[An example of self-evaluation of a sense of achievement by students in 6-year pharmacy school with the model core curriculum of pharmaceutical education].

In March 2012, the first students, finishing the newly introduced 6-year-course of pharmaceutical education, have graduated and gone out into the world. At this point, the Ministry of Education, Culture, Sports, Science and Technology (MEXT) is going to revise the model core curriculum of pharmaceutical education to be more suited for educating students to achieve their goal of becoming the clinical pharmacist standard defined by the revised School Education Act. Here we report the self-evaluation study based on the survey using questionnaire about a sense of achievement with Visual Analog Scales, regarding the fundamental quality as a pharmacist standard proposed by the Professional Activities Committee in the MEXT. The sample size of survey was about 600 of students studying in the Faculty of Pharmaceutical Sciences in Josai International University (JIU) and the survey was carried out during the period of March-April in 2012. The study suggested that the majority of graduates were satisfied with the new education system and marked as a well-balanced quality to be a pharmacist standard, after completing the 6-year pharmaceutical education based on "the model core-curriculum". It would be worthwhile to perform this kind of survey continuously to monitor the student's self-evaluation of a sense of achievement to verify the effectiveness of 6-year-course pharmaceutical education based on the newly establishing core curriculum in Japan.

Synergistic effect of isoflavone glycosides and fructooligosaccharides on postgastrectomy osteopenia in rats.

Fructooligosaccharides stimulate the growth of Bifidobacteria, which cleave isoflavone glycosides to yield corresponding aglycones, and convert metabolites by enhancing enterohepatic recirculation of isoflavones in rats. In the present study, we determined the synergistic effect of dietary isoflavone glycosides and fructooligosaccharides on postgastrectomy osteopenia in rats. Nine-week-old male Sprague-Dawley rats were gastrectomized (n = 20) or sham operated, (control, n = 5) and then randomly assigned to 5 diet groups: sham-a purified diet control, gastrectomized-control, gastrectomized-isoflavone (0.2% isoflavone glycosides), gastrectomized-fructooligosaccharides (7.5% fructooligosaccharides), and isoflavone and fructooligosaccharides (0.2% isoflavone glycosides + 7.5% fructooligosaccharides). After 6 weeks, the rats were killed and biological samples were collected. In gastrectomized rats, fructooligosaccharides prevented femoral bone fragility, but isoflavone without fructooligosaccharides did not inhibit postgastrectomy osteopenia. Isoflavone and fructooligosaccharides exhibited a synergistic in the distal metaphyseal trabecular bone, indicated by peripheral quantitative computed tomography. Moreover, fructooligosaccharides increased calcium absorption and equol production from daidzein in gastrectomized rats. These results indicate that isoflavone alone did not inhibit postgastrectomy osteopenia, but the combination of isoflavone and fructooligosaccharides improved the inhibition of trabecular bone loss by increasing calcium absorption and equol production through fructooligosaccharides supplementation.

The frequency of magnesium consumption directly influences its serum concentration and the amount of elutable bone magnesium in rats.

We investigated the influence of Mg feeding frequency on the variation in serum Mg concentration and tissue Mg levels in Mg-deficient rats. Sprague-Dawley rats, which had been fed a Mg-deficient diet for 14 d, were divided into 3 groups that were kept on 3 diets differing in their Mg content. The rats were fed 0.5-fold (Mg250 group), 1-fold (Mg500 group), or 1.5-fold (Mg750 group) the amounts of recommended Mg in their standard AIN-93G diet (Mg: 478 mg/kg diet) during the recovery period (12 d). The Mg500 and Mg750 groups were intermittently fed (Mg500, every 2 d; Mg750, every 3 d) so that their total intake of Mg during the recovery period could equal the Mg intake of the Mg250 group. The serum Mg concentrations increased in the 3 groups after feeding with a Mg-containing diet. However, serum Mg levels were only maintained within the normal range in the Mg250 group. After feeding on the Mg-deficient diet, in the intermittently fed groups, serum Mg concentrations decreased. Urinary Mg excretion was higher and Mg retention was lower in the Mg500 and Mg750 groups than in the Mg250 group. Moreover, bone Mg, especially elutable bone Mg, was lower in the Mg500 and Mg750 groups than in the Mg250 group. The elutable fraction of bone Mg correlated to the coefficient of variation of serum Mg concentration. In conclusion, for the maintenance of serum Mg concentration, it is important to increase the amount of elutable bone Mg by frequent Mg consumption.

Absence of negative feedback on intestinal magnesium absorption on excessive magnesium administration in rats.

This study aimed to clarify the regulatory mechanism of Mg homeostasis on administration of excessive Mg in rats. Six-week-old male Wistar rats (n=30) were fed a Mg-deficient diet (D) or a control diet (M) in addition to which they received subcutaneous injections of saline (S) or additional Mg (M) for 14 d. Feces and urine were collected from the rats for 4 d every week. Between the MS and MM rats and the DS and DM rats, the injection of additional Mg increased Mg retention, but intestinal Mg absorption did not differ. Urinary Mg excretion in the MM rats was significantly greater than that in the MS rats, but fecal Mg excretion did not increase. Mg retention in the DM rats was approximately 30% of that in the MS rats, and urinary Mg excretion did not differ between the 2 groups, although the serum Mg in DM rats was low. There was no significant difference in the femoral Mg between the MM and MS groups. The physiological Mg pool in the bone appears to be limited. Therefore, there is no physiological Mg pool for the storage of excessive Mg, and there appears to be no negative feedback mechanism on intestinal Mg absorption upon administration of excessive Mg in the rats. In conclusion, it appears that the kidney is the only organ that regulates Mg in the body; apart from this, regulatory mechanisms corresponding to the physiological Mg requirement do not exist or are weak.

[Prevention of osteoporosis by foods and dietary supplements. The effect of fructooligosaccharides (FOS) on the calcium absorption and bone].

Fructooligosaccharides (FOS) are well known as prebiotics which improve intestinal microflaura. FOS also have increasing effect on the intestinal absorption of calcium (Ca), magnesium (Mg) and iron. These effects were inspected by many animal experiments and then by human studies. Especially, FOS clearly prevent the decrease of bone mineral density by gastrectomy in rats. In this report, we mainly explain the preventive effect of FOS on the bone of gastrectomized rats and introduce relationship between another food ingredient or exercise.

Expression of calbindin-D9k, VDR and Cdx-2 messenger RNA in the process by which fructooligosaccharides increase calcium absorption in rats.

We have previously shown, through evidence that the expression of calbindin-D9k protein shows a dose-dependent change in the intestine, that calbindin-D9k plays a role in the ability of a fructooligosaccharide (FOS) diet to increase calcium absorption. This study shows that the regulation of calbindin-D9k expression occurs at the transcriptional level in a segment-specific manner, decreasing in the proximal intestine and increasing in the colorectal segment. To determine the transcriptional regulation of the FOS diet on calbindin-D9k expression, two transcription factors, vitamin D receptor (VDR) and cdx-2, were analyzed during 10 d feeding of the FOS diet. The mRNA expression of VDR and cdx-2 was influenced by the FOS diet and showed a segment-specific change. In the proximal small intestine, there was a significant correlation between the changes in both mRNAs (r = 0.69, p < 0.01), while the expression of calbindin-D9k correlated neither with VDR nor with cdx-2. This means that the transcriptional change induced by the FOS diet was not regulated by VDR and cdx-2. In the colorectal segment, there were significant correlations between gene expressions of calbindin-D9k vs. VDR, r = 0.73, p < 0.01 and calbindin-D9k vs. cdx-2, r = 0.52, p < 0.05. These results suggested that both transcription factors, VDR and cdx-2, were involved in the regulation of calbindin-D9k gene expression in the colorectal segment during the process through which the FOS diet enhanced calcium absorption.

The effect of fructooligosaccharides was analyzed by cDNA expression arrays.

We have already reported that indigestible fructooligosaccharides (FOS) increased calcium absorption in rat large intestines and that calbindin-D9k (CaBP), which is an intestine-specific calcium-binding protein, is involved in that increasing effect. In this study, not only the CaBP gene, every gene that changed expression profiles as the result of FOS feeding was identified by cDNA expression arrays. Sprague-Dawley male rats were fed an experimental diet containing 10% FOS for 10 d. To compare gene expression with rats fed a control diet, total mRNA was extracted from the colorectum and analyzed using a Rat cDNA Expression Arrays filter. This arrays filter contains probes of 588 genes, and 195 of them showed detectable changes in their expression by FOS feeding. There were six genes that increased their expression more than twice that of the control. Among them, genes related to the induction of cell growth such as Map kinase 1 and Max were included. Expressions that decreased to less than half were observed in 20 genes, such as somatostatin and prohibitin, which prohibit cell growth. These results are consistent with the other observation that FOS increases cell growth in the colorectum. This approach has revealed that cDNA array technology is an effective tool for nutritional sciences that involve the regulation of a large number of genes, especially for molecular mechanisms of regulation, by nutritional constituents.

Short-term creatine supplementation does not improve muscle activation or sprint performance in humans.

The purpose of this study was to examine the influence of short-term creatine (Cr) supplementation on exercise-induced transverse relaxation time (T2) and sprint performance during maximum intermittent cycling exercise using the muscle functional magnetic resonance imaging (mfMRI) technique. Twelve men were divided into a Cr supplementation group [the Cr group, taking 4 x (5 g Cr monohydrate + 2.5 g maltodextrin)/day], or a placebo supplementation group (the P group, taking 4 x 7.5 g maltodextrin/day). The allocation to the groups was based on cycling tests and the subject's physical characteristics, and thus was not randomized. A double-blind research design was employed for a 5-day supplementation period. mfMR images of the right thigh were collected at rest and immediately after two, five, and ten 6-s sprint bouts of maximum intermittent cycling exercise with a 30-s recovery interval between sets. Before and after supplementation, blood was taken to calculate lactate accumulation, and the muscle volume of the thigh was determined by MRI. Following supplementation, there was significant body mass gain in the Cr group ( P<0.05), whereas the P group did not change. The exercise-induced T2, blood lactate levels and sprint performance were not affected by Cr supplementation in any sprint bouts. These results suggest that short-term Cr supplementation does not influence short duration repetitive sprint performance and muscle activation and/or metabolic state during sprint cycling evaluated by mfMRI of the skeletal muscle in humans.

Fructooligosaccharides prevent disorders of the femoral neck following gastrectomy in growing rats.

Gastrectomy-evoked osteopenia in the femoral metaphysis of rats can be prevented by the consumption of fructooligosaccharides (FOS). We examined the effect of FOS on the femoral neck. Twenty-eight 5-week-old male Sprague-Dawley rats were divided into two groups, sham-operated (SH) and gastrectomized (GX). One week after each operation, the rats were fed diets containing 0.5% calcium with or without 7.5% FOS for 4 weeks. After dietary treatment, the middle of the femoral neck was cross sectioned. Backscattered electron images of the sections were then taken to calculate the following morphometric parameters: (1) percent trabecular bone volume (%TBV), (2) percent cortical bone volume (%CBV), and (3) percent bone marrow cavity (%MV); all were determined relative to the entire scan area (SC). Calcium, phosphorus, and magnesium (weight percent) were then measured on the cortical bone by energy-dispersive X-ray analysis. Total bone volume (%BV = %TBV + %CBV) and %CBV were almost identical among the groups, except in GX rats. In GX rats, these variables were significantly (approximately 20% and 30%, respectively; P < 0.01) less than those in SH rats, whereas there were no changes in the other groups over the entire scan area. The calcium concentration close to the periosteal surface of cortical bone was markedly reduced by gastrectomy. This reduction was completely prevented by FOS consumption. These results suggest that FOS consumption prevents gastrectomy-evoked osteopenia regarding both volume and calcium concentration of the femoral neck.

Combined intervention of exercise and genistein prevented androgen deficiency-induced bone loss in mice.

There is evidence that estrogen plays an important role in skeletal tissue in males as well as females. We have reported that phytoestrogens, such as genistein, selectively act on bone and exhibit cooperative effects on bone mass when combined with exercise in ovariectomized mice. In this study, we examined whether both interventions exhibit cooperative effects on bone loss in androgen-deficient mice similar to those in estrogen-deficient mice. Male mice aged 7 wk were either sham operated or orchidectomized (ORX) and divided into six groups: 1) sham; 2) ORX; 3) ORX and treated with genistein (0.4 mg/day) subcutaneously; 4) ORX, exercised on a treadmill daily for 30 min/day at 12 m/min; 5) ORX, given genistein, and exercised (ORX+ExG); and 6) ORX and treated with 17beta-estradiol (E(2)). Four weeks after the intervention, seminal vesicle weight strikingly decreased in ORX mice, and it was not affected by administration of genistein or E(2). Bone mineral density of whole femur was significantly reduced by ORX, and bone loss was prevented by the combined intervention. Histomorphometric analysis showed that bone volume and trabecular thickness in the distal femoral cancellous bone were significantly lower in the ORX group than in the Sham group, and they were completely restored in the ORX+ExG group, as in the ORX with E(2) group. These results indicate that the combined intervention of moderate exercise and a low dose of genistein administration shows an additive effect in preventing bone loss in ORX mice similar to that in ovariectomized mice.

Defects in mandibular bone area, enamel iron content and dentine formation following gastrectomy in rats.

Fourteen 5-week-old male Sprague-Dawley rats were equally divided into two groups, sham-operated and gastrectomized. Tetracycline and calcein were given to label dentine. Four weeks after surgery, blood was collected for measurement of serum iron, calcium and parathyroid hormone (PTH) and the mandibles and maxillae were then removed. Sagittal sections of the maxilla or cross-sections of the mandible were prepared and examined. Backscattered electron images of the maxilla were taken and the iron content at the neck of incisors was measured by energy-dispersive X-ray. The dentine apposition rate in maxillary incisors was measured by fluorescence microscopy. Serum iron was significantly decreased, while PTH was significantly elevated without any change in the serum calcium in gastrectomized rats. Gastrectomy caused a gross loss of iron content in superficial enamel. The dentine apposition rate was significantly reduced by 30%. Both cortical and cancellous bone in the mandibula were significantly reduced. However, the total bone area in gastrectomized rats was similar to that in sham-operated rats. These results suggest that bone resorption was enhanced and dentine formation was reduced after gastrectomy.

A combination of dietary fructooligosaccharides and isoflavone conjugates increases femoral bone mineral density and equol production in ovariectomized mice.

Fructooligosaccharides (FOS) stimulate the growth of bifidobacteria, which cleave isoflavone conjugates to yield the corresponding aglycones and metabolites. In a previous study, FOS modified the absorption and enterohepatic recirculation of isoflavones in rats. In the present study, we determined the effect of the combination of dietary FOS and isoflavone conjugates on bone mass in ovariectomized (OVX) and surgical control mice. After undergoing OVX or sham operation, female ddY mice (8 wk old, n = 64) were randomly assigned to four groups: a purified control diet (AIN-93G) group, a FOS diet (AIN-93G + 5% FOS) group, an isoflavone diet (AIN-93G + 0.2% isoflavone conjugates) group, or a FOS and isoflavone diet (AIN-93G + 5% FOS + 0.2% isoflavone conjugates) group. After 6 wk, the mice were killed and the blood and femora were sampled immediately. In OVX mice, both isoflavone conjugates and FOS prevented femoral bone loss. An additive effect of dietary isoflavone conjugates and FOS was observed by dual-energy X-ray absorptiometry in the distal part of the femur and in trabecular bone, by peripheral quantitative computed tomography. Moreover, FOS increased cecal beta-glucosidase activity and equol production from daidzein in both OVX and surgical control mice fed isoflavone conjugates. These results suggest that FOS increase the bioavailability of isoflavones, leading to cooperative effects in the prevention of osteopenia in OVX mice.