Territory: A Collective Case Study of Local Service Provision.

Background: This study evaluates the dementia care system in a local area and aimed to include all specialised services designed to provide health and social services to people with dementia or age-related cognitive impairment, as well as general services with a high or very high proportion of clients with dementia. Methods: The study used an internationally standardised service classification instrument called Description and Evaluation of Services and DirectoriEs for Long Term Care (DESDE-LTC) to identify and describe all services providing care to people with dementia in the Australian Capital Territory (ACT). Results: A total of 47 service providers were eligible for inclusion. Basic information about the services was collected from their websites, and further information was obtained through interviews with the service providers. Of the 107 services offered by the 47 eligible providers, 27% (n = 29) were specialised services and 73% (n = 78) were general services. Most of the services were residential or outpatient, with a target population mostly of people aged 65 or older, and 50 years or older in the case of Aboriginal and Torres Strait Islander Australians. There were government supports available for most types of care through various programmes. Conclusions: Dementia care in the ACT relies heavily on general services. More widespread use of standardised methods of service classification in dementia will facilitate comparison with other local areas, allow for monitoring of changes over time, permit comparison with services provided for other health conditions and support evidence-informed local planning.

The effect of occupational burnout on cognitive failure and general health in industrial workers (case study: Iran Tire Factory).

BACKGROUND: Human resources are the most important organizational resources and play the most important role in the production and productivity cycle. Considering the importance of people's health and the study of their burnout as a possible cause of occupational cognitive failures, this study aimed to investigate the relationship between burnout, cognitive failure, and general health. PARTICIPANTS AND PROCEDURE: A cross-sectional-analytical study was conducted in Iran Tire Factory. The statistical population of this study was 302 personnel who were randomly selected. Data were collected by four valid questionnaires (demographic information, Maslach burnout, cognitive failure, and general health questionnaire). Then data were analyzed using SPSS software. RESULTS: The results of the analysis revealed a significant and direct relationship between burnout and cognitive failure (p < .001), and a significant inverse relationship was found between cognitive failure and physical health (p = .022). The other results showed that emotional exhaustion and depersonalization dimensions are significantly associated with cognitive failure (p < .001, p = .016). CONCLUSIONS: According to the results of this study, burnout causes cognitive failures among factory personnel and on the other hand, cognitive failures affect the physical health of individuals and lead to deterioration of physical health, which in turn can reduce a person's performance and reduce work efficiency.

Reverse expression pattern of sirtuin-1 and histone deacetylase-9 in coronary artery disease.

BACKGROUND: SIRT1 and HDAC 9 genes are related to inflammation and may contribute to the pathogenesis of coronary artery disease (CAD). We aimed to evaluate the expression level, methylation profile and polymorphisms of these genes in CAD patients. METHODS: In this study, 50 CAD patients and 50 healthy individuals were recruited. The expression level change was evaluated using the TaqMan Real-Time PCR method. The methylation of genes promoter and genotyping of polymorphisms were evaluated by the HRM. RESULTS: The expression level of SIRT1 was reduced while the HDAC9 expression level showed a significant elevation (p < .001). The SIRT1 gene promoter was hypomethylated and the HDAC9 gene promoter was hypermethylated in CAD patients. Also, CG + GG genotype in SIRT1 and both genotypes in the HDAC9 gene were associated with expression change. CONCLUSIONS: SIRT1 and HDAC9 genes, expression changes can be suggested as a potential biomarker for CAD detection.

Standardized Systematic Description of Provision of Care for Multiple Sclerosis at a Local Level: A Demonstration Study.

BACKGROUND: This study evaluates and describes the pattern of services provided for people living with multiple sclerosis (MS) in a local area as a starting point for a more global assessment. METHODS: A health care ecosystem approach has been followed using an internationally standardized service classification instrument-the Description and Evaluation of Services and DirectoriEs for Long Term Care (DESDE-LTC)-to identify and describe all services providing care to people with MS in the Australian Capital Territory, Australia. Available services were classified according to the target population into those specifically dedicated to people living with MS and those providing general neurologic services, both public and private, and across both social and health sectors. RESULTS: A limited range of services was available. There were no local facilities providing or coordinating multidisciplinary integrated care specific to people with MS. Subspecialty services specific to MS were limited in number (6 of the 28 services), and use of specialist services provided in neighboring states was frequently reported. Overall, very few services were provided outside the core health sector (4%). CONCLUSIONS: The provision of care to people living with MS in the Australian Capital Territory is fragmented and relies heavily on generic neurology services in the public and private sectors. More widespread use of the DESDE-LTC as a standardized method of service classification in MS will facilitate comparison with other local areas, allow monitoring of changes over time, and permit comparison with services provided for other health conditions (eg, dementia, mental disorders).

Patterns of mental healthcare provision in rural areas: A demonstration study in Australia and Europe.

Introduction: Mental healthcare systems are primarily designed to urban populations. However, the specific characteristics of rural areas require specific strategies, resource allocation, and indicators which fit their local conditions. This planning process requires comparison with other rural areas. This demonstration study aimed to describe and compare specialized rural adult mental health services in Australia, Norway, and Spain; and to demonstrate the readiness of the healthcare ecosystem approach and the DESDE-LTC mapping tool (Description and Evaluation of Services and Directories of Long Term Care) for comparing rural care between countries and across areas. Methods: The study described and classified the services using the DESDE-LTC. The analyses included context analysis, care availability, placement capacity, balance of care, and diversity of care. Additionally, readiness (Technology Readiness Levels - TRL) and impact analyses (Adoption Impact Ladder - AIL) were also assessed by two independent raters. Results: The findings demonstrated the usability of the healthcare ecosystem approach and the DESDE-LTC to map and identify differences and similarities in the pattern of care of highly divergent rural areas. Day care had a greater weight in the European pattern of care, while it was replaced by social outpatient care in Australian areas. In contrast, care coordination was more common in Australia, pointing to a more fragmented system that requires navigation services. The share between hospital and community residential care showed no differences between the two regions, but there were differences between catchment areas. The healthcare ecosystem approach showed a TRL 8 (the tool has been demonstrated in a real-world environment and it is ready for release and general use) and an AIL of 5 (the target public agencies provided resources for its completion). Two experts evaluated the readiness of the use of DESDE-LTC in their respective regional studies. All of them were classified using the TRL. Discussion: In conclusion, this study strongly supports gathering data on the provision of care in rural areas using standardized methods to inform rural service planning. It provides information on context and service availability, capacity and balance of care that may improve, directly or through subsequent analyses, the management and planning of services in rural areas.

Variation in the expression level of MALAT1, MIAT and XIST lncRNAs in coronary artery disease patients with and without type 2 diabetes mellitus.

Background: The MALAT1, MIAT, and XIST long noncoding RNAs (lncRNAs) participate in the pathogenesis of complex diseases and also serve as diagnostic markers. The study aimed to assess their expressions in CAD patients with or without T2DM against diabetic and non-diabetic controls.Methods: The expression levels of three lncRNAs in 50 CAD patients (with or without diabetes) and 50 non-CAD subjects (with or without diabetes) were evaluated by using the TaqMan Assay method.Results: MALAT1 and MIAT were upregulated in CAD patients (p Value= .0008 and .0078, respectively). The expression level of XIST was significantly elevated diabetic compared to non-diabetic CAD patients (p Value= .0003). MALAT1 gene had the highest diagnostic power for discrimination of CAD patients from controls (AUC= 0.682, p Value=.001).Conclusions: The current study supports the participation of lncRNAs in the pathogenesis of CAD and T2DM and highlights their potential as diagnostic biomarkers.

The T Cell Receptor Repertoire Diversity Following Hematopoietic Stem Cell Transplantation.

Immune reconstitution after hematopoietic stem cell transplantation (HSCT) with a conditioning regimen has appeared to be a promising treatment for autoimmune diseases and hematologic malignancies. This study aimed to assess the T cell receptor (TCR) repertoire diversity in CD4+ cells of patients with hematological malignancies who received allogeneic or autologous HSCT. The diversity of the TCR repertoire was evaluated in 13 patients with hematologic malignancies before and four months after HSCT. Amino acid changes in the 25 Vß families were evaluated using Spectratyping and data were presented as Hamming distance (HD). HD more than 20% was considered a change in TCR repertoire after HSCT. The mean HD was significantly changed after transplantation in all Vß gene families, with most amino acid changes in p4 and p22 families. There was a strong negative correlation between the HD as the index of TCR repertoire and age (r = -0.62,). The results revealed no association between HD mean and parameters such as sex, disease, conditioning regimen, and type of transplantation. Our data revealed that commonly used conditioning regimens in Iran could successfully cause TCR repertoire diversity in patients with hematologic malignancies in the short term. The amount of change in TCR repertoire was inversely correlated with the increasing age of patients.

Patterns of service provision in child and adolescent mental health care in Australia.

Standard description of local care provision is essential for evidence-informed planning. This study aimed to map and compare the availability and diversity of current mental health service provision for children and adolescents in Australia. We used a standardised service classification instrument, the Description and Evaluation of Services and DirectoriEs (DESDE) tool, to describe service availability in eight urban and two rural health districts in Australia. The pattern of care was compared with that available for other age groups in Australia. Outpatient care was found to be the most common type of service provision, comprising 212 (81.2%) of all services identified. Hospital care (acute and non-acute) was more available in urban than in rural areas (20 services [9.7%] vs 1 [1.8%]). The level of diversity in the types of care available for children and adolescents was lower than that for the general adult population, but slightly higher than that for older people in the same areas. Standardised comparison of the pattern of care across regions reduces ambiguity in service description and classification, enables gap analysis and can inform policy and planning.

A new general integral transform for solving integral equations.

Introduction: Integral transforms are important to solve real problems. Appropriate choice of integral transforms helps to convert differential equations as well as integral equations into terms of an algebraic equation that can be solved easily.During last two decades many integral transforms in the class of Laplace transform are introduced such as Sumudu, Elzaki, Natural, Aboodh, Pourreza, Mohand, G_transform, Sawi and Kamal transforms. Objectives: In this paper, we introduce a general integral transform in the class of Laplace transform. We study the properties of this transform. Then we compare it with few exiting integral transforms in the Laplace family such as Laplace, Sumudu, Elzaki and G\_transforms, Pourreza, Aboodh and etc. Methods: A new integral transform is introduced. Then some properties of this integral transform are discussed. This integral transform is used to solve this new transform is used for solving higher order initial value problems, integral equations and fractional order integral equation. Results: It is proved that those new transforms in the class of Laplace transform which are introduced during last few decades are a special case of this general transform. It is shown that there is no advantage between theses transforms unless for special problems. Conclusion: It has shown that this new integral transform covers those exiting transforms such as Laplace, Elzaki and Sumudu transforms for different value of p(s) and q(s). We used this new transform for solving ODE, integral equations and fractional integral equations. Also, we can introduce new integral transforms by using this new general integral transform.

Development of Novel Adhesive Bilayer Lyophilized Wafer of Moxifloxacin as a Modern Wound Dressing.

 Wound healing is a complex process and is influenced by different factors. Aimed to enhance the wound healing procedure, the Moxifloxacin bilayer wafer was designed, optimized and evaluated as an advanced wound healing dressing. The wafers were prepared by the lyophilization and casting method. Optimization was done according to the results of bioadhesion force, swelling index, release rate, T40 and T90 (the time to reach 40% and 90% of release). The optimized wafer was evaluated against in-vitro and in-vivo efficacy using the disc diffusion method and histologic evaluation after application on the wound. The optimized formulation contained HPMC, MC, gelatin and PVP with mounts of 50 mg, 25 mg, 2 mg and 10 mg respectively. The hydrophilic bilayer wafer is adhered to the wound up to the end of wound healing. Application of optimized formulation led to the healing of wound 6 days faster without any sign of infection. The application of this wafer promoted wound healing and epithelium regeneration without any inflammation.

Cloning and Expression of N-CFTX-1 Antigen from Chironex fleckeri in Escherichia coli and Determination of Immunogenicity in Mice.

BACKGROUND: Most jellyfish species are poisonous. Human victims of jellyfish sting each year are 120 million. Chironex fleckeri is a venomous box jellyfish that inflicts painful and potentially fatal stings to humans. The CfTX-1 is one of the antigenic proteins of venom that is suggested to stimulate the immune system for treatment and vaccine. This study aimed to clone and express the CfTX-1 antigen in E. coli and then to determine the synthesis of related antibody in the mice. METHODS: The study was performed in the Persian Gulf and Oman Sea Ecology Research Center, Bandar Abbas, Iran in autumn 2016. The synthetic CfTX-1 gene in PUC57 plasmid was purchased from Nedaye Fan Company. The 723 bp fragment of N-CfTX-1 was amplified by PCR, PUC57 plasmid containing CfTX-1 with BamHI SalI restriction enzyme sites were subcloned in pET28a [+] expression vector and transformed into E. coli BL21 (DE3). The CfTX-1 gene expression was induced by IPTG. Then antibody produced from the mice serum were isolated and confirmed by ELISA. After protein purification, resulted antigen was injected to mice in 4 repeats and then evaluated the rate of antibody in mice serum. Mice were challenged by the Carybdea alata. RESULTS: The 726 bp of N-CfTX-1 were cloned in a vector of expression pET28a [+] and confirmed by PCR, sequencing and enzymatic analysis. Moreover, the recombinant protein was confirmed by SDS-PAGE and Western blotting. Then the antibody was isolated from mice serum and confirmed by ELISA test. The results showed that immunized mice tolerated 50x LD501 of jellyfish venom. CONCLUSION: The CfTX-1 recombinant protein was able to protect the BALB/c mice against jellyfish venom. The produced protein can be used as a candidate for vaccine against jellyfish venom.