Nutritional Supplementation during Resistance Training Improved Skeletal Muscle Mass in Community-Dwelling Frail Older Adults.

OBJECTIVE: Sarcopenia, the age-related loss of skeletal muscle mass, is highly prevalent in older adults. The aim of this study was to investigate the effects of the combination of resistance training and multinutrients supplementation (including vitamin D and protein) on muscle mass and physical performance in frail older adults. METHODS: This trial was conducted in Japanese frail older adults (n=77), which underwent a standardized protocol of a 3-month physical exercise intervention. The sample population was divided into two groups, according to the adoption (S/Ex: n = 38) or not (Ex: n = 39) of the additional multinutrient supplementation. The outcome measures of interest for the present analyses were the skeletal muscle mass index (SMI) and several physical performance tests. RESULTS: Participants in S/Ex group had significant improvements for the outcome measures, including SMI and maximum walking time (P<0.05), compared to those in Ex group. The prevalence of sarcopenia decreased from 65.7% to 42.9% in S/Ex group, while that in Ex group remained unchanged (68.6% to 68.6%) (relative risk = 1.60, 95% CI: 1.03-2.49). CONCLUSION: The results of this study suggest that the combination of resistance training and multinutritional supplementation may be more effective at improving muscle mass and walking speed than an intervention only based on resistance training.

Development of a phantom compatible for MRI and hyperthermia using carrageenan gel--relationship between dielectric properties and NaCl concentration.

A phantom has previously been developed containing carrageenan, agarose and gadolinium chloride (called CAG phantom) for MRI with 1.5 T. T(1) and T(2) relaxation times of this phantom are independently changeable by varying concentrations of relaxation-time modifiers to simulate relaxation times of the various types of human tissues. The CAG phantom has a T(1) value of 202-1904 ms and a T(2) value of 38-423 ms, when the GdCl(3) concentration is varied from 0-140 micromol/kg and the agarose concentration is varied from 0-1.6%. A new phantom has now been developed (called CAGN phantom), made by adding an electric conductive agent, NaCl, to the CAG phantom for use in the areas of MRI and hyperthermia research. Dielectric properties of the CAGN phantom were measured and the results of experiments were expressed by the Cole-Cole equation in the frequency range of 5-130 MHz. The relationship between the conductivity of the CAGN phantom and the concentration of NaCl was expressed by a linear function in the frequency range of 1-130 MHz. The linear function involves a parameter of frequency and, when the frequency is 10 MHz, the conductivity of the CAGN phantom can be changed from 0.27-1.26 Sm(-1) by increasing the NaCl concentration from 0-0.7%. The CAGN phantom developed can be employed in basic experiments for non-invasive temperature measurement using MRI and as a loading phantom for MRI with up to 3 T.

Development of a phantom compatible for MRI and hyperthermia using carrageenan gel-relationship between T1 and T2 values and NaCl concentration.

The authors developed a phantom, designated as the CAGN phantom, compatible for MRI and hyperthermia that is useful in the fundamental studies of non-invasive MR thermometry. The ingredients of this phantom are carrageenan, GdCl3 as a T1 modifier, agarose as a T2 modifier, NaCl as a conductivity modifier, NaN3 as an antiseptic and distilled water. Another phantom that was developed, the CAG phantom, has relaxation times that are adjustable to those of any human tissue. To use this phantom for electromagnetic heating, NaCl was added to change the conductivity of the phantom and clarified the relationship between the conductivity and NaCl concentration. This study examined the relationship between relaxation times and NaCl concentration of the CAGN phantom. The results showed that both T1 and T2 values were affected by NaCl and the experimental results led to the empirical formulae expressing the relationship between the relaxation rates (1/T1, 1/T2) and the concentrations of GdCl3, agarose and NaCl. The appropriate concentrations of T1 and T2 modifiers were calculated from these empirical formulae when making a specified phantom that has the required relaxation times and NaCl concentration.

Microwave thermotherapy for benign prostatic hyperplasia with the Dornier Urowave: response durability and variables potentially predicting response.

OBJECTIVES: To evaluate the clinical efficacy and durability of transurethral microwave thermotherapy (TUMT) in the treatment of benign prostatic hyperplasia. The clinical variables useful in predicting outcome were identified. METHODS: From October 1996 to March 2000, 58 patients with symptomatic benign prostatic hyperplasia were treated with TUMT using the Urowave device. Treatment outcome was evaluated by the International Prostate Symptom Score (IPSS), quality-of-life assessment score, and urodynamic investigation. The patients were divided into those having a good and poor response on the basis of the degree of IPSS decrease at 3 months. RESULTS: The mean IPSS significantly decreased from 19.2 at baseline to 13.3 at 3 months (P <0.0001). The mean quality-of-life score changed from 4.6 at baseline to 2.9 at 3 months (P <0.0001). No statistically significant differences in peak flow rate, postvoid residual volume, Schäfer's obstruction scale, or detrusor pressure at peak flow were noted before or after TUMT. The pretreatment IPSS of the good response group was significantly higher than that of the poor response group (P=0.017). A more significant difference was obtained for the obstructive score (P = 0.002), and no difference was observed in the irritative score (P = 0.631). The Schäfer grading scale score of the good response group was significantly smaller than that of the poor response group (P = 0.047). CONCLUSIONS: TUMT with the Urowave was effective in eliminating symptoms associated with benign prostatic hyperplasia, but did not markedly improve the objective voiding parameters. Patients with urodynamically less obstructive symptoms but subjectively more obstructive symptoms are therefore probably good candidates for TUMT.

Molecular cloning of ring finger protein 21 (RNF21)/interferon-responsive finger protein (ifp1), which possesses two RING-B box-coiled coil domains in tandem.

We have cloned the full length of a novel cDNA, named ring finger protein 21 (RNF21), composed of the RING finger-B box-coiled coil (RBCC) domain and the B30.2 domain, which are characteristic of the RBCC-B30.2 family. As a structural feature, the RNF21 cDNA possessed at least three kinds of isoforms, due to alternative splicing, consisting of the long form with the RBCC-RBCC-B30.2 domain, the medium form with the RBCC-B30.2 domain, and the short form with only the RBCC domain. Moreover, respective transcripts corresponding to the three isoforms were detected in various human organs by reverse transcription-PCR and Northern blot analyses. Interestingly, the medium form of the RNF21 mRNA expressed most predominantly was dramatically up-regulated within 8-16 h by interferon stimulation of HeLa cells. These findings suggest that RNF21 is a downstream gene that may mediate interferon's biological action.

Anticachectic effects of Coptidis rhizoma, an anti-inflammatory herb, on esophageal cancer cells that produce interleukin 6.

Herbs as alternative cancer therapies have attracted a great deal of recent attention due to their low toxicity and costs. In this study, the antitumor activity and anticachectic effect of Coptidis rhizoma, an anti-inflammatory herb, were investigated in nude mice carrying a human esophageal cancer cell line YES-2, which constitutively secretes interleukin-6 (IL-6) and induces cachexia when injected into these mice. In this study, in vivo growth of YES-2 cells was not affected by an oral supplement containing the extract powder of C. rhizoma at a final concentration of 1% (CR supplement). However, in comparison with normal diet, CR supplement significantly attenuated weight loss of tumor-bearing mice without a change in food or water intake. Tumor IL-6 levels were significantly lower in mice treated with CR supplement than in control mice (P<0.001). Serum IL-6 was detectable in four (50%) of eight control mice; IL-6 was not detected in mice treated with CR supplement. We also confirmed that berberine (8-32 microM), a major component of C. rhizoma, dose-dependently inhibited secretion of IL-6 by YES-2 cells in vitro. Moreover, reverse transcription-PCR assay showed that treatment of YES-2 cells with berberine (8-32 microM) for 24 h reduced IL-6 mRNA expression. Our results suggest that C. rhizoma may have an anticachectic effect on esophageal cancer and an effect is associated with the ability of berberine to down-regulate tumor IL-6 production.

Molecular characterization and physiological role of ascorbate peroxidase from halotolerant Chlamydomonas sp. W80 strain.

A cDNA clone encoding an ascorbate peroxidase was isolated from the cDNA library from halotolerant Chlamydomonas W80 by a simple screening method based on the bacterial expression system. The cDNA clone contained an open reading frame encoding a mature protein of 282 amino acids with a calculated molecular mass of 30,031 Da, preceded by the chloroplast transit peptide consisting of 37 amino acids. In fact, ascorbate peroxidase was localized in the chloroplasts of Chlamydomonas W80 cells; the activity was detected in the stromal fraction but not in the thylakoid membrane. The deduced amino acid sequence of the cDNA showed 54 and 49% homology to chloroplastic and cytosolic ascorbate peroxidase isoenzymes of spinach leaves, respectively. The enzyme from Chlamydomonas W80 cells was purified to electrophoretic homogeneity. The molecular properties of the purified enzyme were similar to those of the other algal ascorbate peroxidases rather than those of ascorbate peroxidases from higher plants. The enzyme was relatively stable in ascorbate-depleted medium compared with the chloroplastic ascorbate peroxidase isoenzymes of higher plants. The presence of NaCl (3%) as well as of beta-d-thiogalactopyranoside was needed for the expression of Chlamydomonas W80 ascorbate peroxidase in Escherichia coli.

[Effect of chorei-to and gorei-san on calcium oxalate crystallization in human urine].

The inhibitory effect of two kinds of Kampou medicine, Chorei-to and Gorei-san, on calcium oxalate crystallization in human urine, was examined. Chorei-to or Gorei-san was administered to 15 healthy male volunteers for three consecutive days under normal dietary conditions and 24-hour urine samples were collected before and after the administration. Urinary parameters including calcium, phosphorus, uric acid, oxalate and citrate were measured on each sample obtained. The inhibitory effect on calcium oxalate crystallization was examined using part of the collected 24-hour urine. There was no significant difference in urinary parameters before and after Chorei-to administration. Gorei-san administration significantly increased magnesium excretion in urine. A significant inhibitory effect on calcium oxalate crystallization in human urine was recognized when Gorei-san was administered, whereas Chorei-to had no inhibitory effect on calcium oxalate crystallization in urine.

Expression of Tamm-Horsfall protein in stone-forming rat models.

OBJECTIVES: To examine the expression of Tamm-Horsfall protein (THP) and calcium oxalate deposition in three rat models to clarify whether THP plays an active role in crystal formation or whether crystals induce the secretion of this protein. MATERIALS AND METHODS: A stone-forming rat model (model 1) with marked tubular dilatation in an entire kidney was produced by rendering Wistar rats (aged 8 weeks) hyperoxaluric and hypercalciuric, through compulsorily feeding with 0.12 mL of 5% ethylene glycol (in two doses daily) and 0.5 microgram of vitamin D3 every other day. Two other rat models were also produced. Model 2 comprised stone-forming rats with minimal tubular dilatation, achieved by giving rats the same dose of ethylene glycol once daily, and model 3 comprised stone-free rats with marked tubular dilatation achieved by unilateral ureteric ligation. The rats' kidneys were resected after 4 weeks and all resected kidneys immunohistochemically stained with an antibody to THP. Simultaneously, the location of calcium oxalate (CaOx) crystals was established with von Kossa staining. The relation between crystals and the secretion of THP was also assessed in vitro. Cultured renal epithelial cells (NRK-52E) were stained with an antibody to THP after they had been cultured for 72 h in a medium containing CaOx crystals. RESULTS: In model-1 kidneys with both tubular dilatation and many crystals, there was local and intense expression of THP in many renal tubules. CaOx crystals and the intense expression of THP tended to occur in the same renal tubules. In model 2 kidneys with little tubular dilatation, only a few renal tubules expressed THP strongly and the location of the crystals rarely coincided with that of THP expression. In model 3 kidneys with marked tubular dilatation but no crystals, THP was expressed strongly in many renal tubules. The expression of THP in cultured NRK-52E cells was not stimulated by CaOx crystals. CONCLUSIONS: The results from the in vivo models suggest that THP did not initiate crystal formation and the strong expression of THP was induced not by crystals but by renal tubular damage caused by tubular dilatation. From the close association of THP and crystals in model 1 kidneys, this protein might play a secondary role as an adhesive, promoting stone formation.

[Inhibitory effect of oral administration of Takusya on calcium oxalate crystallization in human whole urine].

We previously reported that Takusya had the inhibitory effect on in vitro calcium oxalate crystallization and in vivo stone formation in an animal model and it could be a prophylactic agent against calcium oxalate stone formation. We studied the effect of Takusya on calcium oxalate crystallization in human urine. Takusya (500 mg/day and 1,000 mg/day) was administered to 16 healthy men for 3 days and then 24-hour urine samples were collected to measure the urinary excretion of calcium, phosphate, magnesium, uric acid, creatinine, citric acid and oxalic acid. The urine samples before the administration of Takusya was used as a control. The size and distribution of crystals, which were formed in the urine samples by adding calcium chloride and sodium oxalate, were measured using the Coulter counter technique. Urinary magnesium excretion was significantly reduced by 1,000 mg/day of Takusya compared with the control (p < 0.05). The growth of crystals was significantly inhibited by 500 mg/day of Takusya in the large crystal formers whose urine created crystals more than 3.5 microns before the administration of Takusya (p < 0.05). These findings suggested that Takusya inhibited the growth of crystals formed in human urine.

Alternative mRNA splicing of 3'-terminal exons generates ascorbate peroxidase isoenzymes in spinach (Spinacia oleracea) chloroplasts.

We have isolated two cDNA clones encoding spinach (Spinacia oleracea) stromal and thylakoid-bound ascorbate peroxidase isoenzymes [Ishikawa, Sakai, Yoshimura, Takeda and Shigeoka (1996) FEBS Lett. 384, 289-293]. The gene (ApxII) encoding both chloroplastic ascorbate peroxidase isoenzymes was isolated and the organization of the gene was determined. Alignment between the cDNAs and the gene for chloroplastic ascorbate peroxidase isoenzymes indicates that both enzymes arise from a common pre-mRNA by alternative splicing of two 3'-terminal exons. Genomic Southern-blot analysis supported this finding. The gene spanned nearly 8.5 kbp and contained 13 exons split by 12 introns. The penultimate exon 12 (residues 7376-7530) for the stromal ascorbate peroxidase mRNA consisted of one codon for Asp365 before the TAA termination codon, and the entire 3'-untranslated region, including a potential polyadenylation signal (AATAAA). The final exon 13 (residues 7545-7756) for the thylakoid-bound ascorbate peroxidase mRNA consisted of the corresponding coding sequence of the hydrophobic C-terminal region, the TGA termination codon and the entire 3'-untranslated region, including a potential polyadenylation signal (AATATA). Both exons were interrupted by a 14 bp non-coding sequence. Northern-blot and reverse transcription-PCR analysis showed that the transcripts for stromal and thylakoid-bound ascorbate peroxidase are present in spinach leaves.

[Evaluation of endoscopic pirarubicin-Lipiodol emulsion injection therapy for gastric cancer].

To investigate the effect on gastric cancer and metastatic lymph node, an emulsion made of pirarubicin and lipiodol mixture was injected around the lesion of the gastric cancer using gastrointestinal endoscopy. At the site of emulsion injection and lymph node, the concentration of the THP Lipiodol emulsion was enough despite injection more than 7 days before. This targeting therapy for metastatic lymph nodes was considered effective.

DNA sequence from a fossil pollen of Abies spp. from Pleistocene peat.

DNA was amplified from individual fossil pollen grains of Abies spp. (Pinaceae), which have been detected from Pleistocene peaty deposits (at least 150,000 years old). To identify the species of the fossil pollen by DNA analysis, the region indicating the species-specific sequence was searched among extant Abies species and the spacer region between rrn5 and trnR in chloroplast DNA was sequenced for four grains of the fossil pollen. Three pollen samples produced the same sequence as extant Abies species. The sequence for the remaining sample differed from that of extant Abies by one substitution. This study showed not only a successful DNA analysis from a single grain of fossil pollen but also a new method to identify the species of fossil pollen for the pollen analysis field.

cDNAs encoding spinach stromal and thylakoid-bound ascorbate peroxidase, differing in the presence or absence of their 3'-coding regions.

Two cDNA clones encoding stromal (SAP28) and thylakoid-bound (SAP22) ascorbate peroxidase were isolated from a spinach cDNA library constructed by greening cotyledons. The SAP22 and SAP28 contained an open reading frame encoding mature protein of 295 and 345 amino acids with calculated molecular mass of 32239 Da and 37710 Da, respectively, preceded by the common transit peptides of 70 amino acid residues. Interestingly, the N-terminal 364 amino acids of SAP22 were 100% identical with SAP28 except for one C-terminal amino acid residue (Asp-365), and the C-terminal of SAP22, which is the putative transmembrane segment, was 50 amino acids longer than that of SAP28.

Fasting increases the expression of basic fibroblast growth factor (FGF-2) messenger ribonucleic acid in rat hypothalamus.

Basic fibroblast growth factor (FGF-2) has been isolated from the brain, but the regulation of FGF-2 synthesis in the brain is not yet fully understood. Since exogenously administered FGF-2 has been reported to suppress food intake as well as the secretion of gastric acid and pepsin in rats, we examined the effect of fasting on FGF-2 mRNA levels in the hypothalamus and the cerebral cortex of male rats, using RNase protection assay. Fasting for 72 h resulted in an approximately 2-fold increase in FGF-2 mRNA level in the hypothalamus but did not affect FGF-2 mRNA level in the cerebral cortex significantly. These findings support the hypothesis that FGF-2 plays a significant role in regulation of hypothalamic function.

Expression of basic fibroblast growth factor (FGF-2) messenger ribonucleic acid is regulated by testosterone in the rat anterior pituitary.

Evidence from in vivo studies supports the concept that growth factors are involved in the function of endocrine organs. We studied the effects of target endocrine organs (thyroid, adrenals, and gonads) on levels of basic fibroblast growth factor (FGF-2) messenger ribonucleic acid (mRNA) in the anterior pituitary and the hypothalamus of male rats using RNase protection assays. Castration significantly reduced the levels of FGF-2 mRNA in the anterior pituitary, but not in the hypothalamus. This decrease was restored by testosterone administration. The regulation of pituitary FGF-2 mRNA involves a specific hormone, i.e. testosterone, since neither adrenalectomy nor chemical thyroidectomy affects the expression of the gene for FGF-2.

Preclinical and clinical study of FK506 in uveitis.

Efficacy of a new immunosuppressive agent, FK506, in refractory uveitis was studied in 8 patients: 5 with Behcet's disease and 3 with idiopathic retinal vasculitis. The agent was given by oral administration every 12 hours. The previous therapy with systemic corticosteroids or immunosuppressive agents including cyclosporine failed to subside uveitis in these cases. During the observation period of 21.6 +/- 7.8 weeks (mean +/- SD) under FK506 at doses with 0.05, 0.1, 0.15 or 0.2 mg/kg/day, the visual acuity was increased in 44% of treated eyes, unchanged in 44% and decreased in 12%. The inflammatory activity in the ocular fundus was improved in 69% and unchanged in 6% of treated eyes. The effects of FK506 on uveitis by the criteria of improvement of visual acuity and uveitis activity was dose-dependent: 0.05 and 0.1 mg/kg/day were ineffective but 0.15 and 0.2 mg/kg/day were effective in most cases. One patient with Behcet's disease converted from cyclosporine developed moderate renal impairment in 4 weeks under FK506 and the therapy was discontinued in 8 weeks, though the uveitis activity as well as visual acuity was markedly improved. Other 7 cases had no side effect of FK506. Although the number of cases was small and observation period was short, the present clinical data indicate that FK506 is effective to treat refractory uveitis.