Beta-hydroxy-beta-methylbutyrate associated with low-intensity exercise training improves skeletal muscle regeneration through the IGF-Akt pathway.

The effect of beta-hydroxy-beta-methylbutyrate (HMB) supplementation associated with exercise training at different intensities and frequencies on skeletal muscle regeneration of muscle-injured rats was investigated. Male Wistar rats were divided into sedentary and trained groups. The sedentary groups were subdivided into non-injured (SED-Ct), non-injured supplemented with HMB (SED-Ct-HMB), injured (SED), and injured with HMB (SED-HMB), and the trained groups were injured, supplemented with HMB, and then divided into training three times a week without load (HT3) or with load (HT3L) and training five times a week without load (HT5) and with load (HT5L). The rats received a daily dose of HMB associated with 60 min of swimming with or without 5% body mass load for 14 days. On the 15th day, cryoinjury was performed in the right tibialis anterior muscle (TA), and 48 h later, supplementation and training continued for 15 days. After the last session, the TA was dissected and a cross-sectional area (CSA) of muscle fibers was used to determine the percentage of CSA fibers and connective tissue (%CT), as well as the total and phosphorylated protein contents. SED-HMB showed increased CSA and decreased %CT and TGF-ß when compared to SED. HT3 showed increased CSA and reduced %CT accompanied by increased IGF-1/Akt, myogenin, and MuRF1, and decreased TGF-ß. The CSA of HT5L also increased, but at the cost of a higher %CT compared to the other groups. Our results demonstrated that HMB associated with training without load and with lower frequency per week may be a valuable strategy for skeletal muscle regeneration.

Enhanced in vitro biological activity generated by surface characteristics of anodically oxidized titanium--the contribution of the oxidation effect.

Anodically oxidized titanium surfaces, prepared by spark discharge, have micro-submicron surface topography and nano-scale surface chemistry, such as hydrophilic functional groups or hydroxyl radicals in parallel. The complexity of the surface characteristics makes it difficult to draw a clear conclusion as to which surface characteristic, of anodically oxidized titanium, is critical in each biological event. This study examined the in vitro biological changes, induced by various surface characteristics of anodically oxidized titanium with, or without, release of hydroxyl radicals onto the surface. Anodically oxidized titanium enhanced the expression of genes associated with differentiating osteoblasts and increased the degree of matrix mineralization by these cells in vitro. The phenotypes of cells on the anodically oxidized titanium were the same with, or without, release of hydroxyl radicals. However, the nanomechanical properties of this in vitro mineralized tissue were significantly enhanced on surfaces, with release of hydroxyl radicals by oxidation effects. In addition, the mineralized tissue, produced in the presence of bone morphogenetic protein-2 on bare titanium, had significantly weaker nanomechanical properties, despite there being higher osteogenic gene expression levels. We show that enhanced osteogenic cell differentiation on modified titanium is not a sufficient indicator of enhanced in vitro mineralization. This is based on the inferior mechanical properties of mineralized tissues, without either being cultured on a titanium surface with release of hydroxyl radicals, or being supplemented with lysyl oxidase family members.

Outcomes of immediate perforator flap reconstruction after skin-sparing mastectomy following neoadjuvant chemotherapy.

BACKGROUND: The impact of neoadjuvant chemotherapy (NACT) on immediate free flap breast reconstruction remains controversial. Furthermore, the oncological outcomes of immediate free flap breast reconstruction after skin-sparing mastectomy (SSM) following NACT remain unclear. This study aimed to investigate the surgical complications and oncological outcomes of immediate perforator flap reconstruction after SSM following NACT. METHODS: A total of 201 consecutive patients with indications for immediate perforator flap reconstruction after SSM were included between 2004 and 2012. Surgical and oncological outcomes were compared between patients with and without NACT. RESULTS: There were 38 patients in the NACT group and 163 in the non-NACT control group. The median age of the NACT group was 39.5 years, which was significantly younger than the control group (43.0 years; P < 0.05). Patients in the NACT group also had more advanced and aggressive disease (P < 0.05). There was no significant difference in the frequency of surgical complications between the groups, no difference in the type of complications, and no significant difference in the frequencies of major and minor complications. No patients in the NACT group had delayed adjuvant therapy. Eight patients (4%) developed recurrences, with a median follow-up time of 3.0 years. Local recurrences occurred in three control patients but no patients in the NACT group. CONCLUSION: NACT does not affect short-term or interim outcomes after immediate perforator flap reconstruction and may thus represent a safe and practical treatment option for the multidisciplinary treatment of breast cancer.

Suppression of membrane-type 1 matrix metalloproteinase (MMP)-mediated MMP-2 activation and tumor invasion by testican 3 and its splicing variant gene product, N-Tes.

Using expression cloning to screen a human fetal kidney cDNA library for regulator(s) of pro-matrix metalloproteinase (MMP)-2 processing mediated by membrane-type (MT) 1 MMP, we isolated a cDNA whose product interfered with pro-MMP-2 activation. It encodes the NH(2)-terminal 313-amino acid region of a calcium-binding proteoglycan, testican 3, with a 3-amino acid substitution at the COOH terminus and thus was named N-Tes. N-Tes comprises a signal peptide, a unique domain, a follistatin-like domain, and a Ca(2+)-binding domain but lacks a COOH-terminal thyroglobulin domain and two putative glycosaminoglycan attachment sites of testican 3. Pro-MMP-2 activation by MT3-MMP was also inhibited by the coexpression of N-Tes. Immunoprecipitation analysis demonstrated direct interaction of N-Tes with either MT1-MMP or MT3-MMP. Expression of testican 1 or testican 3 but not testican 2 also inhibited pro-MMP-2 activation by either MT1-MMP or MT3-MMP. Deletion and substitution of amino acids residues in N-Tes revealed that the unique NH(2)-terminal domain of N-Tes is responsible for the inhibition of pro-MMP-2 activation by MT-MMPs. Expression of N-Tes and testican 3 was detected in normal brain but down-regulated in glioma tissues. Transfection of either the N-Tes or testican 3 gene into U251 glioma cells or Madin-Darby canine kidney cells transformed by erbB2 suppressed their invasive growth in collagen gel. These results suggest that both N-Tes and testican 3 would interfere with tumor invasion by inhibiting MT-MMPs.

Vitamin D receptor gene polymorphism detected by digestion with Apa I influences the parathyroid response to extracellular calcium in Japanese chronic dialysis patients.

BACKGROUND: To play its physiological role, 1,25(OH)2D3 must bind to a specific vitamin D receptor (VDR) in the nucleus. We have previously reported that VDR gene polymorphism influences the parathyroid function in patients with end-stage renal disease (ESRD). In the present study, we have investigated the relationship between the parathyroid responsiveness and VDR gene polymorphism, as detected by the Apa I restriction enzyme, by changing the concentration of Ca2+ in the dialysate. METHODS: 58 Japanese ESRD patients undergoing renal replacement therapy in our institution were evaluated. Genomic DNA was extracted from peripheral leukocytes and digested at the intron between exon 8 and exon 9 of the VDR gene using Apa I enzyme. Then alleles were classified into genotype A (undigested allele) and genotype a (digested allele). Extracellular ionized calcium ([Ca2+]e), serum phosphate, and intact parathyroid hormone (PTH) were measured before and after each hemodialysis (HD) session with dialysates having different concentrations of Ca2+ (1.5 or 1.25 mmol/l). The significance of differences in statistical analyses was defined within confidence limits of 5.0%. RESULTS: The AA, Aa, and aa genotypes were observed in 7/58 patients (12.1%), 23/58 patients (39.6%), and 28/58 patients (48.3%), respectively. The PTH reduction after HD with the 1.5-mmol/l Ca dialysate did not differ significantly between group AA+Aa and group aa. On the other hand, the PTH increase was significantly higher in group aa than in group AA+Aa after HD with the 1.25-mmol/l Ca dialysate (p = 0.0107), despite a similar PTH level before HD. Similarly, the percent increase of PTH after HD with the 1.25-mmol/l Ca dialysate was significantly higher (p = 0.0112) in group aa (50.2 +/- 9.4%) than in group AA+Aa (19.7 +/- 7.2%). There were no significant differences between the two groups in [Ca2+]e nor in serum phosphorus (Pi) before and after HD with either dialysate. Group AA+Aa and group aa did not show statistically significant differences in age, female/male ratio, ratio of diabetic nephropathy, or dialysis period. CONCLUSIONS: The study results showed that the patients in group aa were more sensitive to changes in [Ca2+]e than those in group AA+Aa. Moreover, they suggested that the VDR gene polymorphism may affect parathyroid responsiveness to changes in [Ca2+]e, which in turn may influence onset and progression of hyperparathyroidism in ESRD patients.

Increased oxidative stress in childhood atopic dermatitis.

Atopic dermatitis (AD) is a chronic inflammatory skin disease of unknown etiology. To examine the involvement of impaired homeostasis of oxygen/nitrogen radicals in childhood AD, we compared the levels of urinary 8-hydroxy-2'-deoxyguanosine (marker of oxidative stress), nitrite/nitrate (marker of nitric oxide synthesis) and selenium (marker of selenium store) in 27 children with AD to those of 25 healthy control children. Urinary 8-hydroxy-2'-deoxyguanosine was significantly higher and nitrite/nitrate levels were significantly lower in patients with AD than in the control. Urinary selenium levels were similar in both groups. Our findings suggest that impaired homeostasis of oxygen/nitrogen radicals and increased oxidative stress are involved in the pathophysiology of childhood AD, and indicate that suppression of oxidative stress might be a potentially useful strategy for the treatment of AD.

Differences in the recognition of glucan elicitor signals between rice and soybean: beta-glucan fragments from the rice blast disease fungus Pyricularia oryzae that elicit phytoalexin biosynthesis in suspension-cultured rice cells.

Partial acid/enzymatic hydrolysis of the beta-(1-->3, 1-->6)-glucan from the cell walls of the rice blast disease fungus Pyricularia oryzae (Magnaporthe grisea) released elicitor-active fragments that induced phytoalexin biosynthesis in suspension-cultured rice cells. From the digestion of the glucan by an endo-beta-(1-->3)-glucanase, one highly elicitor-active glucopentaose was purified as a reduced compound, tetraglucosyl glucitol. The structure of this tetraglucosyl glucitol as well as two other related tetraglucosyl glucitols was elucidated as follows: (1) Glcbeta(1-->3)Glcbeta(1-->3)(Glcbeta(1-->6)) Glcbeta(1-->3)Glucitol (most active fragment); (2) Glcbeta(1-->3)(Glcbeta(1-->6))Glcbeta(1-->3)Glcbeta (1-->3)Glucitol; and (3) Glcbeta(1-->6) Glcbeta(1-->3)Glcbeta(1-->3)Glcbeta(1-->3)Glucitol. However, a synthetic hexa-beta-glucoside, known as a minimal structural element for the phytoalexin elicitor for soybean cotyledon cells, did not induce phytoalexin biosynthesis in the rice cells. Conversely, the beta-glucan fragment from P. oryzae did not induce phytoalexin biosynthesis in the soybean cotyledon cells, indicating differences in the recognition of glucooligosaccharide elicitor signals in these two plants. Because rice cells have been shown to recognize chitin fragments larger than pentamers as potent elicitors, these results also indicate that the rice cells can recognize at least two types of oligosaccharides from fungal cell walls as signal molecules to initiate defense response.

[A case of hepatocellular carcinoma with tumor thrombus in the right atrium successfully treated by arterial administration of lipiodol-SMANCS].

In February, 1996, a 73-year-old male with liver dysfunction was admitted to our hospital for further examination and treatment of liver tumor. The liver tumor was revealed by imaging examination, which was mainly in the S4-S8 of liver with a thrombus growing from the right anterior branch to the right branch of the portal vein, and from the right hepatic vein to the inferior vena cava and right atrium. The serum AFP and PIVKA-II levels were elevated to 3.610 ng/ml and 54 AU/ml, respectively. The patient was diagnosed as having hepatocellular carcinoma, and was treated by arterial administration of anticancer drugs (epirubicin hydrochloride, mitomycin C and carboplatin) and TAE. Though the main tumor (S4-S8 of liver) was reduced by TAE, the portal and atrial tumor thrombus did not respond. One month after TAE (20 May, 1996), the first arterial administration of Lipiodol-SMANCS was given, followed by 4 successive procedures with an interval of about 1.5 months (total dose 15 mg), resulting in remarkable tumor thrombus shrinkage and reduction of AFP levels to 80 ng/ml. This case shows that arterial administration of SMANCS may be one of the effective treatments for hepatocellular carcinoma, even with tumor thrombus of hepatic vein, IVC and right atrium.

Pharmacological properties of traditional medicines. XXIII. Searching for active compounds in the blood and bile of rats after oral administrations of extracts of sansohnin [symbol: see text].

We made a trial of searching for the bioactive substances from Sansohnin [symbol: see text]. The blood and bile of rats after the oral administration of extracts of Sansohnin were analyzed by three-dimensional high-performance liquid chromatography (3D-HPLC). In blood, spinosin and feruloyl spinosin were found after the oral administration of a butanol extract of Sansohnin. In bile, spinosin and feruloyl spinosin were also identified after the oral administration of a water extract of Sansohnin. Spinosin and feruloyl spinosin induced the prolongation of hexobarbital sleeping time in mice at doses of 50 and 100 mg/kg, respectively. We concluded that spinosin and feruloyl spinosin were the bioactive constituents of Sansohnin. It was assumed that spinosin and feruloyl spinosin circulated through the intestine and liver, therefore, these results will provide support for the sedative and hypnotic use of this crude drug in Oriental medicine.

Immunosuppressive effects of gallic acid and chebulagic acid on CTL-mediated cytotoxicity.

Gallic acid (GA) and chebulagic acid (CA) were isolated from the extract of a herbal medicine, kashi (myrobalans: the fruit of Terminalia chebula) as active principles that blocked the cytotoxic T lymphocyte (CTL)-mediated cytotoxicity. GA and CA inhibited the killing activity of CD8+ CTL clone at IC50 values of 30 microM and 50 microM, respectively. Granule exocytosis in response to anti-CD3 stimulation was also blocked by GA and CA at the equivalent concentrations.

Oral administration of a unicellular green algae, Chlorella vulgaris, prevents stress-induced ulcer.

Oral administration of dry powder of Chlorella vulgaris (CVP) showed clear prophylactic effects in water-immersion restraint stress-induced and in cysteamine-induced peptic ulcer models, but not in Shay's rat model. Drugs that enhance the protective factors of ulcer formation are effective in the first two models. CVP may prevent ulcer formation mainly through the "immune-brain-gut" axis and protection of gastric mucosa by its own characteristics.

[Preliminary screening for antiviral AIDS drugs. VIII. Report for fiscal year 1995].

Preliminary screening of antiviral AIDS drugs has been carried out using three different in vitro assay systems. Among 96 samples of different origin tested, two were shown to inhibit the growth of HIV in vitro. One of the positive samples (plant origin) has hopeful signs, as the ranges of effective doses are wider than those of most of positive samples which had been found by us.

Hyperammonemia in a patient with short bowel syndrome and chronic renal failure.

A patient with short bowel syndrome (SBS) and renal failure developed a disturbance of consciousness with hyperammonemia. Abnormally low concentrations of ornithine, citrulline, and arginine were observed on the plasma aminogram. These results suggested that the activities of amino acid synthetase localized in the small intestinal flora were lost. The small intestine is required for arginine synthesis; thus, infusion limited to the essential amino acids to SBS patients will cause a deficiency of the urea cycle intermediates, ornithine, citrulline, and arginine and may lead to hyperammonemia. In addition, the renal insufficiency may have caused decreased excretion of ammonia. In this patient, supplemental arginine improved the symptoms.

[Preliminary screening for antiviral AIDS drugs. VI. Report for fiscal year 1993].

Preliminary screening of antiviral AIDS drugs has been carried out using three different in vitro assay systems. Among 138 samples tested, two were found to inhibit the growth of HIV in vitro. Neither of the positive samples has hopeful signs, as the ranges of effective doses of the samples are very narrow.

[Preliminary screening for antiviral AIDS drugs. VII. Report for fiscal year 1994].

Preliminary screening of antiviral AIDS drugs has been carried out using three different in vitro assay systems. Among 246 samples of different origin tested, six were shown to inhibit the growth of HIV in vitro. Two of the positive samples have hopeful signs, as the ranges of effective doses are wider than those of most of positive samples which had been found by us.

[Successful treatment of advanced gastric cancer (Borrmann 1 type) with FTP chemotherapy after reduction surgery].

A 54-year-old man was diagnosed with Borr 1 type gastric cancer, located just below ECJ with some paraaortic lymph node metastase, during treatment of diabetes mellitus at another hospital. He underwent spleno-total gastrectomy for reduction. The metastatic lymph nodes of the para-aorta were not resected, so the surgery was considered palliative. We administered FTP chemotherapy (CDDP 110 mg/day 1, 5-FU 1,200 mg/day 1-5, THP-ADM 30 mg/day 1) 5 times following surgery. The metastatic lymph nodes were remarkably decreased in size by the initial treatment. The decrement was 52.4% after the initial treatment (PR). After the 4th treatment, there were no lymph nodes detected (CR). After the 5th treatment, CR continued. The PR period was considered to be 5 months, and that of CR 4 months. The patient has no renal or heart dysfunction, and no suppression of bone marrow. His quality of life is satisfactory, and he continues to work as prior to surgery. FTP chemotherapy is considered a successful regimen for postoperative chemotherapy.

[Clinicopathological evaluation of high-range hyperthermia for advanced thoracic esophageal carcinoma].

The effectiveness of high-range hyperthermia over 45 degrees C for advanced esophageal carcinoma was clinicopathologically evaluated. Fifty-eight patients with advanced thoracic esophageal carcinoma were treated with radio-chemotherapy. They were divided into two groups: group I included 32 cases, all of whom received hyperthermia (13 cases: high-range hyperthermia); group II included the other 26 cases. The patients were given 2 Gy/day for a total of 15 sessions in 3 weeks. Bleomycin and cisplatin in combination with 5-fluorouracil have been employed as chemotherapy. Hyperthermia was performed twice a week for a total of 6 sessions. Intraluminal heating was done using Japan Crescent Inc. IH-500 T (RF, 13.56 MHz), with an intraesophageal applicator and two extra-corporeal applicators on the chest and the back. Concerning local effects, the efficacy rate was 81.3% in group I (high-range: 92.9%), and 42.3% in group II. The histologic effectiveness, when Grade 2 and 3 are determined as histologically effective, were 64.3% (high range: 85.7%) and 50.0% in group I and II, respectively.

Enhancement of immunoglobulin A production in Peyer's patches by oral administration of a traditional Chinese medicine, xiao-chai-hu-tang (Shosaiko-to).

The Peyer's patches contain a large number of precursor cells committed to the production of immunoglobulin A (IgA) and play an important role in IgA response in the mucosal immune system. We investigated the induction of IgA producing cells in Peyer's patches by plaque forming cell assay after oral administration of a traditional Chinese herbal medicine, Xiao-chai-hu-tang (Japanese name: Shosaiko-to). The number of total IgA producing cells in the Peyer's patches detected by the protein-A plaque assay was increased about two-fold by Shosaiko-to administration and the numbers of both anti-SRBC and anti-HRBC IgA producing cells were also increased by such a treatment. On the other hand, when SRBC alone were administered orally, only the number of anti-SRBC IgA producing cells was increased. Further, we examined T-cell subpopulations in the gut-associated lymphoid tissues after oral administration of Shosaiko-to by flowcytometry. Marked alternations in T cell subpopulations were not detected in the Peyer's patches, though TcR gamma delta+T-cells in the intraepithelial lymphocytes and Thy1.2-TcR alpha beta+T cells in the mesenteric lymph nodes were slight increased. These results showed that orally administered Shosaiko-to acts as a polyclonal B-cell activator which induces IgA production in the mucosal immune system.

Impact of autologous blood transfusions on patients undergoing radical prostatectomy using hypotensive anesthesia.

We report on a retrospective study consisting of 71 consecutive patients who underwent radical retropublic prostatectomy under controlled hypotensive anesthesia, examining the impact of a preoperative autologous blood collection program on perioperative blood requirements. The population consisted of 34 patients who elected not to use autologous blood (group 1) and 37 patients who underwent radical prostatectomy after storing 1 to 3 units of their own blood (group 2). Median operative time and estimated blood loss were not statistically different. In group 1, 20 homologous blood units were used by 7 patients (21%). In group 2, 37 of 41 units transfused were autologous units and only 3 patients (8%) received homologous blood. However, for every unit of autologous blood deposited preoperatively there was a corresponding decrease in admission blood concentrations and 21 of 37 patients were anemic at hospitalization. In addition, the study suggests that in many patients the anemia produced by preoperative phlebotomy does not resolve preoperatively. In response, the effectiveness of preoperative autologous blood collection is decreased by this preoperative anemia. Although inefficient, we nevertheless conclude that an autologous blood collection program decreases homologous transfusion exposure and efforts should be directed to increase the erythropoietin response to the anemia produced by preoperative phlebotomy. Presently, it is an expensive program that offers only a modest benefit for patients undergoing radical retropubic prostatectomy.

Protective effect of ren-shen-yang-rong-tang (Ninjin-youei-to) in mice with drug-induced leukopenia against Pseudomonas aeruginosa infection.

When 200 mg/kg of cyclophosphamide (CY) or 5-fluorouracil (5-FU) were given subcutaneously to mice, severe reduction of leukocyte numbers in the peripheral blood was observed on day 4 and from day 4 to day 8 after the treatment, respectively. Daily administration of ren-shen-yang-rong-tang (Japanese name: Ninjin-youei-to, NYT), (1 g/kg/day) and recombinant human granulocyte-colony stimulating factor (rhuG-CSF, 2 micrograms/mouse/day) either from day 0 to day 4 after treatment with CY or from day 0 to day 8 after treatment with 5-FU accelerated the recovery of peripheral leukocytes. Administration of NYT and rhuG-CSF inhibited decreases in the number of colony forming units in the spleen (CFU-S) in drug-treated mice. In mice infected intraperitoneally with a virulent strain of Pseudomonas aeruginosa 4 days and 8 days after treatment with CY and 5-FU, respectively, lethal doses were much lower (approximately 1/1000) than that in normal mice. Administration of NYT and rhuG-CSF to drug-treated mice inhibited the enhanced bacterial growth in the peritoneal cavity. Administration of NYT or rhuG-CSF to drug-treated mice enhanced the recovery of accumulation of leukocytes into the infected peritoneal cavity from their depressed state. Administration of NYT was more effective for improving the host resistance of 5-FU-treated mice than for improving that of CY-treated mice in contrast to rhuG-CSF which exhibited stronger effect in CY-treated mice than in 5-FU treated mice.