Synthesis of CS-Fe3O4/GO nanocomposite for adsorption of heavy metal in aqueous environment.

In this study, magnetic material based on graphene oxide (GO) was developed for enhanced adsorption capacity for heavy metals. The Fe3O4nanoparticles were combined with the GO material using a chitosan (CS) binder to obtain the CS-Fe3O4/GO nanocomposite. The adsorption capacity of this nanocomposite was evaluated by removing heavy metals including Ni2+ions. When GO was composed with Fe3O4and CS, the GO films were densely covered with ferromagnetic particles, which were bound and densely distributed on the GO film surface due to the interaction between GO and CS. The optimal conditions for the complexation of Ni2+and 4-(2-pyridyl azo)-rezoxine (PAR) are 1 ml Ni2+, 2 ml PAR 100 mg l-1,pH= 6 (adjusted with 0.7 ml of the 0.1 M K2HPO4solution) and a complexation time of 20 min. After 50 min of adsorption, the Ni2+removal efficiency of the CS-Fe3O4/GO nanocomposite reached 81.21% and the corresponding adsorption capacity was 2.03 mg g-1. The Ni2+removal process followed the first-order model and Freundlich isotherm. This process was spontaneous (ΔGo< 0) and an exothermic process (ΔHo= -1128.875 J·mol-1). In addition, the factors affecting this process were investigated, including thepHsolution, the dosage of the CS-Fe3O4/GO nanocomposite and the initial Ni2+concentration. The CS-Fe3O4/GO nanocomposite showed a potential adsorption capacity in removing Ni2+at low concentrations from wastewater.

Fe3O4/Graphene Oxide/Chitosan Nanocomposite: A Smart Nanosorbent for Lead(II) Ion Removal from Contaminated Water.

A new graphene oxide (GO) nanocomposite that contains chitosan, a biological polymer, combined with a magnetic nanoparticle inorganic material (Fe3O4) was successfully prepared and applied for the adsorption of Pb(II) from aqueous solutions. The structural and morphological properties of the GO/Fe3O4/CS (GFC) nanocomposites were characterized by X-ray diffraction, scanning electron microscopy, and energy-dispersive X-ray spectroscopy. Influent factors for Pb(II) adsorption, including the contacting time, pH of the working medium, working temperature, and adsorbent dosage on the adsorption efficiency, have been optimized. Under optimized conditions, the adsorption isotherm results indicated that the Langmuir model provided a better description for the adsorption of Pb(II) onto the GFC nanosorbent than the Freundlich model. The maximum adsorption capacity (qmax) was 63.45 mg g-1. The pseudo-second-order kinetic model (R2 = 0.999) was fitted with the experimental results, implying that the adsorption of Pb(II) onto GFC is a chemical process. The thermodynamic studies demonstrated the exothermic nature of the adsorption process. Another advantage of the GFC nanosorbent for Pb(II) removal is its capability to be easily recovered under the use of an external magnet and subsequently regenerated. Our work demonstrated that the removal efficiency was stable after several regeneration cycles (i.e., approximately 12% reduction after four successive adsorption-desorption cycles), implying that the GFC nanosorbent exhibits satisfactory regeneration performance. Therefore, with high removal efficiency, high adsorption capacity, and stable reusability, the GFC nanocomposite is a remarkable application potential adsorbent for the in situ treatment of Pb(II) ion-containing aqueous solutions.

Comorbidity Burden and Treatment Patterns of Psoriasis in Vietnam: Real-World Data from the EXPAND Study.

INTRODUCTION: Psoriasis is a multi-faceted, immune-mediated inflammatory disease associated with a wide range of comorbidities. Real-world data on treatment patterns, comorbidities, and economic burden in patients with psoriasis are needed for comprehensive patient care in Vietnam. METHODS: A retrospective chart review study was conducted using secondary data extracted from patients' medical records of two hospitals in Vietnam, with the aim of identifying adult patients with a confirmed diagnosis of psoriasis. The index date was defined as the date of first diagnosis between 1 January 2020 and 31 October 2021. Sociodemographic factors, disease characteristics, comorbidities, medication usage, drug survival, and medication costs were analyzed. RESULTS: A total of 661 patients were identified (mean ± standard deviation [SD] age 43.5 ± 14.8 years). The most prevalent comorbidity was dyslipidemia (49.6% of patients), followed by hypertension (23.4%), and psoriatic arthritis (10.4%). In total, 44% of patients received biologic therapies. Overall, 66.7% and 54.3% of patients receiving biologic and non-biologic therapies, respectively, had ≥ 1 comorbidity. Only 23.2% of patients with psoriasis-related comorbidities stopped therapy with biologics. Biologics had a longer retention time (17.0 months) than non-biologics (6.0 months) in patients with comorbidities. Patients with comorbidities had significantly higher total annual healthcare costs than those without comorbidities (in US dollars: USD901 vs. USD304; p < 0.001), mainly due to the relatively higher costs associated with the use of biologics. CONCLUSION: Patients with psoriasis in Vietnam experience a high disease and economic burden due to comorbidities. Evidence from this real-world study supports the need for routine monitoring of and an appropriate treatment course for psoriasis-related comorbidities.

Investigation of enhanced degradation of the antibiotic under visible in novel B/ZnO/TiO2nanocomposite and its electrical energy consumption.

Both ZnO and TiO2are common semiconducting metal oxides with high mechanical and chemical durability. However, they only have good photocatalytic ability in the UV region, besides the rapid recombination between electrons and holes reduces the efficiency of the decomposition of organic substances. To improve their catalytic efficiency, in this study, ZnO and TiO2were doped with B to produce the novel B/ZnO/TiO2nanocomposites for degrading tetracycline hydrochloride (TCH) in the aqueous solution. The characteristics of samples were analyzed by the diffuse reflectance ultraviolet-visible (DR/UV-vis), scanning electron microscope (SEM), energy-dispersive (EDS), Fourier transform infrared spectroscopy (FT-IR), and x-ray diffraction (XRD) techniques. The 3B/ZnO/TiO2sample had a band gap energy (Eg) of 3.21 eV. Although the B/ZnO/TiO2sample had a tightly aggregated morphology composed of many nanoparticles in 33-137 nm, it still exhibited a higher uniformly and photocatalytic efficiency than ZnO and ZnO/TiO2. At the optimal doped B of 3 wt%, the degradation efficiency (DE) was achieved at 96.33% with a rate constant of 0.067 min-1. The factors that affect the photocatalytic process such as the initial TCH concentration, the catalyst content, and the pH solution were comprehensively investigated. In addition, the stability of 3B/ZnO/TiO2nanocomposite was evaluated via three consecutive cycles and the DE was 69.75% in 3rd cycle. The Z-scheme mechanism was proposed for the photocatalytic mechanism of TCH in the B/ZnO/TiO2catalyst. In addition, electrical energy consumption was estimated that the electrical energy per order only was 29.05 kW.h.l-1.

Preparation of carnation-like Ag-ZnO composites for enhanced photocatalysis under visible light.

Carnation-like ZnO was synthesized by the facile precipitation method (at room temperature and in 120 min) to decompose dyes in an aqueous medium. The carnation-like ZnO had a stratified porous structure with a size of about 2-3µm, its petals had a smooth surface with a thickness of 5-10 nm and a width of about 300-500 nm. Ag-ZnO composites were synthesized using glucose with the assistance of PVP. The morphology of Ag-ZnO composites was almost unchanged compared to ZnO. Where, the Ag nanoparticles in the size range of 5-15 nm were uniformly dispersed on the ZnO petals, improving the catalytic ability of the composites in tartrazine (TA) degradation. The influence of Ag content on catalytic structure and performance of composite was studied. The 5Ag-ZnO sample had the highest BET surface area and pore volume and the lowest gap energy (Eg) among the as-synthesized samples. The 5Ag-ZnO sample proclaimed the degradation efficiency in 70 min of 97.8% and thekapof 0.031 min-1. The influences of catalyst content, solution pH, and concentration of dye on the photodegradation efficiency of the composite were thoroughly studied. Besides, the photocatalytic activity of the composite was demonstrated by degrading various organic substances and reusability. In addition, it was compared to a metal-semiconductor catalyst of Au-ZnO and semiconductor-semiconductor catalysts of MoS2-ZnO, Cu2O-ZnO, and SiO2-ZnO. The catalytic mechanism under visible light was proposed.

Differential Cytotoxicity of Curcumin-Loaded Micelles on Human Tumor and Stromal Cells.

Although curcumin in the form of nanoparticles has been demonstrated as a potential anti-tumor compound, the impact of curcumin and nanocurcumin in vitro on normal cells and in vivo in animal models is largely unknown. This study evaluated the toxicity of curcumin-loaded micelles in vitro and in vivo on several tumor cell lines, primary stromal cells, and zebrafish embryos. Breast tumor cell line (MCF7) and stromal cells (human umbilical cord vein endothelial cells, human fibroblasts, and human umbilical cord-derived mesenchymal stem cells) were used in this study. A zebrafish embryotoxicity (FET) assay was conducted following the Organisation for Economic Co-operation and Development (OECD) Test 236. Compared to free curcumin, curcumin PM showed higher cytotoxicity to MCF7 cells in both monolayer culture and multicellular tumor spheroids. The curcumin-loaded micelles efficiently penetrated the MCF7 spheroids and induced apoptosis. The nanocurcumin reduced the viability and disturbed the function of stromal cells by suppressing cell migration and tube formation. The micelles demonstrated toxicity to the development of zebrafish embryos. Curcumin-loaded micelles demonstrated toxicity to both tumor and normal primary stromal cells and zebrafish embryos, indicating that the use of nanocurcumin in cancer treatment should be carefully investigated and controlled.

Ophthalmologists' and patients' perspectives on treatments for diabetic retinopathy and maculopathy in Vietnam: a descriptive qualitative study.

INTRODUCTION: Globally, diabetic retinopathy (DR) is the leading cause of blindness in working-aged adults. Early detection and treatment of DR is essential for preventing sight loss. Services must be available, accessible and acceptable to patients if we are to ensure they seek such care. OBJECTIVES: To understand patients' knowledge and attitudes towards laser versus antivascular endothelial growth factor (VEGF) injections to treat DR in Vietnam, and to identify factors Vietnamese ophthalmologists consider when making treatment decisions. METHODS: This is a descriptive qualitative study based on semi-structured interviews with 18 patients (12 from Ho Chi Minh City and 6 from Hanoi) plus individual interviews with 24 ophthalmologists working in eye clinics in these cities. Thematic analysis was used to analyse the data. RESULTS: In total, 10/24 (41.7%) ophthalmologists were female, and their median age was 41 years (range 29-69 years). The median age of patients was 56.5 years (range 28-72 years), and 7/18 (38.9%) were female. Briefly, factors that influence DR treatment decisions for ophthalmologists are medical considerations (ie, severity of disease, benefits and risks), availability (ie, treatment and resources) and patient-related factors (ie, costs and adherence). Patient's perceived barriers and facilitators to treatments were based on patient and family related factors (ie, treatment and transportation costs) and previous treatment experiences (ie, positive and negative). Recommendations by all participants included ensuring that both laser and anti-VEGF injections are widely available across the country and controlling costs for patients and the healthcare system. CONCLUSIONS: Reducing DR treatment costs, optimising treatments options, and expanding the network of clinics offering treatment outside metropolitan areas were the main issues raised by participants. These findings can help inform policy changes in Vietnam and may be generalisable to other low-resource settings.

Pyridinedicarboxylate-Tb(III) Complex-Based Luminescent Probes for ATP Monitoring.

The pyridinedicarboxylate-Tb(III) complexes, TbPDC and Tb(PDC)3, as luminescent probes for ATP monitoring have been conveniently prepared and characterized by FT-IR, 1H-NMR, ESI-MS, UV-Vis, excitation, and emission spectroscopy. Interestingly, these two Tb(III) complexes were quenched by ATP by a similar mechanism via π-π stacking interaction between the chelating ligand and adenine moiety. The ability of luminescent probes applied for the determination of ATP in aqueous solution has been investigated. The dynamic ranges for the quantification of ATP are within 10-90 µM and 10-100 µM with detection limits of 7.62 and 11.20 µM for TbPDC and Tb(PDC)3, respectively. The results demonstrated that these luminescent probes would be a potential candidate assay for ATP monitoring in hygiene assessment.

Facile Controlling of the Physical Properties of Zinc Oxide and Its Application to Enhanced Photocatalysis.

In this study, the physical properties of ZnO were facile controlled by the synthesis method with the addition of capping and precipitation agents. As-prepared ZnO samples had different morphologies such as carnation flower-like ZnO (CF-ZnO), rose-flower-like ZnO (RF-ZnO), rod-like ZnO (R-ZnO), and nanoparticle ZnO (N-ZnO) and were characterized by SEM, XRD, N2 adsorption/desorption isotherms, FT-IR, and DR/UV-vis. All samples had a crystallite structure of hexagonal wurtzite type. The CF-ZnO and RF-ZnO samples had the hierarchical structure like a carnation flower and a beautiful rose, respectively. R-ZnO was composed of many hexagonal rods and few spherical particles, while N-ZnO microstructures were made up of nanoparticles with approximately 20-30 nm, exhibiting the largest surface area, pore volume, and pore width among as-prepared samples, and their crystal size and bandgap energy were 17.8 nm and 3.207 eV, respectively. The catalytic performances of ZnO samples were evaluated by degradation of Tartrazine (TA) and Caffeine (CAF) under low UV irradiation (15 W). N-ZnO showed a high photocatalytic activity compared to other samples. Besides, the reaction kinetics was investigated by the first-order kinetic model, and the catalytic performance of ZnO was evaluated through several organic pollutants.

Causes and Clinical Characteristics of Small Bowel Bleeding in Northern Vietnam.

Aim: Causes, clinical features, and diagnostic approaches for small bowel (SB) bleeding were analyzed to derive recommendations in dealing with this clinical condition. Methods: We included 54 patients undergoing surgical treatment for SB bleeding, from January 2009 to December 2019. Detailed clinical data, diagnosis procedures, and causes of bleeding were collected. Results: Among 54 cases with SB bleeding, the most common causes were tumors (64.8%), followed by angiopathy (14.8%), ulcers (9.3%), diverticula (5.6%), tuberculosis (3.7%), and enteritis (1.9%). Most tumors (32/35 cases, 91.4%) and vascular lesions (8/8 cases, 100%) were located in the jejunum. The incidence of tumors was higher in the older (30/41 cases, 73.1%) than that in patients younger than 40 years of age (5/13 cases, 38.5%, P < 0.01). Common initial findings were melena (68.5%) and hematochezia (31.5%). The overall diagnostic yield of computed tomographic enterography (CTE) was 57.4% (31/54 cases), with the figures for tumors, vascular lesions, and inflammatory lesions being 71.4% (25/35 cases), 62.5% (5/8 cases), and 12.5% (1/8 cases), respectively. Double-balloon enteroscopy (DBE) definitively identified SB bleeding sources in 16/22 (72.7%) patients. Conclusion: Tumors, angiopathy, ulcers, and diverticula were the most common causes of SB bleeding in Northern Vietnamese population. CTE has a high detection rate for tumors in patients with SB bleeding. CTE as a triage tool may identify patients before double-balloon enteroscopy because of the high prevalence of SB tumors.

Assessing the Status of Filtering Blebs at 5 Year Post- Trabeculectomy.

BACKGROUND: Glaucoma is a common cause of blindness in the world as well as in Vietnam. It is treated by many different methods but trabeculectomy is still the most popular and highly effective surgical method to treat this condition. AIM: To analyze the status of 5 years filtering blebs following trabeculectomy and to explore multiple factors associated with filtering blebs. METHODS: This is a retrospective, cross-sectional descriptive study. Eyes had been performed trabeculectomy for 5 years were included in these results. The filtering blebs were assessed using slit lamp and OCT. the OCT captured bleb area to evaluate fluid subconjunctival spaces, thickness and height of bleb and to evaluate the related factors. RESULTS: A group of 106 primary glaucoma eyes of 97 patients (88 patients with 1 eye, 9 patients 2 eyes) had been performed trabeculectomy for 5 years were taken OCT anterior image. The proportion of female patients is 1.5 times that of male patients. IOP was controlled with or without topical medication in all eyes. The filtering bleb had a high echo reflection, which accounted for 42.5%, the average echo reflection was 38.6%, the low echo reflection response of 18.9%. 66% of the eyes had the aqueous space under the conjunctiva, 65.1% have the aqueous space under sclera flap, the average height of the bleb on OCT was 0.4 mm ± 0.3 mm. Young patients often have a higher rate of bleb fibrosis and loss of function than older patients. CONCLUSION: OCT is capable of assessing the function of bleb. After 5 years of trabeculectomy, on the OCT image, most cases of blebs are maintaining drainage function.

Evaluation of Anterior Chamber Depth and Anterior Chamber Angle Changing After Phacoemulsification in the Primary Angle Close Suspect Eyes.

BACKGROUND: Phacoemulsification surgery has the ability to deeply alter the segment anterior morphology, especially in eye with shallow anterior chamber (AC), narrow anterior chamber angle (ACA). However, the changes of anterior chamber depth (ACD) and ACA on the close angle suspect eyes after phacoemulsification have not been mentioned in many studies. So, we conduct this research. AIM: To evaluate the alteration in the ACA and ACD after phacoemulsification in the close angle suspect eyes. METHODS: Interventional study with no control group. Subjects were the primary angle closure suspect (PACS) eyes, that were operated by phacoemulsification with intraocular lens (IOL) at Glaucoma Department of VNIO from December 2017 to October 2018. RESULTS: 29 PACS eyes with cataract were operated by phacoemulsification with intraocular lens. After 3 months of monitoring, the average ACD augmented from 2.082 ± 0.244 to 3.673 ± 0.222 mm. AOD500 increase from 0.183 ± 0.088 to 0.388 ± 0.132 µm, AOD750 increased from 0.278 ± 0.105 to 0.576 ± 0.149 µm. The TISA500 enlarged from 0.068 ± 0.033 to 0.140 ± 0.052 mm2, TISA750 enlarged from 0.125 ± 0.052 to 0.256 ± 0.089 mm2 at the third month (p < 0.01). CONCLUSION: Phacoemulsification surgery increases the ACD and enlarged the angle in the PACS eyes.

TSLP expression induced via Toll-like receptor pathways in human keratinocytes.

The skin epidermis and mucosal epithelia (airway, ocular tissues, gut, and so on) are located at the interface between the body and environment and have critical roles in the response to various stimuli. Thymic stromal lymphopoietin (TSLP), a cytokine expressed mainly by epidermal keratinocytes (KCs) and mucosal epithelial cells, is a critical factor linking the innate response at barrier surfaces to Th2-skewed acquired immune response. TSLP is highly expressed in skin lesions of atopic dermatitis patients. Here, we describe on Toll-like receptor (TLR)-mediated induction of TSLP expression in primary cultured human KCs, placing emphasis on experimental methods used in our studies. Double-stranded RNA (TLR3 ligand), flagellin (TLR5 ligand), and diacylated lipopeptide (TLR2-TLR6 ligand) stimulated human KCs to express TSLP and Staphylococcus aureus membranes did so via the TLR2-TLR6 pathway. Atopic cytokine milieu upregulated the TLR-mediated induction of TSLP. Culturing in the absence of glucocorticoid before stimulation enhanced the TSLP expression. Extracellular double-stranded RNA induced TSLP via endosomal acidification- and NF-κB-dependent pathway. Specific measurement of the long TSLP transcript, which contributes to the production of the TSLP protein, rather than total or the short transcript is useful for accurate detection of functional human TSLP gene expression. The results suggest that environment-, infection-, and/or self-derived TLR ligands contribute to the initiation and/or amplification of Th2-type skin inflammation including atopic dermatitis and atopic march through the induction of TSLP expression in KCs and include information helpful for understanding the role of the gene-environment interaction relevant in allergic diseases.

Novel route to size-controlled Fe-MIL-88B-NH2 metal-organic framework nanocrystals.

A new approach for the synthesis of uniform metal-organic framework (MOF) nanocrystals with controlled sizes and aspect ratios has been developed using simultaneously the non-ionic triblock co-polymer F127 and acetic acid as stabilizing and deprotonating agents, respectively. The alkylene oxide segments of the triblock co-polymer can coordinate with metal ions and stabilize MOF nuclei in the early stage of the formation of MOF nanocrystals. Acetic acid can control the deprotonation of carboxylic linkers during the synthesis and, thus, enables the control of the rate of nucleation, leading to the tailoring of the size and aspect ratio (length/width) of nanocrystals. Fe-MIL-88B-NH(2), as an iron-based MOF crystal, was selected as a typical example to illustrate our approach. The results reveal that this approach is used for not only the synthesis of uniform nanocrystals but also the control of the size and aspect ratio of the materials. The size and aspect ratio of nanocrystals increase with an increase in the concentration of acetic acid in the synthetic mixture. The non-ionic triblock co-polymer F127 and acetic acid can be easily removed from the Fe-MIL-88B-NH(2) nanocrystal products by washing with ethanol, and thus, their amine groups are available for practical applications. The approach is expected to synthesize various nanosized carboxylate-based MOF members, such as MIL-53, MIL-89, MIL-100, and MIL-101.

Flagellin induces the expression of thymic stromal lymphopoietin in human keratinocytes via toll-like receptor 5.

BACKGROUND: Thymic stromal lymphopoietin (TSLP), highly expressed by keratinocytes in skin lesions of atopic dermatitis patients and bronchial epithelial cells in asthma, plays a key role in allergic diseases. Information on triggers for the release of TSLP in keratinocytes is still limited. Keratinocytes express Toll-like receptor (TLR) 5, the ligand for which is flagellin, the major structural protein of the flagella of Gram-negative bacteria. IL-4, IL-13 and TNF-α (Th2/TNF) are associated with allergic diseases. TGF-α, one of the ligands for the epidermal growth factor receptor, is overexpressed in keratinocytes in atopic dermatitis. We investigated the induction of TSLP expression in keratinocytes stimulated with flagellin and its modulation by the Th2/TNF cytokines and TGF-α. METHODS: Primary human keratinocytes were stimulated with flagellin with or without cytokines. The TSLP released was measured by ELISA. Gene expression was analyzed by quantitative real-time PCR. RESULTS: Stimulation of keratinocytes with flagellin induced the release of TSLP protein and upregulation of the gene expression of TSLP and other pro-inflammatory molecules. The flagellin-induced release of TSLP was enhanced by the Th2/TNF cytokines or TGF-α. Small interfering RNA-mediated knockdown of TLR5 expression suppressed the flagellin-induced TSLP gene expression. CONCLUSIONS: Flagellin induces TSLP expression in keratinocytes via TLR5 and the expression can be upregulated by a cytokine milieu with Th2/TNF or TGF-α, suggesting that exposure of barrier-defective skin to Gram-negative bacteria or environmental flagellin contributes to the initiation and/or amplification of Th2-type skin inflammation including atopic dermatitis through the induction of TSLP expression in keratinocytes.

Staphylococcus aureus membrane and diacylated lipopeptide induce thymic stromal lymphopoietin in keratinocytes through the Toll-like receptor 2-Toll-like receptor 6 pathway.

BACKGROUND: Staphylococcus aureus heavily colonizes the lesions of patients with atopic dermatitis (AD) and is known to trigger a worsening of AD. However, the exact mechanism by which S. aureus promotes AD is unknown. Thymic stromal lymphopoietin (TSLP), which is highly expressed by keratinocytes in skin lesions of patients with AD and bronchial epithelial cells in asthmatic patients, represents a critical factor linking responses at interfaces between the body and the environment to allergic type 2 immune responses. OBJECTIVES: We sought to examine the ability of synthetic lipopeptides and S. aureus to induce TSLP expression in human keratinocytes and identify the pathway of induction. METHODS: We stimulated primary human keratinocytes with lipopeptides and S. aureus-derived materials. The release and gene expression of TSLP were measured by means of ELISA and quantitative PCR, respectively. RESULTS: Diacylated lipopeptide upregulated the expression of TSLP and other proinflammatory molecules. Heat-killed S. aureus and the subcellular fractions of S. aureus induced TSLP's release, with the membranous fraction having the greatest activity. Small interfering RNA-mediated knockdown of either Toll-like receptor (TLR) 2 or TLR6 inhibited the diacylated lipopeptide- and S. aureus membrane-induced TSLP gene expression. S. aureus membrane- and diacylated lipopeptide-induced release of TSLP was enhanced by T(H)2/TNF-α cytokines and partially suppressed by IFN-γ and TGF-ß. CONCLUSIONS: The results suggest that ligands for the TLR2-TLR6 heterodimer in S. aureus membranes, including diacylated lipoproteins, could promote T(H)2-type inflammation through TSLP production in keratinocytes, providing an overall picture of the vicious cycles between colonization by S. aureus and AD in the T(H)2-skewed sensitization process, exacerbation of the disease, or both.

Glucocorticoids inhibit double-stranded RNA-induced thymic stromal lymphopoietin release from keratinocytes in an atopic cytokine milieu more effectively than tacrolimus.

BACKGROUND: Thymic stromal lymphopoietin (TSLP), highly expressed by keratinocytes in skin lesions in atopic dermatitis and bronchial epithelial cells in asthma, plays a key role in allergic diseases. Double-stranded RNA (dsRNA) stimulates keratinocytes to release TSLP in vitro. OBJECTIVE: To examine the potential of glucocorticoids and calcineurin inhibitors to suppress dsRNA-induced release of TSLP from keratinocytes. METHODS: Primary human kerarinocytes were stimulated with dsRNA in the presence of IL-4, IL-13 and TNF-alpha. TSLP release was measured by ELISA. The effects of glucocorticoids and 2 calcineurin inhibitors, cyclosporin A and FK506/tacrolimus, were analyzed. RESULTS: The glucocorticoids inhibited dsRNA-induced release of TSLP. The inhibitory effect became saturated (50-70% reduction) at concentrations higher than 10(-10)M. Cyclosporin A inhibited the release of TSLP by 50-60% at 10(-5) and 10(-4)M. FK506 had no effect at 10(-5)M or less, but almost completely inhibited the release of TSLP at 10(-4)M. No synergistic effect was obtained with a glucocorticoid plus either of the calcineurin inhibitors. An additive inhibitory effect was obtained with a glucocorticoid plus 10(-5)M cyclosporin A. Glucocorticoid inhibited dsRNA-induced TSLP transcription in the absence of Th2/TNF cytokines. CONCLUSIONS: Glucocorticoids inhibited the dsRNA-induced release of TSLP in the atopic cytokine milieu at much lower concentrations than calcineurin inhibitors, suggesting that they could be effective in the treatment of atopic dermatitis when exogenous or endogenous dsRNA is involved in the pathogenesis. In addition, the in vitro system established in this study would be useful for screening of therapeutic reagents which target TSLP expression.

Cytokine milieu modulates release of thymic stromal lymphopoietin from human keratinocytes stimulated with double-stranded RNA.

BACKGROUND: Thymic stromal lymphopoietin (TSLP) plays a key role in allergic diseases, such as atopic dermatitis (AD) and asthma. TSLP is highly expressed by keratinocytes in skin lesions of patients with AD, but environmental triggers for its release from keratinocytes with endogenous factors are not well understood. Patients with AD, in whom allergic sensitization is already established, are susceptible to viral dissemination. OBJECTIVES: We investigated TSLP's release from primary human keratinocytes stimulated with a Toll-like receptor (TLR) 3 ligand, polyinosinic-polycytidylic acid, which mimics viral double-stranded RNA (dsRNA), and its modulation by cytokines. METHODS: Primary human keratinocytes were stimulated with TLR ligands, cytokines, or both. TSLP released into culture supernatants was measured by means of ELISA. RESULTS: Stimulation of keratinocytes with dsRNA induced release of TSLP and upregulated gene expression of TSLP and other cytokines and chemokines. The release of TSLP was enhanced by the addition of IL-4, IL-13, and/or TNF-alpha. With or without the T(H)2/TNF cytokines, the dsRNA-induced release of TSLP was upregulated by IFN-alpha and IFN-beta and suppressed by IFN-gamma, TGF-beta, or IL-17. CONCLUSIONS: The effect of the TLR3 ligand on keratinocytes suggests contribution of viral dsRNA to skin inflammations under the influence of a cytokine milieu. The results imply that viral dsRNA and a T(H)2 cytokine milieu might promote T(H)2-type inflammation through an induction of TSLP expression, suggesting that a vicious cycle exists between AD with T(H)2-type inflammation and viral infections and a possible blockade of this cycle by other cytokine milieus provided by cells, such as T(H)1, regulatory T, and T(H)17 cells.

CD8+ T lymphocytes specific for glutamic acid decarboxylase 90-98 epitope mediate diabetes in NOD SCID mouse.

During the past decade, glutamic acid decarboxylase (GAD) has been considered a crucial beta-cell autoantigen involved in type 1 diabetes in the NOD mouse and human. Recently, the etiological role of GAD has remained controversy. In the NOD mouse, some previous studies argued in favor of a regulatory role for GAD-specific CD4+ T cells, and no diabetogenic CD8+ T cells specific for GAD have been identified so far, discrediting the importance of GAD in beta-cell injury. Here, we identified, in the NOD model, a relevant GAD CD8+ T cell epitope (GAD(90-98)) using immunization with a plasmid encoding GAD, a protocol relying on in vivo processing of peptides from the autoantigenic protein. In pancreatic lymph nodes of naïve female NOD mice, CD8+ T lymphocytes recognizing GAD(90-98) peptide were detected during the initial phase of invasive insulitis (between 4 and 8 weeks of age), suggesting an important role for these cells in the first stage of the disease. GAD(90-98) specific CD8+ lymphocytes lysed efficiently islet cells in vitro and transferred diabetes into NOD(SCID) mice (100%). Finally, diabetes was accelerated greatly in 3-week-old female NOD mice injected i.p. with GAD(90-98), strengthening the role of GAD-specific CTLs in diabetes pathogenesis.

Gene-gun biolistic immunization encoding glutamic acid decarboxylase: a model for studying Langerhans cell abnormalities and mimicry in the nonobese diabetic mouse.

Plasmid-DNA gene-gun immunization may be an efficient approach for investigating the role of skin dendritic cells (DCs) in type 1 diabetes (T1D) pathogenesis and the significance of the presentation of peptides that mimic autoantigenic epitopes in aggravating or modulating the autoimmune reaction. Gene-gun immunization has been described as producing long-lasting immune responses elicited by skin DCs, especially Langerhans cells (LCs). Therefore, we tested the immune response and diabetes modulation in nonobese diabetic (NOD) mice and in control BALB/c mice, by gene-gun administration of plasmid-DNA encoding (1) human 65 kDa glutamic acid decarboxylase (hGAD65) mimicking the crucial mouse autoantigen GAD65 (similarity of 95.7%) or (2) beta-galactosidase (betaGAL) as a negative control. Expression of GAD and betaGAL in skin of pc-GAD- and pc-LacZ-injected mice, respectively, was confirmed. It was surprising that both pc-LacZ-injected BALB/c and NOD mice exhibited a betaGAL-specific Th1 immune response: spleen cells of pc-LacZ mice proliferated specifically to betaGAL (P < 10(-4)) and secreted significant amounts of IFNgamma (P < 10(-4)). pc-LacZ mice also developed a betaGAL-specific Th1-related (IgG2a/2c) and Th2-related (IgG1) humoral response. Although pc-GAD BALB/c mice showed Th2-related GAD-specific IgG1 production and a significant secretion of IL4 (P < .03), pc-GAD NOD mice did not generate either an antibody response or a T cell response specific to GAD. Moreover, gene-gun immunization encoding hGAD65 did not clearly modulate diabetes onset in NOD mice. This absence of detectable GAD-specific response may implicate skin DC deficiencies in NOD mice. The gene-gun technique could thus provide an interesting model for studying skin DC abnormalities in NOD mice and their potential implication of presenting mimetic peptides that modulate the autoimmune response in T1D.