Core cell cycle regulatory genes in rice and their expression profiles across the growth zone of the leaf.

Rice (Oryza sativa L.) as a model and crop plant with a sequenced genome offers an outstanding experimental system for discovering and functionally analyzing the major cell cycle control elements in a cereal species. In this study, we identified the core cell cycle genes in the rice genome through a hidden Markov model search and multiple alignments supported with the use of short protein sequence probes. In total we present 55 rice putative cell cycle genes with locus identity, chromosomal location, approximate chromosome position and EST accession number. These cell cycle genes include nine cyclin dependent-kinase (CDK) genes, 27 cyclin genes, one CKS gene, two RBR genes, nine E2F/DP/DEL genes, six KRP genes, and one WEE gene. We also provide characteristic protein sequence signatures encoded by CDK and cyclin gene variants. Promoter analysis by the FootPrinter program discovered several motifs in the regulatory region of the core cell cycle genes. As a first step towards functional characterization we performed transcript analysis by RT-PCR to determine gene specific variation in transcript levels along the rice leaves. The meristematic zone of the leaves where cells are actively dividing was identified based on kinematic analysis and flow cytometry. As expected, expression of the majority of cell cycle genes was exclusively associated with the meristematic region. However genes such as different D-type cyclins, DEL1, KRP1/3, and RBR2 were also expressed in leaf segments representing the transition zone in which cells start differentiation.

Mycotoxin-degradation profile of Rhodococcus strains.

Mycotoxins are secondary fungal metabolites that may have mutagenic, carcinogenic, cytotoxic and endocrine disrupting effects. These substances frequently contaminate agricultural commodities despite efforts to prevent them, so successful detoxification tools are needed. The application of microorganisms to biodegrade mycotoxins is a novel strategy that shows potential for application in food and feed processing. In this study we investigated the mycotoxin degradation ability of thirty-two Rhodococcus strains on economically important mycotoxins: aflatoxin B1, zearalenone, fumonisin B1, T2 toxin and ochratoxin A, and monitored the safety of aflatoxin B1 and zearalenone degradation processes and degradation products using previously developed toxicity profiling methods. Moreover, experiments were performed to analyse multi-mycotoxin-degrading ability of the best toxin degrader/detoxifier strains on aflatoxin B1, zearalenone and T2 toxin mixtures. This enabled the safest and the most effective Rhodococcus strains to be selected, even for multi-mycotoxin degradation. We concluded that several Rhodococcus species are effective in the degradation of aromatic mycotoxins and their application in mycotoxin biodetoxification processes is a promising field of biotechnology.

Synovial sarcomas usually metastasize after >5 years: a multicenter retrospective analysis with minimum follow-up of 10 years for survivors.

BACKGROUND: Synovial sarcoma (SS) is a malignant soft tissue sarcoma with a poor prognosis because of late local recurrence and distant metastases. To our knowledge, no studies have minimum follow-up of 10 years that evaluate long-term outcomes for survivors. PATIENTS AND METHODS: Data on 62 patients who had been treated for SS from 1968 to 1999 were studied retrospectively in a multicenter study. Mean follow-up of living patients was 17.2 years and of dead patients 7.7 years. RESULTS: Mean age at diagnosis was 35.4 years (range 6-82 years). Overall survival was 38.7%. The 5-year survival was 74.2%; 10-year survival was 61.2%; and 15-year survival was 46.5%. Fifteen patients (24%) died of disease after 10 years of follow-up. Local recurrence occurred after a mean of 3.6 years (range 0.5-14.9 years) and metastases at a mean of 5.7 years (range 0.5-16.3 years). Only four patients were treated technically correctly with a planned biopsy followed by a wide resection or amputation. Factors associated with significantly worse prognosis included larger tumor size, metastases at the time of diagnosis, high-grade histology, trunk-related disease, and lack of wide resection as primary surgical treatment. CONCLUSIONS: In SS, metastases develop late with high mortality. Patients with SS should be followed for >10 years.

[Giant cell tumor of bone with rapid malignant course]. / Riesenzelltumor des Knochens mit rapid malignem Verlauf.

The case of a 28-year-old male patient with a locally aggressive lesion of the distal tibia is presented. Following the diagnosis of giant cell tumor of bone (GCT) on biopsy and curettage, a rapid malignant course was observed with recurrence 2.5 months later. Multiple metastases appeared 6 months after initial presentation. Following initial chemotherapy according to the COSS protocol and later with carboplatin and VP-16, therapy was changed to Adriamycin and later gemcitabine due to progressive disease. Good palliation was achieved, and the patient felt well with less shortness of breath on exertion and was ambulatory with walking aids. The malignant nature of the tumor was not detected in the initial pathologic examinations. Review of the pathologic material provided histologic clues permitting the diagnosis of a primary malignant GCT with a fibrohistiocytic/fibrosarcomatous component. Malignancy in a giant cell tumor is a much debated diagnostic dilemma when a frank sarcomatous component is lacking. Cytologic atypias and flame-like tufts of infiltration of soft tissue are important clues. Surgical treatment should be commensurate. Monotherapy with Adriamycin or gemcitabine can be considered in order to inhibit the disease progression.

Preoperative radiotherapy without femoral shielding for prevention of heterotopic ossification in hydroxyapatite-coated hip prostheses.

Twenty-seven consecutive patients at high risk of developing heterotopic ossifications (HO) after implantation of a hydroxyapatite (HA)-coated hip prosthesis were irradiated with a single dose of 7 Gy, at least 4 h before the operation. The femoral stem was not shielded during radiotherapy (RT). After a median follow-up of 14.8 months, no clinically significant HO could be found, while 12 (52%) patients in this high-risk population had only minor HO (grade I). No reoperation was needed, and no evidence of prosthesis migration was observed. We conclude that single-dose, preoperative RT for HA-coated hip prosthesis can effectively inhibit HO. Not blocking the femoral stem does not result in prosthesis migration.

Zürich experience with preoperative, high dose methotrexate-containing chemotherapy in patients with extremity osteosarcomas (OSA).

From 1979 to 1990, 37 patients with extremity osteosarcomas, 22 of them males and 15 females, median age 19 years, received pre- and postoperative chemotherapy. The period of observation, calculated from after the primary operation, ranged from 1-114 months, median 25 months. After preoperative chemotherapy, 9 (24%) underwent a primary amputation, in 28 (76%) a limb salvage procedure was possible, 4/29 (14%) later developed local recurrences, metastases were diagnosed in 8/9 amputees and 5/28 after limb-sparing surgery. The five-year disease-free and overall survivals after amputation are 11% and 33%, respectively, compared to 68% and 75%, respectively (p = 0.001 and 0.018, respectively, for long rank). Results of histologic assessment after preoperative chemotherapy are of significant prognostic impact. The earlier prognostic groups of 0%-49% necrosis versus 50%-100% necrosis were statistically no longer suitable for distinguishing useful prognostic groups. In this second analysis, patients with 0%-79% necrosis versus 80%-100% necrosis had 5-year disease-free survivals of 81% versus 44% (p = 0.032) and 5-year overall survivals of 88% In summary, 24% of our patients with extremity osteosarcomas, most of them with large primaries close to joints, had to undergo primary amputation and had only a 33% 5-yr-survival, whereas limb salvage procedures with pre-and postoperative chemotherapy were associated with a 5-yr survival of 75% and thus had no adverse impact. Careful selection of patients for successful management of osteosarcomas is important; necrosis of 80% and more after preoperative chemotherapy is a prerequisite for a favourable outcome.

A case of congenital non-progressive sensory neuropathy with tonic pupils.

One case of a non-progressive congenital neuropathy is reported. Clinical findings included subtotal analgesia, and diminished temperature, vibration and proprioceptive sense in arms and legs. The sensory nerve action potentials were absent. Autonomic dysfunctions were restricted to tonic pupils. Sural nerve biopsy taken at the age of 8 1/2 showed fascicular hypoplasia, subtotal loss of myelinated nerve fibers and severe loss of unmyelinated nerve fibers with a pathological size distribution. In the skin biopsy, including several cutaneous nerves, no myelinated nerve fibers were present, the unmyelinated nerve fibers were severely reduced and the collagen density was increased. Perivascular and periglandular innervation was significantly reduced. The findings in this case are suggestive of a malformative or fetally acquired lesion and further illustrate the difficulties in classification of the hereditary sensory neuropathies.

[Value of punch and puncture drill biopsy in the diagnosis of tuberculous spondylitis]. / Die Wertigkeit der Stanz- und Punktionsbohrbiopsie in der Diagnostik der Spondylitis-Tbc.

Thanks to a specially designed guide instrumentarium, punch and drill aspiration biopsy for the vertebrae and intervertebral spaces have become routine diagnostic procedures which any patient can be expected to tolerate. With the authors' puncture technique material can be obtained from a single vertebra at various heights or simultaneously from different segments with the drilling cannula. For an experienced osteopathologist histologic assessment of the tissue cylinders obtained or of aspirated material presents no problems. Percutaneous removal of samples by this method avoids an open diagnostic intervention with its considerably greater attendant risks, or enables a therapeutic intervention to be prepared with greater precision. The authors report on their own experience of drill aspiration biopsy in 35 fully documented patients and discuss considerations regarding indication and confirmation of the diagnosis.

Effects of leukocyte interferon (E. coli) on human bone sarcoma growth in vitro and in the nude mouse.

Effects of highly purified human leukocyte interferon (rIFN-alpha 2) on colony formation, DNA synthesis and proliferation in nude mice of tumor cells from eight bone sarcomas have been studied. rIFN-alpha 2 produced a dose-dependent inhibition of [3H]thymidine incorporation by sarcoma cells. Even at high doses (10(4) U/ml), however, [3H]thymidine uptake could not be completely blocked by rIFN-alpha 2. In a cloning assay three established sarcoma cell lines and five other sarcoma samples obtained after short-term in vitro culture were found to be sensitive to various degrees to rIFN-alpha 2, complete inhibition being seen only at 10(4) U/ml. Three sarcomas were sensitive in the nude mouse model. Scheduling experiments revealed that rIFN-alpha 2 produces a delay in tumor growth only when administered either before or shortly after tumor implantation. Therefore rIFN-alpha 2 appears to be most active when tumor size is small and growth not exponential, indicating that rIFN-alpha 2 may play a role in an adjuvant setting. Growth sarcomas strongly suppressed by rIFN-alpha 2 in the cloning assay was markedly inhibited in the nude mouse. One sarcoma which was only moderately sensitive in the cloning assay was resistant in the animal experiment, confirming the predictive value of the clonogenic assay. Although the present findings demonstrate strong antitumor activity of rIFN-alpha 2 against human bone sarcoma cells they should be interpreted with caution mainly because the high rIFN-alpha 2 levels used in the experiments cannot be maintained in patients over a prolonged period.

High-dose ifosfamide in advanced osteosarcoma.

In a prospective study, 18 evaluable patients with recurrent osteosarcoma were treated with ifosfamide, 1.8 g/m2 daily for 5 consecutive days. Courses were repeated every 4 weeks. Additional mesna (2-mercaptoethane sulfonate) was given to prevent urotoxicity. All patients had measurable lung deposits and all but one had been pretreated with various cytotoxic agents. Six patients (33%) showed therapeutic response, two complete and four partial, with a median duration of 5.5 months (range, 3-47+). Toxicity included myelosuppression, alopecia, nausea, and vomiting. No severe urotoxicity or central nervous system toxicity was observed. Thus, high-dose ifosfamide in combination with mesna seems to be a safe and effective agent for the chemotherapy of osteosarcoma.

Clinical experience with preoperative chemotherapy for osteosarcoma.

Since 1978 preoperative chemotherapy has been administered to 15 consecutive patients with osteosarcomas in Zurich. Preoperative chemotherapy was acceptably well tolerated and did not impair surgical procedures. Our retrospective analysis confirmed that the extent of necrosis after preoperative chemotherapy is of biological importance for the further course of the disease. Patients with extensive necrosis had better relapse-free survival and longer overall survival than those with little necrosis.

[Occurrence of pannus in arthrosis]. / Das Vorkommen von Pannus bei Arthrose.

Eight human femoral heads, surgically removed from patients with the clinical diagnosis: coxarthrosis or necrosis, were examined. In six of these we found severe arthrosis combined with synovitis and pannus formation. The pannus appeared as granulation tissue or fibrous connective tissue covering the surface of the cartilage or growing into it. In most cases the pannus showed a destructive appearance, although newly formed cartilage repair tissue was also observed. There was no indication for a pannus originating from the bone marrow. We therefore assume that we are dealing with a pannus of synovial origin. The possibility of a local transformation of chondrocytes into fibroblasts, is also discussed. The earlier reports concerning the occurrence of arthrosis in combination with pannus, are contradictory. We suggest that coxarthrosis can occur both with and without pannus. These two possibilities may represent different forms of arthrosis.

[The Zurich experience with preoperative high-dose methotrexate in osteosarcomas]. / Zürcher Erfahrungen mit präoperativem, hochdosiertem Methotrexat bei Osteosarkomen.

15 consecutive patients with osteosarcoma underwent preoperative chemotherapy with high dose methotrexate (HDMTX) containing regimens according to the T7 or T10 protocols of ROSEN, Preoperative chemotherapy was well tolerated and did not impair surgical procedures. 67% of the patients responded clinically with reduction of pain and tumor size. Histologic examination of the tumor after preoperative chemotherapy revealed extensive necrosis in 53% of patients. In a retrospective analysis, patients with extensive necrosis (group B) were compared with those with little or no necrosis (group A). Patients from group B had a longer relapse free and overall survival period than group A. In addition, patients of group A had significantly higher initial levels of alkaline phosphatase than group B. The incidence of a 2.5-fold increase of the transaminases 2-3 days after HDMTX was significantly greater in patients of group B compared to group A. In the absence of documented necrosis after chemotherapy according to the T7 or T10 protocols, further use of HDMTX is not indicated. New aspects on the treatment of osteosarcoma, derived from recent publications, are discussed.

Evaluation of magnification radiography in bone tumors.

The additional information obtained by direct radiographic magnification is examined in comparison to standard exposures of 50 bone tumors. A diagnostically significant increase in information is obtained in 84% of these cases. This is distributed at an irregular rate over the various decisive phases of the tumor diagnosis. In 24% of the cases the magnification exposure was helpful excluding other diseases, in 40% it permitted a differentiation between benign and malignant lesions and in 20% it permitted a classification of the type. The clinical importance of the magnification technique is relevant.

Histomorphological investigations of coxa femoral ends following double-cup arthroplasty according to Freeman.

Of 20 shell prostheses inserted in cases of early coxarthrosis in our clinic during the period from mid-1976 to the end of 1978, three had to be removed. Morphological examinations, employing different techniques, were made of the coxa femoral ends which were removed. The foreign body reaction of the bone tissue beneath the cement within the prosthetic shell and the biomechanical response were investigated on the basis of macroscopic and microscopic structural analyses. Dependent upon the time elapsed, the exposed spongiosa surface was observed to have become remodeled to a thin cortical protective layer. Independent of time between insertion and removal of the prosthetic shell, a continuous wide layer, consisting of cellular and fibrous tissue, was formed between the bone and the cement (which remained intact) even after the formation of the above-mentioned cortical bone substrate.