Cross-resistance of cadmium-resistant cells to manganese is associated with reduced accumulation of both cadmium and manganese

The mechanism of cellular entry of cadmium remains unclear. We have previously established cadmium-resistant cells from mouse embryonic cells of metallothionein (MT)-null mice, and demonstrated that the down-regulation of a zinc transporter, Zrt/Irt-related protein (ZIP) 8, was responsible for the reduced cadmium incorporation into cells. In the present study, we developed cadmium-resistant cells (A+70 and B+70) from mouse embryonic cells of MT-expressing wild-type mice. The LC?? values of CdCl? for A+70 and B+70 cells were about 200 μM while that of the parental cells was 30 μM. We found that the cadmium resistance of these cells was conferred not only by enhanced expression of MT, but also by a decrease in cadmium accumulation. Since the uptake rates of cadmium into A+70 and B+70 cells were lowered, we determined the expression levels of the metal transporters and channels potentially involved in the cellular uptake of cadmium. We found a down-regulation of multiple transport systems, including ZIP8, divalent metal transporter 1 (DMT1), and α? subunits of L-type (Ca(V)1.2) and T-type (Ca(V)3.1) voltage-dependent calcium channels, in A+70 and B+70 cells. Furthermore, A+70 and B+70 cells exhibited cross-resistance to cytotoxicity of MnCl?, probably due to a marked decrease in manganese uptake in these cells. These results suggest that the suppressed expression of ZIP8 and DMT1, which are known to have affinities for both cadmium and manganese, may be responsible for the reduction in the uptake, and consequently the cytotoxicity, of cadmium and manganese in A+70 and B+70 cells.     

Lack of chronic toxicity and carcinogenicity of dietary administrated catechin mixture in Wistar Hannover GALAS rats

Chronic toxicity and carcinogenicity of catechin mixture were examined in Wistar Hannover GALAS rats. Administration was in the diet at concentrations of 0, 0.02, 0.3, 1 or 3%. Slight increases in relative liver weight and centrilobular hypertrophy of hepatocytes associated with induction of CYP3A2 were found at the 3% in males of both studies. However, because there were no signs indicative of hepatotoxicity on serum biochemical and histopathological examinations, the changes observed in the liver were regarded as adaptation, and not adverse effects. The slight depressions of body weights at the 3% in females of the chronic toxicity study and in both sexes of the carcinogenicity study were observed. These decreases were because the diet at the highest concentration was frangible and nominal food consumption may not have reflected the actual food consumption resulting in decrease in caloric intake, rather than toxic effects. Thus it was concluded that catechin mixture had no toxicity. In addition, tumor incidences and types were comparable between treated and control groups. Based on the results, the no observed adverse effect levels estimated from the chronic toxicity study were 3% in both sexes equal to 1922.9 in males and 2525.7 mg/kg/day in females. Catechin mixture has no carcinogenic potential in male and female rats.     

Chronic exposure to arsenite induces S100A8 and S100A9 expression in rat RBL-2H3 mast cells

To investigate the effects of chronic exposure to arsenite on the gene expression profiles of mast cells, microarray analysis was performed on rat basophilic leukemia RBL-2H3 cells exposed to arsenite for 28 days. Upregulated genes include calcium-binding S100 proteins such as S100A9, S100A10, S100A6, and S100A13, and granzymes B and C. Among S100 proteins, S100A9 showed the highest expression (8.62-fold of untreated cells) after 4-weeks of exposure to arsenite. As S100A8 and S100A9 comprise a heterodimer called calprotectin, and are implicated in the development of atherosclerosis and cancer, mRNA levels of both S100A8 and S100A9 were analyzed. The results demonstrated that exposure of RBL-2H3 cells to arsenite for a few weeks induces marked increases in mRNA levels of S100A8 and S100A9.     

Cannabidiol-2',6'-dimethyl ether as an effective protector of 15-lipoxygenase-mediated low-density lipoprotein oxidation in vitro

15-Lipoxygenase (15-LOX) is one of the key enzymes responsible for the formation of oxidized low-density lipoprotein (ox-LDL), a major causal factor for atherosclerosis. Both enzymatic (15-LOX) and non-enzymatic (Cu(2+)) mechanisms have been proposed for the production of ox-LDL. We have recently reported that cannabidiol-2',6'-dimethyl ether (CBDD) is a selective and potent inhibitor of 15-LOX-catalyzed linoleic acid oxygenation (Takeda et al., Drug Metab. Dispos., 37, 1733-1737 (2009)). In the LDL, linoleic acid is present as cholesteryl linoleate, the major fatty acid esterified to cholesterol, and is susceptible to oxidative modification by 15-LOX or Cu(2+). In this investigation, we examined the efficacy of CBDD on i) 15-LOX-catalyzed oxygenation of cholesteryl linoleate, and ii) ox-LDL formation catalyzed by 15-LOX versus Cu(2+)-mediated non-enzymatic generation of this important mediator. The results obtained demonstrate that CBDD is a potent and selective inhibitor of ox-LDL formation generated by the 15-LOX pathway. These studies establish CBDD as both an important experimental tool for characterizing 15-LOX-mediated ox-LDL formation, and as a potentially useful therapeutic agent for treatment of atherosclerosis.     

Efficacy and safety of lisinopril for mild childhood IgA nephropathy: a pilot study

Even in children with mild immunoglobulin (Ig)A nephropathy (IgA-N) showing minimal/focal mesangial proliferation, persistent proteinuria seems to be a risk factor for progression of the disease, indicating the need for an effective and safe treatment even in such cases. Studies carried out to date have indicated that angiotensin-converting enzyme inhibitors (ACEIs) reduce urinary protein excretion and preserve renal function in adult IgA-N. However, no prospective study of ACEI only for childhood IgA-N has yet been carried out. In this prospective single-arm pilot trial, we administered lisinopril (0.4 mg/kg per day) as therapeutic treatment to 40 children with mild IgA-N with proteinuria [morning urinary protein/creatinine ratio (uP/Cr) ≥ 0.2 g/g]. Thirty-three patients reached the primary endpoint (uP/Cr < 0.2) during the 2-year treatment period. The cumulative disappearance rate of proteinuria determined by the Kaplan–Meier method was 80.9%. Mean uP excretion was reduced from 0.40 to 0.18 g/m²/day (p < 0.0001). Of the 40 patients treated, five (12.5%) showed dizziness, and four of these five needed the lisinopril dose reduced. However, lisinopril therapy was continued in all patients during the 2-year treatment period. No other side effect, such as cough, was observed. We conclude that the efficacy and safety of lisinopril is seemingly acceptable for the treatment of children with mild IgA-N. The participants in the Japanese Pediatric IgA Nephropathy Treatment Study Group are listed in the Appendix.     

Localization of the low-density lipoprotein receptor-related protein regions involved in binding to the A2 domain of coagulation factor VIII

Catabolism of coagulation factor VIII (FVIII) is mediated by low-density lipoprotein receptor-related protein (LRP). The ligand-binding sites of LRP are formed by complement-type repeats (CR), and CR clusters II and IV bind most of LRP ligands. FVIII contains two major LRP-binding sites located in the A2 and A3 domains. This study was aimed to identify specific complement-type repeats of LRP involved in interaction with the A2 site and to probe their functional importance in A2 catabolism. We generated individual LRP clusters II, III and IV, along with nine overlapping CR triplets encompassing clusters II and IV in a baculovirus expression system and studied their interaction with isolated A2. In surface plasmon resonance (SPR) assay, A2 bound to clusters II and IV with KDs 22 and 39 nM, respectively, and to the majority of CR triplets with affinities in the range of KDs 25-90 nM. Similar affinities were determined for A2 interaction with a panel of CR doublets overlapping cluster II (CR 3-4, 4-5, 5-6, 6-7 and 7-8). These LRP fragments inhibited the binding of 125I-A2 to LRP in solid-phase assay, LRP-mediated internalization of 125I-A2 in cell culture and 125I-A2 clearance from the mouse circulation. Point mutations of critical A2 residues of the LRPbinding site resulted in differential reduction or abolishment of its binding to LRP fragments. We conclude that A2 interacts with LRP via multiple binding sites spanning CR 3-8 in cluster II and CR 23-29 in cluster IV, and the minimal A2-binding unit of LRP is formed by two adjacent CR.     

Comparison of Patient's Preference,Pain Perception,and Usability between Micro Fine Plus? 31-Gauge Needle and Microtapered NanoPass? 33-Gauge Needle for Insulin Therapy

Background

How smoothly insulin is injected is one of the major concerns when patients commence insulin injection therapy. Improving its usability may be important in initiation therapy and adherence, resulting in clinical benefits to the patient.

Methods

In a single-center, open-label and randomized two-period crossover trial, the effect of the tapered needle of NanoPass® (33 gauge, 5 mm) on usability in comparison with the standard needle of Micro Fine Plus® (31 gauge, 5 mm) was examined using a questionnaire. Patients with insulin-dependent diabetes (n = 40, self-injecting insulin four times daily for more than 3 months) were randomized to use NanoPass or Micro Fine Plus needles for 1 week and then use the alternative for 1 week. Patients completed the questionnaire before and after each test week. Each evaluation was scored from −100 (worst) to +100 (best) by a visual analogue scale. A higher score indicated a more favorable outcome compared with the other needle.

Results

The NanoPass needle was significantly less painful to insert and caused less bruising than the Micro Fine Plus needle. However, there was no significant difference in the overall patient satisfaction score between the two needles. Meanwhile, the NanoPass needle, which had less resistance in insertion with a new lubricant coating method, had a significantly superior (P < 0.001) overall patient''s satisfaction score, including less frightening use, less bleeding, and less dribbling of injected insulin in comparison with the former evaluation.

Conclusions

For overall patient satisfaction in using an insulin needle, developing a thinner needle and improving other factors, such as lubricity coating the needle, are important.     

Functional map of orientation tuning width and specificity

To map the orientation-tuning property based on optical signals, we propose using correlation coefficients that can determine the correlation between the measured optical intensities and a Gaussian function at each pixel. The correlation coefficients at each pixel were computed by setting the spread (or SD) of the Gaussian function at 20 and 40 degrees. Comparison between the coefficients showed that, in the region representing cardinal lines, there was a significantly larger area that had a higher correlation with the 40 degree Gaussian function, in contrast to the region representing oblique lines, which had a larger area that displayed a higher correlation for the 20 degree Gaussian function. These results demonstrate that the cardinal orientation-sensitive cortical area tuned over a broader spectrum than the oblique orientation-sensitive cortical area did.