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Background: Anterior cruciate ligament deficiency (ACL-D) causes dysfunction in the quadriceps femoris muscle, and this dysfunction hampers a safe return to sports. However, how the dysfunctional quadriceps femoris muscle affects instantaneous re-programming of motor command in response to unpredictable events remains unknown. This study aimed to examine the effects of ACL-D on re-programming of preparatory muscle activity during an unpredictable landing task., Methods: Eighteen patients with ACL-D and 20 healthy participants (controls) performed normal landing and surprise landing tasks. In the surprise landing task, a false floor, designed to dislodge easily under load, was positioned in the middle of the descent path. This setup causes participants to unpredictably fall through the false floor onto the actual landing surface. Electromyography data collected during the period after passing through the false floor until landing was segmented into two equal halves. The average electromyography amplitude for each muscle in each period was compared between patients and controls., Results: In the vastus medialis and rectus femoris during the surprise landing task, the average electromyography amplitude during only the second half period in patients with ACL-D was significantly smaller than that in controls (p = 0.011 and 0.004, respectively)., Conclusions: Abnormalities were detected in the re-programming of preparatory muscle activation during an unpredictable landing task in the vastus medialis and rectus femoris of patients with ACL-D. The surprise landing task used in the present study has the potential to become a diagnostic tool to evaluate readiness for safely returning to sports., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.)

Anthropogenic activities' impacts over 140 years were studied at West Nanao Bay using a variety of geochemical techniques on sedimentary records. The bay is influenced by the Ninomiya River which is fed by a small watershed at which Tatzuruhama Town is located. Sedimentation rate was calculated using 210 Pb-excess and 137 Cs activities. C/N decreased after 1975, indicating a decrease in lignin-rich organic matter. From δ 13 C, δ 15 N and biogenic silica it was indicated that the population increased sewage-discharges until the construction of waste-water treatment plant in 1986. Several recorded changes in the landuse matched with the variation of the particle size. Total PAHs concentration was 1.17-62.78 μg g -1 , being highest during Japan's fastest economic growth period (1946-1975). Using diagnostic ratios and PCA analysis, PAHs' sources were identified as pyrogenic for all depths, varying from coal combustion (90.7 %) before 1946 to a mixture of biomass and vehicle combustion after 1961., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2023. Published by Elsevier Ltd.)

From August 2019 to August 2020, particulate and dissolved polycyclic aromatic hydrocarbons concentrations (PAHs) were analyzed in the water body of West Nanao Bay, Japan, to determinate their levels, environmental pathways, and ecological risks at this remote but shallow and semi-enclosed bay. The 14 targeted PAHs were analyzed by high performance liquid chromatography-fluorescence detector. Even when water column stratifies, the summatory of 14 targeted dissolved PAHs did not follow significantly change with depth. Results agreed with our previous findings in the surface distribution at the bay and can be attributed to long retention time of the water mass of the semi-enclosed bay. Suspended solids start precipitating according to their size; with biggest particles rapidly settling in the proximities of river mouths. Partition coefficients (Kp) varied from 10 3 to 10 7 , according to molecular weights. In general, highest Kp were found in the nepheloid layer. The risk quotients, RQ ∑14 PAHs (NCs) (1.04-174.08), indicated that PAHs represented a very low to low environmental risks., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2022. Published by Elsevier Ltd.)

Background: The risk of locomotive syndrome (LS) has been proposed as a criterion for evaluating physical ability. The expression levels of circulating miRNAs (c-miRNAs) are predictors of various diseases. This preliminary study aimed to evaluate the relationship between serum levels of several miRNAs and LS., Methods: We enrolled 423 participants in whom we conducted a survey with the 25-question Geriatric Locomotive Function Scale (GLFS-25) and measured the serum levels of 21 c-miRNAs. The relationship between the GLFS-25 and each c-miRNA was evaluated with a linear regression analysis, and independent associations between the GLFS-25 and each c-miRNA were assessed with a multiple regression analysis using various independent variables., Results: Only the serum level of miR-199 was significantly associated with LS after adjustment for age, BMI, sex, and all comorbidities. The receiver operating characteristics curve for the predictive value of the miR-199 level to indicate the presence or absence of LS risk had an area under the curve (AUC) of 0.576 (95% confidence interval: 0.501-0.651)., Conclusion: The expression level of miRNA-199 was associated with the risk of LS in community-dwelling Japanese people., Competing Interests: Declaration of competing interest None., (Copyright © 2021 The Japanese Orthopaedic Association. Published by Elsevier B.V. All rights reserved.)

The mechanistic/mammalian target of rapamycin (mTOR) regulates various cellular processes, in part through incorporation into distinct protein complexes. The mTOR complex 1 (mTORC1) contains the Raptor subunit, while mTORC2 specifically contains the Rictor subunit. Mouse genetic studies, including ours, have revealed a critical role for mTOR in skeletogenesis through its expression in undifferentiated mesenchymal cells. In addition, we have recently revealed that mTORC1 expression in chondrocytes is crucial for skeletogenesis. Recent work indicates that mTOR regulates cellular functions, depending on the context, through both complex-dependent (canonical pathway) and complex-independent roles (noncanonical pathway). Here, we determined that mTOR regulates skeletal development through the noncanonical pathway, as well as the canonical pathway, in a cell-type and context-specific manner. Inactivation of Mtor in undifferentiated mesenchymal cells or chondrocytes led to either severe hypoplasia in appendicular skeletons or a severe and generalized chondrodysplasia, respectively. Moreover, Rictor deletion in undifferentiated mesenchymal cells or chondrocytes led to mineralization defects in some skeletal components. Finally, we revealed that simultaneous deletion of Raptor and Rictor in undifferentiated mesenchymal cells recapitulated the appendicular skeletal phenotypes of Mtor deficiency, whereas chondrocyte-specific Raptor and Rictor double-mutants exhibited milder hypoplasia of appendicular and axial skeletons than those seen upon Mtor deletion. These findings indicate that mTOR regulates skeletal development mainly through the canonical pathway in undifferentiated mesenchymal cells, but at least in part through the noncanonical pathway in chondrocytes., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Please note that all Biochemical and Biophysical Research Communications authors are required to report the following potential conflicts of interest with each submission. If applicable to your manuscript, please provide the necessary declaration in the box above., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Background: Pulmonary thromboembolism (PTE) and deep vein thrombosis (DVT) are serious complications after total hip arthroplasty (THA). Aspirin has been considered a safe and cost-effective prophylaxis for venous thromboembolism (VTE), and there have been some reports about the incidence of PTE (0%-0.57%) and DVT (0.1%-0.35%) with low-dose aspirin for prophylaxis after THA. The aim of this study was to investigate the incidence of postoperative symptomatic VTE in our hospital and to evaluate the clinical efficacy of our prophylactic regimen., Patient and Methods: We retrospectively reviewed the medical records of consecutive patients who underwent THA in our hospital between 2011 and 2016. A total of 3295 hips (male: 337 patients, 365 hips; female: 2527 patients, 2930 hips) were enrolled in this study. Patients were divided into low-risk and high-risk groups. Low-risk patients were administered aspirin (100 mg/day) for 28 days postoperatively. High-risk patients, such as those diagnosed with obesity and/or with a history of VTE, received anticoagulants (enoxaparin or edoxaban) for 5 days postoperatively, followed by a dose of aspirin for 28 days. Based on our criteria, 218 of 3295 hips were considered high risk., Results: No VTE-related mortality was observed. One patient developed symptomatic PTE, and one patient developed symptomatic DVT. Both were successfully treated. Postoperative fatal bleeding or bleeding from any organ such as gastrointestinal and cerebral hemorrhage were not observed. A low incidence (0.03%) was observed for symptomatic DVT and PTE., Conclusions: This study demonstrated that the hospital's risk-stratified protocol using low-dose aspirin or anticoagulants was clinically effective in preventing symptomatic VTE. These results were considerably better than those reported from Western countries. However, all patients in this study were the Japanese. It was unclear whether similar results were given to non-Japanese patients. Therefore, this protocol needs severe carefulness to be applied to non-Japanese populations., (Copyright © 2019 The Japanese Orthopaedic Association. Published by Elsevier B.V. All rights reserved.)

Background: The circadian clock is modulated by the timing of ingestion or food composition, but the effects of specific nutrients are poorly understood. Objective: We aimed to identify the amino acids that modulate the circadian clock and reset the light-induced circadian phase in mice and humans. Methods: Male CBA/N mice were orally administered 1 of 20 l-amino acids, and the circadian and light-induced phase shifts of wheel-running activity were analyzed. Antagonists of several neurotransmitter pathways were injected before l-serine administration, and light-induced phase shifts were analyzed. In addition, the effect of l-serine on the light-induced phase advance was investigated in healthy male students (mean ± SD age 22.2 ± 1.8 y) by using dim-light melatonin onset (DLMO) determined by saliva samples as an index of the circadian phase. Results: l-Serine administration enhanced light-induced phase shifts in mice (1.86-fold; P < 0.05). Both l-serine and its metabolite d-serine, a coagonist of N -methyl-d-aspartic acid (NMDA) receptors, exerted this effect, but d-serine concentrations in the hypothalamus did not increase after l-serine administration. The effect of l-serine was blocked by picrotoxin, an antagonist of γ-aminobutyric acid A receptors, but not by MK801, an antagonist of NMDA receptors. l-Serine administration altered the long-term expression patterns of clock genes in the suprachiasmatic nuclei. After advancing the light-dark cycle by 6 h, l-serine administration slightly accelerated re-entrainment to the shifted cycle. In humans, l-serine ingestion before bedtime induced significantly larger phase advances of DLMO after bright-light exposure during the morning (means ± SEMs-l-serine: 25.9 ± 6.6 min; placebo: 12.1 ± 7.0 min; P < 0.05). Conclusion: These results suggest that l-serine enhances light-induced phase resetting in mice and humans, and it may be useful for treating circadian disturbances., Competing Interests: Author disclosures: SY, AI, S-iL, SO, RH, SS, MF, and SH, no conflicts of interest. NU, CT, and SM are employees of FANCL Co., (© 2017 American Society for Nutrition.)

Background: There are reports that the maternal diet during pregnancy may affect development of babies' eczema. We sought to investigate the association between the maternal diet during pregnancy and the risk of eczema in infancy in Japan., Methods: A birth cohort was set up at 2 hospitals in Chiba city. Dietary habits concerning fish, butter, margarine, yogurt and natto during pregnancy was obtained from mothers just after delivery. The intake frequencies of these foods were classified into four groups: 1) daily, 2) 2-3 times a week, 3) once a week and 4) once a month or less. Diagnosis of eczema at 6 months of age was made by the presence of an itchy rash that persisted more than two months., Results: Valid data on 650 mother-baby pairs were obtained. No relationship between frequencies of the maternal intake of fish, margarine and yogurt during pregnancy and the onset rate of the babies' eczema were observed. For butter consumption, the incidence of babies' eczema was significantly higher in the group with daily intake than in those with an intake 2-3 times a week or less (p = 0.044). For natto, incidence of babies' eczema was significantly lower in the group with everyday intake than those eating it 2-3 times a week or less (p = 0.020)., Conclusions: High frequency intake of natto during pregnancy possibly reduces the incidence of eczema in children at 6 months of age.

Splenic langerin(+) CD8α(+) dendritic cells (DCs) have exhibited a critical role in cross-priming CD8(+) T cell responses. To further study the roles of this DC subset, a protocol for the continuous depletion of langerin(+) CD8α(+) DCs was established using the pre-existing lang-DTREGFP mouse model. Due to the fast turnover rate of splenic CD8α(+) DCs, maintaining the depletion of langerin(+) CD8α(+) DCs required multiple diphtheria toxin (DT) treatments. We found that prolonged treatment with DT did not cause weight loss, or neutrophilia, as reported in some DT-based depletion models. Therefore, the in vivo depletion of murine langerin(+) CD8α(+) DCs can be maintained over time to analyse their function during the full course of an immune response., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Novel 316 L stainless steel (SS) foam with 85% porosity and an open pore diameter of 70-440 μm was developed for hard tissue application. The foam sheet with a 200-μm diameter had superior cell proliferation and penetration as identified through in vitro experiments. Calcification of human osteosarcoma cells in the SS foam was observed. Multi-layered foam preparation is a potential alternative technique that satisfies multi-functional requirements such as cell penetration and binding strength to the solid metal. In tensile tests, Young's modulus and the strength of the SS foam were 4.0 GPa and 11.2 MPa respectively, which is comparable with human cancellous bone., (Copyright © 2013 Elsevier B.V. All rights reserved.)

The major functions required for load-bearing orthopaedic implants are load-bearing and mechanical or biological fixation with the surrounding bone. Porous materials with appropriate mechanical properties and adequate pore structure for fixation are promising candidates for load-bearing implant material. In previous work, the authors developed a novel titanium (Ti) foam sheet 1-2mm thick by an original slurry foaming method. In the present work, novel Ti foam is developed with mechanical properties compatible with cortical bone and biological fixation capabilities by layer-by-layer stacking of different foam sheets with volumetric porosities of 80% and 17%. The resulting multilayer Ti foam exhibited a Young's modulus of 11-12GPa and yield strength of 150-240MPa in compression tests. In vitro cell culture on the sample revealed good cell penetration in the higher-porosity foam (80% volumetric porosity), which reached 1.2mm for 21 days of incubation. Cell penetration into the high-porosity layers of a multilayer sample was good and not influenced by the lower-porosity layers. Calcification was also observed in the high-porosity foam, suggesting that this Ti foam does not inhibit bone formation. Contradictory requirements for high volumetric porosity and high strength were attained by role-sharing between the foam sheets of different porosities. The unique characteristics of the present multilayer Ti foam make them attractive for application in the field of orthopaedics., (Copyright © 2012 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

Objective: This study demonstrated pterygomaxillary disjunction patterns and elucidated factors related to unfavorable pterygomaxillary junction fractures in Le Fort I osteotomy without using an osteotome., Study Design: Clinical and anatomical data obtained from computed tomography images (100 sides) were analyzed for their ability to predict patterns of pterygomaxillary disjunction., Results: Separation of the pterygomaxillary junction was most frequently performed at the maxillary tuberosity (48.0%). Twenty-eight pterygoid plates were fractured. Male gender, increased age, thickness of the pterygomaxillary junction, and length of the maxillary tuberosity were significant risk factors for pterygoid process fractures. We also identified that a pterygomaxillary junction thickness less than 2.6 mm and a maxillary tuberosity length of more than 11.5 mm indicated a statistically significant risk of pterygoid process fractures., Conclusions: Prediction of frangible pterygoid plates by preoperative quantitative evaluation of morphometric values provides useful information for selecting safe procedures., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Endoscopic-assisted surgery has gained widespread popularity as a minimally invasive procedure, particularly in the field of maxillofacial surgery. Because the surgical field around the mandibular angle is extremely narrow, the surrounding tissues may get caught in sharp rotary cutting instruments. In piezosurgery, bone tissues are selectively cut. This technique has various applications because minimal damage is caused by the rotary cutting instruments when they briefly come in contact with soft tissues. We report the case of a 33-year-old man who underwent resection of an osteoma in the region of the mandibular angle region via an intraoral approach. During surgery, the complete surgical field was within the view of the endoscope, thereby enabling the surgeon to easily resection the osteoma with the piezosurgery device. Considering that piezosurgery limits the extent of surgical invasion, this is an excellent low-risk technique that can be used in the field of maxillofacial surgery., (Copyright © 2013 Elsevier Inc. All rights reserved.)

We report a patient with intraarticular nodular fasciitis who developed mechanical symptoms in the knee, limiting knee extension. A 24 year-old man presented with mechanical restriction of extension and pain at maximum extension. MRI revealed an intraarticular mass within the knee joint. The mass was excised arthroscopically. Histological examination of the excised tissue led to a diagnosis of nodular fasciitis. Pain and limitation of motion of the knee disappeared from one day after surgery. No recurrence was found after 1 year. Intraarticular nodular fasciitis is rare, and although this lesion within the knee has been reported previously, it has not been associated with mechanical symptoms. This case indicates that intra-articular fasciitis should be considered in the differential diagnosis of a patient presenting with mechanical limitation of knee extension and a mass lesion inside the knee joint., (Copyright © 2009 Elsevier B.V. All rights reserved.)

Recent studies clearly demonstrated that several types of pathogenic amyloid proteins acted as agents that could transmit amyloidosis by means of a prion-like mechanism. Systemic AA amyloidosis is one of the most severe complications of chronic inflammatory disorders, particularly rheumatoid arthritis. It is well known that, similar to an infectious prion protein, amyloid-enhancing factor (AEF) acts as a transmissible agent in AA amyloidosis. However, how AEF transmits AA amyloidosis in vivo remained to be fully elucidated. In the present study, we focused on finding cell-free forms of AEF and its carriers in circulation by using the murine transfer model of AA amyloidosis. We first determined that circulating cell-free AEF existed in blood and plasma in mice with systemic AA amyloidosis. Second, we established that plasma exosomes containing AA amyloid oligomers derived from serum amyloid A had AEF activity and could transmit systemic AA amyloidosis via a prion-like mechanism. These novel findings should provide insights into the transmission mechanism of systemic amyloidoses., (Copyright © 2010 Elsevier Inc. All rights reserved.)

Lactic acidosis has been considered to be a side effect of some biguanides, after phenformin was withdrawn from the market because of its association with lactic acidosis. The potential of lactic acidosis induced by biguanides at human therapeutic exposure levels, however, has not been examined. Then, we compared the risk of lactic acid at doses providing exposure levels comparable to human therapeutic doses. Metformin and phenformin were orally administered to rats for up to 28 days, and plasma drug concentrations and blood lactic acid levels were examined. Metformin did not elevate lactic acid levels at the dose corresponding to higher systemic drug exposure than human therapeutic level, even for repeated doses. In contrast, phenformin elevated lactic acid levels at the dose corresponding to lower exposure than human therapeutic level, and sustained high levels were observed up to 24h post-dose; furthermore, these changes were enhanced by repeated doses. Direct comparison at each rat equivalent dose clearly indicated that lactic acid levels of phenformin were higher than those of metformin. These non-clinical findings suggest that metformin dose not increase lactic acid levels like phenformin does, and therefore may not increase the risk for lactic acidosis at human therapeutic exposure level., (Copyright (c) 2010 Elsevier Inc. All rights reserved.)

Neu2 mRNA from the mouse thymus, as we have reported [K. Kotani, A. Kuroiwa, T. Saito, Y. Matsuda, T. Koda, S. Kijimoto-Ochiai, Cloning, chromosomal mapping, and characteristic 5'-UTR sequence of murine cytosolic sialidase, Biochem. Biophys. Res. Commun. 286 (2001) 250-258], has a novel sequence at the 5' terminus that shows the ability to encode 6 extra amino acids in the N-terminus than that of the muscle. In this paper, we analyzed the cDNA and EST database and found the five types of alternative splicing of Neu2 mRNA: A, B, C, D and N. We studied the expression of these types in the immune tissues and found that the thymus expressed only type B. We constructed 2 types of plasmid that encode long (B) or short (C) form of Neu2 protein, and transfected them into COS7 cells to study them under the same conditions. We found that 30-40% of the both forms of Neu2 activity was located in the crude membrane-fraction, and hydrolyzed ganglioside effectively, while both soluble fraction showed particular behavior with substrate specificity. Microscopic study by active staining with X-NANA showed that they located not only in the cytoplasm but also in areas surrounding the nucleus and in the peripheral ruffled spot.

Alpha-synuclein is a major component of several pathological lesions diagnostic of specific neurodegenerative disease such as Parkinson's disease. This study focuses on the non-amyloid beta component of Alzheimer's disease amyloid, a key region for the aggregation and fibril formation of alpha-synuclein. Several mutations were introduced in an attempt to repress beta-strand formation and hydrophobic interaction-based aggregation. Although reducing the hydrophobicity drastically decreased fibril formation, the Val70Thr and Val70Pro mutations resulted in an unstable secondary structure thereby increasing non-structural aggregation, instead of fibril formation. Therefore, the stabilization of non-structural natively unfolded status is important to prevent alpha-synuclein fibril formation. Mixing the Val70Thr/Val71Thr double mutant, which has inherently low potential, with the fibril forming alpha-synucleins, WT and Ala53Thr, greatly reduced their fibril formation and aggregation. This double mutant has great potential for further therapeutic approaches.

We have totally sequenced a cytosolic sialidase [EC 3.2.1.18] by RT-PCR from the murine thymus (murine thymic sialidase, MTS) which has a 1844-base length (encoding 385 amino acids including two sialidase motifs) and is the longest cytosolic sialidase ever reported. MTS has high and relatively low homologies with those of mammalian cytosolic sialidases from the mouse brain (99%), rat (91%), and human skeletal muscle (75%), and those of the mouse lysosomal (47%) and membrane-bound (51%) sialidases, respectively. Chromosomal mapping, being the first report of mouse cytosolic sialidase gene, showed that the MTS gene is localized to the distal part of mouse chromosome 1D and to rat chromosome 9q36. RT-PCR with the site-specific primers revealed that the coding region was expressed in all organs tested, but expressions including the 5'-UTR were barely detectable except for in the upper-thymic fraction. Also, soluble sialidase activity in the thymus was the highest of these organs. There were mRNA instability signals and AT-rich regions in 143 bp of MTS 5'-end., (Copyright 2001 Academic Press.)

Objectives: Endothelial function in the brachial arteries is impaired in smokers. However, little is known about this condition in young adult men. The relationship between nitric oxide(NO) production and the endothelial function was investigated in young smokers and compared with non-smokers., Methods: Flow-mediated vasodilation of the brachial artery during reactive hyperemia was examined in 10 young smokers(mean age 31 years) and 12 control subjects(mean age 28 years). The vasodilator response in the brachial artery was measured by ultrasonography, and blood samples were obtained from the right cephalic vein. Blood samples were taken at baseline, 30 sec after cuff deflation, and before and 5 min after 0.3 mg of nitroglycerin administration. Blood flow was calculated by multiplying mean flow velocity and vessel cross-sectional area. Plasma NOx(nitrate + nitrite) levels were measured, and the percentage change of NOx production(delta NOx) was calculated as follows: delta NOx(%) = [(NOx concentration at peak flow-mediated vasodilation or after 0.3 mg nitroglycerin administration) - baseline NOx concentration)] x 100/baseline NOx concentration., Results: Percentage changes in diameter of the brachial artery, NOx production and delta NOx in response to nitroglycerin were not statistically different between the two groups(smokers: 27.6 +/- 8.0 mumol/l, control subjects: 34.0 +/- 8.7 mumol/l). However, percentage change of flow-mediated vasodilation during reactive hyperemia in the young smokers was significantly smaller than that in the control subjects(4.8 +/- 2.7%, 9.1 +/- 5.3%, respectively, p < 0.05). Moreover, delta NOx during reactive hyperemia in the smokers was significantly smaller than that in the control subjects(388.8 +/- 90.2%, 738.0 +/- 284.5%, respectively, p < 0.05)., Conclusions: The impaired response to reactive hyperemia in young smokers might be associated with decreases in flow-dependent NO production.

The human CD23 molecule (low affinity receptor for IgE) has a C-type lectin domain, a reversed Arg-Gly-Asp (RGD) sequence near the C-terminus, and an "RGD-binding inhibitory peptide" at the root of the N-sugar chain. Three peptides were synthesized to determine their functions, i.e., #1, including an inverse RGD sequence near the C-terminus; #2, RGD-binding inhibitory peptides in the gpIIIa chain of platelet integrin gpIIb/IIIa; and #3, the inverse sequence located at the root of the N-sugar chain of CD23 which has homology to peptide 2. Among the three peptide, only peptide 3 inhibited aggregation of L-KT9 cells. Isotope-labeled peptides 1 and 3 bound to MHC class II molecules but peptide 1 did not bind to CD23 molecules. Peptide 3 showed a higher affinity to MHC class II than did peptide 1. Both peptides in CD23, therefore, seem to have interesting and important functions in relation to MHC class II molecules and also to CD23 molecules when CD23 on EBV-transformed B cells acts as a lectin in homotypic cell aggregation. The physiological function of CD23 was discussed from an evolutional point of view., (Copyright 2000 Academic Press.)

The change of acid cholesteryl ester hydrolase activity in mononuclear leukocyte following treatment of diabetes mellitus was studied in 21 patients with non-insulin-dependent diabetes mellitus (NIDDM). Enzyme activity before treatment in the patients was significantly lower than that in 14 age-matched healthy subjects (1.20 +/- 0.15; mean +/- S.E. vs. 2.20 +/- 0.17 nmol/mg protein/h, P less than 0.01). Enzyme activity before treatment in the patients was significantly increased (P less than 0.05) after 4-8 weeks of treatment. However, enzyme activity of 1.43 +/- 0.14 nmol/mg protein/h observed after treatment in the patients was significantly lower (P less than 0.01) than that in the healthy subjects. There was a significant negative correlation between enzyme activity before treatment and the increase in enzyme activity following treatment (rs = -0.555, P less than 0.01, n = 21). These results indicate that low level of enzyme activity may be insufficiently improved by the treatment of diabetes, and the risk for the development of atherosclerosis as viewed from the enzyme activity may persist even after the treatment in NIDDM.

A rapid and convenient method was established for analysis of the N-linked carbohydrate chains of glycoproteins on nitrocellulose sheets. Proteins were separated by polyacrylamide gel electrophoresis, transferred to nitrocellulose sheets, reacted with peroxidase-coupled lectins, and detected by color development of the enzyme reaction. Four glycoproteins having N-linked oligosaccharide chains were used as test materials: Taka-amylase A (which has a high-mannose-type chain), ovalbumin (high-mannose-type chains and hybrid-type chains), transferrin (biantennary chains of complex type), and fetuin (triantennary chains of complex type and O-linked-type chains). Concanavalin A interacted with Taka-amylase A, transferrin, and ovalbumin but barely interacted with fetuin. After treatment of the glycoproteins on a nitrocellulose sheet with endo-beta-N-acetylglucosaminidase H, transferrin reacted with concanavalin A but Taka-amylase A and ovalbumin did not. Wheat germ agglutinin interacted with Taka-amylase A but not ovalbumin; therefore, they were distinguishable from each other. Fetuin and transferrin were detected by Ricinus communis agglutinin or peanut agglutinin after removal of sialic acid by treatment with neuraminidase or by weak-acid hydrolysis. Erythroagglutinating Phaseolus vulgaris agglutinin detected fetuin and transferrin. Thus, the combined use of these procedures distinguished the four different types of N-linked glycoproteins. This method was also applied to the analysis of membrane glycoproteins from sheep red blood cells. The terminally positioned sugars of sialic acid, alpha-fucose, alpha-galactose, and alpha-N-acetylgalactosamine were also detected with lectins from Limulus polyphemus, Lotus tetragonolobus, Maclura pomifera, and Dolichos biflorus, respectively.