Scoliosis, diabetes mellitus and total laminectomy at the 4th lumbar vertebra are independent risk factors for post-laminectomy fracture around the isthmus.

OBJECTIVE: We aimed to identify the risk factors and clinical outcomes for post-laminectomy fracture around the isthmus, which can cause back pain or radiculopathy. METHODS: We performed a retrospective cohort study involving all patients who underwent laminectomy splitting the spinous process for lumbar spinal stenosis between 2010 and 2014. The primary outcome measure was post-laminectomy fracture around the isthmus. Clinical outcomes were evaluated based on reoperation rate. To evaluate risk factors for fracture, the following parameters were collected: (1) patient characteristics and concomitant diabetes mellitus, (2) lumbar scoliosis and sagittal alignment parameters, and (3) surgical data, such as rate of total laminectomy. Logistic regression analysis was performed to identify the independent risk factors for post-laminectomy fracture. RESULTS: Twelve of the 92 patients suffered a post-laminectomy fracture around the isthmus. Logistic regression analysis revealed that diabetes mellitus (odds ratio [OR]: 15.41; 95% confidence interval [CI]: 2.93-80.98; P=0.001), L4 total laminectomy (OR: 14.68; 95% CI: 1.51-142.76; P=0.021), and lumbar scoliosis (OR: 5.72; 95% CI: 1.16-28.21; P=0.032) were independent risk factors. The fracture group included 2 patients (16.7%) who required reoperation at the decompression level for recurrent leg pain, whereas the non-fracture group included 2 (2.5%) who underwent reoperation at a level different from the index procedure. CONCLUSIONS: Post-laminectomy fractures around the isthmus were significantly associated with scoliosis, diabetes mellitus, and total laminectomy at L4. Total laminectomy at L4 is best avoided to reduce the risk of post-laminectomy fracture in patients with scoliosis or diabetes mellitus.

Temperature distributions of electron beam-irradiated samples by scanning electron microscopy.

An electron beam (EB) generated by a scanning electron microscope (SEM) was used to irradiate two samples having different thermal conductivities, and the resulting temperatures of the EB-irradiated areas as well as the temperature distributions within the samples were then measured using a thermal camera. These measurements showed overall increases in sample temperatures, as well as revealed temperature rises at the EB-irradiated areas that had little difference with one of the theoretical predictions. Differences between the actual and the predicted temperature measurements were analysed in terms of the accuracy with which parameters could be estimated. The temperature distributions of the samples were measured and, On the basis of the results, it was hypothesized that the temperature differential over an irradiated sample will be inversely correlated with its thermal conductivity.

Differences between subtotal corpectomy and laminoplasty for cervical spondylotic myelopathy.

OBJECTIVE: This study aimed to obtain guidelines for choosing between subtotal corpectomy (SC) and laminoplasty (LP) by analysing the surgical outcomes, radiological changes and problems associated with each surgical modality. STUDY DESIGN: A retrospective analysis of two interventional case series. SETTING: Department of Orthopaedic Surgery, Kagawa University, Japan. METHODS: Subjects comprised 34 patients who underwent SC and 49 patients who underwent LP. SC was performed by high-speed drilling to remove vertebral bodies. Autologous strut bone grafting was used. LP was performed as an expansive open-door LP. The level of decompression was from C3 to C7. Clinical evaluations included recovery rate (RR), frequency of C5 root palsy after surgery, re-operation and axial pain. Radiographic assessments included sagittal cervical alignment and bone union. RESULTS: Comparisons between the two groups showed no significant differences in age at surgery, preoperative factors, RR and frequency of C5 palsy. Progression of kyphotic changes, operation time and volumes of blood loss and blood transfusion were significantly greater in the SC (two- or three-level) group. Six patients in the SC group required additional surgery because of pseudoarthrosis, and four patients underwent re-operation because of adjacent level disc degeneration. In the LP group, the problem of elimination of postoperative axial symptoms remains to be solved. CONCLUSIONS: The merit of SC is the low frequency of axial symptoms. One-level SC can be considered to have similar degree of invasiveness as LP. Compared with SC, LP is more suitable for elderly patients with multilevel stenosis.

Collagen maturity, glycation induced-pentosidine, and mineralization are increased following 3-year treatment with incadronate in dogs.

UNLABELLED: Collagen cross-linking is a determinant of bone quality. A three-year treatment of bisphosphonate-incadronate disodium-in beagles increased degree of mineralization, collagen maturity, and pentosidine, a compound with advanced glycation end products. The treatment had no effect on the total amount of enzymatic cross-link formation. INTRODUCTION: Collagen cross-linking is a determinant of bone quality. Recently, we reported that long-term treatment with bisphosphonate increased microdamage accumulation. The aim of this study was to clarify the effect of a three-year treatment with bisphosphonate on degree of mineralization and immature and mature enzymatic cross-links and non-enzymatic collagen cross-link, pentosidine, in cortical bone in the same dogs. METHODS: Twenty-nine 1-year-old beagles (15 males, 14 females) were divided into three groups that daily were given vehicle or incadronate at doses of 0.3 or 0.6 mg/kg/day orally for three years. A cortex of a rib was fractionated into low- and high-density portions. The contents of calcium, phosphorus, enzymatic immature and mature cross-links, and the non-enzymatic glycation product pentosidine were determined in each fraction. RESULTS: Calcium, phosphorus, and pentosidine contents and the ratio of mature to immature cross-links increased significantly with incadronate in a dose-dependent manner, but the total amount of enzymatic cross-links was unchanged. The pentosidine content correlated inversely with cortical activation frequency (p < 0.01). CONCLUSION: Long-term suppression of bone remodeling by bisphosphonate increases degree of mineralization, collagen maturity, and non-enzymatic cross-linking.

TMC-69, a new antitumor antibiotic with Cdc25A inhibitory activity, produced by Chrysosporium sp. TC1068. Taxonomy, fermentation and biological activities.

A new antibiotic designated TMC-69 has been isolated from the fermentation broth of a fungal strain Chrysosporium sp. TC 1068. TMC-69 exhibited moderate in vitro cytotoxic activity. TMC-69-6H, a derivative of TMC-69 prepared by hydrogenation, possessed more potent in vitro cytotoxicity than TMC-69, and exhibited in vivo antitumor activity against murine P388 leukemia and B16 melanoma. TMC-69-6H was found to specifically inhibit Cdc25A and B phosphatases.

Aurantiamide acetate, a selective cathepsin inhibitor, produced by Aspergillus penicilloides.

Aurantiamide acetate was isolated from the fermentation broth of Aspergillus penicilloides for the first time. Aurantiamide acetate inhibited cysteine proteinases, in particular, cathepsin L (3.4.22.15) and B (3.4.22.1) with IC50 of 12 microM and 49 microM, respectively. In the adjuvant-arthritic rat model, subcutaneously administered 10 mg/kg body weight of this compound suppressed hind paw swelling.

Long and accurate PCR with a mixture of KOD DNA polymerase and its exonuclease deficient mutant enzyme.

DNA polymerase from Thermococcus kodakaraensis KOD1 (previously Pyrococcus sp. KOD1) is one of the most efficient thermostable PCR enzymes exhibiting higher accuracy and elongation velocity than any other commercially available DNA polymerase [M. Takagi et al. (1997) Appl. Environ. Microbiol. 63, 4504-4510]. However, when long distance PCR (>5 kbp) was performed with KOD DNA polymerase, amplification efficiency (product yield) becomes lower because of its strong 3'-5' exonuclease activity for proof-reading. In order to improve a target length limitation in PCR, mutant DNA polymerases with decreased 3'-5' exonuclease activity were designed by substituting amino acid residues in conserved exonuclease motifs, Exo I (Asp141-Xaa-Glu), Exo II (Asn210-Xaa-Xaa-Xaa-Phe-Asp), and Exo III (Tyr311-Xaa-Xaa-Xaa-Asp). Exonuclease activity and amplification fidelity (error rate) of the DNA polymerases were altered by mutagenesis. However, long and accurate PCR by a single-type of mutant DNA polymerase was very difficult. The wild-type DNA polymerase (WT) and its exonuclease deficient mutant (N210D) were mixed in different ratio and their characteristics in PCR were examined. When the mixed enzyme (WT and N210D) was made at the ratio of 1:40, long PCR (15 kbp) at lower mutation frequency could be efficiently achieved.

Selective isolation and distribution of the genus Planomonospora in soils.

For the screening of bioactive compounds and study of global distribution, a selective isolation method for Planomonospora strains by centrifugation from soil is examined. Planomonospora strains produced characteristic sporangia on the humic acid-trace salts gellan gum medium (pH 9.0) so that this genus was readily recognized on the isolation plate. High yields of motile spores were obtained by using a flooding solution containing 0.1% skim milk in 5 mM N-cyclohexyl-2-amino-ethanesulfonic acid (pH 9.0) followed by incubating the preparation at 32 degreesC for 90 min, centrifuging it at 1000 x g for 10 min, and further incubation at 32 degreesC for 60 min after centrifugation. By combining the techniques described above, we isolated 246 Planomonospora strains from 137 of the 1200 soil samples examined. Ninety-four percent of these strains were recovered from neutral to slightly alkaline soils (pH 7.0 to 9.0). Strains of P. venezuelensis group were obtained from 13 soil samples (1.1%), which were collected in Bolivia, Cyprus, Egypt, Greece, India, Japan, New Caledonia, and Turkey. Strains of this group appear widely distributed in the soil of tropical to temperate regions. To our knowledge, this is the first record that strains of this group have been isolated from a location other than Venezuela.

Primary subclavian venous thrombosis which developed after sleeping with the arm in an outstretched position: report of a case.

Primary subclavian venous thrombosis is more rare than secondary thrombosis. This type of thrombosis is called "effort thrombosis" or Paget-Schroetter syndrome, and develops after a strenuous effort of the superior limb. A day after a 55-year-old man got drunk and slept in the left lateral position in combination with an abducted and elevated position of the left superior limb, he became aware of swelling and an oppressive feeling in his left superior limb and was admitted 9 days later. Thrombus of the left axillary-subclavian vein was confirmed by venography, and thrombolytic therapy with urokinase was performed immediately. The left arm symptoms improved for the most part. Venography after the therapy revealed thrombolysis at the site of the axillary vein, while the subclavian vein enhanced the collateral vessel pathway. The patient was discharged on the seventh hospital day, and anticoagulant therapy with oral warfarin sodium has since been continued. This is considered to be a rare case of subclavian venous thrombosis caused by sleeping in an abnormal position with the arm outstretched.

Isolation and analysis of lipase-overproducing mutants of Serratia marcescens.

We have isolated a lipase-overproducing mutant, GE14, from Serratia marcescens 8000 after three rounds of N-methyl-N'-nitro-N-nitrosoguanidine mutagenesis. The mutant GE14 produced 95 kU/ml of extracellular lipase in the lipase medium, which was about threefold higher than that of produced by the original strain 8000. Enzymatic characteristics including specific activity of purified lipases from culture supernatants of GE14 and 8000 were almost same. The lipase gene (lipA) of GE14 contained two base substitutions; one in the promoter region and another in the N-terminal region of the lipA gene without an amino acid substitution. Promoter analysis using lipA-lacZ fusion plasmids revealed that these substitutions were responsible for the increase in the lipA expression level, independently. In contrast, no base substitution was found in the genes encoding the lipase secretion device, the Lip system. In addition, the genes coding for metalloprotease and the cell surface layer protein which are both secreted through the Lip system and associated with extracellular lipase production, also contained no base substitution. The strain GE14 carrying a high-copy-number lipA plasmid produced a larger amount of the extracellular lipase than the recombinant strains of 8000 and other mutants also did, indicating that GE14 was not only a lipase-overproducing strain, but also an advantageous host strain for overproducing the lipase by a recombinant DNA technique. These results suggest that the lipase-overproducing mutant GE14 and its recombinant strains are promising candidates for the industrial production of the S. marcescens lipase.

Selective isolation and study on the global distribution of the genus Planobispora in soils.

Planobispora strains, including the type strain, produced abundant sporangia on humic acid-vitamin gellan gum medium when more than 50 colonies grew on a plate. However, these strains did not produce sporangia on the aerial mycelium, a diagnostic characteristic of this genus, when fewer than 20 colonies grew on a plate. Trace salts, such as FeSO4 x 7H2O, MnCl2 x 4H2O, ZnSO4 x 7H2O, and NiSO4 x 6H2O, stimulated sporangium formation of Planobispora strains. Better sporangium formation of Planobispora strains was observed on the humic acid-trace salts gellan gum medium at pH 9.0 than at neutral pH. Moreover, an alkaline medium repressed the growth of three out of six Streptomyces strains so that this condition was effective for selective isolation of Planobispora strains. An alkaline flooding solution of skim milk, five antimicrobial agents, and dry heat treatment at 90 degrees C for 60 min were effective for selective isolation. Using these techniques, we examined the distribution of Planobispora strains by using 1467 soil samples collected from Japan and other parts of the world. One hundred and nineteen Planobispora strains were isolated from 51 soil samples (3.5% of the samples tested) that were collected in Ecuador, Egypt, French Guiana, India, and Madagascar. Planobispora strains were recovered only in tropical and subtropical soils. To our knowledge, this is the first record that Planobispora strains have been isolated from locations other than Venezuela, Namibia, or South Africa.

Pancreatitis induced by valproic acid: report of a case.

A 22-year-old woman with intellectual impairment, who had been taking valproic acid continuously for 19 years since being diagnosed with epiloia at the age of 3 years, presented to our hospital following the sudden development of epigastric pain. An abdominal computed tomography scan revealed acute exacerbation of chronic pancreatitis. Conservative treatment was initiated, despite which the pancreatitis became exacerbated, necessitating resection of the pancreatic head and duodenum. Histological examination of the resected specimens revealed a large number of pancreatic calculi in the main pancreatic duct, suggesting chronic pancreatitis with fibrosis at the periphery. The incidence of pancreatitis developing in association with valproic acid is unclear; however, only 40 such cases have been reported in the English literature. Most of the patients previously described presented with acute pancreatitis in the initial stage. However, the clinical course of our patient, with acute exacerbation following a relatively chronic course, was different from those previously described, suggesting the presence of chronic pancreatitis related to valproic acid.

[Treatment of metastatic seminoma by chemotherapy.: an experience].

PURPOSE: To describe the outcome of chemotherapy using cisplatin-based regimen, and experimental combination with carboplatin and ifosfamide to treat advanced seminoma. METHODS: From 1981 to Jan. 1999, 15 patients with Stage IIA, IIB, IIIA or IIIC metastatic seminoma and one patient with lung disease, who suffered a relapse of his primary mediastinal lesion were treated. Three of these patients had relapsed, following surveillance for Stage I testicular cancer, and another had received prophylactic radiotherapy to the retroperitoneal lymph nodes in advance. The first patient's regimen consisted of cisplatin and cyclophosphamide. Since 1983, cases have been treated with the same regimen as that used to treat non-seminomatous germ-cell tumors; cisplatin/vinblastine/bleomycin (PVB); cisplatin/vinblastine/actinomycin D/cyclophosphamide/bleomycin (VAB-6); cisplatin/etoposide/bleomycin (BEP). From 1993, six patients with non-bulky metastatic seminoma participated in a trial involving 3 courses of carboplatin (400 mg/m2) and ifosfamide (2,000 mg/m2, 3 days). RESULTS: Of the entire group, 10 patients (62.5%) achieved a CR after chemotherapy alone. Four cases who received radiation, following chemotherapy, produced CR. Surgical resection of residual tumors were performed on 2 patients. Resected tumors were fibrous and no evidence of malignancy. All those individuals who participated in this study, are alive and disease-free today, from 11 months to 18 years. Carboplatin and ifosfamide demonstrated only mild toxicity, during a 4-week cycle, with subjects being treated on an outpatient basis. CONCLUSION: As expected, the type of chemotherapy we used, to treat non-seminomatous germ-cell tumors proved to be highly effective for seminomatous types, as well. Carboplatin and ifosfamide performed well and safe, in the treatment of non-bulky metastatic seminoma. Comparative studies of long-term treatment results and QOL, using either radiotherapy or low-toxicity chemotherapy for Stage IIA disease should be undertaken.

Selective isolation and distribution of Actinobispora strains in soil.

A simplified enrichment method for selective isolation of Actinobispora strains from soil is described. Actinobispora spores were tolerant to dry-heat treatment at 110 degrees C for 15 min. Actinobispora was more resistant to 1 microgram/mL leucomycin, 1 microgram/mL novobiocin, and 0.5 microgram/mL tunicamycin than Streptomyces dominant in soil, which prevents selective isolation of Actinobispora. Percentages of Actinobispora colonies on the isolation plate were increased by addition of antibiotics and dry-heat treatment of the soil samples. By combining the techniques described above, this genus was isolated from 105 out of 574 soil samples (18% of the samples tested). It was recovered from the soil samples with pH values ranging 5.0 to 8.9, and 78% of strains were isolated from neutral soil (pH 6.0-8.0). A number of Actinobispora strains were isolated from various soils around the world. Actinobispora strains are widely distributed in the world at relatively high frequency.

Structures of TMC-95A-D: novel proteasome inhibitors from Apiospora montagnei sacc. TC 1093.

Four novel proteasome inhibitors, TMC-95A-D (1-4) have been isolated from the fermentation broth of Apiospora montagnei Sacc. TC 1093, isolated from a soil sample. All of the molecular formulas of 1-4 were established as C(33)H(38)N(6)O(10) by high-resolution FAB-MS. Their planar structures were determined on the basis of extensive analyses of 1D and 2D NMR, and degradation studies. Compounds 1-4 have the same planar structures to each other, and are unique highly modified cyclic peptides containing L-tyrosine, L-aspargine, highly oxidized L-tryptophan, (Z)-1-propenylamine, and 3-methyl-2-oxopentanoic acid units. The absolute configuration at C-11 and C-36 of 1-4 was determined based on chiral TLC and HPLC analyses of their chemical degradation products. The ROESY analysis along with (1)H-(1)H coupling constants clarified the absolute stereochemistry at C-6, -7, -8, and -14 of the cyclic moieties. These studies revealed the relationships of 1-4 to be diastereomers at C-7 and C-36.

TMC-95A, B, C, and D, novel proteasome inhibitors produced by Apiospora montagnei Sacc. TC 1093. Taxonomy, production, isolation, and biological activities.

In our course of screening for novel proteasome inhibitors, TMC-95A and its diastereomers, TMC-95B to D, were isolated from the fermentation broth of Apiospora montagnei Sacc. TC 1093. TMC-95A inhibited the chymotrypsin-like (ChT-L), trypsin-like (T-L), and peptidylglutamyl-peptide hydrolyzing (PGPH) activities of 20S proteasome with IC50 values of 5.4nM, 200nM, and 60nM, respectively. TMC-95B inhibited these activities to the same extent as TMC-95A, while the inhibitory activities of TMC-95C and D were 20 to 150 times weaker than that of TMC-95A and B. TMC-95A did not inhibit m-calpain, cathepsin L, and trypsin at 30 microM, suggesting its high selectivity for proteasome. Taxonomy of the producing strain is also described.