Oral administration of antigen has long been used in the induction of immune tolerance in various animal models of autoimmune diseases including rheumatoid arthritis (RA). Alleveation of arthritogenic symptoms has been reported from RA patients who received oral administration of type II collagen (CII) without side effects, however its rather inconsistent therapeutic efficacy and variation among patients calls for more detailed investigation on the mechanism of oral tolerance to be settled as regular treatment for RA. In an attempt to understand the immunogenic processes underpinning tolerance induction by orally administered CII, we analyzed changes in the expression of costimulatory molecules and STAT/SOCS signaling messengers in the mouse model of collagen induced arthritis (CIA). We found thatin the spleen of CIA mice, that has been undergone repeated oral feeding of CII prior to the induction of arthritis, showed increased promortion of CTLA4 expressing lymphocytes than in the spleen of PBS fed control. On the other hand, cells expressing CD28 or ICOS were decreased in the spleen of tolerized mice. Tolerance induction by oral CII administration also enhanced the expression of STAT6 in both RNA and protein level, while not affecting the expression of STAT3. The expression of SOCS3, which hasbeen known to transmit STAT-mediated signals from Th2 type cytokines, remained unchanged in the spleen of tolerized mice. Interestingly transcript of SOCS1, which has been associated with Th1 related pathways, was only visible in the spleen of tolerized but not of control mice, suggesting that as in the case of IL-6 signaling, it may exert a feed back inhibition toward the Th1 type stimulation.
Administration, Oral* ; Animals ; Arthritis* ; Arthritis, Rheumatoid ; Autoimmune Diseases ; Collagen ; Collagen Type II* ; Cytokines ; Hand ; Humans ; Immune Tolerance ; Interleukin-6 ; Lymphocytes ; Mice* ; Models, Animal ; RNA ; Spleen

BACKGROUND: To determine the molecular structure of type II collagen-specific T-cell receptors associated with rheumatoid arthritis (RA). METHODS: We generated CII-specific T-cell lines of 8 RA patient s by prolonged in vitro culture with bovine CII (bCII) and the immunogenic peptide (256-270) of human CII. The proliferation response towards CII stimulation was measured from the uptake of 3 H-thymidine. Changes in the secretion of Th 1 and Th2 cytokines in the culture supernatent were measured by ELISA. The TCR clonotypes of these T-cells were examined by RT-PCR/ SSCP analyses of all 22 V beta chains. RESULTS: T-cells from patients' tissue exhibited strong proliferation index upon CII stimulation, which was maintained up to 6 months in the culture. The secretion of INF-gamma from these T-cells increased along with the duration of culture time, while the amount of IL-4 production did not show significant changes. The SSCP band patterns of patients' T-cells appear as discrete bands unlike the smeary streak produced from normal samples. Some SSCP bands, each representing selected expansion of a TCR containing certain subtype of V beta peptides, appeared to be identical in more than one patients. Among these, the expansion of SSCP band representing the V beta 14 CDR3 region persisted after switching the antigen to the immunogenic human peptide (256-270). CONCLUSION: CII-reactive T-cells expressing distinct CDR3 motifs are selectively expanded in the peripheral blood and synovial fluid of RA patients, and their persistent proliferation upon CII stimulation, as well as the production Th 1-type cytokines, may play pivotal roles in RA pathogenesis.
Arthritis, Rheumatoid* ; Cytokines ; Enzyme-Linked Immunosorbent Assay ; Humans ; Interleukin-4 ; Molecular Structure ; Peptides ; Polymorphism, Single-Stranded Conformational ; Receptors, Antigen, T-Cell ; Synovial Fluid ; T-Lymphocytes*

We report three cases of cystinuria, presenting with urinary stones. A 2-year-old girl presented with urinary difficulty, hematuria, dysuria of sudden onset, and her 7-month-old younger brother also was presented with urinary difficulty, irritability on urination & stone passage. Other 6-month-old boy was admitted due to sudden onset anuria. They had radioopague renal & ureter stones and stone analysis revealed mixed cystine stones. The diagnosis of cystinuria was confirmed metabolic studies and stone analysis. Lrinary amino acid analysis showed excessive excretion of dibasic amino acids(cystine, ornithine, lysine, arginine). And they all had hypercalciuria and hyperuricosuria. They were treated with combination of percutaneous lithotripsy for large obstructing senes a nd an oral drug therapy with sodium bicarbonate for rendering the urine more alkaline, and alpha-mercaptopropionylglycine(ThiolaR). This form of treatment was sucessful in our three cases with elimination of recurrent nephrolithiasis, but in one patient, nephrotic syndrome possibly caused by ThiolaR was developed. The nephrotic syndrome was recovered spontaneously after cessation of Thiola. A review of literatures was also attempted briefly.
Anuria ; Child, Preschool ; Cystine ; Cystinuria* ; Diagnosis ; Drug Therapy ; Dysuria ; Female ; Hematuria ; Humans ; Hypercalciuria ; Infant ; Lithotripsy ; Lysine ; Male ; Nephrolithiasis ; Nephrotic Syndrome ; Ornithine ; Siblings ; Sodium Bicarbonate ; Tiopronin ; Ureter ; Urinary Calculi ; Urination

Recent evidence has strongly suggested that the CAP/TC10 pathway is involved in the trafficking, docking,and fusion of vesicles containing the insulin- responsive glucose transporter Glut4 to the plasma membrane. However, little is known about how the genes employed in the CAP/TC10 pathway are associated with the development of type 2 diabetes mellitus. In this study, we sequenced 4 genes of the CAP/TC10 pathway [SORBS1, CBL, CRK, and RHOQ] in 24 individuals to identify genetic variations in these loci. A total of 48 sequence variants were identified, including 23 novel variations. To investigate the possible association with type 2 diabetes mellitus, 3 single nucleotide polymorphisms from SORBS1, 3 from CBL , and 4 from RHOQ were genotyped in 1122 Korean type 2 diabetic patients and 1138 nondiabetic controls. Using logistic regression analysis, 1 significant association between SNP rs1376405 in RHOQ and type 2 diabetes mellitus [OR = 8.714 (C.I. 1.714-44.29), p = 0.009] was found in the recessive model. Our data demonstrate a positive association of the RHOQ gene in the CAP/TC10 pathway with T2DM in the Korean population.
Cell Membrane ; Diabetes Mellitus, Type 2 ; Genetic Variation ; Glucose Transport Proteins, Facilitative ; Humans ; Insulin ; Logistic Models ; Polymorphism, Single Nucleotide ; Signal Transduction

The earliest stages of mammalian embryogenesis are governed by the activity of maternally inherited transcripts and proteins. Cytoplasmic polyadenylation of selected maternal mRNA has been reported to be a major control mechanism of delayed translation during preimplantation embryogenesis in mice. The presence of cis-elements required for cytoplasmic polyadenylation (e.g., CPE) can serve as a useful tag in the screening of maternal genes partaking in key functions in the transcriptionally dormant egg and early embryo. However, due to its relative simplicity, UA-rich sequences satisfying the canonical rule of known CPE consensus sequences are often found in the 3'-UTR of maternal transcripts that do not actually undergo cytoplasmic polyadenylation. In this study, we developed a method to confirm the validity of candidate CPE sequences in a given gene by a multiplex comparison of 3'-UTR sequences between mammalian homologs. We found that genes undergoing cytoplasmic polyadenylation tend to create a conserved block around the CPE, while CPE-like sequences in the 3'-UTR of genes lacking cytoplasmic polyadenylation do not exhibit such conservation between species. Through this cross-species comparison, we also identified an alternative CPE in the 3'-UTR of tissue-type plasminogen activator (tPA), which is more likely to serve as a functional element. We suggest that verification of CPEs based on sequence conservation can provide a convenient tool for mass screening of factors governing the earliest processes of mammalian embryogenesis.
Animals ; Consensus Sequence ; Cytoplasm ; Embryonic Development ; Embryonic Structures ; Female ; Mass Screening ; Mice ; Ovum ; Polyadenylation ; Pregnancy ; RNA, Messenger, Stored ; Tissue Plasminogen Activator

BACKGROUND: Volatile agents have been reported to protect myocardium against ischemia. But, there were a few clinical reports about the myocardial protection of inhalation agents. So we investigated the cardiac protection of sevoflurane in comparison with total intravenous anesthesia (TIVA). The study is a retrospective unrandomized study via the medical record review. METHODS: The records of 102 patients who received off-pump CABG were reviewed. One patient group received TIVA by midazolam and sufentanil continuous infusion (TIVA group, n = 68), and the other patient group received an inhalational anesthesia by sevoflurane (sevoflurane group, n = 34). Except maintenance of anesthesia, two groups of patients received an identical surgical, anesthetical, and postoperative care. At arrival in the intensive care unit, and after 1, 2, 3 and 5 days, serum cardiac enzyme levels were measured. RESULTS: All the median values of cardiac enzyme concentrations were lower in the sevoflurane group than TIVA group. Moreover, there were the significant differences between groups at the immediate postoperative CK-MB (median 4.7 ng/ml versus 5.9 ng/ml (P = 0.049)), 1-5 days postoperative LD (1 day 271.5 U/L versus 292 U/L (P = 0.045), 2 day 227.5 U/L versus 270 U/L (P = 0.009), 3 day 215 U/L versus 250 U/L (P = 0.030), 5 day 218 U/L versus 231 U/L (P = 0.005)), and 1, 3 postoperative troponin I level (0.485 ng/ml versus 1.12 ng/ml [P = 0.029], 0.090 ng/ml versus 0.235 ng/ml [P = 0.047] respectively). CONCLUSIONS: Sevoflurane lowered cardiac enzyme levels in comparison with TIVA after off-pump CABG anesthesia. These data suggest a cardioprotective effect of sevoflurane during CABG.
Anesthesia ; Anesthesia, Intravenous ; Coronary Artery Bypass, Off-Pump ; Humans ; Inhalation ; Intensive Care Units ; Ischemia ; Medical Records ; Methyl Ethers ; Midazolam ; Myocardium ; Postoperative Care ; Retrospective Studies ; Sufentanil ; Troponin I

PURPOSE: The interactive effects of genetic polymorphisms of cytochrome P4502E1 (CYP2E1) & N-acetyltransferase 1 (NAT1) and smoking on lung cancer development were evaluated in hospital based case-control study. MATERIALS AND METHODS: Male lung cancer patients (N= 157) and the male patients with no present or previous history of systemic illnesses who visited the urology department (N=138) were recruited (1998-1999). CYP2E1 & NAT1 genotypes were determined by PCR-RFLP method using RsaI and MboII digestion, respectively. RESULTS: CYP2E1 c2 or NAT1 *10 allele did not increased the risk of lung cancer. Heavy smokers (35 Alleles ; Case-Control Studies ; Cytochrome P-450 CYP2E1* ; Cytochromes ; Digestion ; Gene-Environment Interaction ; Genotype ; Humans ; Lung Neoplasms* ; Lung* ; Male ; Polymorphism, Genetic* ; Smoke* ; Smoking* ; Urology

Oral administration of antigen has long been considered as a promising alternative for the treatment of chronic autoimmune diseases including rheumatoid arthritis (RA), and oral application of type II collagen (CII) has been proven to improve pathogenic symptoms in RA patients without problematic side effects. To further current understandings about the immune suppression mechanisms mediated by orally administered antigens, we examined the changes in IgG subtypes, T-cell proliferative response, and proportion of interleukin (IL)-10 producing Th subsets in a time course study of collagen induced arthritis (CIA) animal models. We found that joint inflammation in CIA mouse peaked at 5 weeks after first immunization with CII, which was significantly subdued in mice pre-treated by repeated oral administration of CII. Orally tolerized mice also showed increase in their serum level of IgG1, while the level of IgG2a was decreased. T-cell proliferation upon CII stimulation was also suppressed in lymph nodes of mice given oral administration of CII compared to non-tolerized controls. When cultured in vitro in the presence of CII, T-cells isolated from orally tolerized mice presented higher proportion of CD4+ IL-10+ subsets compared to non-tolerized controls. Interestingly, such increase in IL-10 producing cells were obvious first in Peyer's patch, then by 5 weeks after immunization, in mesenteric lymph node and spleen instead. This result indicates that a particular subset of T-cells with immune suppressive functions might have migrated from the original contact site with CII to inflamed joints via peripheral blood after 5 weeks post immunization.
Administration, Oral ; Animals* ; Arthritis* ; Arthritis, Rheumatoid ; Autoimmune Diseases ; Collagen ; Collagen Type II ; Humans ; Immunity, Cellular* ; Immunization ; Immunoglobulin G ; Inflammation ; Interleukin-10 ; Interleukins ; Joints ; Lymph Nodes ; Mice ; Models, Animal* ; Spleen ; T-Lymphocytes

Background: In Korea, during the early phase of the coronavirus disease 2019 (COVID-19) pandemic, we responded to the uncertainty of treatments under various conditions, consistently playing catch up with the speed of evidence updates. Therefore, there was high demand for national-level evidence-based clinical practice guidelines for clinicians in a timely manner. We developed evidence-based and updated living recommendations for clinicians through a transparent development process and multidisciplinary expert collaboration. Methods: The National Evidence-based Healthcare Collaborating Agency (NECA) and the Korean Academy of Medical Sciences (KAMS) collaborated to develop trustworthy Korean living guidelines. The NECA-supported methodological sections and 8 professional medical societies of the KAMS worked with clinical experts, and 31 clinicians were involved annually. We developed a total of 35 clinical questions, including medications, respiratory/critical care, pediatric care, emergency care, diagnostic tests, and radiological examinations. Results: An evidence-based search for treatments began in March 2021 and monthly updates were performed. It was expanded to other areas, and the search interval was organized by a steering committee owing to priority changes. Evidence synthesis and recommendation review was performed by researchers, and living recommendations were updated within 3–4 months. Conclusion We provided timely recommendations on living schemes and disseminated them to the public, policymakers and various stakeholders using webpages and social media.Although the output was successful, there were some limitations. The rigor of development issues, urgent timelines for public dissemination, education for new developers, and spread of several new COVID-19 variants have worked as barriers. Therefore, we must prepare systematic processes and funding for future pandemics.