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Introduction: Autologous bone grafting or various bone-regenerating materials are used to treat bone defects caused by tumor resection and accident trauma. Octacalcium phosphate, a reasonable bone regenerative material, activates osteoblasts. We fabricated a composite material, octacalcium phosphate/weakly denatured collagen, as a new scaffold. We aimed to investigate the osteoregenerative effect of the octacalcium phosphate/weakly denatured collagen scaffold (compared with that of weakly denatured collagen) in skull defects in a canine model. Methods: Atelocollagen was extracted from porcine skin via pepsin treatment. The weakly denatured collagen scaffold was fabricated with a freeze-dried and thermally crosslinked atelocollagen suspension at pH 7.4. Octacalcium phosphate was synthesized using Ca-acetate and NaH2PO4. Octacalcium phosphate particles (diameter, 199–298 µm) were mixed with a collagen matrix to fabricate an octacalcium phosphate/weakly denatured collagen scaffold. Bilateral defects (diameter, 10 mm; full-thickness) were induced in dog skulls, and the octacalcium phosphate/weakly denatured collagen and weakly denatured collagen scaffolds were implanted into the defects. Results: Eight weeks after implantations, bone regeneration was evaluated via histopathological analysis. It revealed osteoblast infiltration and osteoregeneration in all defects treated with the octacalcium phosphate/weakly denatured collagen scaffold. The defects treated with weakly denatured collagen scaffold or without any scaffold mostly contained connective tissue, with no neo-osteogenesis. Discussion: The novel octacalcium phosphate/weakly denatured collagen scaffold better promotes osteoregeneration than the weakly denatured collagen scaffold; this “in situ tissue engineering” approach is potentially clinically applicable for bone reconstruction.

This study introduces the latest progress on the study of artificial sensory organs, with a special emphasis on the clinical results of artificial nerves and the concept of in situ tissue engineering. Peripheral nerves have a strong potential for regeneration. An artificial nerve uses this potential to recover a damaged peripheral nerve. The polyglycolic acid collagen tube (PGA-C tube) is a bio-absorbable tube stuffed with collagen of multi-chamber structure that consists of thin collagen films. The clinical application of the PGA-C tube began in 2002 in Japan. The number of PGA-C tubes used is now beyond 300, and satisfactory results have been reported on peripheral nerve repairs. This PGA-C tube is also effective for patients suffering from neuropathic pain.

Objective. Although surgery using a polyglycolic acid–collagen (PGA-c) tube is effective for peripheral nerve injury-induced chronic hand pain, it has not been applied to trigeminal nerve lesions because of the difficult approach. We used a PGA-c tube during surgery for trigeminal neuropathy and evaluated its prognosis based on the outcomes. Design. Case report. Setting and Patients. In the dental anesthesia division of a university hospital, 11 patients with severe dysesthesia underwent surgical repair of a damaged lingual nerve (LN) or inferior alveolar nerve (IAN). One patient was lost to follow-up. Changes in quantitative sensory testing (QST) and the presence of dysesthesia as a treatment outcome were compared preoperatively and postoperatively in 10 patients. Two surgical treatments, bridging or encircling peripheral nerves, were applied. Bridging of both stumps was selected when neurotmesis was detected or the nerve was lacerated during surgery (N = 4). Otherwise, a longitudinal PGA-c tube was used to encircle the lesion (N = 6). Outcomes were evaluated 2 months to 8 years postoperatively. Results. Both methods improved the patients’ condition based on QST results (brush stroke perception, mechanical touch threshold, sensitivity to cold/hot stimuli). Preoperative allodynia or dysesthesia was resolved in six patients and greatly reduced in four. Two patients (one with inflammation-induced pain, one with implant-related pain) developed prolonged postoperative allodynia requiring pain-relief medication. Conclusions. Use of a PGA-c tube for surgical treatment of intractable pain due to LN or IAN neuropathy helps alleviate sensory impairment. The possibility of new dysesthesias emerging postoperatively, however, should be noted.

Case A 65-year-old man with a completely transected left sciatic nerve at the buttock received an implant with a bioabsorbable nerve conduit over a 20-mm gap. The conduit was filled with collagen to facilitate nerve regeneration. At 4 years after implantation, reinnervation potentials were detected in the muscles, and there was sensory recovery in the reinnervated areas. Thereafter, motor and sensory function recovered gradually over a 14-year period. Conclusion Mixed nerve regeneration after conduit-based reconstruction was demonstrated objectively. Although it took >4 years for motor and sensory functions to be regained, the nerve connection between the buttock and the foot was restored.

The mechanical properties of nerve guide tubes must be taken into consideration when they are being developed. We previously reported the feasibility of using 50:50 tubes in a canine 40mm peroneal nerve defect model, where 50:50 represents the proportion of poly(L-lactic) acid (PLLA) and polyglycolic acid (PGA). The aim of the current study was to show that 50:50 tubes have suitable mechanical properties for repairing long nerve defects. Four types of nerve guide tubes made with PLLA to PGA fiber ratios of 100:0 (i.e. 100% PLLA) (100:0 tube), 50:50 (50:50 tube), 10:90 (10:90 tube), and 0:100 (0:100 tube) were designed and created using a tubular braiding machine. Their mechanical properties were examined in vitro (up to 16 weeks). In compression testing, 50:50 tubes had the highest normalized force value, followed in order by the 100:0, 10:90, and 0:100 tubes up to 8 weeks after immersion. From the point of view of biomechanics and bioresorbability, out of the 4 tube types tested, 50:50 tubes appeared to have the optimal mechanical properties for longer nerve defects.

One of the ways to mitigate global warming is to reduce carbon dioxide emissions. In Japan, carbon dioxide emissions from vehicles, which run on fossil fuels, account for about 1/4 of the total carbon dioxide emissions. In this study, we applied a commercial engine, and remodeled the engine which could work using bioethanol as a fuel. We subsequently examined the influence of exhaust emission of the engine with the bioethanol and documented the entire study as teaching materials, including the remodeling method. Next, we began our mechanical engineering courses at the university by using the teaching materials in this study as introductory education, with the aim of stimulating the interest of the students. We then developed an education program for the students.

Purpose Chronic lunotriquetral (LT) instability (persisting >3 months after injury) can be treated with a variety of surgical methods (LT fusion, ligament repair or reconstruction, or arthroscopic debridement), with varying degrees of success. We retrospectively evaluated the results of dorsal capsulodesis using the dorsal radiocarpal ligament in an attempt to reinforce the dorsal LT interosseous ligament in patients with chronic dynamic LT instability. Methods A total of 11 patients (mean age, 37 years) with persistent posttraumatic wrist pain and weakness were diagnosed with dynamic LT instability (positive LT ballotment test, disruption of Gilula's lines or volar intercalated segment instability deformity on stress radiography, arthroscopic findings of Geissler grade 3, or 4 LT tears). They were treated consecutively with dorsal capsulodesis. We used subjective and functional outcome measures to evaluate the results. Results The mean follow-up period was 31 months (range, 12–65 months). The postoperative visual analog pain score and Mayo wrist score were significantly improved compared with their preoperative values. The average grip strength increased by 5 kg, and the average range of wrist flexion was reduced by 3°. Conclusions In this small series, in the short term, dorsal radiocarpal ligament capsulodesis reduced pain and instability and improved function in patients with chronic dynamic LT instability. Type of study/level of evidence Therapeutic IV.

Adipose-derived stromal cells (ASCs) include stem cells, which have the potential to differentiate into a variety of cell lineages. The regeneration of central nerves was examined using ASCs and a collagen scaffold. A cerebral cortex defect (3 × 4 × 3 mm3) was created in the left frontal lobe of 16 male rats. In one group (n = 8), collagen (3 × 4 × 3 mm3) seeded with DiI-labeled ASCs was implanted in the defect. In order to seed the ASCs, a combination of the rotary cell culture system and pressing the collagen scaffold gently several times with a glass rod was applied. In the control group (n = 8), collagen was implanted without ASCs. The rats were sacrificed at 1 month after the scaffold implantation. Histologically, 0.2% of the implanted ASCs were positive for anti-human/rat microtubule-associated protein 2 (MAP2) antibody and microvessels were present at a density of 4.6 ± 1.2/mm2 within the collagen scaffold-implanted area in each coronal section. In the control group, no MAP2-positive cells were detected and the microvessel density was 0.6 ± 0.4/mm2. These data suggest that ASCs seeded into a collagen scaffold may have the potential to promote regeneration of nervous tissue after cerebral cortex injury.

Summary Over the last 20 years, an increasing number of research articles have reported on the use of artificial nerve tubes to repair nerve defects. The development of an artificial nerve tube as an alternative to autogenous nerve grafting is currently a focus of interest for peripheral nerve repair. The clinical employment of tubes as an alternative to autogenous nerve grafts is mainly justified by the limited availability of donor tissue for nerve autografts and the related morbidity. Numerous studies indicate that short-distance defects in humans can be successfully treated by implantation of artificial nerve guides. This review provides a brief overview of various preclinical and clinical trials conducted to evaluate the utility of artificial nerve tubes for the regeneration of peripheral nerves. This review is also intended to help update hand surgeons on the rapid advances in tubulization techniques, and to provide them with indications of the various directions toward which future research can proceed. Future studies need to provide us with as much comparative information as possible on the effectiveness of different tubulization techniques, in order to guide the surgeon in choosing the best indications for their optimal clinical employment. Future progress in implant development can be expected from interdisciplinary approaches involving both materials and life sciences, leading to advances in neuro-tissue engineering that will be needed to effectively treat larger nerve defects.

p c f d P r .jp raumatic injury of the lingual nerve is in many cases atrogenic, extraction of the third molar being the ost frequent cause. Deficits in taste, thermal and ouch sensations, and abnormal sensations (eg, pareshesia and dysesthesia) are frequently caused by peipheral nerve damage, leading to difficulties in eating nd/or speech. In contrast to natural improvement fter slight or moderate sensory impairment in the rofacial region, several studies have reported the use f surgical intervention with either direct anastomosis r autograft to reconstruct nerve gap and/or remove

Marrow mesenchymal cells are useful in regenerative medicine because they contain stem cells, but there have been few reports of clinical applications. The authors developed a new wound treatment technique by improving marrow mesenchymal cell culture methods and placing cultured cells in an artificial skin material. This new treatment was useful for tissue regeneration in 20 patients with skin wounds.Marrow mesenchymal cells from a 46-year-old man were cultured and placed in artificial dermis made of collagen sponge. This composite graft was implanted subcutaneously into the back of a nude mouse and removed 10 days later; immunohistological analysis confirmed regeneration of subcutaneous tissue using human marrow mesenchymal cells. Next, in 20 patients (nine men and 11 women; average age, 64.8 years; range, 22 to 91 years) with intractable dermatopathies, 10 to 20 ml of bone marrow fluid was aspirated from the ilium and cultured in medium containing either fetal calf or autologous serum. The resulting cultured cells were placed in artificial dermis made of collagen sponge, and this composite graft was used to treat skin wounds.The wound mostly healed in 18 of the 20 patients; the remaining two patients died of causes unrelated to transplantation. In all patients, autologous marrow mesenchymal cell transplantation was shown to be therapeutically effective.In skin regeneration therapy using a marrow mesenchymal cell/artificial dermis composite graft, skin regeneration is possible with bone marrow aspiration, a minimally invasive procedure. Compared with existing skin grafting techniques, the present technique is practical and much less invasive.

The F(1) hybrid of autoimmune hemolytic anemia-prone NZB and nonautoimmune NZW strains of mice has been studied as a murine model of systemic lupus erythematosus. Both NZB and F(1) hybrid mice show age-dependent spontaneous activation of peripheral CD4(+) T cells as reflected by the elevated frequencies of CD4(+) T cells positive for CD69 early activation marker. Both strains also show age-dependent abnormal decrease of the frequencies of CD62L(+) naive CD4(+) T cells and/or NTA260(+) memory CD4(+) T cells in the spleen. We studied the multigenic control of these abnormal features of peripheral CD4(+) T cells in (NZB x NZW) F(1) x NZW backcross mice by quantitative trait loci mapping and by association rule analysis. The abnormally elevated frequencies of CD69(+)CD4(+) T cells and decreased frequencies of CD62L(+) naive and/or NTA260(+) memory CD4(+) T cells were under the common genetic control, in which the interaction between MHC and a hitherto unknown locus, designated Sta-1 (spontaneous T-cell activation) on chromosome 12, plays a major role. The allelic effects of these loci likely predispose CD4(+) T cells to the loss of self-tolerance, and are responsible for the accelerated autoimmune phenotypes of (NZB x NZW) F(1) hybrid mice.

An arginine specific protease, Sp-protease, was purified by column chromatography from freeze-dried Spirulina platensis using a five-step process. Purified Sp-protease has a molecular weight of 80 kDa. It hydrolyzed the synthetic substrates containing arginine residue in the P1 position but did not hydrolyze synthetic substrates containing other amino acid residues, including lysine residue in the P1 position. Among the synthetic substrates tested, a substrate of plasminogen activator (Pyr-Gly-Arg-MCA) was hydrolyzed most effectively with the enzyme (Km = 5.5 x 10(-6) M), and fibrin gel was solubilized via activation of intrinsic plasminogen to plasmin with the enzyme. Activity was inhibited completely with camostat mesilate (Ki = 1.1 x 10(-8) M) and leupeptin (Ki = 3.9 x 10(-8) M) but was not inhibited with Nalpha-tosyl-L-lysine chloromethyl ketone (TLCK). The optimum pH of the enzyme has a range of pH 9.0 to pH 11.0. The optimum temperature was 50 degrees C; the enzyme was stable at 0-50 degrees C.

Biliverdin, the precursor of bilirubin, was adsorbed onto the synthetic clay mineral, smectite, in a mixed solution of benzene and methyl alcohol (7:3 v/v) to form a biliverdin–smectite conjugate. The adsorption of biliverdin to smectite followed the Freundlich equation with an adsorption coefficient of 1.0. The water-soluble conjugate had a green color with the absorbance maxima at 679 and 378 nm. The biliverdin–smectite conjugate became photostable in water to irradiation with UV, although the biliverdin in benzyl alcohol was rapidly decolorized.

Phycocyanin with blue color, phycocyanobilin bound to protein, with the absorption maximum at 614 nm is quite unstable against urea-treatment because of the denaturation of the protein conformation. To prevent the bleaching of phycocyanin by urea, amino groups of lysine residues in the phycocyanin molecule were modified with DSP [dithiobis(succinimidyl propionate)] to cross-link amino groups in the protein molecule in order to maintain the high order structure of phycocyanin. The color development of DSP-modified phycocyanin became stable, but unmodified phycocyanin was markedly decreased to 20% of the original absorbance at 614 nm against urea-treatment. Splitting of the S–S linkage of DSP-modified phycocyanin molecule by 2-mercaptoethanol caused the reduction of color development of phycocyanin under urea-treatment. The color development of phycocyanin is closely associated with its high order structure of protein.

OBJECTIVE: The aim of this study was to report by means of objective methods on the effectiveness of a nerve reconstruction procedure using a bioresorbable tube in two patients. Our previous successes in regenerating canine peripheral nerves across long distances (80-mm gaps) using a bioabsorbable tube have led us to investigate the value of such a tube for the treatment of human patients with chronic nerve injuries. METHODS: The device was made from a cylindrically woven polyglycolic acid tube filled with a collagen sponge. It was designed to be resurbed after nerve regeneration. Peripheral sensory nerve defects in two patients with neuroma and pain were reconstructed using this tube. Patient 1 (a 62-year-old man) had a 20-mm defect of the proper digital nerve, and Patient 2 (a 56-year-old woman) had a 65-mm defect of the superficial peroneal nerve. RESULTS: After surgery, both patients recovered from the unpleasant sensations and intolerable pain. In Patient 1, furictional recovery was objectively identified at 2 months, and conduction velocity of the nerve recovered to 49.1 m/s, in patient 2, conduction velocity of the nerve was determined to be 16.9 m/s at 5 months, Current perception threshold testing indicated that sensory nerve function had been recovered by 65 days after surgery. CONCLUSION: This work represents the first precise clinical evaluation, performed under objective evaluation criteria, of sensory recovery achieved using a nerve tube, suggesting that the use of a polyglycofic acid-coflagen tube has the potential to become a viable alternative to conventional autografting for the repair of peripheral nerve defects.

Chlorophyll a molecules were adsorbed to mesoporous silica (FSM: folded-sheet mesoporous material) to form a chlorophyll−FSM conjugate capable of nanometer-scale interaction between chlorophyll a molecules as seen in a living plant leaf. The chlorophyll a molecules introduced into nanopores of FSM-22 (with a pore diameter of 4 nm) exhibited a red shift in the absorption peak and showed efficient excitation energy transfer from the shorter to the longer wavelength forms as characterized by the fast transient phase of 38 ps detected in the decay-associated spectrum (DAS) analysis of the fluorescence decay. Illumination of the chlorophyll−FSM-22 conjugate with visible light generated a stable chlorophyll a cation radical in the electron spin resonance (ESR) measurement, although the illumination of free chlorophyll a in solution produced almost no cation signal. The physiological function of the chlorophyll−FSM conjugate intercalated with ruthenium oxide (RuO2) was explored as chlorophyll−FSM exhibited the ...

The aim of this study was to investigate the feasibility of transferring the free dorsoulnar perforator flap nourished by the cutaneous perforator branched dorsoulnar artery to reconstruct severely injured fingers under upper arm anesthesia. Between April of 2001 and April of 2002, 13 free dorsoulnar perforator flaps were used in 13 patients. There were 11 men and two women ranging in age from 18 to 64 years, with an average age of 38 years. The affected fingers were one thumb, four index fingers, five middle fingers, two ring fingers, and one little finger. All cases were performed under upper arm anesthesia combined with intravenous local anesthesia. The operative time ranged from 103 to 140 minutes, with an average time of 120 minutes. The flap size ranged from 1 x 3 to 3 x 4 cm, and was transferred from the same forearm of the injured finger. All donor sites were closed primarily without a skin graft. The aim of reconstruction for fingers was to repair a traumatic defect (five cases), partial necrosis following replantation (two cases), and soft-tissue defects resulting from resection of a scar (three cases) and to revascularize ischemic fingers (three cases). All flaps survived completely. After repair of the flow-through circulation of the common digital artery and ischemic finger, a postoperative angiogram showed the vascular patency and hypervascularity of the reconstructed fingers, and the patients' complaints were reduced. The free dorsoulnar perforator flap under regional anesthesia is first reported; it may become one valuable option as a very small flap for the treatment of repairing intercalated or segmental defects as a flow-through flap for soft-tissue defects and ischemic fingers.

Eleven patients (12 ankles) who had Pirogoff ankle disarticulation were followed up for an average of 5.6 years (range, 2.1-8.9 years). After disarticulation of the Chopart joint, the talus was removed and the calcaneus was cut off in the coronal plane. The heel then was rotated 90 degrees to the tibial plafond. The postoperative result was evaluated using a scoring sheet. Excellent results were seen in four feet: two in patients with trauma, one in a patient with a metastatic tumor, and one in a patient with osteomyelitis. Good results were seen in two feet: one in a patient with trauma and one in a patient with diabetes mellitus. A fair result was seen in one foot in a patient with diabetes mellitus. Poor results were seen in four feet in three patients with arteriosclerosis and one foot in a patient with diabetes mellitus. In Pirogoff ankle disarticulation, the leg-length discrepancy is compensated for by using the calcaneus. The patients only had a leg-length discrepancy of 2.8 cm on average, which enabled them to walk outdoors wearing only a shoe orthosis. The postoperative results were unfavorable in patients with vascular diseases and were satisfying in patients without vascular diseases.

Biopterin-α-glucoside purified from Spirulina platensis is stable to irradiation by visible and ultraviolet light. Its absorption spectrum shows maxima at 268 nm and 340 nm and a fluorescence band appears at 447 nm. Although cyanidin-3-galactoside with an absorption maximum at 514 nm (red colour) is unstable and is decolourized by UV light, the pigment becomes stable in the presence of biopterin-α-glucoside. Similar photostabilization phenomenon was observed for phycocyanin (λ max 614 nm) consisting of phycocyanobilin as chromophore and protein subunits α 3 β 3 in the presence of biopterin-α-glucoside. Although the chromophore of phycocyanin is related to phycocyanobilin and protein conformation, the chromophore can be stabilized to UV light in the presence of biopterin-α-glucoside.

Hemin (Fe(3+)) was adsorbed onto synthetic smectite (clay mineral) intercalated with a quaternary alkenylammonium compound, dioleyldimethylammonium chloride (DOA), to form a hemin-smectite-DOA conjugate. The hemin-smectite-DOA conjugate was soluble in organic solvents such as benzene and toluene to form a transparent colloidal solution with a light yellow color. Its absorption spectrum in benzene showed two bands, 600 and 568 nm, in the visible region and a sharp Soret band at 400 nm with the molar extinction coefficient of 7.5 x 10(4) M(-1) cm(-1). The formation of the conjugate of smectite and DOA was confirmed by X-ray diffraction analysis: the basal spacing, d(001), of hemin-smectite-DOA conjugate was 19 A which is an expansion of the interlayer space by 5 A based upon the basal spacing of smectite of 14 A. Hemin-smectite-DOA conjugate catalyzed the peroxidase-like reaction in organic solvents using benzoyl peroxide as the hydrogen acceptor and leucocrystal violet as the hydrogen donor. The temperature-dependent peroxidase-like activity of the conjugate was compared with peroxidase activity of horseradish peroxidase. The hemin-smectite-DOA conjugate exhibited higher activity as the temperature was increased from 30 to 70 degrees C, while horseradish peroxidase activity was reduced as the temperature was increased.

Brachial plexus injury is very rare in neurosurgical practice, so many neurosurgeons have never experienced this problem in Japan. This study describes a clinical analysis of 16 patients aged 5 to 62 years (mean 32.9 years) who presented at our institution with brachial plexus injuries. Nine patients presented with paralysis and seven with paresis. Head injury was the most common associated injury in eight of 16 patients. Six patients were managed conservatively. All patients with C8-T1 paresis spontaneously recovered to a useful level. Surgery was performed in 10 patients: six neurolysis, two neurotization, and three nerve grafting procedures. All six patients who underwent neurolysis of the brachial plexus attained useful recovery. Four of five patients achieved useful recovery after nerve repair. Nerve grafting achieved a better outcome than neurotization in this study. The difference of outcome was attributed to the graft length. The management of brachial plexus injury is a great challenge, but surgical outcome can be improved if the optimal repair procedure is selected for brachial plexus injury.

A reversed pedicled venous flap containing perivenous areolar tissue and/or nerve was used to cover traumatic skin defects of seven fingers in six patients. The series consisted of six men, ranging in age from 20 to 57 years (average: 39 years). The reconstructed sites were four dorsal skin defects and three volar skin defects of the finger. The flap was designed on the dorsum of the hand, in such a way as to place a vein at the centre of the flap and not to involve the dorsal metacarpal artery. The flap contained a dorsal vein, perivenous areolar tissue and fascia of the interosseous muscle. Cutaneous nerves were present in three of the seven flaps. The pedicle of the flap was dissected distally to the finger web space and the flap was transferred to the skin defect. The size of the flap ranged from 1.4x4.5 cm to 6.0x7.0 cm. The average length of the pedicle was 1.6 cm. Skin grafting was needed at the donor site in one case (flap size: 6.0x7.0 cm), but primary closure was possible in the remaining cases. Postoperatively, the largest flap showed superficial necrosis, although it survived. The remaining flaps survived completely. This suggests that in a large flap the skin should be attached to the pedicle to prevent congestion. The flap can be elevated without reference to the dorsal metacarpal artery at the ulnar side of the dorsum of the hand. The flap is an effective option to reconstruct skin defects of the finger, especially the little finger.

Hemin (Fe(3+)) was adsorbed onto synthetic smectite (clay mineral) in acetone to form a hemin-smectite conjugate. The hemin-smectite conjugate became soluble in water to form a transparent colloidal solution with a dark brown color. Its absorption spectrum in water showed a sharp Soret band at 398 nm with the molar extinction coefficient as epsilon(398nm) = 11.6 x 10(4) M(-1) cm(-1), which is in good agreement with epsilon(398nm) = (12.2 +/- 3) x 10(4) M(-1) cm(-1) of monomeric hematin (1). Hemin (Fe(3+))-smectite conjugate had a peroxidase-like activity in the presence of hydrogen peroxide (a hydrogen acceptor) and guaiacol (a hydrogen donor) in aqueous solution and its activity was higher than that of hematin. Hemin (Fe(3+))-smectite conjugate in water was reduced by adding sodium dithionite to form a heme (Fe(2+))-smectite conjugate which is also a transparent colloidal solution in water. Its absorption spectrum in aqueous solution was surprisingly in close agreement with that of oxyhemoglobin. Its peak positions of alpha, beta, and Soret bands were located in only a 9--3 nm shift to shorter wavelengths in comparison with those of oxyhemoglobin. Therefore, heme (Fe(2+))-smectite conjugate was bound to O(2) to form O(2)-heme (Fe(2+))-smectite conjugate. The addition of carbon monoxide, CO, to O(2)-heme (Fe(2+))-smectite conjugate caused the formation of CO-heme (Fe(2+))-smectite conjugate with a similar absorption spectrum of carboxyhemoglobin (HbCO) accompanied by shifting 8--10 nm to shorter wavelength. Therefore, the transformation of O(2)-heme (Fe(2+))-smectite conjugate to CO-heme (Fe(2+))-smectite conjugate was accompanied by shifting of 7, 4, and 3 nm to shorter wavelengths in the alpha, beta, and Soret bands respectively, which are similar to the spectral change from oxyhemoglobin to carboxyhemoglobin. Also the ratio (1:1.6) of the molar extinction coefficient of Soret band of O(2)-heme (Fe(2+))-smectite conjugate and CO-heme (Fe(2+))-smectite conjugate was surprisingly agreement with ratio (1:1.5) of oxyhemoglobin and carboxyhemoglobin. The phenomenon shown above was unexpectedly found during the course of study of bioconjugate of a bioactive substance, hemin (Fe(3+)) or heme (Fe(2+)), and a clay mineral, smectite, in place of the protein of globin in hemoglobin.

Objectives/hypothesisDue to its anatomical features, the chorda tympani nerve (CTN) is sometimes sacrificed during middle ear surgery, resulting in taste dysfunction. We examined the effect of placing an artificial nerve conduit, a polyglycolic acid (PGA)-collagen tube, across the gap in the section of the resected chorda tympani nerve (CTN) running through the tympanic cavity.MethodsThe CTN was reconstructed with a PGA-collagen tube in three patients with taste disturbance who underwent CTN resection. To evaluate the effect of the reconstruction procedure on the patients' gustatory function, we measured the patients' electrogustometry (EGM) thresholds. The patients were followed-up for at least two years.ResultsGustatory function was completely restored in all of the patients after the reconstruction. The patients' EGM thresholds exhibited early improvements within one to two weeks and had returned to their normal ranges within three months. They subsequently remained stable throughout the two-year follow-up period. In a patient who underwent a second surgical procedure, it was found that the PGA-collagen tube used in the first surgical procedure had been absorbed and replaced by new CTN fibers with blood vessels on their surfaces.ConclusionThese results suggest that reconstruction of the CTN with an artificial nerve conduit, a PGA-collagen tube, allows functional and morphological regeneration of the nerve and facilitates the recovery of taste function. PGA-collagen tubes might be useful for repairing CTNs that are resected during middle ear surgery. Further research is required to confirm these preliminary results although this is the first report to describe the successful regeneration of a nerve running through an aerial space.

The authors performed anastomoses of small vessels with the Nd-YAG laser, comparing these with conventional suture anastomoses. Some arteries were sutured with 10-0 nylon with about eight suture sites, and others were held by stay sutures at three points with laser irradiation performed between them. Two surgeons performed the anastomoses in the same way. The first was well-trained and experienced, and the second was technically inexperienced. The authors evaluated clamping time, patency rate, and endoscopic and histologic findings. The patency rate of the first surgeon was 100 percent (30/30) in the suture anastomoses and was 97 percent (29/30) in the laser anastomoses. For the second surgeon, the rates were 60 percent (18/30) in the suture group and 80 percent (24/30) in the laser group. On histologic examination, the suture group showed inflammatory cells around the suture site at the fourth week after the operation. In the laser group, an inflammatory reaction around the suture material was observed, but the other areas recovered. For the experienced surgeon, the patency rate and clamping time of the laser anastomosis provided no statistically significant difference to those of the suture anastomosis. On the other hand, for the inexperienced surgeon, the patency rate of the laser anastomosis was superior to that of the suture anastomosis, and the clamping time of the laser anastomosis was shorter than that of the suture anastomosis. Therefore, the authors concluded that the Nd-YAG laser anastomosis is useful for small vessels.

A 65 year-old female was admitted to a regional hospital with a complaint of ileus following a total hysterectomy performed 14 years ago. She was treated conservatively by total parental nutrition (TPN). Nine days later she underwent resection of the intestine due to rebound tenderness in the lower abdomen. Infusion toward TPN was resumed the following day after the operation. On the 4th postoperative day she developed restlessness and stupor. She was treated as a hyperosmolar hyperglycemic nonketotic coma (HHNC). She was referred to our intensive care unit (ICU) due to coma and hypotension. Her conscious level was a score of 5 on the Glasgow Coma Scale (GCS). Continuous hemodiafiltration (CHDF) was introduced intermittently due to oliguria. During these treatments, she fell into a deep coma and almost stopped spontaneous respiration on the 10th hospital day. CT scans showed a marked brain stem edema and acute obstructive hydrocephalus. She underwent an emergency ventricular drainage. After these treatments, her consciousness level gradually recovered and subsequently she was transferred to the general ward with a score of 13 on the GCS. Although she had serious complication of renal failure, rabdomyolysis and also hydrocephalus in this case, she was recovered in intensive care. It is advisable to maintain the patients on clinical parameters (monitoring therapy) using CHDF.

Polyethylene glycol (PEG) conjugates of protein antigens induce antigen-specific immune tolerance of helper T (Th)-cells. However, the mechanism of this Th-cell tolerance has remained unelucidated. Using transgenic mice with ovalbumin (OVA)-specific T-cell receptor (TCR) genes, we examined the response of OVA-specific Th-cells towards tolerogenic PEG-conjugate of OVA in vitro and in vivo. When stimulated with PEG--OVA in vitro, transgenic OVA-specific Th-cells proliferated and produced interleukin 2, the levels of which were comparable to those induced by unmodified OVA. In contrast, PEG--OVA administered into the circulation of transgenic mice induced unresponsiveness in peripheral OVA-specific Th-cells. Moreover, in the thymus of these transgenic mice, the frequency of immature CD4+CD8+ (double positive) thymocytes was reduced. A similar phenomenon was not observed in transgenic mice treated with unmodified OVA. As autoreactive T-cells are known to be clonally deleted at the immature double positive stage in the thymus. Th-cell tolerance induced by PEG--protein antigens is at least in part mediated by central tolerance in the thymus, and is likely caused by the markedly enhanced stability of PEG--protein conjugates in the circulatory system.

New Zealand Black (NZB) mice spontaneously produce anti-erythrocyte autoantibodies (AEA) in association with splenomegaly, thus serving as a model for autoimmune hemolytic anemia. Although these autoimmune traits are inherited as a dominant fashion, expression in F(1) hybrids of NZB and most non-New Zealand strains is suppressed due to the contribution of wild-type modifying genes present in the latter strains. Using chromosomal microsatellite markers in the (C57BL/6 x NZB)F(1) x NZB backcross progeny, we mapped C57BL/6 modifying loci for AEA production and splenomegaly. Generation of AEA was found to be down-regulated by a combined effect of two major independently segregating dominant alleles-one linked to D7MIT30 on chromosome 7 and the other linked to D10MIT42 on chromosome 10. Splenomegaly was modified mainly by a single C57BL/6 allele linked to D4MIT58 on chromosome 4. Thus, the autoimmune hemolytic anemia in the NZB strain is under multigenic control and a combined action of not only susceptibility but also modifying alleles with suppressive activities affects the outcome of disease features in the progeny. There are potentially important candidate genes which may be linked to the regulation of AEA and splenomegaly.

不安定性骨盤骨折に対するIliosacral screw(以下,IS screw)による固定方法は,欧米を中心に広く用いられてきているが,その刺入には技術的に困難を伴い,合併症の発生率は1-6%と報告されている。われわれは,独自の工夫として,鈍的刺入法と運動機能モニタリング(motor-evoked potential monitoring,以下MEPM)を用いて,その合併症の発生頻度を抑止できるか否かを検討した。症例は9症例の不安定性骨盤骨折に対して,12本のIS screwを刺入し,術中,MEPMを使用したのは6症例,7 screwであった。最終的に全例が神経損傷を回避でき,疼痛なく独歩可能となった。IS screw刺入に際しての挿入の困難さは鈍的刺入法とMEPMの組合せにより,軽減できるものと考えられた。術中,MEPMはIS screwの締めすぎによって57%に低電位を示し,神経圧迫を感知する可能性が示唆されたが,screwを緩めることで低電位は回復した。

The morphology of progenitor endothelial cells on vascular graft surfaces is addressed in this report. Such cells were seen to attach to intima-expressed CD34 and Flk-1 antigen and showed positive 5-bromo-2-deoxyuridine (BrdU) uptake. We examined CD34 and Flk-1 antigen-expressing endothelial progenitor cells three-dimensionally using confocal laser scanning microscopy (CLSM). Under detailed CLSM observation, through an ameboid-form cell, these progenitor endothelial cells changed from a globular to a flattened form. We also investigated these morphological changes using scanning electron microscopy. From these results, progenitor endothelial cells were observed not only near the advancing edge of endothelium, but also around the developing intimal site. Their form also changed from globular to flattened as observed in the CLSM results. These morphological changes were seen more frequently near the advancing edge and around the developing intimal site. They attached directly to vascular prosthesis fibers and likewise covered the graft luminal surface. Progenitor endothelial cells in any form had a common surface structure. We conclude from our results that progenitor endothelial cells can attach to graft fibers directly without clotting and directly cover the graft luminal surfaces.

Chlorophyllin a was conjugated with alpha-(3-aminopropyl)-omega-methoxypoly(oxyethylene), PEG-NH(2), to form the PEG-chlorophyllin conjugate through acid-amide bonds. The PEG-chlorophyllin conjugate was stable toward light illumination under anaerobic condition in comparison with chlorophyllin a. The conjugate catalyzed the reduction of methyl viologen in the presence of 2-mercaptoethanol and the evolution of hydrogen gas in the presence of methyl viologen (an electron carrier), 2-mercaptoethanol (an electron donor) and hydrogenase (Scheme 1). Furthermore, the PEG-chlorophyllin conjugate catalyzed the photoreduction of NADP(+) or NAD(+) in the presence of ascorbate as an electron donor and ferredoxin-NADP(+) reductase as the coupling enzyme. Utilizing the reducing power of NADPH generated by the PEG-chlorophyllin conjugate under the illumination, CO(2) fixation was accomplished by the synthesis of malate (C(4)) from pyruvate (C(3)) and CO(2) in the presence of malic enzyme (Scheme 2). These reactions mentioned above did never proceed in dark or without each enzyme.

Objective After previous success in regenerating canine peripheral nerves over 80 mm gaps using a bioabsorbable nerve guide tube, we have extended this method to the treatment of patients experiencing various types of nerve injury. This report describes the treatment of two cases of motor nerve disorder. Methods The bioabsorbable nerve tube was a cylindrically woven polyglycolic acid (PGA) tube filled with collagen. A peripheral motor nerve defect (the frontalis branch of the facial nerve) was reconstructed using this PGA-collagen tube in two patients who experienced posttraumatic unilateral eyebrow ptosis for 3 months. Results Five months after surgery, both patients regained their ability to voluntarily lift their eyebrows symmetrically. Electrophysiological examination at 5 months revealed recovery of compound muscle action potential and disappearance of distal latency on the affected side. Conclusion This is the first clinical report of motor nerve recovery achieved using the PGA-collagen nerve guide tube. The results suggest that use of a PGA-collagen tube is a promising option for the repair of motor nerve defects.

Polyethylene glycolated horseradish peroxidase (PEG-HRP) can catalyze one- and two-electron oxidation reactions in organic solvents as well as in aqueous buffer. Even though the oxidation of guaiacol in benzene and chlorobenzene is 5 orders of magnitude slower than in phosphate buffer, compounds I and II are involved in the catalytic cycle in organic media. Factor analysis and global fittings of rapid scan data set reveal that the formation of compound I of PEG-HRP in organic media consists of two steps (the first fast and the second slow) and suggest the involvement of a H2O2−HRP complex in the catalytic cycle. The labile precursor of compound I is stabilized when PEG-HRP reacts with hydrogen peroxide in chlorobenzene at −20 °C. The absorption spectrum of the precursor does not exhibit the features of hyperporphyrin spectrum but has a normal Soret as previously observed in R38L HRP. More importantly, compound I of PEG-HRP can be maintained for more than an hour at −20 °C in chlorobenzene.

Chlorophyll a was adsorbed to a synthetic smectite intercalated by poly(vinylpyrrolidone) (PVP) to form the chlorophyll-PVP-smectite conjugate (Chl-PVP-SME) having an absorption maximum at 677 nm. The conjugate was found to be stable toward light illumination in comparison with chlorophyll-smectite, chlorophyll-PVP, and free chlorophyll a. Chl-PVP-SME had a photoinduced activity for catalyzing the reduction of methyl viologen. Furthermore, the evolution of hydrogen gas was observed when an aqueous suspension containing Chl-PVP-SME, methyl viologen (an electron carrier), 2-mercaptoethanol (an electron donor), and hydrogenase was illuminated by visible light.

The authors used a delayed arteriovenous (A-V) shunt method to prevent congestion and necrosis of arterialized venous flaps. Two weeks before elevating the flap, an A-V shunt between the central artery and vein was created at the more proximal flap end. Twelve rabbits (24 flaps) were used for this experiment. Flaps were followed for 4 weeks. Twelve of 24 flaps survived completely; another 10 flaps showed partial necrosis; and two flaps became completely necrotic. These results demonstrated that this method is superior to a simple A-V shunt in preventing congestion and necrosis in the arterialized venous flap.