Synthesis and unusual response to potassium of bipyridinium-benzocrown ether conjugate

A bipyridinium derivative appending a benzocrown ether, in which the phenyl unit in the benzocrown ether was directly bounded to the N-position of the bipyridinium unit, has been synthesized. The compound showed a yellow color associated with an intramolecular charge transfer (CT), which was affected by the presence of alkali and alkaline earth metal ions. An unusual CT response to K⁺ for 1 was observed and could be applicable for K⁺ sensing.     

Structural development of 1,2,3,4-tetrahydroisoquinoline-type positive allosteric modulators of prostacyclin receptor (IPPAMs) to improve metabolic stability,and investigation of metabolic fate

We synthesized a series of 1,2,3,4-tetrahydroisoquinoline-type positive allosteric modulators of prostacyclin receptor (IPPAMs), aiming to improve the metabolic stability of the previously identified hit compound IPPAM-3 (2). Our results indicated that the 3-position of the 2-substituted phenyl ring in this series of IPPAM-3 derivatives is a hot spot for metabolism catalyzed by human hepatic microsomes. This conclusion was confirmed by the finding that 8, in which the 3-position is blocked by a fluorine substituent, exhibited superior metabolic stability (t_1/2 21 min versus 7 min for parent compound 2). The primary route of metabolism of 8 was found to be oxidative defluorination, i.e., ipso-substitution of the fluorine atom to a hydroxyl group, affording catechol derivative 12. The primary metabolite 12 underwent further hydroxylation mainly on the 1,2,3,4-tetrahydroisoquinoline moiety. These findings should be helpful for design of IPPAMs with longer duration of action.     

A novel method for protecting slow-release sachets of predatory mites against environmental stresses and increasing predator release to crops

Slow-release sachets of predatory mites are widely employed for controlling small pest arthropods in protected crops. However, environmental stresses can adversely affect the performance of such sachets. To solve this problem, we developed plant-attached shelters that hold sachets of Neoseiulus californicus (McGregor) or Amblyseius swirskii (Athias-Henriot). We conducted laboratory experiments to reveal whether sheltered sachets can protect predators against pesticides and drenching. The numbers of each predator in unsheltered sachets were drastically decreased after spraying with a non-selective pesticide (methidathion), and after continuous spraying (four days) with water, whereas the numbers in the sheltered sachets were not seriously affected by these factors. We also found that more predators (at least for N. californicus) were released from sheltered sachets at different temperatures (25 and 17 °C) than from unsheltered sachets. These results indicate sheltered sachets to be potentially useful for protecting predatory mites against environmental stresses and enhancing their release to crops.     

Use of frequency domain analysis of skin conductance for evaluation of mental workload

A specially designed mental task was performed by 16 subjects for frequency domain analysis of skin conductance (SC) to evaluate mental workload. The task was to memorize target letters, detect them within a 4 x 4 alphabet arrangement, and answer whether the number of targets contained in the arrangement corresponded to a randomly displayed number. As the number of target letters increased, the score for the card-sort NASA Task Load Index (CSTLX) and task performance increased significantly. The traditional parameter for the number of transient wave forms of SC response (SCR) and the height of its wave did not show any significant effect of task difficulty. In addition, SCRs were subjected to Fourier transformation and integration of the spectrum from 0.03 to 0.5 Hz. This frequency domain analysis enabled detection of small differences in mental workload that could not be detected by traditional amplitude domain analysis. Frequency-based analysis enables easy processing of physiological signals and is very effective in evaluating mental stress using SC recorded under actual environmental conditions such as the driving of a vehicle.     

Analysis of area-specific expression patterns of RORbeta, ER81 and Nurr1 mRNAs in rat neocortex by double in situ hybridization and cortical box method

Background

The mammalian neocortex is subdivided into many areas, each of which exhibits distinctive lamina architecture. To investigate such area differences in detail, we chose three genes for comparative analyses, namely, RORbeta, ER81 and Nurr1, mRNAs of which have been reported to be mainly expressed in layers 4, 5 and 6, respectively. To analyze their qualitative and quantitative coexpression profiles in the rat neocortex, we used double in situ hybridization (ISH) histochemistry and cortical box method which we previously developed to integrate the data of different staining and individuals in a standard three-dimensional space.

Principal Findings

Our new approach resulted in three main observations. First, the three genes showed unique area distribution patterns that are mostly complementary to one another. The patterns revealed by cortical box method matched well with the cytoarchitectonic areas defined by Nissl staining. Second, at single cell level, RORbeta and ER81 mRNAs were coexpressed in a subpopulation of layer 5 neurons, whereas Nurr1 and ER81 mRNAs were not colocalized. Third, principal component analysis showed that the order of hierarchical processing in the cortex correlates well with the expression profiles of these three genes. Based on this analysis, the dysgranular zone (DZ) in the somatosensory area was considered to exhibit a profile of a higher order area, which is consistent with previous proposal.

Conclusions/Significance

The tight relationship between the expression of the three layer specific genes and functional areas were revealed, demonstrating the usefulness of cortical box method in the study on the cerebral cortex. In particular, it allowed us to perform statistical evaluation and pattern matching, which would become important in interpreting the ever-increasing data of gene expression in the cortex.     

Dynamic functional relay between insulin receptor substrate 1 and 2 in hepatic insulin signaling during fasting and feeding

Insulin receptor substrate (Irs) mediates metabolic actions of insulin. Here, we show that hepatic Irs1 and Irs2 function in a distinct manner in the regulation of glucose homeostasis. The PI3K activity associated with Irs2 began to increase during fasting, reached its peak immediately after refeeding, and decreased rapidly thereafter. By contrast, the PI3K activity associated with Irs1 began to increase a few hours after refeeding and reached its peak thereafter. The data indicate that Irs2 mainly functions during fasting and immediately after refeeding, and Irs1 functions primarily after refeeding. In fact, liver-specific Irs1-knockout mice failed to exhibit insulin resistance during fasting, but showed insulin resistance after refeeding; conversely, liver-specific Irs2-knockout mice displayed insulin resistance during fasting but not after refeeding. We propose the concept of the existence of a dynamic relay between Irs1 and Irs2 in hepatic insulin signaling during fasting and feeding.     

Expression and function of tight junctions in the crypt epithelium of human palatine tonsils.

The human palatine tonsils have surface and crypt stratified epithelium and may be initiated via the epithelium to mount immune responses to various presenting antigens. Here we investigated the expression and function of tight junctions in the epithelium of human palatine tonsils from patients with tonsillar hypertrophy or recurrent tonsillitis. Occludin, ZO-1, JAM-1, and claudin-1, -3, -4, -7, -8, and -14 mRNAs were detected in tonsillar hypertrophy. Occludin and claudin-14 were expressed in the uppermost layer of the tonsil surface epithelium, whereas ZO-1, JAM-1, and claudin-1, -4, and -7 were found throughout the epithelium. In the crypt epithelium, claudin-4 was preferentially expressed in the upper layers. In freeze-fracture replicas, short fragments of continuous tight junction strands were observed but never formed networks. In the crypt epithelium of recurrent tonsillitis, the tracer was leaked from the surface regions where occludin and claudin-4 disappeared. Occludin, ZO-1, JAM-1, and claudin-1, -3, -4, and -14, but not claudin-7, mRNAs were decreased in recurrent tonsillitis compared with those of tonsillar hypertrophy. These studies suggest unique expression of tight junctions in human palatine tonsillar epithelium, and the crypt epithelium may possess an epithelial barrier different from that of the surface epithelium.     

Propagation of senescent mice using nuclear transfer embryonic stem cell lines

Senescent mice are often infertile, and the cloning success rate decreases with age, making it almost impossible to produce cloned progeny directly from such animals. In this study, we tried to produce offspring from such "unclonable" senescent mice using nuclear transfer techniques. Donor fibroblasts were obtained from the tail tips of mice aged up to 2 years and 9 months. Although most attempts failed to produce cloned mice by direct somatic cell nuclear transfer, we managed to establish nuclear transfer embryonic stem (ntES) cell lines from all aged mice with an establishment rate of 10-25%, irrespective of sex or strain. Finally, cloned mice were obtained from these ntES cells by a second round of nuclear transfer. In addition, healthy offspring was obtained from all aged donors via germline transmission of ntES cells in chimeric mice. This technique is thus applicable to the propagation of a variety of animals, irrespective of age or fertile potential.     

Small and valid species of Trichiurus brevis Wang and You, 1992 and T. russelli Dutt and Thankam, 1966, defined as the “T. russelli complex” (Perciformes: Trichiuridae)

 The taxonomic status of three small [probably less than ca. 600 mm in the largest total length (TL)] and poorly known species, Trichiurus brevis Wang and You, 1992 and T. minor Li, 1992, both from South China Sea, and T. russelli Dutt and Thankam, 1966, from Bengal Bay, was reexamined. Trichiurus brevis and T. russelli, being valid species, are redescribed and a neotype is designated for the latter. Trichiurus brevis is considered to be a senior synonym of T. minor, there being no significant differences in their type specimens. Both valid species, i.e., T. brevis and T. russelli, are similar to T. lepturus Linnaeus, 1758 (larger than ca. 1000 mm TL) in general appearance, but clearly differ from the latter in having the top of the supraoccipital crest situated directly above the posterior margin of eye (vs. well behind posterior margin), lower number of total vertebrae (147–155 and 149–153, respectively, in the former two species vs. 168–173 in T. lepturus), longer dorsal fin base (mean 87% and mean 84% vs. mean 76% of TL), and shorter caudal peduncle length (6% and 8% vs. 13% of TL). Furthermore, the former two species are characterized by having the anal fin origin situated below about the 32nd–35th dorsal fin ray base, whereas in T. lepturus it is situated below the 37th–41st dorsal fin ray base. We recognize that T. brevis and T. russelli together comprise a species group, defined as the “T. russelli complex,” in the genus Trichiurus, the two species differing in snout shape (strongly pointed in T. brevis vs. moderately pointed in T. russelli), snout length [mean 35% vs. mean 30% of head length (HL)], preopercle length (22% vs. 19% HL), predorsal length (70% vs. 63% HL), and dermal eye opening (16% vs. 18% HL). Received: April 26, 2001 / Revised: January 19, 2002 / Accepted: February 1, 2002     

Calcineurin is critical for sodium-induced neointimal formation in normotensive and hypertensive rats

It is well known that excessive intake of sodium chloride (sodium) is a risk factor for cardiovascular disease because it raises blood pressure. However, sodium loading reportedly promotes cardiovascular disease independently of its effect on blood pressure. To examine the mechanisms by which sodium loading promotes vascular inflammation independently of its effect on blood pressure, we examined the role of calcineurin in sodium loading-induced vascular inflammation using a wire injury model of the rat femoral artery. Calcineurin mRNA expression in the wire-injured femoral artery was significantly higher in sodium-loaded normotensive rats, such as Wistar-Kyoto (WKY) rats, than that in control WKY rats. Neointimal formation was also significantly enhanced in sodium-loaded WKY rats compared with control WKY rats. Gene transfer of an adenovirus expressing a dominant negative mutant of calcineurin (AdCalADeltaC92Q) significantly suppressed neointimal formation in sodium-loaded WKY rats to a level similar to that observed in control WKY rats. Calcineurin expression and neointimal formation were more significantly enhanced in hypertensive rats, such as spontaneously hypertensive rats (SHRs), than those in control WKY rats. AdCalADeltaC92Q infection significantly suppressed neointimal formation in SHRs to a level similar to that observed in control WKY rats. These results suggest that sodium loading promotes neointimal formation, even in normotensive rats, and that hypertension further stimulates neointimal formation. These results also suggest that calcineurin plays a pivotal role in this process.