Impact of oesophagectomy on physical fitness and health-related quality of life in patients with oesophageal cancer.

Although several studies have reported the impact of oesophagectomy on health-related quality of life (HRQOL), none has objectively assessed its impact on physical fitness. This study aimed to evaluate the impact of oesophagectomy on physical fitness and HRQOL in patients with oesophageal cancer. In this prospective study, we investigated 30 consecutive patients with newly diagnosed resectable oesophageal cancer who were scheduled to receive either neoadjuvant chemotherapy followed by surgery or surgery alone. The primary end-points were change from baseline in two measures of physical fitness (knee-extensor muscle strength and 6-min walking distance) after oesophagectomy. The secondary end-point was change from baseline in HRQOL measured with the European Organization for the Research and Treatment of Cancer Quality of Life Core Questionnaire with 30 items (EORTC QLQ-C30). Physical fitness was significantly lower after oesophagectomy than before oesophagectomy (P < 0.001). With regard to HRQOL, there was a significant pre- to post-oesophagectomy change in most of the scales. In the multiple regression analysis, the change in 6-min walking distance was the only significant variable affecting the change in physical functional score on the EORTC QLQ-C30 (P = 0.032). In conclusion, oesophagectomy adversely affects physical fitness and HRQOL in patients with oesophageal cancer. Six-minute walking distance may be a good indicator of HRQOL shortly after oesophagectomy.

Extracellular volume fraction using contrast-enhanced CT is useful in differentiating intrahepatic cholangiocellular carcinoma from hepatocellular carcinoma.

Objectives: To evaluate whether tumor extracellular volume fraction (fECV) on contrast-enhanced computed tomography (CT) aids in the differentiation between intrahepatic cholangiocarcinoma (ICC) and hepatocellular carcinoma (HCC). Methods: In this retrospective study, 113 patients with pathologically confirmed ICC (n = 39) or HCC (n = 74) who had undergone preoperative contrast-enhanced CT were enrolled. Enhancement values of the tumor (Etumor) and aorta (Eaorta) were obtained in the precontrast and equilibrium phase CT images. fECV was calculated using the following equation: fECV [%] = Etumor/Eaorta × (100 - hematocrit [%]). fECV values were compared between the ICC and HCC groups using Welch's t-test. The diagnostic performance of fECV for differentiating ICC and HCC was assessed using receiver-operating characteristic (ROC) analysis. fECV and the CT imaging features of tumors were evaluated by two radiologists. Multivariate logistic regression analysis was performed to identify factors predicting a diagnosis of ICC. Results: Mean fECV was significantly higher in ICCs (43.8% ± 13.2%) than that in HCCs (31.6% ± 9.0%, p < 0.001). The area under the curve for differentiating ICC from HCC was 0.763 when the cutoff value of fECV was 41.5%. The multivariate analysis identified fECV (unit OR: 1.10; 95% CI: 1.01-1.21; p < 0.05), peripheral rim enhancement during the arterial phase (OR: 17.0; 95% CI: 1.29-225; p < 0.05), and absence of washout pattern (OR: 235; 95% CI: 14.03-3933; p < 0.001) as independent CT features for differentiating between the two tumor types. Conclusions: A high value of fECV, peripheral rim enhancement during the arterial phase, and absence of washout pattern were independent factors in the differentiation of ICC from HCC.

Translin binds to the sequences adjacent to the breakpoints of the TLS and CHOP genes in liposarcomas with translocation t(12;6).

Myxoid and round-cell liposarcomas share the translocation t(12;16)(q13;p11) creating the TLS-CHOP fusion gene as a common genetic alteration. We previously reported several unique characteristics of genomic sequences around the breakpoints in the TLS and CHOP loci, and among them was the presence of consensus recognition motifs of Translin, a protein that associates with chromosomal translocations of lymphoid neoplasms. We further extended our search for Translin binding motifs in sequences adjacent to breakpoints and investigated whether Translin binds to these sequences in vitro by mobility-shift assay. Computer-assisted search found sequences highly homologous (>70%) with Translin binding motifs adjacent to the breakpoints in 10 out of 11 liposarcomas with the TLS-CHOP fusion genes. All of 13 oligonucleotides corresponding to the putative binding sequences in these cases bind to Hela cell extract and also recombinant Translin protein, although the binding affinity of each motif showed considerable differences. The DNA-protein complex formation was inhibited by non-labeled competitor or anti-Translin antibody, suggesting the specificity of the complex formation. Considering the high incidence and specific binding property, the presence of Translin binding motif may be one of the important determinants for the location of breakpoints in the TLS and CHOP genes in liposarcomas.

Characteristics of genomic breakpoints in TLS-CHOP translocations in liposarcomas suggest the involvement of Translin and topoisomerase II in the process of translocation.

Fusion of TLS/FUS and CHOP gene by reciprocal translocation t(12;16)(q32;q16) is a common genetic event found in myxoid and round-cell liposarcomas. Characterization of this genetic event was performed by three methods, Southern blot, RT-PCR, and genomic long-distance PCR in nine myxoid and three round-cell liposarcomas. All but one tumors showed genetic alternations indicating the fusion of TLS/FUS and CHOP gene. Two novel types of fusion transcripts were found, of which one lacked exon 2 sequence of CHOP gene, and the other lacked 3' half of exon 5 of TLS gene. The latter case was caused by a cryptic splicing site which was created by the genomic fusion. Detailed analyses genomic fusion points revealed several sequence characteristics surrounding the fusion points. Homology analyses of breakpoint sequences with known sequence motifs possibly involve in the process of translocation uncovered Translin binding sequences at both of TLS/ FUS and CHOP breakpoints in two cases. Translocations were always associated with other genetic alterations, such as deletions, duplications, or insertions. Short direct repeats were almost always found at both ends of deleted or duplicated fragments some of which had apparently been created by joining of sequences that flank the rearrangement. Finally, consensus topoisomerase II cleavage sites were found at breakpoints in all cases analysed, suggesting a role of this enzyme in creating staggered ends at the breakpoint. These data suggested that sequence characteristics may play an important role to recruit several factors such as Translin and topoisomerase II in the process of chromosomal translation in liposarcomas.

1 alpha,25-dihydroxyvitamin D3 inhibits cell growth and chondrogenesis of a clonal mouse EC cell line, ATDC5.

Here we report the effects of 1 alpha,25-dihydroxyvitamin D3 [1,25(OH)2D3] in vitro on the growth and chondrogenesis of a chondroprogenitor-like clonal mouse EC cell line, 10(-10) to 10(-7) M ATDC5. 1,25(OH)2D3 inhibited [3H]thymidine incorporation in undifferentiated chondroprogenitor-like ATDC5 cells in time- and dose-dependent manners. 1,25(OH)2D3 suppressed cartilage-nodule formation and the accumulation of cartilage-specific proteoglycan in ATDC5 cells in a dose-dependent manner. The 1,25(OH)2D3-induced inhibition of cartilage-nodule formation was reversible and direct, unrelated to the antiproliferative action of the hormone on the undifferentiated ATDC5 cells. ATDC5 cells even in the precartilaginous stage expressed 4.4 kb vitamin D receptor (VDR) mRNA as assessed by northern blot analysis. The equilibrium saturation binding experiment revealed the presence of a single class of saturable and high-affinity binding sites for 1,25(OH)2D3 in the cytosols. These results provide evidence for the hypothesis that both recruitment and chondrogenesis of chondroprogenitors are negatively regulated by 1,25(OH)2D3 via a VDR-mediated process in vivo.

Synovial fluids from patients with osteoarthritis and rheumatoid arthritis contain high levels of parathyroid hormone-related peptide.

High levels of immunoreactive and biologically active parathyroid hormone-related peptide (PTHrP) were detected in synovial fluids from patients with osteoarthritis (OA) and rheumatoid arthritis (RA). The levels of PTHrP immunoreactivity in synovial fluids, measured by a two-site immunoradiometric assay (IRMA) which detects hPTHrP(1-72) or longer peptides and a radioimmunoassay (RIA) specific to the carboxy-terminal portion of hPTHrP, were 3.2 +/- 0.3 pmol of hPTHrP(1-86)/l and 61 +/- 7.0 pmol of hPTHrP(109-141)/l in OA patients (mean +/- SE, n = 23), and 4.8 +/- 0.8 pmol of hPTHrP(1-86)/l and 164 +/- 30 pmol of hPTHrP(109-141)/l in RA patients (n = 26). Synovial fluid PTHrP levels distributed above the normal plasma reference ranges in each assay (0.7-2.6 pmol of hPTHrP(1-86)/l; 16-60.6 pmol of hPTHrP(109-141)/l). After concentration using sequential cation-exchange and reverse-phase chromatography, synovial fluid exhibited the activity that stimulated cyclic adenosine monophosphate (cAMP) accumulation in osteoblastic ROS 17/2.8 cells expressing PTH/PTHrP receptors. The cAMP accumulation activity in synovial fluid was sensitive to coincubation with excess hPTHrP(3-40), a PTH/PTHrP receptor antagonist, and was completely neutralized by preincubation with a monoclonal antibody specific to hPTHrP but not PTH. Immunohistochemical analysis of RA synovium revealed that PTHrP was localized in fibroblast-like cells in the synovial pannus invading articular cartilage. Our data show that PTHrP is produced locally by the diseased synovial tissue and released into synovial fluid at high concentrations, allowing us to hypothesize that PTHrP plays a novel role as a paracrine/autocrine factor in the pathology of OA and RA.

Chromosome 13 locus, Pbd2, regulates bone density in mice.

Bone density is inherited as a complex polygenic trait. Previously, we identified two quantitative trait loci (QTLs) specifying the peak relative bone mass (bone mass corrected by bone size) on chromosomes (Chrs) 11 and 13 by interval mapping in two mouse strains: SAMP2 and SAMP6. The latter strain is an established murine model of senile osteoporosis and exhibits a significantly lower peak relative bone mass than SAMP2 mice. In this study, we report the effects of the Chr 13 QTL on peak bone density (Pbd2). First, we constructed a congenic strain P6.P2-Pbd2b, which carried a single genomic interval from the Chr 13 of SAMP2 on an SAMP6-derived osteoporotic background, to dissect this polygenic trait into single gene factors. This congenic strain had a higher bone density than the background strain using three measurement methods with different principles for bone density. Next, we measured the peak relative bone mass of the AKR/J strain and the 13 senescence-accelerated mouse (SAM) strains, which are considered to be a series of recombinant-like inbred (RI) strains derived from the AKR/J strain and other unspecified strains. We then determined the microsatellite marker haplotypes of these strains around the Pbd2 locus, in which three strains with a high relative bone mass shared the same haplotype over the 26-centimorgan (cM) region. In the Pbd2 locus, a high relative bone mass was associated with alleles of the unknown strain, whereas a low relative bone mass was associated with the alleles from the AKR/J strain. These results confirmed the existence of a Pbd2 locus regulating bone density in the SAM strains.

Spontaneous spongy degeneration of the brain stem in SAM-P/8 mice, a newly developed memory-deficient strain.

A spontaneous spongy degeneration of the brain stem and spinal cord was discovered in a murine model of accelerated senescence (SAM), cared for under both conventional (SAM-P/8) and specific pathogen-free (SAM-P/8/Ta) conditions. SAM-P/8 and SAM-P/8/Ta showed no clinical neurological abnormalities, yet there was a deterioration in learning and memory abilities. Light microscopic examination revealed a spongy degeneration in the brain stem and spinal cord, in the reticular formation, and proliferation of hypertrophic astrocytes in the spongy area. The spongiform degeneration progressed with advancing age from four to eight months, after which the entire brain was involved. Astrocytosis increased with advancing degeneration. Ultrastructurally, mild dendritic swelling occurred at one month of age. At two months of age, moderate postsynaptic swelling and a widening of intracellular membrane structure were observed, and at age five months there were large vacuoles circumscribed by membranous lamellae, identifiable as myelin. Vacuoles in SAM-P/8 proved to be swollen neuronal processes and oligodendroglial processes. These SAM-P/8 and SAM-P/8/Ta strains of mice are new memory-deficient strains with spontaneous spongy degeneration associated with aging.

Expression of the c-fos gene induced by parathyroid hormone in the bones of SAMP6 mice, a murine model for senile osteoporosis.

SAMP6 mice are a murine model for senile osteoporosis, characterized by low peak bone mass seen at 4 or 5 months of age. Parathyroid hormone (PTH)-induced c-fos expression was examined in the bones, bone-marrow cells and kidney tissues of 2-month-old male SAMP6 mice. SAMP2 mice, which have a higher peak bone mass, were used as controls. The expression of c-fos in the bone peaked at 30 min after 60 microg/kg of human PTH(1-34) administration. After peaking, the expression fell quickly in SAMP2 mice. This decrease in expression was delayed in SAMP6 mice and the expression was higher at 1 h than in SAMP2 mice. The phenomenon observed in the bone appears to be tissue specific as it was not seen in the bone-marrow cells or kidney tissue. Immunohistochemical studies showed that c-Fos protein was localized to the nuclei of some of the osteocytes and a few of the osteoblasts in the cortical bone, and that osteocytes expressing c-Fos protein increased after PTH treatment. These results suggest that osteocytes might contribute to the maintenance of higher levels of c-fos expression in the bones of SAMP6 mice and may be related to cortical osteopenia in these mice by modulating bone remodeling and/or modeling.

Modification of strain-specific femoral bone density by bone marrow chimerism in mice: a study on the spontaneously osteoporotic mouse (SAM-P/6).

The role of marrow-derived cells in determining strain differences in femoral bone density in mice was investigated by inducing marrow chimerism in a newly developed, osteoporotic strain of mouse: SAM-P/6. As a normative study prior to the chimeric experiment, bone density of F1 hybrids between SAM-P/6 and SAM-P/2 (strain with a high bone density) was assessed. Microdensitometrically, the F1(P2P6F1) exhibited a significantly higher bone density than did the SAM-P/6. P2P6F1 marrow cells, injected into SAM-P/6 neonates, increased the bone density at age 60 days while the syngeneic cells were without effect. The successful induction and maintenance of chimerism was confirmed by flow cytometry in which a donor-specific H-2K haplotype was used as the marker. This study shows that, at least in certain combinations of strains of mice, strain-specific bone density can be modified by bone marrow chimerism. Also, it suggests that the low bone density observed in the SAM-P/6 might reflect functional characteristics of the marrow-derived cells.

Effective intervention of low peak bone mass and bone modeling in the spontaneous murine model of senile osteoporosis, SAM-P/6, by Ca supplement and hormone treatment.

SAM-P/6 is a recently developed strain of osteoporotic mice. In this study we tried to determine whether calcium, vitamin D3, parathyroid hormone (PTH), and estrogen modified the peak bone mass of young SAM-P/6 mice, and whether the effect of these medications persisted after treatment had been discontinued. Calcium supplement, PTH, and estrogen treatment increased the peak bone mass of SAM-P/6 mice. To clarify the process by which bone mass was increased in these treated mice, we evaluated their bone formation and resorption by histomorphometry and measured the levels of ions and serum enzymes relevant to bone metabolism. We found that bone formation was increased by calcium supplementation, and bone resorption was decreased by estrogen treatment. Furthermore, the effectiveness of calcium supplement on peak bone mass was retained after treatment had been discontinued, but the effect of estrogen treatment on peak bone mass was reduced after estrogen treatment had been discontinued. The results of this study indicate that calcium supplementation and estrogen and PTH treatment each increased peak bone mass at the midpoint of the femur of SAM-P/6, and that the effect of calcium supplementation, but not that of estrogen treatment, persisted after treatment was discontinued.

Definitive intraoperative very high-dose radiotherapy for localized osteosarcoma in the extremities.

PURPOSE: To evaluate the outcome and adverse effects in patients with osteosarcoma treated with very high-dose definitive intraoperative radiotherapy (IORT), with the intention of saving the affected limb. METHODS AND MATERIALS: Thirty-nine patients with osteosarcoma in their extremities were treated with definitive IORT. The irradiation field included the tumor plus an adequate wide margin and excluded the major vessels and nerves. Forty-five to 80 Gy of electrons or X-rays were delivered. The median follow-up of the surviving patients was 124 months. RESULTS: The cause-specific and relapse-free 5-year survival rate was 50% and 43%, respectively. Distant metastasis developed in 23 patients; 19 died and 4 were alive for >10 years. Nine local recurrences were found 4-29 months after IORT in the affected limb. No radiation-induced skin reaction or nerve palsy was observed in the patients treated with X-rays. Experiments using phantoms also confirmed that the scatter dose was below the toxic level in the IORT setting with X-rays. CONCLUSIONS: Very high-dose definitive IORT combined with preventive nailing and chemotherapy appeared to be a promising quality-of-life-oriented alternative to treating patients with osteosarcomas in the extremities, although the problem of recurrences from the surrounding unirradiated soft tissue remains to be solved.

Effect of bone cylinder length on distraction osteogenesis in the rabbit tibia.

The viability and osteogenic potential of bone cylinders for bone transport was investigated in one tibia of 18 mature male New Zealand rabbits. The length of the bone cylinder was equal to or twice that of the diameter of the tibia. The cylinder was cut subperiosteally with an externally cooled oscillating saw from a lateral approach, after a specific unilateral external fixator had been applied. To simulate bone transport, one end of the cylinder was fixed to the distal bone stump by a cerclage wire and healing and revascularization was prevented by an interposed expanded polytetrafluoroethylene membrane. The periosteum was re-adapted and sutured, and distraction began 10 days postoperatively at 0.25 mm/12 hours for 22 days. New bone formation in the distraction gap was quantified by dual energy x-ray absorptiometry and by computer-assisted histomorphometry of polyfluorochrome-labeled undecalcified bone sections and corresponding microradiographs. In half of the animals with each size cylinder, osseous bridging occurred, so the findings on distraction osteogenesis are reported only for the remaining nine animals. Generally, centripetal mineralization of the gap with two distinct zones of ripening bone structures and a central radiolucent fibrocartilaginous zone could be distinguished. Neither absorptiometry nor histomorphometry showed significant differences in the overall amount of this new bone formation for the bone cylinders of two different lengths. However, osteogenesis was significantly greater at the proximal end than at the cylinder. New bone was formed predominantly from endosteal sites in the smaller cylinders and from periosteal sites in the larger cylinders. Histologically, there was complete necrosis of both sizes of cylinders, followed by revitalization through newly formed vascular channels.(ABSTRACT TRUNCATED AT 250 WORDS)

Recorticalization after bifocal internal bone transport in the double-plated sheep femur.

Distraction osteogenesis and bone remodelling after the end of bone transport were investigated in one femur each of six adult male sheep. A newly designed internal distraction device was used. A custom-made osteosynthesis plate was fixed on the lateral side of the femur, and two transporting plates driven by a transcutaneously inserted screwdriver moved two bone cylinders simultaneously over a 40 mm defect. An additional plate was applied ventrally to stabilize the device. Bone transport was begun 2 weeks postoperatively at 1 mm/day at each transporting plate until they came into contact. New bone formation within the distraction gaps was evaluated by computed tomography scans and was quantified at 4 and 6 months by dual energy x-ray absorptiometry as well as histomorphometry on polyfluorochrome-labelled undecalcified ground sections and microradiographs. At 4 months, all distraction gaps were bridged by abundant newly formed bone trabeculae, which were reduced and condensed to cortex-like layers of new bone at 6 months. Less new bone was always found under the lateral device than in the unplated medical and dorsal segments, but the amount of new bone under the ventral plate was comparable with that in the unplated regions. The results of this pilot study show that distraction osteogenesis can be achieved with an internal device such as this one and that recorticalization and restoration of a medullary canal occur despite the relatively rigid internal stabilization by the plates.

Amplification of the CDK4 gene in sarcomas: tumor specificity and relationship with the RB gene mutation.

Amplification of the CDK4 gene, which encodes a key molecule in the cell cycle, has been shown in some types of human neoplasms, including bone and soft tissue tumors. It is also reported that the CDK4 gene is coamplified with other sequences in the 12q13-15 region, including the MDM2 and SAS genes. Using 146 DNA samples derived from a variety of bone and soft tissue tumors, we have studied the pattern of amplification of these three genes, CDK4, MDM2, and SAS, to investigate whether there are any tumor type specific patterns of amplification. Amplification of at least one of these three genes was found in 18 tumors, and five different patterns of amplification were observed. Amplification of all of these three genes was detected in 9 cases. Amplification of the CDK4 gene without MDM2 amplification was observed in osteosarcomas and a chondrosarcoma but not in soft tissue tumors, whereas amplification of MDM2 gene alone was observed in malignant fibrous histiocytomas (MFHs), liposarcomas, and lipomas, but not in bone tumors. These results suggested that the CDK4 region is the primary target for amplification in bone tumors, whereas the MDM2 region is in soft tissue tumors. We also investigated the relationship of CDK4 amplification with retinoblastoma (RB) gene mutations in osteosarcomas, for which we have already performed the mutation analyses in detail. Interestingly, contrary to the prevailing theory that CDK4 amplification is an alternative mechanism for RB gene mutation, we found that three of four cases with amplification of the CDK4 gene showed loss of expression of the RB protein, one of which was proved to have an gross DNA alteration in the RB locus. This redundancy of mutations may indicate that the amplification of CDK4 may have some roles other than the inactivation of the RB protein in the development of osteosarcomas.

Uptake of pentavalent technetium-99m dimercaptosuccinic acid in idiopathic synovial chondromatosis.

We reported two Tc-99m(V) DMSA scintigrams in patients with idiopathic synovial chondromatosis which affected the metacarpo-phalangeal joint and shoulder joint. Tc-99m(V) DMSA accumulated markedly and diffusely in the tumor. Tc-99m(V) DMSA scintigraphy would be valuable for deciding the optimal site for biopsy.

MRI and scintigraphic features of extraabdominal desmoid tumors.

To determine whether extraabdominal desmoid can be correctly diagnosed using both magnetic resonance imaging (MRI) and scintigraphy with pentavalent technetium-99m dimercaptosuccinic acid and gallium-67 citrate, MRI (T1- and T2-weighted images) and scintigraphy were performed in 18 patients with 27 histologically proved extraabdominal desmoid tumors. The extraabdominal desmoid tumors were characterized by positive uptake of pentavalent technetium-99m dimercaptosuccinic acid and lack of uptake of gallium-67 citrate on scintigraphy. These tumors were isointense to skeletal muscle on T1-weighted MRIs and hyperintense on T2-weighted images. They also displayed septum-like internal inhomogeneity and surrounding hypointense capsular band. The combination of scintigraphy and MRI is of value in correctly diagnosing extraabdominal desmoid tumors except in the rare case of fibrotic verylow-grade sarcoma.

3D-spiral CT of multiple exostoses.

We report pre- and post-operative three-dimensional (3D)-spiral CT images in a patient with multiple exostoses. Images of 3D-CT, which were performed using the integrated 3D software of the CT system, showed the exact shapes and locations of the individual tumors around the knee joint in comparison with the surgical findings and resected specimen. 3D-spiral CT images of multiple exostoses would be useful for the planning of surgical procedure.

Solitary muscular involvement by tuberculosis: CT, MRI, and scintigraphic features.

The authors reported computed tomography (CT), magnetic resonance imaging (MRI) and scintigraphic findings of a patient with solitary muscular tuberculosis in the forearm. All these findings resembled those of other granulomatous inflammatory lesions in the soft tissue such as muscular sarcoidosis.

Diagnostic value of Tc-99m (V) DMSA for chondrogenic tumors with positive Tc-99m HMDP uptake on bone scintigraphy.

Technetium-99m (V) DMSA scintigraphy was performed in 17 patients with 37 chondrogenic tumors (13 osteochondromas, 14 enchondromas, and 10 chondrosarcomas) that had previously shown uptake of Tc-99m HMDP. Technetium-99m (V) DMSA showed high uptake by all chrondrosarcomas, but low or no uptake always indicated benign chondrogenic tumors. Technetium-99m (V) DMSA scintigraphy may be superior to Tc-99m HMDP scintigraphy for distinguishing benign and malignant chondrogenic tumors, and could also be useful for diagnosing the malignant transformation of chondrogenic tumors.