Search

Introduction: The Seamless Treatment of Osteoporosis against Fractures (STOP-Fx) study was initiated to provide and continue therapeutic interventions for registered patients with osteoporotic fractures., Materials and Methods: Women who visited six hospitals in the western Kitakyushu area for osteoporotic fractures between October 2016 and December 2018 were included in the study. Data collection for primary and secondary outcomes was conducted from October 2018 to December 2020, 2 years after STOP-Fx study enrollment. The primary outcome included the number of surgeries for osteoporotic fractures after the STOP-Fx study intervention, while secondary outcomes were the intervention rate of osteoporosis treatment, incidence and timing of secondary fractures, and factors associated with secondary fractures and loss to follow-up., Results: Concerning the primary outcome, the number of surgeries for osteoporotic fractures decreased since the STOP-Fx study initiation: 813 in 2017, 786 in 2018, 754 in 2019, 716 in 2020, and 683 in 2021. Regarding the secondary outcome, of the 805 patients enrolled, 445 were available for follow-up at 24 months. Of the 279 patients who were untreated for osteoporosis at enrollment, 255 (91%) were on treatment at 24 months. There were 28 secondary fractures, which were associated with increased tartrate-resistant acid phosphatase-5b and decreased lumbar spine bone mineral density during enrollment in the STOP-Fx study., Conclusion: As the demographics and medical area served by six hospitals in the western Kitakyushu region have not changed significantly since the STOP-Fx study initiation, the STOP-Fx study may have contributed in decreasing the number of osteoporotic fractures., (© 2023. The Japanese Society Bone and Mineral Research.)

The role of steric hindrance and charge interactions in governing ionic transport through reduced graphene oxide (rGO) and commercial (DOW-Filmtec NF270) membranes was elucidated by a comprehensive study of experimental and established mathematical analysis based on Nernst-Planck equation. A charge-dominated salt exclusion mechanism was observed for the rGO membranes, which exhibited retention from low (7%) to moderate (70%) extent depending on the nature of ions (5 mM). Swelling of GO (1.2 nm interlayer distance) in water beyond the hydrated diameter of ions was attributed as a primary cause for lowering steric hindrance effects. The influence of parameters affecting charge interactions, such as pH and ionic strength, on the extent of salt rejection was modelled. The potential impact of the membrane's charge density, GO loading and interlayer spacing on salt retention was quantified by performing sensitivity analyses. For a high TDS produced water sample, the rGO membranes partially retained divalent cations (Ca:13%) and exhibited high dissolved oil rejection. The membranes were found to be suitable for the treatment of high TDS water with the goal of selectively removing organic impurities, and thus minimizing the impact of osmotic pressure effect. Performance of the membranes was also investigated for retention of water remediation related organic anions, using perfluoro octanoic (PFOA) acid as a model compound. rGO membranes exhibited a charge-dominated exclusion mechanism for retention (90%) of PFOA (1 ppm)., Competing Interests: Declaration of competing interest The authors have no competing interests to declare.

Pore functionalized membranes with appropriate ion exchange/chelate groups allow toxic metal sorption under convective flow conditions. This study explores the sorption capacity of ionic mercury in a polyvinylidene fluoride-poly(acrylic acid) (PVDFs-PAA) functionalized membrane immobilized with cysteamine (MEA). Two methods of MEA immobilization to the PVDF-PAA membrane have been assessed: (i) ion exchange (IE) and (ii) carbodiimide cross-linker chemistry using 1-(3-dimethylaminopropyl)-3-ethyl carbodiimide hydrochloride (EDC) and N -hydroxysuccinimide (NHS), known as EDC/NHS coupling. The ion exchange method demonstrates that cysteamine (MEA) can be immobilized effectively on PVDF-PAA membranes without covalent attachment. The effectiveness of the MEA immobilized membranes to remove ionic mercury from the water was evaluated by passing a dissolved mercury(II) nitrate solution through the membranes. The sorption capacity of mercury for MEA immobilized membrane prepared by the IE method is 1015 mg/g PAA. On the other hand, the sorption capacity of mercury for MEA immobilized membrane prepared by EDC/NHS chemistry is 2446 mg/g PAA, indicating that membrane functionalization by EDC/NHS coupling enhanced mercury sorption 2.4 times compared to the IE method. The efficiencies of Hg removal are 94.1 ± 1.1 and 99.1 ± 0.1% for the MEA immobilized membranes prepared by IE and EDC/NHS coupling methods, respectively. These results show potential applications of MEA immobilized PVDF-PAA membranes for industrial wastewater treatment specifically from energy and mining industries to remove mercury and other toxic metals.

Objectives: To assess the vitamin D status in postmenopausal women with osteoporotic fractures, determine its concentration by fracture site at the clinical setting, and compare the proportion of vitamin D deficiency with that reported in literature., Methods: The prospective study included 317 postmenopausal women with osteoporotic fractures who were treated consecutively from 2016 to 2018. After obtaining informed consent for participation in the seamless treatment of osteoporosis against fractures study, which is our initiative to prevent secondary osteoporotic fractures, we registered the patients, examined bone mineral density (BMD) at the unfractured femoral neck and lumbar spine, serum 25-hydroxyvitamin D (25(OH)D) concentration, blood chemistry, and bone turnover markers., Results: The mean age of the patients was 80.7 years. Moreover, 78% of patients of all fractures had 25(OH)D concentration < 20 ng/mL, whereas 12% of patients had 25(OH)D concentration ≥ 30 ng/mL 25(OH)D concentration in hip fractures was significantly lower than that in vertebral or distal radius fractures (P < 0.05). Multiple regression analysis revealed that 25(OH)D concentration is significantly associated with femoral neck BMD (β = 0.16; 95% confidence interval [CI], 0.78-12.17, P = 0.03) and serum albumin concentration (β = 0.21; 95% CI, 0.62-2.96, P < 0.001) in patients with 25(OH)D concentration < 30 ng/mL., Conclusions: The results of this study show that the proportion of postmenopausal women with osteoporotic fractures who had vitamin D deficiency was higher than the proportion in previous reports that examined general postmenopausal women (35.2%-52.0%)., Competing Interests: No potential conflict of interest relevant to this article was reported., (© 2019 The Korean Society of Osteoporosis. Publishing services by Elsevier B.V.)

Microtubule severing is essential for reorganization of microtubules during neuronal migration and process elongation. Katanin is a microtubule-severing enzyme, of which the major catalytic subunits are katanin A1 (KATNA1) and katanin A-like 1 (KATNAL1). The domain organization of the two subunits are almost the same; however, little is known about their functional difference. Here, we compared the expression pattern, microtubule-severing activity, intracellular degradation and knockdown phenotype in cultured cells of the two subunits. While KATNA1 was expressed ubiquitously among tissues of young adult mice, KATNAL1 was highly expressed in the brain and the testis. Neurons expressed almost only KATNAL1. When introduced into Neuro2a cells, KATNAL1 showed higher microtubule-severing activity. Cycloheximide chase analysis revealed that KATNAL1 is more stable in cells. To elucidate which part of the molecules are responsible for these characteristics, we generated chimeric molecules by swapping the amino-terminal and carboxyl-terminal halves between the two subunits. Experiments using these chimeras revealed that the amino-terminal half region is the determinant for their characteristics. Furthermore, KATNAL1 knockdown in Neuro2a cells resulted in enhancement of process elongation, while KATNA1 knockdown showed no effect. These data suggest that more active and more stable katanin subunit, KATNAL1, plays more important role in process elongation., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Oil industries generate large amounts of produced water containing organic contaminants, such as naphthenic acids (NA) and very high concentrations of inorganic salts. Recovery of potable water from produced water can be highly energy intensive is some cases due to its high salt concentration, and safe discharge is more suitable. Here, we explored catalytic properties of iron oxide (Fe x O y nanoparticles) functionalized membranes in oxidizing NA from water containing high concentrations of total dissolved solids (TDS) using persulfate as an oxidizing agent. Catalytic decomposition of persulfate by Fe x O y functionalized membranes followed pseudo-first order kinetics with an apparent activation energy of 18 Kcal/mol. Fe x O y functionalized membranes were capable of lowering the NA concentrations to less than discharge limits of 10 ppm at 40 °C. Oxidation state of iron during reaction was quantified. Membrane performance was investigated for extended period of time. A coupled process of advanced oxidation catalyzed by membrane and nanofiltration was also evaluated. Commercially available nanofiltration membranes were found capable of retaining NA from water containing high concentrations of dissolved salts. Commercial NF membranes, Dow NF270 (Dow), and NF8 (Nanostone) had NA rejection of 79% and 82%, respectively. Retentate for the nanofiltration was further treated with advanced oxidation catalyzed by Fe x O y functionalized membrane for removal of NA.

We developed a rapid and useful routine screening assay for total L-carnitine content in various infant formulas and materials by liquid chromatography coupled with a tandem mass spectrometric method (LC-MS/MS) and alkaline hydrolysis. For separation of L-carnitine, a multi-mode octadecylsilane (ODS) column was used that contained ODS ligands, anion ligands, and cation ligands to avoid using ion-pairing agents. The stable isotope L-carnitine-d3 (m/z 165 → 103/85) was used in electrospray MS/MS in the multiple reaction monitoring mode with the ion transitions of m/z 162 → 103/85 for detection and quantitation of L-carnitine. Alkaline hydrolysis of short/medium chain (C2 - C15) acyl-L-carnitines in infant formula was analyzed by LC with quadrupole time-of-flight mass spectrometry (QTOF-MS). The majority of short/medium chain acyl-L-carnitines were hydrolyzed to free L-carnitine. The overall standard deviations for L-carnitine in infant formula, follow-up formula and raw materials ranged from 2.1 to 4.0. The overall mean recoveries ranged from 90.2 to 94.2%.

The bifidogenic effect of human milk oligosaccharides (HMOs) has long been known, yet the precise mechanism underlying it remains unresolved. Recent studies show that some species/subspecies of Bifidobacterium are equipped with genetic and enzymatic sets dedicated to the utilization of HMOs, and consequently they can grow on HMOs; however, the ability to metabolize HMOs has not been directly linked to the actual metabolic behavior of the bacteria. In this report, we clarify the fate of each HMO during cultivation of infant gut-associated bifidobacteria. Bifidobacterium bifidum JCM1254, Bifidobacterium longum subsp. infantis JCM1222, Bifidobacterium longum subsp. longum JCM1217, and Bifidobacterium breve JCM1192 were selected for this purpose and were grown on HMO media containing a main neutral oligosaccharide fraction. The mono- and oligosaccharides in the spent media were labeled with 2-anthranilic acid, and their concentrations were determined at various incubation times using normal phase high performance liquid chromatography. The results reflect the metabolic abilities of the respective bifidobacteria. B. bifidum used secretory glycosidases to degrade HMOs, whereas B. longum subsp. infantis assimilated all HMOs by incorporating them in their intact forms. B. longum subsp. longum and B. breve consumed lacto-N-tetraose only. Interestingly, B. bifidum left degraded HMO metabolites outside of the cell even when the cells initiate vegetative growth, which indicates that the different species/subspecies can share the produced sugars. The predominance of type 1 chains in HMOs and the preferential use of type 1 HMO by infant gut-associated bifidobacteria suggest the coevolution of the bacteria with humans.

We have already reported the nucleotide sequences of the NP, P/V, M, F and HN genes of human parainfluenza virus type 4A (hPIV-4A) and type 4B (hPIV-4B). Here, we have determined the sequences of the L protein genes as well as the gene start, intergenic and end sequences, thereby completing the full-length genome sequence of hPIV-4A and 4B. hPIV-4A and 4B have 17,052 and 17,304 nucleotides, respectively. The end sequence of hPIV-4, especially 4B, was extraordinarily long. In a comparison with members of the genus Rubulavirus, the hPIV-4 L proteins were closely related to those of mumps virus (MUV) and hPIV-2, less closely related to those of Menangle virus and Tioman virus, and more distantly related to those of Mapuera virus and porcine rubulavirus.

The Carnivora include the superfamilies Canoidea and Feloidea. In species of Canoidea other than the domestic dog, Canis lupus, the milk contains only traces of lactose and much larger concentrations of oligosaccharides. In this study, lactose was found to be the dominant saccharide in the milk or colostrum of two species of Feloidea, namely the African lion (Panthera leo) and the clouded leopard (Neofelis nebulosa). In addition to lactose, the following oligosaccharides were characterized in the milk of a lion; Neu5Gc(α2-3)Gal(β1-4)Glc (3'-NGc-SL), Fuc(α1-2)Gal(β1-4)Glc (2'-fucosyllactose) and GalNAc(α1-3)[Fuc(α1-2)]Gal(β1-4)Glc (A-tetrasaccharide). The colostrum of a clouded leopard contained 3'-NGc-SL, Gal(α1-3)Gal(β1-4)Glc (isoglobotriose) and A-tetrasaccharide. These oligosaccharides differ in some respects from those previously identified in another species of Feloidea, the spotted hyena (Crocuta crocuta). These milks contained 3'-NGc-SL and A-tetrasaccharide, while spotted hyena colostrum did not; however, it contained Neu5Ac(α2-3)Gal(β1-4)Glc (3'-NAc-SL) and Gal(α1-3)[Fuc(α1-2)]Gal(β1-4)Glc (B-tetrasaccharide)., (© 2010 The Authors. Journal compilation © 2010 Japanese Society of Animal Science.)

To attempt a risk assessment of the excess intake of trivalent chromium (Cr), tissue Cr accumulation and urinary Cr excretion were examined in weanling rats fed experimental diets containing graded levels of Cr chloride (CrCl3) or Cr picolinate (CrPic). Thirty-six male weanling 4-weeks-old Wistar rats were divided into six groups and fed a casein-based semi-purified diet (Cr content: <0.02 microg/g) supplemented with 1, 10, or 100 microg Cr/g as CrCl3 or CrPic for 28 days. Among the experimental groups, no significant difference was observed in body weight; however, supplementation of 100 microg Cr/g to the diets caused a significant low liver weight irrespective of the chemical species of Cr. Activities of serum aspartate aminotransferase and alanine aminotransferase were significantly elevated in rats given CrPic at 100 microg Cr/g. In the liver, kidney and femur, Cr accumulation increased with elevation of the dietary Cr level. No influence of the difference in the chemical species of supplemented Cr was observed in the liver and kidney, but CrCl3 caused significantly higher Cr accumulation than CrPic in the femur of rats given 100 microg Cr/g. Daily urinary Cr excretion elevated with the increase of the dietary Cr level. Rats given CrPic showed significantly higher daily urinary Cr excretion than those given CrCl3, particularly at a dietary Cr level of 100 microg/g. The rate of urinary Cr excretion in rats given CrPic was constant, irrespective of the dietary Cr level, but that of rats given CrCl3 fell with the increase of the dietary Cr level. These results indicate that the lowest adverse effect level of dietary Cr is less than 100 microg/g, irrespective of the chemical species of Cr.

In the present study, we isolated and identified an active component from the Driselase-treated fraction and investigated its effect by acute and chronic oral administration on hypertension, lipid, and glucose metabolism in stroke-prone spontaneously hypertensive rats. The active component was identified as adenosine and improves hypertension after single oral administration. Rats who were 10 weeks old were divided into control and adenosine groups and were administered water or water with adenosine (10 mg/L), respectively, for 3 weeks. Hypertension and plasma lipid, nitric oxide, insulin, leptin, adiponectin levels, and glucose metabolism were significantly improved in the adenosine group. The mRNA expression levels of genes involved in lipid and glucose metabolism were altered in the adenosine group. Single oral administration of adenosine (10 mg/kg body weight) improved hypertension and plasma triglyceride, glucose, and nitric oxide levels 2 h after administration. In conclusion, oral acute and chronic administration of adenosine are beneficial and improve the metabolic syndrome-related disease parameters.

A multiresidue method was developed for determination of nine aminoglycoside antibiotics (streptomycin, dihydrostreptomycin, spectinomycin, neomycin, kanamycin, gentamicin, destomycin A, apramycin, and tobramycin) in milk by LC/MS/MS. The drugs were extracted with 0.01 mol/L potassium dihydrogen phosphate containing 2% trichloroacetic acid, and the extracted solution was cleaned on cation-exchange cartridge columns (Oasis WCX and Oasis MCX). LC separation was performed on a TSK-gel VMpak25 column (50 mmx2.0 mm i.d.) using gradient elution with 0.1% formic acid and acetonitrile containing 0.1% formic acid as the mobile phase. Recoveries of the drugs spiked at 0.01 or 0.1 microg/g in milk ranged from 66.1 to 110.8%, with a coefficient of variation of less than 17.1%. Limits of quantification of the drugs in milk were 0.001 approximately 0.01 microg/g. This method was used for analysis of milk from a lactating cow treated for clinical mastitis with two intramammary infusions of kanamycin (KM). Milk samples were analyzed during the withdrawal times at 12-hour intervals. KM concentrations were lower than the Japanese provisional MRLs (0.4 microg/g) at 60 hours after infusion, and subsequently fell to 0.01 microg/g.

A rapid multiresidue method was developed for determination of 98 veterinary drugs in livestock products and fish by LC/MS/MS. The drugs were extracted with methanol, and the extracted solution was diluted with water. The methanol concentration was adjusted to 50%, and finally the diluted solution was filtered through a microfiltration membrane (0.02 microm diameter pore size) prior to LC/MS/MS. Recoveries of 87 drugs from 4 foods (milk, egg, rainbow trout and cattle muscle) fortified at 0.2 microg/g were in the range of 50-150% with a coefficient of variation (%) of less than 20%. The values obtained by this method from livestock products containing antibiotics were similar to those obtained by the official methods. This proposed method is expected to be useful as a multiresidue analysis method for screening of veterinary drugs in livestock products and fish.

The purpose of this study is to investigate the difference of cerebral activity in the prefrontal region between young adults and elderly subjects during driving. The procedure of the experiment was explained to the subjects and informed consent was obtained. Fourteen young male adults (21.6+/-0.76 yrs), seven elderly males (69.9+/-4.91 yrs), and seven elderly females (66.6+/-6.02 yrs) volunteered as subjects for the experiments. Non-invasive measurement of regional cerebral activity was estimated by measuring the deoxygenated hemoglobin, oxygenated hemoglobin, and total hemoglobin of both sides of the prefrontal region using near-infrared spectroscopy (NIRS) and time-resolved spectroscopy (TRS). The distance to the vehicle in front, speed, and braking were recorded and the behavior of the drivers was obtained using a CCD camera and video recorder. Temperature and relative humidity in the experimental car were 23-25 degrees Centigrade and 30-40%RH respectively. Background noise in the car was 50-65 dB (A). The less experienced young adults display a greater increase in prefrontal cerebral activity than do the experienced young adults during driving. Prefrontal cerebral activity in elderly subjects is lower than that in young adults at rest and shows little variation compared with young adults during driving. Less experienced young adults and elderly males display similar behavior patterns in driving, such as not observing the door mirror carefully when changing lane. The less experienced young adults are considered to be less adapted to driving. It is possible to evaluate adaptability for driving by means of measuring cerebral hemodynamic changes while driving.

A method for simultaneous determination of multiple pesticide residues in agricultural products was developed by using a pretreatment with ultrafiltration, followed by liquid chromatography-tandem mass spectrometry (LC/MS/MS). The pretreatment process (extraction of pesticides from agricultural products with methanol, dilution of the extract with water, and ultrafiltration) gave recoveries in the range of 50-150% for 63 of 83 pesticides spiked at 0.25 microg/ g into 6 agricultural products. The detection limits of pesticides by LC/MS/MS were below 0.0005-0.05 micro/g. This method is useful for screening purposes and for multiresidue analysis of pesticides in agricultural products. Pesticide residues in 50 domestic crops were investigated by this method, and residues of 14 pesticides were detected in 30 crops.

Our new environment can afford one PC (personal computer) for every two students in the course of PC classes as well as for research. It has been clarified that the students acquired greater information literacy through this new environment and PC classes.

This paper investigates the applicability of cerebral blood flow in evaluating the technological adaptability for operating industrial products. The procedure of the experiment was explained to the subjects and informed consent was obtained from them. Twenty male and twenty female subjects (19-22 yrs) operated the destination setting task of a car navigation system. Subjects were divided into two sub groups to operate tasks of model A and model B of a car navigation system. Operation time of tasks and cerebral blood flow of frontal region were measured during tasks. Non-invasive measuring of regional cerebral blood flow was estimated by measuring deoxygenated hemoglobin, oxygenated hemoglobin, and total haemoglobin using the time resolved spectroscopy (TRS). Females were faster than males in operating the task of setting the destination searched by street address. Total haemoglobin of male subjects was significantly higher than that of females during resting and tasks. Changes of cerebral blood flow were observed during operating a car navigation system. In this paper we discussed the possibility of physiological evaluation for technological adaptability by means of the performance and brain hemodynamics measurement.

This study aims to clarify the effects of sample hardness on chewing force of humans. Silicone rubber was chosen as a food model, and three silicone rubber samples of varying hardness were chewed between the upper and lower incisors or molars in a natural way. Chewing force, contact area between a specimen and teeth, and pressure applied into the specimen were dynamically measured with a multiple-point sheet sensor. The multiple-point measurement revealed that a harder sample evoked higher peak force, longer duration, higher impulse, and higher active pressure values, whereas the chewing cycle and time to peak were not affected by sample hardness. The contact area between the sample and teeth decreased for the harder sample during incisor chewing. The peak force and force-related parameters, contact area, time to peak and bite force duration showed higher values for molar bites than for incisor bites, while the chewing cycle was similar in both cases. The measurement with the sheet sensor system clearly indicated that sample hardness modified chewing force of humans. We hypothesise that the effect of sample hardness is mainly mediated by mechanoreceptors in the periodontal ligament.

The effects of the cross-sectional area of food samples on bite force with molar teeth were investigated using raw carrots and surimi gels. We evaluated human bite force for food samples with different sizes between the upper and lower molars using a multiple-point sheet sensor and electromyography (EMG). The bite force curve and EMG clearly showed textural characteristics of the carrot and gel. In particular, the first peak in the bite curves corresponded to breaking point in the compression test. With increasing cross-sectional area of both foodstuffs, the bite force and contact area increased and the average stress to which the specimen was subjected (mean stress) tended to decrease, while the stress produced between the teeth and the specimen (active stress) did not change. Chewing rhythm and EMG activities were not greatly influenced by sample size. These findings suggest that higher bite force might cause difficulty in biting food with a larger cross-sectional area.

Background: Growth factors, cytokines, and the renin-angiotensin system (RAS) are involved in chronic allograft dysfunction. However, it is unclear whether clinical evaluations of TGFBeta1 and the RAS in longterm stable transplant patients can predict the development of chronic allograft dysfunction., Methods: Urinary TGFBeta1 excretion and the response of plasma renin activity (PRA) to angiotensin I converting-enzyme inhibition (ACE-I) were prospectively examined in transplant patients who had had stable graft function (n = 16) for at least 1 year after renal transplantation. Four-year follow-up studies were undertaken to evaluate the impact of these parameters on the development of chronic allograft dysfunction., Results: Urinary TGFBeta1 excretion and PRA response to ACE-I in renal transplant patients who developed chronic allograft nephropathy 4 years after the evaluations (n = 7) were significantly higher and greater, respectively, than these values in those who did not ( n = 9; P < 0.01). If the cutoff level for urinary TGFBeta1 excretion was 250 pg/min, the 4-year positive predictive value (PPV) with respect to the development of chronic allograft nephropathy was 83% and the negative predictive value (NPV) was 78% (sensitivity [sen.], 71%; specificity [sp.], 88%). If the cutoff level for PRA at 60 min after ACE-I was 4.0 ng/ml per h, the 4-year PPV was 71% and NPV was 75% (sen., 70%; sp., 75%). The stable transplant patients with high TGFBeta1 excretion and exaggerated PRA response showed significantly higher rates of chronic allograft dysfunction than those with low TGFBeta1 excretion and weak PRA response., Conclusions: This study demonstrates that some transplant patients with longterm stable graft function show increases in the activities of the TGFBeta system and the RAS. Evaluations of urinary TGFBeta1 excretion and PRA response to ACE-I present a possibility for predicting the development of chronic allograft dysfunction, with significant 4-year predictive values.

Two series of combinatorial tripeptide libraries were constructed, based on an antioxidative peptide isolated from a soybean protein hydrolysate. One was a library of 108 peptides containing either His or Tyr residues. Another was a library of 114 peptides related to Pro-His-His, which had been identified as an active core of the antioxidative peptide. The antioxidative properties of these libraries were examined by several methods, such as the antioxidative activity against the peroxidation of linoleic acid, the reducing activity, the radical scavenging activity, and the peroxynitrite scavenging activity. Two Tyr-containg tripeptides showed higher activities than those of two His-containing tripeptides in the peroxidation of linoleic acid. Tyr-His-Tyr showed a strong synergistic effects with phenolic antioxidants. However, the tripeptide had only marginal reducing activity and a moderate peroxynitrite scavenging activity. Cysteine-containing tripeptides showed the strong peroxynitrite scavenging activity. Change of either the N-terminus or C-terminus of Pro-His-His to other amino acid residues did not significantly alter their antioxidative activity. Tripeptides containing Trp or Tyr residues at the C-terminus had strong radical scavenging activities, but very weak peroxynitrite scavenging activity. The present results allow us to understand why protein digests have such a variety of antioxidative properties.

Background: The role of activated T cells in graft arteriosclerosis, which is observed in chronic renal allograft nephropathy, and the involvement of major histocompatibility complex (MHC) incompatibility remain to be determined. We examined the effect of T lymphocytes that were obtained from renal transplant patients undergoing chronic rejection treated with cyclosporine (CsA) on platelet-derived growth factor (PDGF)-induced proliferation of cultured human vascular smooth muscle cells (SMC) and compared the proliferation activity of T lymphocytes with MHC incompatibility, especially DRB 1 mismatch., Methods: Renal transplant patients with continued allograft function, who survived more than 1 year after transplantation, were recruited. Chronic rejection was documented by graft-biopsy findings together with increasing serum creatinine levels (10-20% per year). After the incubation of supernatant (conditioning medium) of cultured T cells from CsA-treated renal transplant patients with chronic rejection (n=18) and with normal renal function (n=14) as control, normal subjects (n=11) and chronic hemodialysis (HD) patients (n=5) with cultured SMC in the presence or absence of PDGF, DNA synthesis (3H-thymidine uptake) of SMC was examined. The in vitro effects of CsA on DNA synthesis of cultured SMC were also evaluated., Results: The supernatant of cultured T cells from renal transplant patients with chronic rejection stimulated PDGF-induced DNA synthesis of SMC in a dose-dependent manner, showing significant enhancement as compared with control transplant patients, normal subjects, and chronic HD patients. The supernatant itself did not significantly stimulate DNA synthesis of SMC. No significant in vitro stimulation of CsA on DNA synthesis was observed. The supernatant of T cells obtained from recipients undergoing chronic rejection with two DRB 1 mismatches showed significantly higher enhanced activity of PDGF-induced DNA synthesis than the supernatant from those recipients without mismatch of DRB 1. On the other hand, no significant correlation of the enhanced activity by T cell supernatant to HLA A and B mismatch numbers was observed., Conclusions: Growth factor-promoting factors(s) derived from activated T cells associated with MHC class II DR expression, which promotes PDGF-induced proliferation of SMC, exists in renal transplant patients with chronic renal allograft nephropathy, and is probably involved in arteriosclerosis of the graft kidney.

Protein hydrolysates, prepared by enzymatic digestion of soybean protein and egg white albumin using several proteases, inhibited the crystal growth of calcium carbonate. Each hydrolysate showed different inhibitory activities, suggesting the key role of peptide structures in the inhibition. The deamidation of protein hydrolysates by glutaminase increased not only the inhibitory activity toward the crystal growth of calcium carbonate but also the resistance of the hydrolysates against peptic digestion. Furthermore, the addition of sodium chloride, citric acid, or lactose into the reaction mixture enhanced the inhibitory activity. The protein hydrolysates inhibited both nucleation and crystal growth of calcium carbonate and also affected the crystal morphology.

Cardiovascular disease is one of the most common complications of dialysis and renal transplant patients, and high levels of AGE are present in end-stage renal failure. To address the potential involvement of AGE and growth factors in the pathophysiology of cardiovascular complications, we performed immunostaining using cardiac tissues from autopsy cases of patients on maintenance dialysis (10 cases), long-term surviving renal transplant patients with functioning grafts (8 cases), control subjects with normal renal function (7 cases) and non diabetic subjects with mild renal insufficiency (8 cases). We used two types of AGE-antibodies, 6D12 [monoclonal anti-AGE antibody, recognizing N epsilon-(carboxymethyl) lysine(CML)-modified AGE] (oxidative AGE) and non-CML-PA [polyclonal, not recognizing CML], and antibodies against PDGFs, PDGF receptors and TGF beta. Positive 6D12 staining was observed in the coronary arterial walls and in macrophages. The accumulation of 6D12-reactive AGE in the coronary arterial walls of maintenance dialysis patients was significantly greater than that of control subjects (p < 0.05). Renal transplantation significantly reduced this accumulation (p < 0.05). On the other hand non-CML-PA mainly detected AGE in intracardiac arterioles and neural tissues. There was little difference in the accumulation of non-CML-AGE among the four groups. PDGFs and PDGF receptors were mainly detected in vascular endothelial cells and infiltrating cells of cardiac tissues of renal transplant patients, but not of maintenance dialysis patients. TGF beta was not detected in cardiovascular tissue of transplant patients. Our results indicated that the accumulation of oxidative AGE (CML-AGE) in the cardiac vascular tissue is one of the factors for cardiovascular complications of maintenance dialysis patients, and also that renal transplantation has a reducing effect on CML-AGE accumulation. PDGFs may be involved in the cardiovascular complications after renal transplantation.

In patients with diabetic renal failure, attention must be paid to the prevention of atherosclerosis as well as the preservation of renal function. Insulin resistance is one of the important risk-factors of atherosclerosis and the involvement of tumor necrosis factor-alpha (TNF-alpha) has been shown in the pathogenesis of insulin resistance in some diseases. A low-protein diet (LPD) is recommended for patients with advanced renal disease, but a large proportion of the total caloric intake is supplied from carbohydrates and fat in LPD. Therefore, we designed a study to determine: (1) the effect of TNF-alpha on insulin sensitivity, and (2) the effect of LPD on the TNF-alpha response and the risk factors of atherosclerosis, such as insulin sensitivity and lipid metabolism, in patients with diabetic renal failure. Insulin sensitivity was measured by an euglycemic hyperinsulinemic clamp technique and serum TNF-alpha level and in vitro release of TNF-alpha from peripheral blood mononuclear cells (PBMCs) was measured in patients with diabetic renal failure. A significant negative correlation was observed between lipopolysaccharide-stimulated TNF-alpha release from PBMCs and insulin sensitivity (r = -0.58, p < 0.05). Secondly, risk factors of atherosclerosis were measured before and two weeks after the introduction of LPD in patients with diabetic renal failure. LPD did not have any significant effect on insulin sensitivity, the production of TNF-alpha by PBMCs, lipid metabolism and glucose metabolism. These results indicate that: (1) TNF-alpha derived from PBMCs might affect insulin sensitivity in patients with diabetic renal failure, and (2) LPD does not have any significant effect on the risk factors of atherosclerosis.

We examined the relation of diurnal alteration of platelet-free Ca2+ to blood pressure and plasma vasoactive substances in 6 subjects with normal renal function (N group) (with both normal GFR and normal urine concentration, and with the renal biopsy finding of minor glomerular change) and 7 patients undergoing CAPD (CAPD group), then evaluated the pathophysiological difference in diurnal variations between both groups and the effect of native kidney function. Diurnal values of platelet basal-free Ca2+ concentration in N showed a positive correlation with the corresponding PRA levels. On the other hand, in CAPD they showed a positive correlation with the corresponding levels of plasma AVP. The larger increases in platelet-free Ca2+ concentration due to thrombin stimulation were observed in the daytime and the smaller increases in the nighttime in the N groups. The thrombin-induced changes showed a positive correlation with the diurnal levels of mean blood pressure in N (p < 0.004). However, in contrast, in CAPD the increase in platelet-free Ca2+ due to thrombin tended to be larger in the nighttime and showed a negative correlation with the daily mean blood pressure levels (p < 0.004). These findings suggest that there might be differences in the regulation of intracellular-free Ca2+ dynamics and the diurnal variation between subjects with normal renal function and patients undergoing CAPD. Those differences could affect the progression of vascular disturbance in CAPD patients.