The characteristic of the key aroma-active components in white tea using GC-TOF-MS and GC-olfactometry combined with sensory-directed flavor analysis.

BACKGROUND: White tea has become more and more popular with consumers due to its health benefits and unique flavor. However, the key aroma-active compounds of white tea during the aging process are still unclear. Thus, the key aroma-active compounds of white tea during the aging process were investigated using gas chromatography-time-of-flight-mass spectrometry (GC-TOF-MS) and gas chromatography-olfactometry (GC-O) combined with sensory-directed flavor analysis. RESULTS: A total of 127 volatile compounds were identified from white tea samples with different aging years by GC-TOF-MS. Fifty-eight aroma-active compounds were then determined by GC-O, and 19 of them were further selected as the key aroma-active compounds based on modified frequency (MF) and odor activity value (OAV). CONCLUSION: Aroma recombination and omission testing confirmed that 1-octen-3-ol, linalool, phenethyl alcohol, geraniol, (E)-ß-ionone, α-ionone, hexanal, phenylacetaldehyde, nonanal, (E, Z)-(2,6)-nonadienal, safranal, γ-nonalactone and 2-amylfuran were the common key aroma-active compounds to all samples. Cedrol, linalool oxide II and methyl salicylate were confirmed peculiar in new white tea, while ß-damascenone and jasmone were peculiar in aged white tea. This work will offer support for further studies on the material basis of flavor formation of white tea. © 2023 Society of Chemical Industry.

Overall survival and toxicity of Y90 radioembolization for hepatocellular carcinoma patients in Barcelona Clinic Liver Cancer stage C (BCLC-C).

INTRODUCTION: National Comprehensive Cancer Network HCC guidelines recommend Y90 to treat BCLC-C patients only in select cases given the development of systemic regimens. We sought to identify ideal candidates for Y90 by assessing survival and toxicities in this patient group. MATERIALS AND METHODS: The Radiation-Emitting Selective Internal radiation spheres in Non-resectable tumor registry is a prospective observational study (NCT: 02,685,631). Patients with advanced HCC were stratified into 3 groups based on tumor location, Eastern Cooperative Oncology Group (ECOG) performance status, and liver function. Group 1: liver isolated HCC, ECOG 0 and Child Pugh (CP) A (n = 12, 16%), Group 2: liver isolated HCC, ECOG ≥ 1 or CP B/C (n = 37, 49%), and Group 3: extrahepatic HCC with any ECOG or CP score (n = 26, 35%). Patients in any group could have macrovascular invasion. Overall survival (OS) and progression-free survival (PFS) with 95% confidence intervals (95% CI) were calculated. Grade 3 + toxicities were tracked using Common Terminology Criteria for Adverse Events v5. Cox proportional hazard model was performed to determine factors affecting OS. RESULTS: Seventy-five BCLC-C patients treated between 2015 and 2019 were reviewed. The groups were similar in age, sex, race, and ethnicity (all p > 0.05). Bilobar disease was least common in Group 1 (p < 0.001). Median OS of the entire cohort was 13.6 (95% CI 7.5-16.1) months. Median OS of Groups 1-3 were 21.8, 13.1 and 11.5 months respectively (p = 0.6). Median PFS for the cohort was 6.3 (4.8-14.7) months. Median PFS for group 1 was not reached. Mean PFS for Group 1 was 17.3 ± 4.8 months. Median PFS for Groups 2 and 3 was 6.8 and 5.9 months (X2 = 1.5, p = 0.5). Twenty-four Grade 3 or greater toxicities developed, most commonly hyperbilirubinemia (8/75, 11%) and thrombocytopenia (2/75, 3%). The incidence of toxicities between groups was similar (all p > 0.05). Cox Proportional Hazard analysis predicted shorter OS with CP class B/C (X2 = 6.7, p = 0.01), while macrovascular invasion (X2 = 0.5, p = 0.5) and ECOG score of ≥ 1 (X2 = 2.1, p = 0.3) was not associated with OS. CONCLUSIONS: OS of CPA patients with advanced HCC and performance status of 0 was 21.8 months following Y90. CP A cirrhosis is the best predictor of prolonged OS in advanced (BCLC-C) HCC.

Survival and Toxicities after 90Y Transarterial Radioembolization of Metastatic Colorectal Cancer in the RESIN Registry.

Background Patients with unresectable, chemorefractory hepatic metastases from colorectal cancer have considerable mortality. The role of transarterial radioembolization (TARE) with yttrium 90 (90Y) microspheres is not defined because most reports are from a single center with limited patient numbers. Purpose To report outcomes in participants with colorectal cancer metastases treated with resin 90Y microspheres from a prospective multicenter observational registry. Materials and Methods This study treated enrolled adult participants with TARE using resin microspheres for liver-dominant metastatic colorectal cancer at 42 centers, with enrollment from July 2015 through August 2020. TARE was used as the first-, second-, or third-line therapy or beyond. Overall survival (OS), progression-free survival (PFS), and toxicity outcomes were assessed by line of therapy by using Kaplan-Meier analysis for OS and PFS and Common Terminology Criteria for Adverse Events, version 5, for toxicities. Results A total of 498 participants (median age, 60 years [IQR, 52-69 years]; 298 men [60%]) were treated. TARE was used in first-line therapy in 74 of 442 participants (17%), second-line therapy in 180 participants (41%), and third-line therapy or beyond in 188 participants (43%). The median OS of the entire cohort was 15.0 months (95% CI: 13.3, 16.9). The median OS by line of therapy was 13.9 months for first-line therapy, 17.4 months for second-line therapy, and 12.5 months for third-line therapy (χ2 = 9.7; P = .002). Whole-group PFS was 7.4 months (95% CI: 6.4, 9.5). The median PFS by line of therapy was 7.9 months for first-line therapy, 10.0 months for second-line therapy, and 5.9 months for third-line therapy (χ2 = 8.3; P = .004). TARE-attributable grade 3 or 4 hepatic toxicities were 8.4% for bilirubin (29 of 347 participants) and 3.7% for albumin (13 of 347). Grade 3 and higher toxicities were greater with third-line therapy for bilirubin (P = .01) and albumin (P = .008). Conclusion Median overall survival (OS) after transarterial radioembolization (TARE) with yttrium 90 microspheres for liver-dominant metastatic colorectal cancer was 15.0 months. The longest OS was achieved when TARE was part of second-line therapy. Grade 3 or greater hepatic function toxicity rates were less than 10%. Clinical trial registration no. NCT02685631 Published under a CC BY 4.0 license. Online supplemental material is available for this article. See also the editorial by Liddell in this issue.

Characterization of the key aroma-active compounds in high-grade Dianhong tea using GC-MS and GC-O combined with sensory-directed flavor analysis.

Dianhong tea (DHT) is popular for its pleasant caramel-like aroma. In this study, the aroma profile of high-grade DHT have been studied using gas chromatography-mass spectrometry (GC-MS) and gas chromatography-olfactometry (GC-O) combined with headspace solid phase microextraction (HS-SPME). A total of 52 aroma-active compounds were identified by GC-O coupled with aroma extract dilution analysis (AEDA) and odor specific magnitude estimation (Osme). Among them, quantification of 21 aroma-active compounds indicated that the content of linalool (5928 µg/kg) was the highest in high-grade DHT, followed by phenylethanol (3923 µg/kg) and phenylacetaldehyde (1801 µg/kg). Sensory-directed aroma recombination and omission tests further verified that phenylacetaldehyde, linalool, geraniol and 3-ethyl-2,5-dimethylpyrazine were important contributors to the overall sensory characteristics of high-grade DHT which dominated mainly by floral, sweet and caramel-like odors. This work will provide a theoretical reference for comprehensively understanding the aroma characteristic of DHT.

Synthesis of a Removable Cytoprotective Exoskeleton by Tea Polyphenol Complexes for Living Cell Encapsulation.

Cell encapsulation is a chemical tool for endowing living cells with exogenous properties and enhancing their in vitro tolerance against lethal factors, which has shown promising prospects and potential applications in many fields such as cell transplantation, drug delivery, and tissue engineering. One-pot precipitation of a polyphenol-metal complex on cells protects cells from UV irradiation and lytic enzymes. However, the involvement of metal ions brings side effects on cell viability and growth. Moreover, an external removal agent is needed for cell division and growth. Herein, a polymer shell composed of hydrogen bonded constituents without affecting cell viability and growth by the precipitation of tea polyphenol and polyvinyl pyrrolidone is reported. The formation of the polymer shell was verified by the Au nanoparticle's laser scanning confocal reflectance and quartz crystal microbalance measurement. The thickness of the shell was managed by the concentration of the complex. When exposed to UV irradiation for 15 or 30 min, polymer-coating-protected Saccharomyces cerevisiae (yeast) had much higher cell viability than the native one. Exposed to a high temperature environment (60 °C), most of the coated yeasts survived in contrast to uncoated ones. For the cell division and growth curve, the polymer coating with various thicknesses had no difference to the native one, which indicated no suppression of cell growth and no external side effects involved. As applied to mammalian HeLa cells under UV irradiation for 15 min, the coated cells had an obvious higher cell viability than that of untreated ones. Therefore, the tea polyphenol-poly(vinylpyrrolidone) shell is a versatile tool for chemically controlling the external properties of cells without side effects on cell viability and growth.

Higher 25-hydroxyvitamin D level is associated with increased risk for Behçet's disease.

BACKGROUND & AIM: Previous studies showed a vitamin D deficiency in patients with Behçet's disease, suggesting potential benefits of vitamin D supplementation in the prevention and treatment of Behçet's disease. Interpretation of these studies may be limited by reverse causality or confounding bias. We aim to determine the causal association between serum 25-hydroxyvitamin D [25(OH)D] and the risk of Behçet's disease by Mendelian randomization. METHODS: An allele score formed by four variants (rs2282679, rs10741657, rs12785878 and rs6013897) that were associated with serum 25(OH)D level, was examined using data of genome-wide association study (GWAS) on 999 Behçet's disease and 4417 healthy individuals of Chinese ancestry and validated using data of GWAS on 1215 Behçet's disease and 1278 controls of Turkish ancestry. The primary outcome was the risk of Behçet's disease, evaluated by an inverse variance weighted average of the associations with genetically determined 25(OH)D levels. RESULTS: The inverse variance weighted estimate showed that genetically increased 25(OH)D level was associated with a higher risk of Behçet's disease. In the Chinese cohort, the odds ratio for Behçet's disease in one standard deviation increase of natural log-transformed 25(OH)D level was 3.82 (95% CI: 1.27-11.42). Data from Turkish cohort confirmed the association with Behçet's disease (OR, 95% CI: 4.18, 1.15-15.12). In overall combination of Chinese and Turkish cohorts, the odds ratio for Behçet's disease per standard deviation increase of natural log-transformed 25(OH)D level was estimated to be 3.96 (95% CI: 1.72-9.13; P = 0.001). No significant evidence of pleiotropy and heterogeneity was detected. CONCLUSIONS: On the basis of evidence in 7909 human beings, this study provides the newest indication that a lifelong higher 25(OH)D level is associated with an increased risk of Behçet's disease. Special attention should be paid to the potential harm of long-term or high-dose use of vitamin D supplements in clinical practice.

[Pharmacogenomic Research in Direct Oral Anticoagulants].

Oral anticoagulants play an important role in the prevention and treatment of thromboembolic diseases.Warfarin,a traditional oral anticoagulant,is limited in clinical use due to its limitations such as narrow therapeutic window and requirements on frequent monitoring and dose adjustment.Direct oral anticoagulants(DOACs)such as dabigatran,rivaroxaban,apixaban,and edoxaban are increasingly used to prevent and treat venous thrombosis or thrombus formation.However,recent studies have documented inter-individual variability in plasma drug levels of DOACs.This article summarizes the recent advances in the pharmacogenomics of DOACs.

Sensory and instrumental analysis-guided exploration of odor-active compounds recovery with oil during the water-boiling extraction of Pu-erh tea.

The objective of this study was to identify the released behavior of odor-active compounds of Pu-erh tea during the water-boiling extraction, as well as to explore the feasibility of aroma recovery using absorption oils. The behavior of odor-active compounds was identified by using piecewise recovered condensed water combined with sensory, GC-O, and GC-MS analysis. The power-function type kinetic model was the model that best fitted odor intensities, alcohols, methoxy-phenolics, esters, aldehydes, ketones, and total compounds of condensed water. The sensory quality of the condensed water was the best when 16% of added water was evaporated. Six absorption oils were evaluated, and paraffin oil (35 cSt) performed the best in terms of both absorption efficiency and sensory quality of the essential oil. Undertheoptimizedconditions, the absorption and recovery efficiency of odor compounds reached 54.49% and 39.52%, respectively. The alcohols, aldehydes, and methoxy-phenolics exhibited higher absorption and recovery efficiency.

Adenocarcinoma Ex-Goblet Cell: a Retrospective Experience.

PURPOSE: Adenocarcinoma ex-goblet cell carcinoids (AGCCs) are rare appendiceal tumors with mixed neuroendocrine and glandular features. They tend to behave more aggressively than typical carcinoid tumors, affect younger patients, and have a greater predilection for spreading to the peritoneum. Outcomes of AGCC patients treated with chemotherapy, extrapolated from colon cancer regimens, in the adjuvant or metastatic setting have not been explicitly reported. We sought to explore outcomes of AGCC patients with either local disease treated with adjuvant FOLFOX or metastatic disease treated with FOLFOX/FOLFIRI post-cytoreductive debulking (or CRS plus HIPEC in the peritoneal-limited setting). METHODS: We performed a single-institution retrospective analysis of 23 pathologically identified AGCC patients from Vanderbilt University Medical Center treated with chemotherapy in either the adjuvant or metastatic settings. Each patient's tumor was categorized as group B or group C based on the criteria from Tang et al. Median progression-free survival (PFS) or disease-free survival (DFS) (in the curative setting) and overall survival (OS) were determined for each patient and specified patient subgroup. RESULTS AND CONCLUSION: AGCC patients who were treated with FOLFOX chemotherapy in the adjuvant setting or FOLFOX/FOLFIRI in the metastatic setting experienced prolonged PFS, DFS, and OS. Five patients with peritoneal-limited disease treated with CRS plus HIPEC have not yet reached median PFS or OS. While small sample size, patient selection, and retrospective nature limit the generalizability of findings from our analysis, the efficacy signals we observed suggest prospective evaluation with chemotherapy and CRS plus HIPEC is warranted in AGCC patients.

A comparative study of volatile components in Dianhong teas from fresh leaves of four tea cultivars by using chromatography-mass spectrometry, multivariate data analysis, and descriptive sensory analysis.

Dianhong teas produced from fresh leaves of different tea cultivars (YK is Yunkang No. 10, XY is Xueya 100, CY is Changyebaihao, SS is Shishengmiao), were compared in terms of volatile compounds and descriptive sensory analysis. A total of 73 volatile compounds in 16 tea samples were tentatively identified. YK, XY, CY, and SS contained 55, 53, 49, and 51 volatile compounds, respectively. Partial least squares-discriminant analysis (PLS-DA) and hierarchical cluster analysis (HCA) were used to classify the samples, and 40 key components were selected based on variable importance in the projection. Moreover, 11 flavor attributes, namely, floral, fruity, grass/green, woody, sweet, roasty, caramel, mellow and thick, bitter, astringent, and sweet aftertaste were identified through descriptive sensory analysis (DSA). In generally, innate differences among the tea varieties significantly affected the intensities of most of the key sensory attributes of Dianhong teas possibly because of the different amounts of aroma-active and taste components in Dianhong teas.

Determination of phthalate esters in teas and tea infusions by gas chromatography-mass spectrometry.

Phthalate esters (PAEs), a group of environmental pollutants which are carcinogenic to human body, have been detected in teas. In this work, five PAEs in teas and tea infusions were quantitatively determined by a modified simultaneous distillation extraction (SDE) coupled with gas chromatography-mass spectrometry. After the optimization of SDE, the proposed method afforded a wide range of linearity and high linear regression coefficients with the limits of detection range of 0.24-3.72 µg/kg. The average recoveries were 79.83-116.67% for tea samples and 78.22-101.64% for tea infusions with all the relative standard deviations below 20%. The total content of five PAEs in teas was 1.135-3.734 mg/kg and the total dissolving ratio of five PAEs from tea to infusion was 19.05-28.07% for the selected tea samples. The risk assessment result of all the selected tea samples demonstrated that the population with the habit of drinking tea won't cause risk to human health.

Likelihood approach for evaluating bioequivalence of highly variable drugs.

Bioequivalence (BE) is required for approving a generic drug. The two one-sided tests procedure (TOST, or the 90% confidence interval approach) has been used as the mainstream methodology to test average BE (ABE) on pharmacokinetic parameters such as the area under the blood concentration-time curve and the peak concentration. However, for highly variable drugs (%CV > 30%), it is difficult to demonstrate ABE in a standard cross-over study with the typical number of subjects using the TOST because of lack of power. Recently, the US Food and Drug Administration and the European Medicines Agency recommended similar but not identical reference-scaled average BE (RSABE) approaches to address this issue. Although the power is improved, the new approaches may not guarantee a high level of confidence for the true difference between two drugs at the ABE boundaries. It is also difficult for these approaches to address the issues of population BE (PBE) and individual BE (IBE). We advocate the use of a likelihood approach for representing and interpreting BE data as evidence. Using example data from a full replicate 2 × 4 cross-over study, we demonstrate how to present evidence using the profile likelihoods for the mean difference and standard deviation ratios of the two drugs for the pharmacokinetic parameters. With this approach, we present evidence for PBE and IBE as well as ABE within a unified framework. Our simulations show that the operating characteristics of the proposed likelihood approach are comparable with the RSABE approaches when the same criteria are applied.

Populus euphratica XTH overexpression enhances salinity tolerance by the development of leaf succulence in transgenic tobacco plants.

Populus euphratica is a salt-tolerant tree species that develops leaf succulence after a prolonged period of salinity stress. In the present study, a putative xyloglucan endotransglucosylase/hydrolase gene (PeXTH) from P. euphratica was isolated and transferred to tobacco plants. PeXTH localized exclusively to the endoplasmic reticulum and cell wall. Plants overexpressing PeXTH were more salt tolerant than wild-type tobacco with respect to root and leaf growth, and survival. The increased capacity for salt tolerance was due mainly to the anatomical and physiological alterations caused by PeXTH overexpression. Compared with the wild type, PeXTH-transgenic plants contained 36% higher water content per unit area and 39% higher ratio of fresh weight to dry weight, a hallmark of leaf succulence. However, the increased water storage in the leaves in PeXTH-transgenic plants was not accompanied by greater leaf thickness but was due to highly packed palisade parenchyma cells and fewer intercellular air spaces between mesophyll cells. In addition to the salt dilution effect in response to NaCl, these anatomical changes increased leaf water-retaining capacity, which lowered the increase of salt concentration in the succulent tissues and mesophyll cells. Moreover, the increased number of mesophyll cells reduced the intercellular air space, which improved carbon economy and resulted in a 47-78% greater net photosynthesis under control and salt treatments (100-150 mM NaCl). Taken together, the results indicate that PeXTH overexpression enhanced salt tolerance by the development of succulent leaves in tobacco plants without swelling.

Optimization of headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry for detecting methoxyphenolic compounds in pu-erh tea.

A method based on headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry was developed for the analysis of volatile methoxyphenolic compounds in pu-erh tea. Six fibers with different polarities were initially evaluated. The 75 µm carboxen/polydimethylsiloxane fiber exhibited the highest extraction efficiency and was selected for further optimization. A Plackett-Burman design was used to screen for the brewing proportion of tea and water, amount of pu-erh tea, ionic strength, extraction time, extraction temperature, desorption time, rate of agitation, and equilibrium time. A Box-Behnken design was then applied to optimize the significant factors. Under optimal conditions, the proposed method affords a wide range of linearity, high linear regression coefficients (0.996-0.999), less than 9.0% repeatability of relative standard deviation, and limits of detection ranging from 2.31 to 21.80 ng/g. The proposed method has satisfactory accuracy, with recoveries of 79.08-113.9%. This method was successfully applied for the analysis of pu-erh tea samples.

Regulatory effects of IFN-ß on the development of experimental autoimmune uveoretinitis in B10RIII mice.

BACKGROUND: Experimental autoimmune uveoretinitis (EAU) serves as a model for human intraocular inflammation. IFN-ß has been used in the treatment of certain autoimmune diseases. Earlier studies showed that it ameliorated EAU; however, the mechanisms involved in this inhibition are still largely unknown. METHODOLOGY/PRINCIPAL FINDINGS: B10RIII mice were immunized with interphotoreceptor retinoid-binding protein (IRBP) peptide 161-180 in Complete Freund's adjuvant. Splenocytes from different time points after immunization were used to evaluate the expression of IFN-ß. An increased expression of IFN-ß was observed during EAU and its highest expression was observed on day 16, 3 days after the peak of intraocular inflammation. Splenocytes and draining lymph node cells from mice immunized with IRBP(161-180) on day 13 and control mice were activated with anti-CD3/anti-CD28 antibodies or IRBP(161-180) to evaluate the production of IFN-γ and IL-17. The results showed that IFN-γ and IL-17 were significantly higher in immunized mice as compared to the control mice when exposed to anti-CD3/anti-CD28 antibodies. However, the production of IFN-γ and IL-17 was detected only in immunized mice, but not in the control mice when stimulated with IRBP(161-180). Multiple subcutaneous injections of IFN-ß significantly inhibited EAU activity in association with a down-regulated expression of IFN-γ, IL-17 and an enhanced IL-10 production. In an in vitro system using cells from mice, IFN-ß suppressed IFN-γ production by CD4(+)CD62L(-) T cells, IL-17 production by CD4(+)CD62L(+/-) T cells and proliferation of CD4(+)CD62L(+/-) T cells. IFN-ß inhibited the secretion of IL-6, but promoted the secretion of IL-10 by monocytes. IFN-ß-treated monocytes inhibited IL-17 secretion by CD4(+)CD62L(+/-) T cells, but did not influence IFN-γ expression and T cell proliferation. CONCLUSIONS/SIGNIFICANCE: IFN-ß may exert its inhibitory effect on EAU by inhibiting Th1, Th17 cells and modulating relevant cytokines. IFN-ß may provide a potential treatment for diseases mediated by Th1 and Th17 cells.

Increased Regulatory T Cells in Spleen during Experimental Autoimmune Uveoretinitis.

PURPOSE: To investigate the role of Foxp3-positive regulatory T cells in the development of experimental autoimmune uveoretinitis (EAU). METHODS: B10RIII mice were immunized with 50 microg IRBP(161-180) in complete Freund's adjuvant (CFA) to induce EAU. EAU was evaluated clinically and pathologically on days 0, 7, 14, 21, and 28. Foxp3 mRNA levels were detected using reverse transcription-PCR (RT-PCR) and the frequencies of CD4(+)Foxp3(+) T cells and CD4(+)CD25(+)Foxp3(+) T cells in splenocytes were assessed by flow cytometry at the aforementioned time points. RESULTS: The first clinical signs of EAU were observed on day 8-9, worsened up to day 14, and then gradually resolved. Histopathologic results showed that inflammatory signs occurred on day 7, reached their peak on day 14, and then gradually decreased. The levels of Foxp3 mRNA and the frequencies of CD4(+)Foxp3(+) T cells and CD4(+)CD25(+)Foxp3(+) T cells in splenocytes increased on day 7, reached a peak on day 14, and then maintained at a high level until day 28. CONCLUSION: An upregulation of Foxp3 expression is induced in EAU and paralleled with disease activity, suggesting a role for this lymphocyte subpopulation in the regression of this experimental uveitis model.

Contribution of CD4+CD25+ T cells to the regression phase of experimental autoimmune uveoretinitis.

PURPOSE: To investigate the role of CD4(+)CD25(+) Treg cells in the development of experimental autoimmune uveoretinitis (EAU). METHODS: EAU was induced in B10RIII mice by immunization with IRBP(161-180) in complete Freund's adjuvant and evaluated clinically and pathologically on days 0, 7, 14, 21, and 28. Lymphocytes from draining lymph nodes (LNs) were subjected to flow cytometry to analyze the frequency of CD4(+)CD25(+) Treg cells. CD4(+)CD25(+) Treg cells and CD4(+)CD25(-) T cells were separated by means of magnetic-assisted cell sorting and cocultured or crossover cultured for 3 days. Proliferation of CD4(+)CD25(-) T cells was measured using a modified MTT assay. The levels of IFN-gamma and IL-17 in the supernatants were determined by enzyme-linked immunosorbent assay. RESULTS: Clinical and histopathologic results showed a severe intraocular inflammation in the immunized mice. The frequency of CD4(+)Foxp3(+) T cells and CD4(+)CD25(+)Foxp3(+) T cells in the draining LN lymphocytes was increased on day 7, reached its peak on day 14, and maintained a high level up to day 42. CD4(+)CD25(+) Treg cells obtained from mice on days 14 and 28 after immunization showed a stronger inhibitory effect on the proliferation of CD4(+)CD25(-) T cells and the production of IFN-gamma by CD4(+)CD25(-) T cells compared with those obtained from control mice. CD4(+)CD25(+) Treg cells did not affect IL-17 production. Transfer of CD4(+)CD25(+) Treg cells obtained from EAU mice was able to suppress EAU induction by IRBP(161-180) that was not observed after transfer of cells from mice that had received CFA alone, suggesting antigen specificity of the Treg response. CONCLUSIONS: A significantly increased frequency and immunoregulatory action of CD4(+)CD25(+) Treg cells is associated with the development and regression of EAU, suggesting that CD4(+)CD25(+) Treg cells are induced during EAU and may be involved in its regression.

The synthesis of MP-CDCA conjugates and dissolution kinetics of model cholesterol gallstones.

The comb-like copolymers of polycarboxylic acid were synthesized and then reacted with chenodeoxycholic acid (CDCA) to obtain a series of conjugates, MPn-CDCA, where n is the number of the groups of oxyethylene in each graft chain. This was confirmed by infrared spectroscopy and thin-layer chromatography. We investigated the effects of dissolving model cholesterol gallstones with the MPn-CDCA conjugates in phosphate-buffered saline at pH 7.4. The dissolution rates of CDCA, MP40-CDCA, MP30-CDCA, MP20-CDCA and MP10-CDCA were 5.33, 5.717, 17.59, 6.868 and 9.615x10(-7)kgm(-2)s(-1), micellar solubilities were 0.2431, 3.095, 12.972, 5.248 and 5.790kgm(-3) and total resistances were 5.33, 5.717, 17.59, 6.868 and 9.615x10(-7)kgm(-2)s(-1), respectively. These studies suggested that the interfacial resistance was the dominant rate-determining factor in dissolving model cholesterol gallstones. Model cholesterol gallstones could be more effectively dissolved by increasing the steric interactive potential energy of side chains and ensuring that the hydrophilic-lipophilic properties of MP-CDCA are within an appropriate range. The micellar dissolution rates of model cholesterol gallstones by MP20-CDCA were significantly faster than by the other conjugates.