Association between nutritional level, menstrual-related symptoms, and mental health in female medical students.

OBJECTIVE: Research suggests that medical students as well as women are at greater risk of depression compared to the general population and men. This suggests that female medical students are crucial targets requiring specific monitoring for mental health disorder prevention and intervention. However, little is known regarding the risk factors for poor mental health among this population. Therefore, we investigated whether menstrual symptoms and nutritional status are associated with psychological distress in Japanese female medical students. METHODS: This cross-sectional study assessed 326 female medical students who attended a school medical check-up, which included blood sampling in 2018. The levels of psychological distress were evaluated using the Japanese General Health Questionnaire (J-GHQ)-30. We defined high GHQ scores as GHQ-30 ≥7. We checked dysmenorrhea levels and assessed menstrual symptoms according to the presence of premenstrual syndrome (PMS). Dysmenorrhea was evaluated according to quartiles of the sum of the Menstrual Distress Questionnaire (MDQ). PMS was assessed using the Premenstrual Symptoms Questionnaire (PSQ). We evaluated levels of serum albumin, hemoglobin, ferritin, and lipid metabolite as nutritional factors. A multivariate logistic regression analysis was used to identify the association between menstrual-related symptoms or nutritional factors and the levels of psychological distress. RESULTS: A total of 45 female medical students (15%) experienced psychological distress. Serum albumin levels were associated with psychological distress, while lipid metabolite levels were not. The intensity of dysmenorrhea and the presence of PMS were associated with psychological distress, independent of nutritional status. CONCLUSION: Both menstrual symptoms and nutrition markers were associated with the levels of psychological distress in Japanese female medical students. School doctors and nurses can help improve the mental health of young female medical students by encouraging a healthy diet and checking for the presence of menstrual symptoms.

Association between dyslipidemia and plasma levels of branched-chain amino acids in the Japanese population without diabetes mellitus.

BACKGROUND: Branched-chain amino acids (BCAAs) play a key role in energy homeostasis. OBJECTIVE: We aimed to investigate the association between plasma BCAA levels and dyslipidemia in the Japanese population without diabetes mellitus. METHODS: This cross-sectional study included 4952 participants without diabetes mellitus, enrolled in the Tsuruoka Metabolomic Cohort Study. Plasma BCAA levels were measured by capillary electrophoresis-mass spectrometry. Correlations between lipid and BCAA profiles were evaluated by sex-stratified multiple linear regression analyses, after adjusting for confounders. Logistic regression was used to identify associations between BCAAs and metabolic dyslipidemia (MD) defined as triglyceride levels ≥150 mg/dL, high-density lipoprotein cholesterol levels ≤40 mg/dL for men and ≤50 mg/dL for women, or low-density lipoprotein cholesterol (LDL-C) levels ≥140 mg/dL. RESULTS: In both sexes, the levels of individual BCAAs and the total BCAA levels correlated positively with triglyceride levels and negatively with high-density lipoprotein cholesterol levels. Valine, leucine, and total BCAA levels were weakly and positively correlated with LDL-C levels. Increased BCAA levels showed positive associations with MD. However, associations between BCAAs and elevated LDL-C levels were unclear. Furthermore, the associations between BCAA levels and MD regardless of fasting blood sugar (FBS) levels (high or low). Although valine, leucine, and total BCAA levels were weakly associated with elevated LDL-C levels in the high-FBS group, no such association was observed in the low-FBS group. CONCLUSIONS: BCAAs might be associated with MD independently of the FBS level and might play an important role in lipid metabolism and dyslipidemia.

Association of aortic root dilatation with left ventricular function in patients with postoperative ventricular septal defect.

Proximal aortic enlargement is associated with an increased risk of heart failure and all-cause mortality. Recently, aortic root dilatation (ARD) was reported in postoperative patients with ventricular septal defects (VSDs). However, the impact of ARD on left ventricular (LV) function in postoperative VSD patients remains unclear. Thus, the aim of this study was to investigate the effect of ARD on LV function in patients with postoperative VSD. One hundred and thirty-five patients (> 15 years of age) with surgically repaired isolated ventricular defects and who underwent transthoracic echocardiography in our institution between 2009 and 2013 were identified. ARD was defined as an observed aortic root diameter/body surface area > 2.1 cm/m2. The propensity score estimating the probability of having ARD adjusted for anatomical and clinical characteristics was calculated. Forty-four patients (32.6%) had ARD. In unadjusted analyses, right ventricular systolic pressure, Tei index, and E/e' were significantly (p < 0.05) higher in patients with ARD than in those without ARD (31.3 ± 7.5 vs. 35.4 ± 13.7 mmHg, 0.32 ± 0.10 vs. 0.44 ± 0.15, and 7.1 ± 1.7 vs. 9.5 ± 2.9, respectively). In the propensity score-adjusted analysis, significant differences in the Tei index and E/e' were confirmed between the two groups (Tei index difference: 0.11, 95% confidence interval 0.05-0.17; E/e' difference: 2.4, 95% confidence interval 1.3-3.5). However, there were no differences in the other echocardiographic measurements. The presence of ARD in patients with postoperative VSD was significantly associated with LV diastolic dysfunction. Thus, surgically repaired VSD patients require careful screening for aortic enlargement and LV function.

Factors associated with aortic root dilatation after surgically repaired ventricular septal defect.

BACKGROUND: Aortic root dilatation (ARD) in congenital heart disease is related to aortic aneurysm, rupture, and dissection. This study aimed to investigate the factors associated with ARD in patients with surgically repaired ventricular septal defect (VSD). METHODS: This cross-sectional study included 152 patients with surgically repaired VSD. Two definitions of ARD were used: (1) observed Valsalva diameter to body surface area (BSA) ratio >2.1 cm/m2 and (2) absolute value of Valsalva diameter ≥4.0 cm. Odds ratios (ORs) and 95% confidence intervals (CIs) of ARD presence were calculated using multivariate logistic regressions. RESULTS: The prevalence of ARD ranged between 8.6% and 32.9%. Using the definition of observed aortic root diameter/BSA >2.1 cm/m2 , patients with nonsubarterial VSD type were more likely to have ARD (OR 5.65, 95% CI; 1.83-17.44, P=.003) than those with subarterial type, and patients with preoperative right- or noncoronary cusp prolapse (R/NCCP) were more likely to have ARD (OR 3.68, 95% CI; 1.20-11.23, P=.022) than patients without preoperative R/NCCP after adjustment for sex, age at repair (ie, shunt duration), VSD size, and postoperative follow-up period. Using the definition of absolute Valsalva diameter ≥4 cm, nonsubarterial VSD type and presence of R/NCCP were also significantly associated with ARD after adjustment for the same covariates. CONCLUSIONS: Anatomical and morphological features (nonsubarterial type and presence of preoperative R/NCCP) are independently correlated with ARD in patients with VSD regardless of the different definitions of ARD. Patients with surgically repaired VSD may need careful monitoring for potential ARD development.

Correlation between left atrial appendage morphology and flow velocity in patients with paroxysmal atrial fibrillation.

AIMS: Reduction of left atrial appendage (LAA) flow velocity (FV) is a risk factor for thrombus formation and increases the risk of stroke in patients with atrial fibrillation (AF). Furthermore, LAA morphology is correlated with stroke in patients with AF. The aim of this study was to correlate LAAFV with LAA morphology in patients with AF. METHODS AND RESULTS: We studied 96 patients (age 59.0 ± 10.2 years, 75% male) referred for radiofrequency catheter ablation for paroxysmal AF. All patients underwent computed tomography (CT) and transthoracic and transoesophageal echocardiography during sinus rhythm. LAA morphology was classified as one of the four types (chicken wing, windsock, cactus, and cauliflower) on CT images. There were significant differences in LAAFV among LAA morphologies (chicken wing 73.7 ± 21.9 cm/s, windsock 61.9 ± 19.6 cm/s, cactus 55.3 ± 14.1 cm/s, cauliflower 52.7 ± 18.1 cm/s, P = 0.008). Post hoc multiple comparisons showed that LAAFV was higher in patients with chicken wing than in those with cactus (P = 0.006, vs. chicken wing) and cauliflower (P = 0.006, vs. chicken wing), but not with windsock (P = 0.102). After adjustment for clinical and LAA anatomical covariates (orifice area, volume, and trabeculation), multiple linear regression analyses revealed that LAA morphology was an independent determinant of LAAFV [chickens wing: standardized partial regression coefficients (ß) = 0.317, P = 0.0014; windsock: ß = 0.303, P = 0.038]. CONCLUSION: LAA morphology is a significant determinant of LAAFV, suggesting an underlying mechanism for the association between LAA morphology and embolic events.

Impact of an Empiric Isolation of the Superior Vena Cava in Addition to Circumferential Pulmonary Vein Isolation on the Outcome of Paroxysmal Atrial Fibrillation Ablation.

The safety and efficacy of an empiric superior vena cava isolation (SVCI) in addition to circumferential pulmonary vein isolation (CPVI) in patients with paroxysmal atrial fibrillation (PAF) have not been clarified. A total of 186 consecutive patients who underwent catheter ablation of PAF were included. All patients underwent a CPVI. Patients in the first half underwent an additional SVCI only if SVC-triggered AF or rapid SVC activity was observed during the procedure (n = 93, as-needed SVCI, group I), and those in the second half underwent an empirical SVCI after the CPVI (n = 93, empiric SVCI, group II). The CPVI was successfully performed in all patients. An SVCI was performed in 8 of 93 patients (9%) in group I and 81 of the 93 patients (87%) in group II. In the remaining 12 patients in group II, an SVCI was not performed because of the lack of SVC potentials. During a mean follow-up of 27 ± 12 months, the atrial tachyarrhythmia recurrence rate after a single ablation procedure in the patients in group II was lower than that in group I (44% vs 23%, p = 0.035). A Cox regression multivariate analysis demonstrated that an empiric SVCI was an independent predictor of an atrial tachyarrhythmia recurrence after a single ablation procedure (odds ratio: 0.57, 95% confidence interval 0.31 to 0.999; p = 0.049). Neither sinus node injury nor any injury to the phrenic nerve was observed. In conclusion, an empiric SVCI in addition to the CPVI improved the outcome of AF ablation in patients with PAF without any additional adverse effects.

Left atrial appendage flow velocity and time from P-wave onset to tissue Doppler-derived A' predict atrial fibrillation recurrence after radiofrequency catheter ablation.

BACKGROUND: Atrial fibrillation (AF) is associated with atrial remodeling. We investigate the abilities of preprocedural echocardiographic parameters reflecting atrial remodeling to predict AF recurrence after radiofrequency catheter ablation (RFCA) for paroxysmal AF (PAF). METHODS: Preprocedural echocardiographic parameters were measured during sinus rhythm in 105 patients with PAF undergoing RFCA. Electrical remodeling was assessed by the time from the onset of the P-wave to the peak A'-wave on the tissue Doppler imaging (PA-TDI), functional remodeling was assessed by the left atrial appendage flow velocity (LAAFV), and structural remodeling was assessed by the left atrial volume index (LAVI). PA-TDI, LAAFV, and LAVI values were divided into tertiles, and their abilities to predict AF recurrence were assessed using Cox regression analysis. RESULTS: AF recurrence occurred in 39/105 (37.1%) patients. After adjustment for confounders, the rate of AF recurrence was significantly higher in the highest tertile of PA-TDI compared with the lowest tertile (≥151.3 msec vs. <131.0 msec; hazard ratio [HR]: 2.477, 95% confidence interval [CI]: 1.031-5.950; P = 0.042), and in the lowest tertile of LAAFV compared with the highest tertile (<48.5 cm/sec vs. ≥64.9 cm/sec; HR: 2.680, 95% CI: 1.136-6.318; P = 0.024). The risk of AF recurrence was also higher in the highest tertile of LAVI (≥34.2 mL/m(2) ) compared with the lowest tertile, but this difference was not significant (HR: 2.146, 95% CI: 0.834-5.523; P = 0.113). CONCLUSIONS: LAAFV (reflecting functional remodeling) and PA-TDI (reflecting electrical remodeling) are independent predictors of AF recurrence after RFCA for PAF.

Impact of atrial remodeling on the outcome of radiofrequency catheter ablation of paroxysmal atrial fibrillation.

BACKGROUND: Both the left atrial volume index (LAVI) and estimated total atrial conduction time measured using tissue Doppler imaging of the atria (PA-TDI duration) are echocardiographic parameters reflecting atrial remodeling. We investigated their prognostic value for atrial tachyarrhythmia (AF/AT) recurrence after radiofrequency catheter ablation (RFCA) of paroxysmal atrial fibrillation (PAF). METHODS AND RESULTS: We analyzed the data for 100 consecutive patients with drug-refractory PAF who underwent RFCA. The correlation between the LAVI and PA-TDI was extremely weak (r=0.26, P<0.01). We categorized the patients into 4 groups based on the median LAVI and PA-TDI duration: group 1 (LAVI <29ml/m(2)/PA-TDI duration <143ms), group 2 (LAVI ≥29ml/m(2)/PA-TDI duration <143ms), group 3 (LAVI <29ml/m(2)/PA-TDI duration ≥143ms), and group 4 (LAVI ≥29ml/m(2)/PA-TDI duration ≥143ms). With a mean follow-up of 20.2±8.9 months after a single RFCA procedure, 60 patients (60%) were in sinus rhythm without any antiarrhythmic drugs. Multivariate analysis using a Cox proportional hazards model demonstrated that the group was an independent predictor of AF/AT recurrence after RFCA (P=0.0017). The patients in groups 2, 3, and 4 had a 4.0-fold (P=0.048), 6.8-fold (P=0.0034) and 10.9-fold (P=0.0001) increase, respectively, in the probability of recurrent AF/AT as compared with group 1. CONCLUSIONS: Preprocedural echocardiographic estimation of atrial remodeling was a useful predictor of AF/AT recurrence following a single RFCA of PAF.

Impact of myocardial reperfusion status on QT dispersion after successful recanalization of the infarct-related artery in acute myocardial infarction.

OBJECTIVE: To determine whether adequate myocardial perfusion status after transluminal recanalization is associated with prompt improvement of QT dispersion (QTd). BACKGROUND: Transluminal recanalization of the infarct-related coronary artery in acute myocardial infarction aims to promptly restore myocardial perfusion, to maximize electrical and mechanical recovery. QTd represents the heterogeneity of ventricular repolarization, which may affect electrical stability. METHODS: Forty patients who underwent primary percutaneous coronary intervention for their first anterior acute ST-elevation myocardial infarction were prospectively enrolled. Myocardial reperfusion status was assessed by myocardial blush grade (MBG) on the final angiogram after successful recanalization (Thrombolysis In Myocardial Infarction Grade 3 flow). RESULTS: Preprocedural QTd was similar in patients with final MBG 0-1, 2, and 3 (76 ± 24, 67 ± 13, and 69 ± 13 milliseconds, respectively; P = 0.661). After recanalization, QTd decreased in patients with MBG 3 (39 ± 16 milliseconds, P < 0.001) but not in patients with MBG 0-1 (74 ± 20 milliseconds) or MBG 2 (82 ± 16 milliseconds). Multivariate analysis showed that postprocedural MBG was an independent predictor of QTd after recanalization (standardized regression coefficient = -0.628, P < 0.001). CONCLUSIONS: Adequate tissue perfusion may be crucial for electrical stability of the myocardium after reperfusion.

Long-term outcome and preprocedural predictors of atrial tachyarrhythmia recurrence following pulmonary vein antrum isolation-based catheter ablation in patients with non-paroxysmal atrial fibrillation.

BACKGROUND: Although various empiric adjunctive ablation techniques are widely performed with pulmonary vein antrum isolation (PVAI) to enhance the procedural efficacy of catheter ablation in non-paroxysmal atrial fibrillation (NPAF) patients, they are not required in all NPAF patients. METHODS AND RESULTS: Eighty consecutive NPAF patients refractory to antiarrhythmic drugs underwent a PVAI-based ablation. Structural heart disease was present in 40% of patients and systolic dysfunction in 21%. After 31 ± 16 months of follow-up, 41% of the patients were free of atrial tachyarrhythmia recurrences after a single procedure. Finally, during a mean follow-up of 25 ± 15 months, 63 of 80 (79%) patients remained in sinus rhythm (SR) after the final procedure (two procedures in 48%, and three in 3%). A Cox regression multivariate analysis revealed that left atrial volume (LAV) was the only independent predictor of atrial tachyarrhythmia recurrences not only after single procedures (p = 0.027), but also after the final procedures (p = 0.001). Ten patients (13%) needed ablation for concomitant atrial tachycardias originating from the left atrium and right atrium other than common atrial flutter. Repeat ablation procedures increased the best cut-off value for predicting recurrences analyzed by receiver operating characteristic curves, from 86 mL (sensitivity 70%, specificity 64%) to 92 mL (sensitivity 71%, specificity 78%). CONCLUSIONS: PVAI-based ablation strategies could achieve SR maintenance in almost 80% of NPAF patients after multiple procedures during long-term follow-up. The preprocedural LAV was an important predictor of the procedural outcome.

Trajectories of anti-islet autoantibodies before development of type 1 diabetes in interferon-treated hepatitis C patients. Case reports and a literature review.

Interferon-alpha (IFN-α) is widely used in the treatment of viral hepatitis, however, it is known that IFN-α therapy may induce type 1 diabetes. We report here on two cases of chronic viral hepatitis C who developed autoimmune type 1 diabetes during Peg-IFN-α plus ribavirin (RBV) therapy. Case 1: a 48-year-old male with chronic hepatitis C with chronic thyroiditis. The patient's plasma glucose level was normal and anti-islet autoantibody tests were negative before Peg-IFN-α+RBV therapy. The emergence of glutamic acid decarboxylase 65 autoantibody (GAD65Ab) was observed after five months of treatment. Autoantibodies to insulin and insulinoma-associated antigen-2 (IA-2) also became positive. Eleven months later, thirst and polydipsia occurred with increased fasting plasma glucose level and the patient was diagnosed with type 1A diabetes. Zinc transporter-8 autoantibody (ZnT8Ab) was not detectable at any point. The patient has type 1 diabetes-susceptible HLA-DRB1-DQB1 haplotypes *0405-*0401 and *0901-*0303. Case 2: a 65-year-old male with chronic hepatitis C with type 2 diabetes on insulin treatment. GAD65Ab and IA-2Ab were negative before Peg-IFN-α+RBV therapy, however, nine months later, a single appearance of GAD65Ab was observed. After twelve months, his plasma glucose control worsened rapidly, and he was diagnosed with type 1A diabetes. IA-2Ab and ZnT8Ab were negative throughout the clinical course. His HLA-DRB1-DQB1 haplotypes were *0410-*0402 and *1407-*0503. Both cases showed a unique GAD65Ab epitope (amino acids 360-442). These clinical courses suggest that IFN-α therapy provoked acute islet autoimmunity and onset of type 1 diabetes. Therefore, during IFN-α therapy, patients should be closely monitored for the occurrence of type 1 diabetes.

alpha1,2-Fucosylated and beta-N-acetylgalactosaminylated prostate-specific antigen as an efficient marker of prostatic cancer.

A prostate-specific antigen (PSA) is widely used as a diagnostic marker for prostate cancer (PC) because of its high specificity. However, elevated serum PSA does not occur only in PC but also in benign prostatic hyperplasia (BPH). Since the structural changes of N-glycans during carcinogenesis are common phenomena, we investigated whether PC-specific N-glycans are linked to PSA. We first analyzed the carbohydrate structures of PSA derived from seminal fluid, serum of BPH and PC patients, and PC cell line, namely, LNCaP using eight lectin-immobilized columns and then with enzyme-linked immunosorbent assay (ELISA). The fraction of serum PSA from PC patients bound to both Fucalpha1-2Gal and betaGalNAc binding Trichosanthes japonica agglutinin-II (TJA-II) column, while that from BPH patients did not exhibit this binding ability, thereby implying that there is elevated expression of alpha1,2-fucosylation and beta-N-acetylgalactosaminylation of PSA during carcinogenesis. We then performed a real-time polymerase chain reaction (PCR) and confirmed that these structural changes were responsible for the elevated expression of fucosyltransferase I (FUT1) and beta-N-acetylgalactosaminyltransferase 4(B4GALNT4). Second, we measured TJA-II-bound PSA contents and the binding ratios of TJA-II column chromatography in serum PSA samples from 40 patients of both PC and BPH. The results indicated that both TJA-II-bound PSA content and TJA-II binding ratios (%) could be used to discriminate between PC and BPH with more than 95% probability, and TJA-II-bound PSA can be regarded as a potential marker of PC.

Neuroblastoma GOTO cells are hypersensitive to disruption of lipid rafts.

GOTO cells, a neuroblastoma cell line retaining the ability to differentiate into neuronal or Schwann cells, were found to be rich in membrane rafts containing ganglioside GM2 and hypersensitive to lipid raft-disrupting methyl-beta-cyclodextrin (MbetaCD); the GM2-rich rafts and sensitivity to MbetaCD were markedly diminished upon their differentiation into Schwann cells. We first raised a monoclonal antibody that specifically binds to GOTO cells but not to differentiated Schwann cells and determined its target antigen as ganglioside GM2, which was shown to be highly concentrated in lipid rafts by its colocalization with flotillin, a marker protein of rafts. Disturbance of normal structure of the lipid raft by depleting its major constituent, cholesterol, with MbetaCD resulted in acute apoptotic cell death of GOTO cells, but little effects were seen on differentiated Schwann cells. Until this study, GM2-rich rafts are poorly characterized and MbetaCD hypersensitivity, which may have clinical implications, has not been reported.

A Caenorhabditis elegans glycolipid-binding galectin functions in host defense against bacterial infection.

Galectins are a family of beta-galactoside-binding proteins that are widely found among animal species and that regulate diverse biological phenomena. To study the biological function of glycolipid-binding galectins, we purified recombinant Caenorhabditis elegans galectins (LEC-1-11) and studied their binding to C. elegans glycolipids. We found that LEC-8 binds to glycolipids in C. elegans through carbohydrate recognition. It has been reported that Cry5B-producing Bacillus thuringiensis strains can infect C. elegans and that the C. elegans Cry5B receptor molecules are glycolipids. We found that Cry5B and LEC-8 bound to C. elegans glycolipid-coated plates in a dose-dependent manner and that Cry5B binding to glycolipids was inhibited by the addition of LEC-8. LEC-8 is usually expressed strongly in the pharyngeal-intestinal valve and intestinal-rectal valve and is expressed weakly in intestine. However, when C. elegans were fed Escherichia coli expressing Cry5B, intestinal LEC-8::EGFP protein levels increased markedly. In contrast, LEC-8::EGFP expression triggered by Cry5B was reduced in toxin-resistant C. elegans mutants, which had mutations in genes involved in biosynthesis of glycolipids. Moreover, the LEC-8-deficient mutant was more susceptible to Cry5B than wild-type worms. These results suggest that the glycolipid-binding lectin LEC-8 contributes to host defense against bacterial infection by competitive binding to target glycolipid molecules.

Bovine lactoferrin potently inhibits liver mitochondrial 8-OHdG levels and retrieves hepatic OGG1 activities in Long-Evans Cinnamon rats.

BACKGROUND/AIMS: To assess the effect of lactoferrin on oxidative liver damage and its mechanism, we used Long-Evans Cinnamon (LEC) rats that spontaneously develop fulminant-like hepatitis and lethal hepatic failure. METHODS: Four-week-old female LEC rats were divided into the untreated and treated groups. The latter was fed bovine lactoferrin at 2% mixed with conventional diet. RESULTS: The cumulative survival rates were 75.0% vs. 100% at 14 weeks, 37.5% vs. 91.7% at 15 weeks, and 12.5% vs. 91.7% at 16 weeks, respectively, for untreated and treated rats (P=0.0008). The 8-OHdG levels in liver mitochondrial DNA and malondialdehyde in plasma and liver tissues were significantly lower in treated than untreated rats (P<0.001, =0.017 and 0.034, respectively). Mitochondrial DNA mutations were more common in untreated rats. OGG1 mRNA and protein expression levels were significantly lower in untreated than treated rats (P=0.003 and 0.007, respectively). Hypermethylation of the second CpG island located upstream of OGG1 gene was observed in untreated rats. CONCLUSIONS: Our findings indicated that lactoferrin inhibits oxidative liver damage in LEC rats. Lactoferrin could be potentially useful for the treatment of oxidative stress-induced liver diseases.

Combined insulin B:9-23 self-peptide and polyinosinic-polycytidylic acid accelerate insulitis but inhibit development of diabetes by increasing the proportion of CD4+Foxp3+ regulatory T cells in the islets in non-obese diabetic mice.

Insulin peptide B:9-23 is a major autoantigen in type 1 diabetes. Combined treatment with B:9-23 peptide and polyinosinic-polycytidylic acid (poly I:C), but neither alone, induce insulitis in normal BALB/c mice. In contrast, the combined treatment accelerated insulitis, but prevented diabetes in NOD mice. Our immunofluorescence study with anti-CD4/anti-Foxp3 revealed that the proportion of Foxp3 positive CD4(+)CD25(+) regulatory T cells (Tregs) was elevated in the islets of NOD mice treated with B:9-23 peptide and poly I:C, as compared to non-treated mice. Depletion of Tregs by anti-CD25 antibody hastened spontaneous development of diabetes in non-treated NOD mice, and abolished the protective effect of the combined treatment and conversely accelerated the onset of diabetes in the treated mice. These results indicate that poly I:C combined with B:9-23 peptide promotes infiltration of both pathogenic T cells and predominantly Tregs into the islets, thereby inhibiting progression from insulitis to overt diabetes in NOD mice.

Humoral immune response to islet autoantigens in Japanese patients with type 1 diabetes.

In this study, we evaluated autoantibodies to IA-2 (IA-2As), glutamic acid decarboxylase 65 (GADAs), and islet cell antibodies (ICAs) in 233 patients with type 1 diabetes (M:F = 90:143, mean duration 4.0 +/- 6.7 yr) as a cross-sectional study. Of 233 patients with type 1 diabetes, IA-2A was detected in 58% of patients with duration within 2 weeks, 61% of patients with duration <1 yr, 41% of patients with diabetes for 1-3 yr, 29% for 4-9 yr, and 21% for >or=10 yr. These prevalences were similar to those of ICA, while the prevalence of GADA was not influenced by duration of diabetes with positivity of 63-74%. Thus, as the duration of diabetes became longer, the frequency of GADA(+)/IA-2A(-) patients increased and the frequency of GADA(+)/IA-2A(+) patients decreased. However, the frequency of GADA(-)/IA-2A(+) patients was not influenced by duration of diabetes. The prevalence of IA-2A was significantly higher in abrupt-onset group (68%, n= 79) compared to the slowly progressive group (23%, n= 22) in new-onset patients (P= 0.0001). However, there was no difference in the IA-2A frequency between these two groups (abrupt-onset 26%, n= 53 vs. slowly progressive 24%, n= 21) in patients with long-standing disease, suggesting that IA-2A positivity might persist in patients with slowly progressive type 1 diabetes. These results emphasize the heterogeneity of humoral autoimmunity to protein tyrosine phosphatase-like molecules, but not to GAD, in patients with type 1 diabetes.

Altered B:9-23 insulin, when administered intranasally with cholera toxin adjuvant, suppresses the expression of insulin autoantibodies and prevents diabetes.

Insulin peptide B:9-23 is a major autoantigen in type 1 diabetes that contains two distinct CD4 epitopes (B:9-16 and B:13-23). One of the two epitopes, B:13-23, overlaps with a CTL epitope (B:15-23). In this study, we report that the elimination of the CTL epitope from the B:9-23 peptide by amino acid substitution (with alanine) at positions B:16 and 19 (A16,19 altered peptide ligand) or truncation of the C-terminal amino acids from the peptide (B:9-21), neither of which stimulated the proliferation of insulin B:15-23 reactive CD8 T cells, provided significant intranasally induced suppression of diabetes when coadministered with a potent mucosal adjuvant cholera toxin (CT). Intranasal treatment with A16,19 resulted in the elimination of spontaneous insulin autoantibodies, significant inhibition of insulitis and remission from hyperglycemia, and prevented the progression to diabetes. Intranasal administration of native B:9-23/CT or B:11-23/CT resulted in a significant enhancement of insulin autoantibody expression and severity of insulitis and failed to prevent diabetes. Our present study indicates that elimination of the CTL epitope from the B:9-23 peptide was critically important for mucosally induced diabetes prevention. The A16,19 altered peptide ligand, but not other native insulin peptides, suppresses insulin autoantibodies associated with protection from and remission of diabetes.

Structural basis for recognition of high mannose type glycoproteins by mammalian transport lectin VIP36.

VIP36 functions as a transport lectin for trafficking certain high mannose type glycoproteins in the secretory pathway. Here we report the crystal structure of VIP36 exoplasmic/luminal domain comprising a carbohydrate recognition domain and a stalk domain. The structures of VIP36 in complex with Ca(2+) and mannosyl ligands are also described. The carbohydrate recognition domain is composed of a 17-stranded antiparallel beta-sandwich and binds one Ca(2+) adjoining the carbohydrate-binding site. The structure reveals that a coordinated Ca(2+) ion orients the side chains of Asp(131), Asn(166), and His(190) for carbohydrate binding. This result explains the Ca(2+)-dependent carbohydrate binding of this protein. The Man-alpha-1,2-Man-alpha-1,2-Man, which corresponds to the D1 arm of high mannose type glycan, is recognized by eight residues through extensive hydrogen bonds. The complex structures reveal the structural basis for high mannose type glycoprotein recognition by VIP36 in a Ca(2+)-dependent and D1 arm-specific manner.