Role of the N-linked glycans of the prM and E envelope proteins in tick-borne encephalitis virus particle secretion

The tick-borne encephalitis (TBE) virus has two membrane glycoproteins (prM and E), which each has one N-linked glycan. Constructs that express prM and E proteins of TBE virus have been shown to produce virus-like particles (VLPs), which have surface properties that are similar to those of infectious viruses. To reveal the function of glycosylation of the TBE virus prM and E proteins in the secretion of VLPs, we expressed glycosylation-mutated prM and E proteins and compared the secretion levels and biological properties of the VLPs. In the prM protein glycosylation-deficient mutant, the level of secreted E protein was reduced to 60% of the wild-type level. On the other hand, in the E or prM-E protein glycosylation-deficient mutant, the level of secreted E protein was reduced to 10% of the wild-type level. Furthermore, the mutant which was glycosylated at positions 66 and 154 in protein E, the level of secreted E protein was four-fold higher than that of the wild-type. However, in the mutant which was glycosylated at position 66 only, E protein secretion was reduced to only 10% of the wild-type level. These data suggest that the glycan associated with the N-linked glycosylation site at position 154 in protein E plays an important role in VLP secretion.     

Impaired glucose tolerance, diabetes mellitus, and gallstone disease: An extended study of male self-defense officials in Japan

Few studies have investigated the relation between glucose tolerance status and ultrasonographically determined gallstone disease. Using a 75-g oral glucose tolerance test, we examined the association of impaired glucose tolerance (IGT) and non-insulin-dependent diabetes mellitus (NIDDM) with gallstone disease in Japanese men. Subjects were men aged 48 to 59 of the Japan Self-Defense Forces who received a preretirement health examination between October 1986 to December 1994. After exclusion of 12 men under insulin treatment in the consecutive series of 7637 men, 174 were found to have gallstones; 103 were at the state of postcholecystectomy, and 6899 had normal gallbladder. IGT and NIDDM were associated with a modestly increased risk of gallstone disease; adjusted odds ratios were 1.3 (95% confidence interval [CI]: 0.9–1.8) for IGT and 1.3 (95% CI: 0.8–2.0) for NIDDM after adjustment for hospital, rank, smoking, alcohol use, and body mass index. Adjusted odds ratio for IGT and NIDDM combined was 1.3 (95% CI: 1.0–1.7, p=0.08). When prevalent gallstones and postcholecystectomy were considered separately, NIDDM showed a significant, positive association with postcholecystectomy, but not with prevalent gallstones. The findings add to evidence that glucose intolerance is associated with a modest increase in the risk of gallstone disease.     

Effect of tea catechins on postprandial plasma lipid responses in human subjects

Epidemiological surveys suggest that a higher intake of tea may be associated with a lower risk of CHD. There is accumulating evidence that postprandial lipaemia makes a substantial contribution to the incidence of CHD. Our aim was, therefore, to evaluate the effect of tea catechins (major ingredients in green tea) on postprandial lipid responses in human subjects after the consumption of test meals. In a randomized triple-crossover design, nine male subjects with mild or borderline hypertriacylglycerolaemia consumed 10 (control), 224 (moderate dose) and 674 mg (high dose) of the assigned tea catechins three times each along with a standardized light meal consisting of a piece of bread spread with 20 g butter. Plasma lipids were measured in the fasting state and 1, 2, 3, 4 and 6 h after consuming the light meal. Results showed that, compared with the control, moderate and high doses of tea catechins reduced the incremental area under the plasma triacylglycerol curves by 15.1 and 28.7%, respectively. Next, the rapid elevation of remnant-like particle cholesterol was significantly inhibited by a high dose of tea catechins 2 h after consuming the light meal (P<0.01). In the range of tea catechin dosages, no significant differences were observed in the postprandial responses for plasma total cholesterol or NEFA at any time point. In conclusion, this trial demonstrated that tea catechins attenuated the postprandial increase in plasma triacylglycerol levels following a fat load. These results may provide evidence for one of the possible mechanisms involved in lowering the incidence of CVD, and may prove useful in further studies on the beneficial health effects of tea drinking.     

A potent apoptosis-inducing activity of a sesquiterpene lactone, arucanolide, in HL60 cells: a crucial role of apoptosis-inducing factor

Six main sesquiterpene lactones (germacranolides) from Calea urticifolia were evaluated for in vitro cytotoxicity against human tumor cell lines HL60 and SW480 cells. Among them, arucanolide and parthenolide displayed marked cytotoxicity against both cell lines. Arucanolide exhibited a low IC(50) in HL60 cells. The cytotoxic activity of arucanolide was observed at lower concentrations compared to that of parthenolide, which has been reported to be a typical and simple germacranolide. The activity was found to be mainly due to apoptosis that was assessed by morphological findings, DNA ladder formation (24 - 36 h), and flow cytometric analysis in HL60 cells. Western blotting and an apoptosis inhibition assay using caspase inhibitors did not demonstrate the activation of any caspases tested. However, the mitochondrial membrane potential of HL60 cells was lost after 24-h treatment with arucanolide, and concurrently apoptosis-inducing factor (AIF) released from mitochondria was detected by Western blot analysis. The inactivation of nuclear factor-kappaB, which has been commonly shown in parthenolide-induced apoptosis, did not occur in arucanolide-induced apoptosis. Taken together, the findings presented here indicate that arucanolide induced marked apoptosis in HL60 cells mainly by dissipating mitochondrial membrane potential, which would trigger AIF-induced apoptosis.     

Relationships between suicidal ideation and psychosocial factors among residents living in Nagano Prefecture of Japan

Objectives

Feeling ashamed for seeking help when distressed is known to be a critical factor promoting suicidal behaviors. The objective of this study was to examine the relationship between suicidal ideation and psychosocial factors, including worries or anxieties, having a person to confide in, feeling ashamed for seeking help when distressed, and K6 score.

Methods

Fourteen out of 77 municipalities from Nagano Prefecture participated in this questionnaire survey. Participants of both sexes over 20 years of age were randomly selected according to age distribution in each municipality. Association between suicidal ideation and sociodemographic and psychosocial factors, including “feeling ashamed for seeking help”, were determined by multiple logistic regression analysis.

Results

Among a total of 11,100 participants, 7394 (66.6 %) returned the questionnaire. 2147 participants responded properly to essential study parameters and were submitted to the final analyses. The adjusted odds ratio of suicidal ideation was 2.09 (95 % CI 1.49–2.94) among participants feeling ashamed for seeking help, compared to those not feeling ashamed. Although the rate of “no person to confide in” was 4.4 %, participants who responded with “no person to confide in” had significantly increased odds ratio of suicidal ideation compared with those who responded with “having a person to confide in” (OR 1.97, 95 % CI 1.12–3.47).

Conclusions

Our findings suggest a need for tailored intervention targeting individuals at risk by gatekeepers to encourage individuals at risk to overcome feeling ashamed for seeking help and to cultivate an appropriate person to confide in.

     

Relationship of toe pinch force to other muscle strength parameters in men with type 2 diabetes

Objective

The aim of this study was to explore the relations between toe pinch force and other muscle strength parameters in male patients with type 2 diabetes mellitus.

Methods

A total of 40 men with type 2 diabetes (age: 53.4 ± 13.1 years, duration of diabetes: 8.5 ± 8.1 years) who needed exercise training were enrolled in this cross-sectional study. We evaluated the clinical parameters and 4 muscle strength parameters, which were toe pinch force, handgrip strength, isometric knee extension force, and isometric ankle dorsiflexion force.

Results

The HbA1c, toe pinch force, handgrip strength, isometric knee extension force, and isometric ankle dorsiflexion force were 10.1 ± 2.4 %, 3.2 ± 1.2 kg, 37.3 ± 7.0 kg, 39.6 ± 11.4 kgf, and 17.0 ± 6.3 kgf, respectively. Toe pinch force was significantly correlated with handgrip strength (r = 0.365, p = 0.0206), isometric knee extension force (r = 0.668, p < 0.0001), and isometric ankle dorsiflexion force (r = 0.514, p = 0.0007). All muscle strength parameters were significantly lower in patients with diabetic polyneuropathy than in those without polyneuropathy.

Conclusion

Although toe pinch force was significantly correlated with the other muscle strength parameters, the correlation was not so strong. However, evaluation of toe pinch force might be recommended for assessment of distal limb muscle strength in patients with type 2 diabetes.

     

Novel Acridine-Based <i>N</i>-Acyl-homoserine Lactone Analogs Induce Endoreduplication in the Human Oral Squamous Carcinoma Cell Line SAS

The cytotoxicity of novel acridine-based N-acyl-homoserine lactone (AHL) analogs was investigated on the human oral squamous carcinoma cell line SAS. One analog induced G2/M phase arrest at 5.3-10.6?μM and induced polyploidy at a higher dose (21.2?μM). Importantly, treatment of SAS cells with a combination of the AHL analog and the Jun N-terminal kinase (JNK) inhibitor, SP600125, prevented mitosis and induced polyploidy. The AHL analog synergized with X-irradiation to inhibit clonogenic survival of SAS cells; however, its radiosensitizing effects were relative to not X-irradiation-induced apoptosis but mitotic failure following enhanced expression of Aurora A and B. These results suggest that the active AHL analog showed growth-suppressive and radiosensitizing effects, which involve polyploidy followed by G2/M accumulation and atypical cell death in the SAS cell line.     

Effect of ethanol pretreatment on skin permeation of drugs

It has been demonstrated that ethanol (EtOH) can enhance skin permeation of drugs when simultaneously applied with drugs. However, only a few studies have reported on the pretreatment effect of EtOH on skin permeations. In this study, the pretreatment effects of EtOH on skin permeation of drugs were investigated by measuring changes in skin permeation and electrical skin resistance. Permeabilities of deuterium oxide (D2O), isosorbide mononitrate (ISMN), isosorbide dinitrate (ISDN), calcein sodium (CA-Na), and fluorescein isothiocyanate-dextran 4?kDa (FD-4, 3.3-4.4?kDa) were evaluated through Yucatan micropig skin pretreated with different concentrations of EtOH solution. From the results, almost constant skin permeabilities of D2O and ISDN were observed independent of EtOH concentration. Skin permeabilities of ISMN, CA, and FD-4 increased with low concentrations of EtOH, but decreased with high concentrations of EtOH. At 99.5% EtOH pretreatment, skin permeabilities of hydrophilic compounds (ISMN, CA, and FD-4) decreased to non-detectable levels. In addition, low molecular ion transports were almost constant at any EtOH concentration. Since molecular (ion) sizes of ISMN, CA, and FD-4 are larger than Na⁺, Cl^－, and D2O, permeation pathway sizes for hydrophilic compounds in the skin barrier may be remarkably decreased by pretreatment with high concentrations of EtOH. However, the permeability coefficient of ISDN was not influenced by any EtOH concentration, since ISDN is a lipophilic, low-molecular compound that permeated through the lipophilic stratum corneum pathway. The present results show useful information for repeatedly and topically applied formulations containing EtOH, and also contribute to the effective use of alcohol formulations.     

Improved intestinal absorption of a poorly water-soluble oral drug using mannitol microparticles containing a nanosolid drug dispersion

A nozzle for a spray dryer that can prepare microparticles of water-soluble carriers containing various nanoparticles in a single step was previously developed in our laboratory. To enhance the solubility and intestinal absorption of poorly water-soluble drugs, we used probucol (PBL) as a poorly water-soluble drug, mannitol (MAN) as a water-soluble carrier for the microparticles, and EUDRAGIT (EUD) as a polymer vehicle for the solid dispersion. PBL–EUD–acetone–methanol and aqueous MAN solutions were simultaneously supplied through different liquid passages of the spray nozzle and dried together. PBL–EUD solid dispersion was nanoprecipitated in the MAN solution using an antisolvent mechanism and rapidly dried by surrounding it with MAN. PBL in the dispersion vehicle was amorphous and had higher physical stability according to powder X-ray diffraction and differential scanning calorimetry analysis. The bioavailability of PBL in PBL–EUD S-100–MAN microparticles after oral administration in rats was markedly higher (14- and 6.2-fold, respectively) than that of the original PBL powder and PBL–MAN microparticles. These results demonstrate that the composite microparticles containing a nanosized solid dispersion of a poorly water-soluble drug prepared using the spray nozzle developed by us should be useful to increase the solubility and bioavailability of drugs after oral administration.     

Pulmonary toxicity of well-dispersed multi-wall carbon nanotubes following inhalation and intratracheal instillation

Multi-walled carbon nanotubes (MWCNTs), dispersed in suspensions consisting mainly of individual tubes, were used for intratracheal instillation and inhalation studies. Rats intratracheally received a dose of 0.2 mg, or 1 mg of MWCNTs and were sacrificed from 3 days to 6 months. MWCNTs induced a pulmonary inflammation, as evidenced by a transient neutrophil response in the low-dose groups, and presence of small granulomatous lesion and persistent neutrophil infiltration in the high-dose groups. In the inhalation study, rats were exposed to 0.37 mg/m(3) aerosols of well-dispersed MWCNTs (>70% of MWCNTs were individual fibers) for 4 weeks, and were sacrificed at 3 days, 1 month, and 3 months after the end of exposure. The inhalation exposures delivered less amounts of MWCNTs into the lungs, and therefore less pulmonary inflammation responses was observed, as compared to intratracheal instillation. The results of our study show that well-dispersed MWCNT can produce pulmonary lesions, including inflammation.