Polarized osteoclasts put marks of tartrate-resistant acid phosphatase on dentin slices--a simple method for identifying polarized osteoclasts

Osteoclasts form ruffled borders and sealing zones toward bone surfaces to resorb bone. Sealing zones are defined as ringed structures of F-actin dots (actin rings). Polarized osteoclasts secrete protons to bone surfaces via vacuolar proton ATPase through ruffled borders. Catabolic enzymes such as tartrate-resistant acid phosphatase (TRAP) and cathepsin K are also secreted to bone surfaces. Here we show a simple method of identifying functional vestiges of polarized osteoclasts. Osteoclasts obtained from cocultures of mouse osteoblasts and bone marrow cells were cultured for 48 h on dentin slices. Cultures were then fixed and stained for TRAP to identify osteoclasts on the slices. Cells were removed from the slices with cotton swabs, and the slices subjected to TRAP and Mayer's hematoxylin staining. Small TRAP-positive spots (TRAP-marks) were detected in the resorption pits stained with Mayer's hematoxylin. Pitted areas were not always located in the places of osteoclasts, but osteoclasts existed on all TRAP-marks. A time course experiment showed that the number of TRAP-marks was maintained, while the number of resorption pits increased with the culture period. The position of actin rings formed in osteoclasts corresponded to that of TRAP-marks on dentin slices. Immunostaining of dentin slices showed that both cathepsin K and vacuolar proton ATPase were colocalized with the TRAP-marks. Treatment of osteoclast cultures with alendronate, a bisphosphonate, suppressed the formation of TRAP-marks and resorption pits without affecting the cell viability. Calcitonin induced the disappearance of both actin rings and TRAP-marks in osteoclast cultures. These results suggest that TRAP-marks are vestiges of proteins secreted by polarized osteoclasts.     

Clinical outcomes and adverse events following transforaminal interbody fusion for lumbar degenerative spondylolisthesis in elderly patients

Transforaminal lumbar interbody fusion (TLIF) procedure is widely used, but the surgical indications for TLIF in elderly patients remain controversial because of potential risks such as inferior bone quality and higher rate of postoperative complications. Clinical efficacy and operative risk of TLIF in elderly patients are unclear. This study investigated the clinical effect and safety of TLIF for lumbar degenerative spondylolisthesis with radiculopathy or neurogenic claudication in patients aged 70 years or older. The clinical records were retrospectively reviewed of 35 consecutive patients aged 70-86 years (mean 74.8 years) who underwent one or two-level TLIF. The preoperative diagnosis included degenerative spondylolisthesis with segmental instability. Clinical outcomes were assessed by the Japanese Orthopaedic Association score, visual analogue scale, Oswestry Disability Index. Radiological fusion rate was also investigated. Clinical and radiological results were compared with those of 43 younger patients. Clinical outcome measures were significantly improved after operation in the elderly patients, but improvement rates were significantly lower than those of younger patients. Fusion rate was similar in both groups. Overall postoperative complications were increased in aged patients, although the prevalence of complications directly related to surgical technique was not significantly increased. Postoperative complications not related to the surgical procedure were factors affecting poor results. TLIF is acceptable for achieving clinical recovery and lumbar fusion with high radiographic fusion success even in elderly patients, although clinical benefits were limited compared with those of younger patients. Postoperative morbidity was mainly related to general or non-operative site complications.     

Long-term culture of Japanese human embryonic stem cells in feeder-free conditions

Human pluripotent embryonic stem cells (hESCs) have great promise for research into human developmental biology, development of cell therapies for the treatment of diseases, toxicology, and drug discovery. Traditionally, undifferentiated hESCs are maintained on mouse embryonic fibroblasts (MEFs), which impede the clinical applications of hESCs. Here we have examined the long-term stability of the Japanese hESC line (KhES-1) in feeder-free culture. KhES-1 cells were cultured with MEF conditioned medium (CM) and different doses of basic fibroblast growth factor (bFGF) in six-well-plates of which the surface was coated with Matrigel. KhES-1 cells were maintained for at least 40 passages. In this culture system, the cells maintained stable proliferation rates and steadily expressed Oct-4, Nanog, and alkaline phosphatase. In addition, KhES-1 cells maintained without direct feeder contact formed embryonic bodies with expression of markers from the three germ layers. Here we demonstrated that Japanese human embryonic stem cells KhES-1 were cultured long term in a feeder-free method, while retaining pluripotency in vitro.     

The Number of Metastatic Lymph Nodes is a Significant Risk Factor for Bone Metastasis and Poor Outcome After Surgery for Linitis Plastica-type Gastric Carcinoma

BACKGROUND: Linitis plastica-type gastric carcinoma remains a disease with poor prognosis despite an aggressive surgical approach. Although a prominent pattern of disease failure is peritoneal carcinomatosis, some patients experience rapid disease progression without signs of the peritoneal disease. METHODS: Clinicopathologic data from 178 patients with linitis plastica-type gastric cancer operated on between 1991 and 2000 were analyzed. Survival stratified by curability of surgery, pN stage, and patterns of failure were evaluated by using the Kaplan-Meier method, and chi(2) test was used to evaluate correlation between the number of metastatic lymph nodes in terms of pN categories and the incidence of various patterns of metastasis and recurrence. Cox regression hazard model was used to identify independent prognostic factors. RESULTS: R0 resection was performed only among 82 patients (46% of those who underwent laparotomy). Node metastasis was frequent with only 22 patients classified as pN0. Peritoneal carcinomatosis was observed in 131 patients and was the commonest pattern of recurrence. Bone metastasis, found in 13 patients, was associated with poor outcome, and its incidence was significantly correlated with the number of metastatic nodes. pT4 status and pN3 status were identified as significant independent prognostic determinants. CONCLUSION: Treatment strategy for the linitis plastica should in general combine surgery with aggressive treatment directed toward peritoneal disease. However, patients with >16 metastatic nodes more often are associated with bone metastasis than those with modest nodal involvement and suffer from poor prognosis.     

Intramuscular hemangioma of the upper extremity in infants and children

PURPOSE: Intramuscular hemangiomas (IMHs) are benign tumors comprising just 0.8% of all hemangiomas and are extremely rare in the upper limbs. These tumors can pose diagnostic as well as therapeutic challenges for orthopaedic surgeons, especially in younger children. We reviewed cases of IMH of the upper extremity in infants and children from our institute. METHODS: Six consecutive patients underwent surgical treatment for IMH in our hospital. There were 4 girls and 2 boys. Long-standing pain and swelling were common symptoms except in a 1-year-old boy. Tumors were evaluated by radiography, computed tomography, magnetic resonance imaging, and angiography. RESULTS: After a mean follow-up of 42 months, all patients except one were free of pain and without tumor recurrence or functional impairment. Minimal symptoms remained in a 6-year-old boy who underwent biopsy only. CONCLUSION: Magnetic resonance imaging is the most useful evaluation for IMH because it not only delineates the extent of tumor but also reveals characteristic structures. For young children with IMH, wide excision is the treatment of choice to prevent local recurrence, but every patient should be treated individually after evaluating the patient's age, tumor location and invasion, and cosmetic considerations. LEVEL OF EVIDENCE: Therapeutic study-level III.     

Reconstruction of the middle hepatic vein tributaries draining segments V and VIII of a right liver graft by using the recipient’s own middle hepatic vein and vascular closure staples

A right liver graft lacking the middle hepatic vein can result in congestion of the anterior segment. We describe a method of reconstructing the middle hepatic vein tributaries by using the recipient’s own middle hepatic vein with vascular closure staples. During a living donor right liver transplantation, the middle hepatic vein tributaries draining segments V (V5) and VIII (V8) of the right lobe graft were reconstructed using the recipient’s own middle hepatic vein and secured with vascular closure staples. Computed tomography showed good venous outflow from the middle hepatic vein and no congestion or atrophy of the anterior segment of the right liver grafts. Thus, using the recipient’s own middle hepatic vein is a suitable option for reconstructing the middle hepatic vein tributaries (V8 and V5) in right-liver living donor transplantation and the application of vascular closure staples helps to accomplish this.     

The original hole-in-one guide for atlantoaxial transarticular screw fixation

Atlantoaxial fusion by transarticular screw fixation provides firm fixation, and good results have been reported. However, there are also problems, such as injury of the vertebral artery at the time of screw insertion. For accurate facet fusion, we developed a new hole-in-one guide for screw fixation and obtained good results with its use. In 60 adult subjects, we measured the antero-posterior and transverse axes and determined the center of the atlantoaxial surface and the insertion point of the screw in three dimensions on computed tomography scans. Based on these values, we measured the optimum screw insertion angle on sagittal and coronal planes, and the distance between the center and the posterior margin of the joint surface; our new hole-in-one guide was produced with these data. When the guide tip is determined to be located at the center posterior margin of the axial joint surface, and the guidewire insertion point is determined to be located at the center of the axial inferior facet immediately above the C2/3 joint, the guidewire is passed through the axial pedicle and the center of the atlantoaxial joint. Since April 1997, we have used this hole-in-one guide technique in eight patients with atlantoaxial instability. The screws passed the pars interarticularis and the center of the atlantoaxial joint in all patients with safety and accuracy. No complications associated with this technique occurred. Received: December 24, 1999 / Accepted: August 25, 2000     

An intraoperative evaluation of the cerebral circulation using the ophthalmic artery Doppler method in a case of brachiocephalic trunk occlusion

A 57-year-old man underwent left-right subclavian artery bypass for brachiocephalic trunk occlusion. The cerebral circulation was evaluated by ophthalmic artery Doppler method during the operation. Before the bypass, maximal flow velocity (Vmax) of the right ophthalmic artery was 6.09 cm x sec(-1), compared to 43.8 cm x sec(-1) of the left. The preoperative flow acceleration (FA) was 8.3 cm x sec(-2) and 500 cm x sec(-2) in the right and left, respectively. Both Vmax and FA of the right ophthalmic artery improved to 17.6 cm x sec(-1) and 96.7 cm x sec(-2) at the end of the bypass. Additional sutures were performed because of difficulty in stopping bleeding at the site of anastomosis. However, this additional procedure blunted the Doppler waveform, with decreases in Vmax to 8.69 cm x sec(-1) and FA to 33.1 cm x sec(-2) Re-anastomosis at the leftsubclavian artery was performed. Anaphylactic shock induced by transfusion was encountered at the end of re-bypass. Vmax of the right ophthalmic artery at that time was only 10.2 cm x sec(-1) However, the fact, that the Doppler waveform was no more blunt and FA had improved up to 116.4 cm x sec(-2), let us conclude that the cerebral circulation had recovered. In conclusion, the ophthalmic artery Doppler method is a useful tool for evaluation of the cerebral circulation.