TLR2 activation by proteosomes promotes uptake of particulate vaccines at mucosal surfaces

Proteosome-based vaccines have TLR2-based adjuvant activity and show promise for mucosal immunization. We examined the effects of proteosomes on mucosal uptake in Peyer's patches in vivo. Proteosomes accelerated transepithelial transport of microparticles by M cells and induced migration of dendritic cells (DCs) into the follicle-associated epithelium (FAE); both effects were dependent on TLR2. Proteosomes induced the release of the DC-attracting chemokine MIP3alpha from Caco-2 epithelial cells in vitro. In HEK cells, proteosome-mediated MIP3alpha release was dependent on TLR2 expression and matrix metalloproteinase activation. Thus, TLR2 activation by proteosomes may promote mucosal uptake of particulate vaccines, and this may contribute to their adjuvanticity.     

In vitro effects of thyroid hormones on gonadotropin-induced estradiol-17 beta secretion by ovarian follicles of rainbow trout, Salmo gairdneri

Ovarian follicles isolated from rainbow trout during early exogenous vitellogenesis were used to study the in vitro effects of thyroid hormones on salmon gonadotropin (GtH)-induced estradiol-17 beta (E2) secretion. Triiodothyronine (T3) alone did not alter E2 secretion but T3 in the presence of GtH (0.5 micrograms/ml) modified E2 secretion according to a biphasic dose-response curve. Maximum E2 secretion occurred at 1.9 x 10(-8) M T3; a concentration of 3.0 x 10(-7) M was inhibitory. T3 was more potent in stimulating GtH-induced E2 secretion than thyroxine. The stimulatory and inhibitory effects of T3 were consistent over a range of GtH concentrations (0.1-1.0 micrograms/ml). Cycloheximide (0.1-10 microM) decreased E2 secretion by GtH-treated follicles in a dose-dependent manner, but failed to overcome all the stimulatory effects of T3. Time course studies with follicles incubated with GtH, GtH + T3, GtH + cycloheximide, or GtH + T3 + cycloheximide indicated that T3 stimulation of GtH-induced E2 secretion occurs within 6 hr. It is concluded that thyroid hormones amplify the effects of GtH on E2 secretion by isolated ovarian follicles; at least a part of this effect does not require de novo protein synthesis.     

Differential effect of phorbol esters and interleukin-3 on protein kinase C isoform content and kinase activity in the FDC-P1 cell line

FD/PMA is a subclone of the interleukin-3 (IL-3)-dependent, FDC-P1 cell line, which proliferates in response to either 12-O- tetradecanoylphorbol-13 acetate (PMA) or IL-3. While several endogenous substrates were phosphorylated in response to protein kinase C (PKC) activation in FDC-P1, phospholipid-dependent phosphorylation in the FD/PMA grown in PMA was not observed. Basal, phosphatidylserine- independent, and diolein-independent phosphorylation of cytosolic substrates with molecular weights of 17, 52, 57, and 105 Kd were enhanced in FD/PMA cells grown in PMA as compared with FDC-P1 cells cultured in IL-3. Phosphorylation of a 105-Kd substrate was enhanced in the particulate fraction of FD/PMA cells maintained in PMA. The 17-Kd substrate in FD/PMA cells comigrated with a substrate phosphorylated in a PKC-dependent manner in FDC-P1 cells. Phosphorylation of the 52- and 57-Kd substrates, but not of the 17-Kd substrate, was inhibited by H-7 and staurosporine. A portion of the PMA-induced cytosolic kinase activity coeluted with PKC on diethyl aminoethyl chromatography. While FD/PMA cells cultured in PMA contained negligible PKC-dependent phosphorylation of endogenous substrates or histone, alpha and epsilon PKC isoforms were detected by Western blot analysis. PKC phosphotransferase activity was observed in FD/PMA cells grown in PMA when peptides corresponding to residues 720 to 737 of PKC-epsilon or residues 4 to 14 of myelin basic protein were used as substrates. These data indicate that maintenance of FD/PMA cells in PMA stimulates proliferation and markedly alters PKC substrate specificity. Generation of at least two phospholipid-independent kinases occurs in PMA-treated cells.     

Seasonal patterns in serum levels of thyroid hormones and sex steroids in relation to photoperiod-induced changes in spawning time in rainbow trout, Salmo gairdneri

Serum levels of thyroid hormones (T4 = L-thyroxine and T3 = 3,5,3'-triiodo-L-thyronine) were measured and correlated with previously published levels of 17 beta-estradiol, testosterone (T), and Ca2+ (index of vitellogenin) in four groups of female trout held for 1-3 years at 8.5-9.0 degrees on a ration of 0.5% of body weight day-1 under different constant photoperiod regimes. In group 1, trout under a regimen of 18L:6D became sexually mature (SM) in April/May and then SM again in September/October; in group 2, trout under a regimen of 6L:18D became SM in January/February; in group 3, trout under a regimen of 6L:18D became SM in March/April; in group 4 trout under a regimen of 18L:6D followed by 10L:14D became SM in September/October. In all groups, regardless of the photoperiod-induced changes in spawning time, serum T3 exceeded T4 and both serum T3 and T4 patterns showed a consistent relationship with the sequence of steroid hormone changes and spawning time. Thyroid hormone levels were high during previtellogenesis but fell as sex steroids and serum Ca2+ increased. T3 and T4 were lowest before spawning when sex steroids were at their peak and then increased sharply following spawning as sex steroid levels declined. Peak serum T coincided with ovulation and usually preceded the postreproductive surge in serum T3 and T4. The hypothesis is discussed that T3 enhances early ovarian development, but as energy-demanding vitellogenesis proceeds T3 formation is suppressed, thereby curtailing growth and favoring energy partition to the ova.     

Food purchased away from home as a predictor of change in BMI z-score among girls

OBJECTIVES: To assess the relationship between eating food purchased away from home (FAH) and longitudinal change in body mass index (BMI) z-score among girls, and to assess the longitudinal tracking of eating FAH from childhood through adolescence. DESIGN: Participants kept 7-day dietary records at two points in time. The records included the place and time for all foods consumed. We recorded how often participants ate FAH, calculated the percent of total energy derived from FAH, and classified foods as quick-service food, coffee-shop food, or restaurant food. PARTICIPANTS: Healthy girls (n=101) between the ages of 8 and 12 y at baseline and 11 and 19 y at follow-up participated in a longitudinal study of growth and development at the Massachusetts Institute of Technology. STATISTICAL ANALYSES: Analysis of variance was used to assess the relationship between change in BMI z-score and both the frequency of eating FAH and energy derived from eating FAH. The participants' baseline BMI z-score was a significant covariate and was controlled for in both models. We used the kappa coefficient to assess FAH tracking from childhood through adolescence. RESULTS: The frequency of eating quick-service food at baseline was positively associated with change in BMI z-score (F=6.49, P<0.01). Participants who ate quick-service food twice a week or more at baseline had the greatest mean increase in BMI z-score compared to those who ate quick-service food once a week or not at all. Quick-service food eating tracked slightly from childhood through adolescence (k=0.17, P<0.05). DISCUSSION: Adolescent girls who eat quick-service food twice a week or more are likely to increase their relative BMI over time.     

Factors Associated With Microalbuminuria in 7,549 Children and Adolescents With Type 1 Diabetes in the T1D Exchange Clinic Registry

OBJECTIVE

To examine factors associated with clinical microalbuminuria (MA) diagnosis in children and adolescents in the T1D Exchange clinic registry.

RESEARCH DESIGN AND METHODS

T1D Exchange participants <20 years of age with type 1 diabetes ≥1 year and urinary albumin-to-creatinine ratio (ACR) measured within the prior 2 years were included in the analysis. MA diagnosis required all of the following: 1) a clinical diagnosis of sustained MA or macroalbuminuria, 2) confirmation of MA diagnosis by either the most recent ACR being ≥30 mg/g or current treatment with an ACE inhibitor (ACEI) or angiotensin receptor blocker (ARB), and 3) no known cause for nephropathy other than diabetes. Logistic regression was used to assess factors associated with MA.

RESULTS

MA was present in 329 of 7,549 (4.4%) participants, with a higher frequency associated with longer diabetes duration, higher mean glycosylated hemoglobin (HbA_1c) level, older age, female sex, higher diastolic blood pressure (BP), and lower BMI (P ≤ 0.01 for each in multivariate analysis). Older age was most strongly associated with MA among participants with HbA_1c ≥9.5% (≥80 mmol/mol). MA was uncommon (<2%) among participants with HbA_1c <7.5% (<58 mmol/mol). Of those with MA, only 36% were receiving ACEI/ARB treatment.

CONCLUSIONS

Our results emphasize the importance of good glycemic and BP control, particularly as diabetes duration increases, in order to reduce the risk of nephropathy. Since age and diabetes duration are important nonmodifiable factors associated with MA, the importance of routine screening is underscored to ensure early diagnosis and timely treatment of MA.Elevated urinary albumin excretion is an early sign of diabetic kidney disease (DKD). The American Diabetes Association (ADA) recommends screening for microalbuminuria (MA) annually in people with type 1 diabetes after 10 years of age and 5 years of diabetes duration, with a diagnosis of MA requiring two of three tests to be abnormal (1). Early diagnosis of MA is important because effective treatments exist to limit the progression of DKD (1). However, although reduced rates of MA have been reported over the past few decades in some (2–4) but not all (5,6) studies, it has been suggested that the development of proteinuria has not been prevented but, rather, has been delayed by ∼10 years and that further improvements in care are needed (7).Limited data exist on the frequency of a clinical diagnosis of MA in the pediatric population with type 1 diabetes in the U.S. Our aim was to use the data from the T1D Exchange clinic registry to assess factors associated with MA in 7,549 children and adolescents with type 1 diabetes.