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We investigated the influence of glycerol esters on oleic acid uptake into IEC-6 cells. Monoolein, especially 2-monoacylglycerol, significantly inhibited the cellular uptake. Although diolein slightly inhibited the oleic acid uptake, triolein, glycerol and monooctanoate had no effect. These results suggest that after lipid digestion in the intestine, long-chain fatty acid uptake may be influenced by another digestive product, 2-monoacylglycerol.

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We investigated the influence of various substrates on the uptake of long-chain fatty acid into IEC-6, rat intestinal epithelial cell line. The uptake of [3H]oleic acid into IEC-6 cells was a saturable function of the oleic acid concentration. Long-chain fatty acids significantly inhibited the oleic acid uptake into IEC-6 cells and shorter-chain fatty acids had little or no effect. Various fatty acid esters suppressed the oleic acid uptake into IEC-6. Fatty alcohols also inhibited oleic acid uptake into IEC-6 and the length of the carbon chain played an important role. These results suggest that long-chain fatty acid uptake was inhibited by the substrates which had a structure similar to long-chain fatty acids, especially those with a long carbon chain. At least two molecules, fatty acid translocase and fatty acid transport protein type 4, which are considered to be involved in the long-chain fatty acid transport into the cell, were expressed on IEC-6 cells, supporting the existence of the carrier-mediated system of long-chain fatty acid transport on IEC-6 cells.

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Long-chain fatty acids are important nutrients, but obesity is the most common nutritional disorder in humans. In this study we investigated the effect of oleyl alcohol on the intestinal long-chain fatty acid absorption in rats. We administered [14C]oleic acid and oleyl alcohol as lipid emulsion intraduodenally in unanesthetized lymph-cannulated rats and measured the lymphatic output of oleic acid. Second, we orally administered lipid emulsion with a stomach tube and measured the luminal and mucosal oleic acid residues. Furthermore, rats were fed oleyl alcohol as a dietary component for 20 days, and fecal lipid and the weight of adipose tissues were measured. In lymph-cannulated rats, triglyceride and [14C]oleic acid output in the lymph were significantly lower in the presence of oleyl alcohol when compared with the absence of oleyl alcohol in a dose-dependent manner. The radioactivity remaining in the intestinal lumen was more strongly detected in rats that had been orally administered oleyl alcohol than in the controls. The feces of rats fed an oleyl-alcohol-added diet contained much higher amounts of lipids, and the weights of their adipose tissues were significantly lower than in the control group. These results suggest that oleyl alcohol inhibits the rat gastrointestinal absorption of long-chain fatty acids in vivo.

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The efficiency of intestinal absorption and metabolic conversion of quercetin aglycone and its glucosides, quercetin-4'-O-beta-D-glucoside (Q4'G), quercetin-3-O-beta-D-glucoside (Q3G), and quercetin-3,4'-di-O-beta-D-glucoside (Q3,4'G), was estimated by using Caco-2 cell monolayers as an intestinal epithelial cell model. Aglycone was significantly lost from the apical side, resulting in the appearance of free and conjugated forms of quercetin and those of isorhamnetin in the cellular extracts. In the basolateral solution, the conjugated form of quercetin was predominant and increased with the elapse of incubation. As compared with quercetin aglycone, none of the quercetin glucosides were absorbed efficiently from apical side. However, Q4'G yielded conjugated quercetin and isorhamnetin in basolateral solution at higher amounts than Q3G or Q3,4'G. Lipophilicity of Q4'G was found to be higher than that of Q3G or Q3,4'G. This suggests that lipophilicity contributes to the relatively efficient absorption of Q4'G. It is likely that the occurrence of hydrolysis enhances the efficiency of intestinal absorption and metabolic conversion of dietary quercetin glucosides.

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The effect of recombinant human granulocyte colony-stimulating factor (rh G-CSF) on bone was evaluated by histomorphometry using Sprague-Dawley rats. rh G-CSF was injected at doses of 0, 50, 150, and 450 microg/kg for 6 weeks. In vivo double fluorochrome labeling was performed before sacrifice. No significant change in body weight was observed. Bone mineral density (BMD) of lumbar vertebrae and femora was significantly decreased in G-CSF-treated groups. In the lumbar vertebra, osteoid surface, osteoid thickness, trabecular thickness, and labeled surface in G-CSF-treated groups were also significantly lower. In addition, osteoclast number and osteoclast surface were significantly higher in the G-CSF-treated groups. The endocortical surface at the mid-tibia showed lower labeled surface and mineral apposition rate in G-CSF-treated groups, without significant changes at the periosteal surface. Furthermore, numerous granulocytes fully occupied the bone marrow area. We conclude that proliferating granulocytes in the bone marrow may inhibit bone-forming cells from contacting the bone surface, resulting in reduction of bone formation; and increased osteoclastic bone resorption induced by G-CSF treatment contributed to the reduction of BMD.

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In order to evaluate the ability of a guanidine extract of demineralized bone to repair osteochondral defects in articular cartilage, plugs made of this extract were implanted into defects in rabbit knees. The repair tissue was examined macroscopically, histologically, and immunohistochemically at 4, 8, 12, and 30 weeks. Controls (defects that were left empty) showed no cartilage formation. Four weeks after implantation of a guanidine extract plug, histological examination showed a nonhomogeneous metachromatically stained region extending from the surface of the repair tissue down to cancellous bone. This region also was labeled by an anti-type-II collagen antibody, indicating that cartilage-like tissue had been induced. At 8 weeks, the newly formed cartilage in the subchondral and cancellous bone had been partially replaced by bone. At 12 weeks, the thickness of the newly formed cartilage layer had decreased, and most of the newly formed cartilage in the subchondral and cancellous bone had been replaced by bone. In addition, a tidemark was observed. At 30 weeks, the repair tissue was a mixture of cartilage and fibrocartilage, and there was severe degeneration of the cartilage surrounding the repaired defects. These findings indicate that osteochondral defects of articular cartilage can be partially repaired by the implantation of a guanidine extract and that the newly formed cartilage-like tissue is not permanent.

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In open femoral bone fractures osteomyelitis may develop as a complication. Many difficulties are experienced in the treatment of those fractures because an extended bone defect may be formed after repeated operations, and then amputation of the femoral bone becomes necessary. Since 1981 the present authors have performed the vascularized double fibula grafts in the treatment of 18 patients with successful results. With this grafting method, both vascularized double fibula grafts are collected at the same time, one as an intramedullary graft and the other as an onlay graft. The most important point in carrying out grafting by this method is to prepare the recipient bone bed adequately. In many cases, it is necessary to carry out the primary operation to curette the focus, and resect necrotic tissues and sequestrated bone before the vascularized double fibula grafts, and then grafting is performed as a second operation after infection has been controlled following the initial operation. Although differences in time required for recovery cannot be eliminated completely, it becomes possible for 16 out of 18 patients to walk without the use of a brace and crutches within 13 months on average.

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Congenital kyphosis and atlantoaxial dislocation in a 13-year-old boy was treated by a C1 laminectomy and C2-C5 laminoplasty with fusion from the occiput to C2. This resulted in postoperative neurologic deterioration, but a secondary anterior C3 vertebrectomy followed by a C2-C5 fusion helped restore neural function. In the presence of congenital cervical kyphosis, anterior rather than posterior decompression and fusion is recommended, particularly in the presence of a stenotic spinal canal.

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A solution of 0.5 g of meropenem (MEPM) in 100 ml of saline was administered to adult patients before the orthopaedic surgery via intravenous drip infusion for 30 minutes. Eleven samples of bone marrow blood, 13 bone, 8 joint fluid and 8 joint tissues obtained from 15 clinical cases were analyzed for MEPM levels. At the same time, blood samples were taken from peripheral veins and serum served as control was analyzed. The concentration of MEPM in the marrow blood at 30 minutes after administration of MEPM reached 15.4 micrograms/ml, which was above 50% of the maximum concentration in serum. Ratios of concentration in bone marrow to that in serum were between 93% and 105% at that time. MEPM levels were 5.74-0.40 micrograms/g in bones, 20.3-4.55 micrograms/ml in joint fluid and 18.2-4.05 micrograms/g in joint tissues at 30 to 75 minutes after administration. In joint fluid and joint tissues, the concentration above 50% of maximum level in serum were maintained for more than an hour. MEPM is thought to be useful for the treatment of bone and joint infections and also prophylaxis of infections in cases of major surgery of skeletal tissue in the field of orthopaedic surgery.

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We present our technique for reconstruction of the posterior cruciate ligament (PCL) using the semitendinosus and gracilis tendons with the Kennedy ligament augmentation device (LAD). The safe and excellent exposure of the posterior aspect of the knee allowed us to identify the most isometric position in the intercondylar notch of the femur. In addition to this advantage, firm fixation of the LAD-augmented tendons with staples prevented the tibia from sagging posteriorly during early protected motion of the knee. Evaluation of 12 patients followed for more than 2 years showed 9 (75%) good results. In this small series no correlation was found between clinical results and the number of major structures injured, indicating that postoperative care is as important as isometric placement of the PCL in obtaining satisfactory results.

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An improved enzyme-linked immunosorbent assay (ELISA) for the detection of anticollagen antibodies in human serum has been developed. With the use of this method, antibodies against native human Type II collagen were detected in 22.7% of sera from 480 patients with rheumatoid arthritis (RA). The antibodies were found to be collagen type specific, showing no reaction with human Type I and Type III collagens. The antibodies appeared in high incidence during the early phase of the disease, and RA patients with involvement of a single joint, mono-articular arthritis, were often positive for anti-Type II collagen antibodies. In most of these patients, anti-Type II collagen antibodies preceded the appearance of rheumatoid factors. The antibodies were all negative in sera from patients with gout, osteoarthritis (OA) and non-arthritic diseases. Thus, anti-Type II collagen antibody assay may have diagnostic significance for RA patients, especially those in whom laboratory and clinical findings provide only minimal help in diagnosis.

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In order to elucidate the relationship between the severity of osteoporosis and the fixation strength of a pedicle screw, screw pull-out tests were performed using cadaveric lumbar vertebrae. The severity of osteoporosis was evaluated by the Jikei osteoporosis grading scale (Jikei method), bone mineral density, and microdensitometry. When a 7.0-mm screw was used, the pull-out force of the screw was 1,056.4 N in the normal group (as determined by the Jikei method), while it was 495.6 N in the Grade I osteoporosis and 269.5 N in the Grade II osteoporosis groups, respectively. There were also positive correlations between the pull-out force and bone mineral density and each parameter of the microdensitometry method. When bone cement was used in an osteoporotic vertebra, twofold stronger pull-out force was obtained in comparison to that obtained without bone cement.

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In two cases with a giant cell tumor of the axis, the mandible and tongue-splitting approach permitted excision of the tumor and anterior fusion of the spine from the atlas to the third cervical vertebra. With this approach, there is no need to extend or rotate the cervical spine during the operation. In addition, the operative field extends from the clivus to the fourth cervical vertebra, and safe and sufficient anterior decompression is possible. Although this approach has some disadvantages, contemporary techniques of intravenous hyperalimentation and postoperative respiratory management provide a solution to these problems. Thus, this approach should always be considered for patients requiring extensive anterior decompression of the craniovertebral junction or upper cervical spine.

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An improved enzyme-linked immunosorbent assay (ELISA) for the determination of anti-collagen antibodies in human serum has been developed. The method is based on the use of serum samples diluted to 1/50 with heat-inactivated normal rabbit serum adjusted to pH 8.0 with solid Tris (0.05 M), NaCl (0.15 M) and 2 M HCl. The use of normal rabbit serum minimizes non-specific adsorption of immunoglobulin G onto the plastic surface of microtiter plate. The applicability of the method for the quantitation of anti-collagen antibodies in human serum is demonstrated with 290 specimens of sera from normal controls (194) and patients with rheumatoid arthritis (96).

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Three major NSAID (tiaprofenic acid, indomethacin and acetylsalicylic acid) were studied with regard to their effects on the biosynthesis and gene expression of collagen and proteoglycan of chondrocytes. We found that the biosynthesis of type 11 collagen was suppressed in 60-70% of the controls by either indomethacin or acetylsalicylic acid; tiaprofenic acid did not suppress collagen synthesis. Although the biosynthesis of proteoglycan was suppressed to 80-90% of the control by indomethacin or acetylsalicylic acid, tiaprofenic acid did not suppress proteoglycan synthesis. In order to investigate the gene expression of type 11 collagen and proteoglycan, dot blot hybridization was performed using type 11 collagen and proteoglycan cDNA. The effects of NSAID on the amount of type 11 collagen and proteoglycan mRNA were consistent with those on collagen and proteoglycan biosynthesis. Thus, we concluded that tiaprofenic acid has less effect on cultured chondrocytes.

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In conclusion, the data obtained from the present study show that TA has less effect than indomethacin or ASA on cultured chondrocytes in gene expression and synthesis of collagen. These findings cannot be directly interpolated to clinical application, but attention to pharmacologic activities of NSAIDs on chondrocytes may have future applications. Two major NSAIDs, TA, indomethacin, and ASA, were examined for their effects on the biosynthesis and gene expression of articular cartilage collagen. The biosynthesis of type II collagen was suppressed to 70% to 80% of the control by indomethacin and ASA. TA did not suppress collagen synthesis. To know the gene expression of type II collagen, dot blot hybridization was performed using type II collagen cDNA. The effects of NSAIDs on the amount of type II collagen mRNA were consistent with those on collagen biosynthesis. Thus, TA had the least effect on cultured chondrocytes.

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The effect of temperature on the antibacterial activity of lidocaine to Staphylococcus aureus and Pseudomonas aeruginosa was investigated in vitro. At 10 C at which S. aureus organisms do not grow and might be metabolically inactive, the antibacterial activity of lidocaine to S. aureus was not observed in a concentration of 1%, which was quite antibacterial to S. aureus at 37 C. On the other hand, at 40 C a conspicuously increased antibacterial activity to S. aureus of lidocaine was observed in a concentration of 0.25% which was not antibacterial to S. aureus organisms at 37 C. Similar results were obtained when P. aeruginosa organisms were examined in place of S. aureus, although P. aeruginosa was found to be less susceptible to lidocaine than S. aureus. The clinical significance of the thermal effect on the antibacterial activity of lidocaine was discussed in brief.

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