Substituted indanylacetic acids as PPAR-alpha-gamma activators

A series of oxazole-substituted indanylacetic acids were prepared which show a spectrum of activity as ligands for PPAR nuclear receptor subtypes.     

AT1 receptor blocker azilsartan medoxomil normalizes plasma miR-146a and miR-342-3p in a murine heart failure model

Our study measured circulating microRNA (miRNA) levels in the plasma of calsequestrin (CSQ)-tg mouse, a severe heart failure model, and evaluated whether treatment with angiotensin II type 1 receptor blocker, azilsartan medoxomil (AZL-M) influenced their levels using miRNA array analysis. MiR-146a, miR-149, miR-150, and miR-342-3p were reproducibly reduced in the plasma of CSQ-tg mice. Among them, miR-146a and miR-342-3p were significantly restored by AZL-M, which were associated with improvement of survival rate and reduction of congestion. These results suggest that miRNA, especially miR-146a and miR-342-3p, could be used as potential biomarkers for evaluating the efficacy of anti-heart failure drugs.     

Perlecan-enriched intercellular space of junctional epithelium provides primary infrastructure for leukocyte migration through squamous epithelial cells

The aim of this study was to demonstrate the presence of intraepithelial stroma represented by extracellular matrix (ECM) deposits in the junctional epithelium to clarify its function as a scaffold for leukocyte migration through epithelial cells. Twenty-three biopsy specimens from the gingiva including the junctional epithelium were examined to determine comparative protein and gene level expression profiles for keratin and ECM molecules between the junctional epithelium and the gingival epithelium using immunohistochemistry and in situ hybridization. Intraepithelial leukocyte types and frequencies were also determined and compared between the junctional and gingival epithelia. In the junctional epithelium, which was positive for keratin 19, perlecan was strongly deposited in intercellular space of the whole epithelial layer, while it was faintly positive around the parabasal layer of the gingival epithelium. Perlecan mRNA signals were enhanced to a greater degree in both epithelial and inflammatory cells within the junctional epithelium. In the junctional epithelium, greater numbers of neutrophils and macrophages were found as compared with the gingival epithelium. Our results showed that perlecan is the primary ECM molecule comprising intraepithelial stroma of the junctional epithelium, in which leukocytes may migrate on ECM scaffolds in intercellular space toward the surface of the gingival sulci or pockets.     

Infection of Adult Thymus with Murine Retrovirus Induces Virus-Specific Central Tolerance That Prevents Functional Memory CD8+ T Cell Differentiation

In chronic viral infections, persistent antigen presentation causes progressive exhaustion of virus-specific CD8⁺ T cells. It has become clear, however, that virus-specific naïve CD8⁺ T cells newly generated from the thymus can be primed with persisting antigens. In the setting of low antigen density and resolved inflammation, newly primed CD8⁺ T cells are preferentially recruited into the functional memory pool. Thus, continual recruitment of naïve CD8⁺ T cells from the thymus is important for preserving the population of functional memory CD8⁺ T cells in chronically infected animals. Friend virus (FV) is the pathogenic murine retrovirus that establishes chronic infection in adult mice, which is bolstered by the profound exhaustion of virus-specific CD8⁺ T cells induced during the early phase of infection. Here we show an additional evasion strategy in which FV disseminates efficiently into the thymus, ultimately leading to clonal deletion of thymocytes that are reactive to FV antigens. Owing to the resultant lack of virus-specific recent thymic emigrants, along with the above exhaustion of antigen-experienced peripheral CD8⁺ T cells, mice chronically infected with FV fail to establish a functional virus-specific CD8⁺ T cell pool, and are highly susceptible to challenge with tumor cells expressing FV-encoded antigen. However, FV-specific naïve CD8⁺ T cells generated in uninfected mice can be primed and differentiate into functional memory CD8⁺ T cells upon their transfer into chronically infected animals. These findings indicate that virus-induced central tolerance that develops during the chronic phase of infection accelerates the accumulation of dysfunctional memory CD8⁺ T cells.     

Cry1Aa binding to the cadherin receptor does not require conserved amino acid sequences in the domain II loops

Characterizing the binding mechanism of Bt (Bacillus thuringiensis) Cry toxin to the cadherin receptor is indispensable to understanding the specific insecticidal activity of this toxin. To this end, we constructed 30 loop mutants by randomly inserting four serial amino acids covering all four receptor binding loops (loops α8, 1, 2 and 3) and analysed their binding affinities for Bombyx mori cadherin receptors via Biacore. High binding affinities were confirmed for all 30 mutants containing loop sequences that differed from those of wild-type. Insecticidal activities were confirmed in at least one mutant from loops 1, 2 and 3, suggesting that there is no critical amino acid sequence for the binding of the four loops to BtR175. When two mutations at different loops were integrated into one molecule, no reduction in binding affinity was observed compared with wild-type sequences. Based on these results, we discussed the binding mechanism of Cry toxin to cadherin protein.     

Effects of Different Levels of Intraocular Stray Light on Kinetic Perimetry Findings

PurposeTo evaluate the effect of different levels of intraocular stray light on kinetic perimetry findings.MethodsTwenty-five eyes of 25 healthy young participants were examined by automated kinetic perimetry (Octopus 900) using Goldmann stimuli III4e, I4e, I3e, I2e, and I1e. Each stimulus was presented with a velocity of 3°/s at 24 meridians with 15° intervals. Four levels of intraocular stray light were induced using non-white opacity filter (WOF) filters and WOFs applied to the clear plastic eye covers of the participants. The visual acuity, pupil diameter, isopter area, and kinetic sensitivity of each meridian were analyzed for each WOF density.ResultsVisual acuity deteriorated with increasing WOF densities (p < 0.01). With a visual acuity of 0.1 LogMAR units, the isopter areas for III4e, I4e, I3e, I2e, and I1e decreased by -32.7 degree² (-0.2%), -255.7 degree² (-2.6%), -381.2 degree² (-6.2%), -314.8 degree² (-12.8%), and -59.2 degree² (-15.2%), respectively; kinetic sensitivity for those stimuli decreased by -0.1 degree (-0.1%), -0.8 degree (-1.4%), -1.6 degree (-3.7%), -2.7 degree (-9.7%), and -1.7 degree (-16.2%), respectively. The pupil diameter with each WOF density was not significantly different.ConclusionKinetic perimetry measurements with a high-intensity stimulus (i.e., III4e) were unaffected by intraocular stray light. In contrast, measurements with the I4e, I3e, I2e, and I1e stimuli, especially I2e and I1e, were affected. Changes in the shape of the isopter resulting from opacity must be monitored, especially in cases of smaller and lower-intensity stimuli.     

Indomethacin enhances histamine-induced pulmonary hemodynamic changes

To determine the role of prostaglandins in porcine pulmonary hemodynamic changes caused by histamine, we compared responses to intravenous histamine with and without pre-treatment with the cyclo-oxygenase inhibitor, indomethacin. In anesthetized pigs, pulmonary artery pressure (Ppa), pulmonary arterial wedge pressure (Ppaw), left ventricular end diastolic pressure (Plved) and cardiac output (Q) were measured repeatedly for 30 minutes, following a 1 ml intrajugular injection of histamine 0.6 microM/kg (n = 6), the identical histamine dose after pre-treatment with indomethacin 5 mg/kg (n = 7), or normal saline (n = 5). Pulmonary arterial resistance (Ra) and pulmonary venous resistance (Rv) were calculated as (Ppa-Ppaw)/Q and (Ppaw-Plved)/Q respectively. Indomethacin pre-treatment caused 2-fold greater increases in Ra and Rv with histamine and more prolonged changes. We conclude that inhibition of a vasodilatory prostaglandin released from pulmonary endothelial cells results in unopposed pulmonary vasoconstriction, thereby augmenting pulmonary resistance changes due to histamine.     

Structures of chaperonins from an intracellular symbiont and their functional expression in Escherichia coli groE mutants.

An intracellular symbiont harbored by the aphid bacteriocyte, a specialized fat body cell, synthesizes in vivo substantially only one protein, symbionin, which is a member of the chaperonin-60 family of molecular chaperones. Nucleotide sequence determination of the symbionin region of the endosymbiont genome revealed that it contains the two-cistron operon sym. Just like the Escherichia coli groE operon, the sym operon was dually led by a heat shock and an ordinary promoter sequence. According to the nucleotide sequence, symbionin was 85.5% identical to GroEL of E. coli at the amino acid sequence level. SymS, another protein encoded in the sym operon, which is a member of chaperonin-10, was 79.6% identical to GroES. Complementation experiments with E. coli groE mutants showed that the chaperonin-10 and chaperonin-60 genes from the endosymbiont are expressed in E. coli and that they can function as molecular chaperones together with endogenous GroEL and GroES, respectively.     

Control of foreign polypeptide localization in specific layers of protein body type I in rice seed

Key message

Prolamin–GFP fusion proteins, expressed under the control of native prolamin promoters, were localized in specific layers of PB-Is. Prolamin–GFP fusion proteins were gradually digested from outside by pepsin digestion.

Abstract

In rice seed endosperm, protein body type I (PB-I) has a layered structure consisting of prolamin species and is the resistant to digestive juices in the intestinal tract. We propose the utilization of PB-Is as an oral vaccine carrier to induce mucosal immune response effectively. If vaccine antigens are localized in a specific layer within PB-Is, they could be protected from gastric juice and be delivered intact to the small intestine. We observed the localization of GFP fluorescence in transgenic rice endosperm expressing prolamin–GFP fusion proteins with native prolamin promoters, and we confirmed that the foreign proteins were located in specific layers of PB-Is artificially. Each prolamin–GFP fusion protein was localized in specific layers of PB-Is, such as the outer-most layer, middle layer, and core region. Furthermore, to investigate the resistance of prolamin–GFP fusion proteins against pepsin digestion, we performed in vitro pepsin treatment. Prolamin–GFP fusion proteins were gradually digested from the peripheral region and the contours of PB-Is were made rough by in vitro pepsin treatment. These findings suggested that prolamin–GFP fusion proteins accumulating specific layers of PB-Is were gradually digested and exposed from the outside by pepsin digestion.