Usefulness of multiphase computed tomography angiography in a patient with transient ischemic attack in the hyperacute phase: A case report.

RATIONALE: Patients with transient ischemic attack (TIA) have the possibility of developing stroke in the future. To prevent recurrent TIA or future stroke, identifying the cause of TIA is important. However, about two-third of patients with TIA have negative findings on diffusion-weighted imaging (DWI).We present a case of TIA, the cause of which was identified using multiphase computed tomography angiography (MCTA) in the hyperacute phase of the disease. PATIENT CONCERNS: The patient was a 57-year-old man who was admitted to the emergency department for right-side weakness persisting for 1 hour. DIAGNOSES: Occlusion of the proximal M3 segment of the left middle cerebral artery territory was found on the initial MCTA. OUTCOMES: The weakness completely resolved at 2 hours after symptom onset, and there was no acute infarction on the initial diffusion-weighted magnetic resonance imaging (MRI) on the same day. Follow-up MCTA on the next day showed recanalization of the left M3 segment. Follow-up diffusion-weighted MRI showed focal acute infarction in the left middle cerebral artery territory. LESSONS: MCTA could identify distal occlusion of the anterior circulation in patients with cardioembolic TIA in the hyperacute phase with negative DWI findings.

Characterization of recombinant ß-glucosidase from Arthrobacter chlorophenolicus and biotransformation of ginsenosides Rb1, Rb 2, Rc, and Rd.

The focus of this study was the cloning, expression, and characterization of recombinant ginsenoside hydrolyzing ß-glucosidase from Arthrobacter chlorophenolicus with an ultimate objective to more efficiently bio-transform ginsenosides. The gene bglAch, consisting of 1,260 bp (419 amino acid residues) was cloned and the recombinant enzyme, overexpressed in Escherichia coli BL21 (DE3), was characterized. The GST-fused BglAch was purified using GST·Bind agarose resin and characterized. Under optimal conditions (pH 6.0 and 37°C) BglAch hydrolyzed the outer glucose and arabinopyranose moieties of ginsenosides Rb1 and Rb2 at the C20 position of the aglycone into ginsenoside Rd. This was followed by hydrolysis into F2 of the outer glucose moiety of ginsenoside Rd at the C3 position of the aglycone. Additionally, BglAch more slowly transformed Rc to F2 via C-Mc1 (compared to hydrolysis of Rb1 or Rb2). These results indicate that the recombinant BglAch could be useful for the production of ginsenoside F2 for use in the pharmaceutical and cosmetic industries.

Cryopreservation of mouse spermatogonial stem cells in dimethylsulfoxide and polyethylene glycol.

Assisted reproductive techniques involving isolation, culture, and transplantation of spermatogonial stem cells (SSCs) have the potential to create transgenic livestock and to treat male infertility caused by cancer treatments such as chemotherapy or radiation. Because stem cells may need to be preserved for several years before reintroduction to the patients' testes, efficient SSC cryopreservation techniques need to be developed. SSCs can reinitiate spermatogenesis in recipient testes after freezing; however, optimal cryopreservation protocols have not been identified. The objective of this study was to develop an efficient cryopreservation method for SSCs using permeable cryoprotectant agents (PCAs) or additive cryoprotectant agents (ACAs). To identify an efficient cryopreservation method, populations of mouse testis cells enriched for SSCs were cultured in vitro and frozen using conventional freezing media containing various PCAs or ACAs for 1 wk or 1, 3, 6, 12, or 24 mo. Additionally, various molecular weights and concentrations of polyethylene glycol (PEG) were evaluated. Recovery rate, culture potential, and stem cell activity were significantly greater for cells frozen in 2.5% PEG with a molecular weight of 1000 compared to other treatment groups. These cells also retained the ability to colonize recipient testes, generate normal spermatogenesis, and contribute to viable offspring. The systematic analysis of many cryoprotectant agents indicates that 2.5% PEG (molecular weight 1000) is the most effective agent for efficient SSC cryopreservation.

Dynamic sensor interrogation using wavelength-swept laser with a polygon-scanner-based wavelength filter.

We report a high-speed (~2 kHz) dynamic multiplexed fiber Bragg grating (FBG) sensor interrogation using a wavelength-swept laser (WSL) with a polygon-scanner-based wavelength filter. The scanning frequency of the WSL is 18 kHz, and the 10 dB scanning bandwidth is more than 90 nm around a center wavelength of 1,540 nm. The output from the WSL is coupled into the multiplexed FBG array, which consists of five FBGs. The reflected Bragg wavelengths of the FBGs are 1,532.02 nm, 1,537.84 nm, 1,543.48 nm, 1,547.98 nm, and 1,553.06 nm, respectively. A dynamic periodic strain ranging from 500 Hz to 2 kHz is applied to one of the multiplexed FBGs, which is fixed on the stage of the piezoelectric transducer stack. Good dynamic performance of the FBGs and recording of their fast Fourier transform spectra have been successfully achieved with a measuring speed of 18 kHz. The signal-to-noise ratio and the bandwidth over the whole frequency span are determined to be more than 30 dB and around 10 Hz, respectively. We successfully obtained a real-time measurement of the abrupt change of the periodic strain. The dynamic FBG sensor interrogation system can be read out with a WSL for high-speed and high-sensitivity real-time measurement.

Novosphingobium ginsenosidimutans sp. nov., with the ability to convert ginsenoside.

A Gram-negative, strictly aerobic, non-motile, non-sporeforming, and rod-shaped bacterial strain designated FW-6T was isolated from a freshwater sample and its taxonomic position was investigated by using a polyphasic approach. Strain FW-6T grew optimally at 10-42 degrees C and at pH 7.0 on nutrient and R2A agar. Strain FW-6T displayed beta- glucosidase activity that was responsible for its ability to transform ginsenoside Rb1 (one of the dominant active components of ginseng) to Rd. On the basis of 16S rRNA gene sequence similarity, strain FW-6T was shown to belong to the family Sphingomonadaceae and was related to Novosphingobium aromaticivorans DSM 12444T (98.1% sequence similarity) and N. subterraneum IFO 16086T (98.0%). The G+C content of the genomic DNA was 64.4%. The major menaquinone was Q-10 and the major fatty acids were summed feature 7 (comprising C18:1 omega9c/ omega12t/omega7c), summed feature 4 (comprising C16:1 omega7c/iso- C15:0 2OH), C16:0, and C14:0 2OH. DNA and chemotaxonomic data supported the affiliation of strain FW-6T to the genus Novosphingobium. Strain FW-6T could be differentiated genotypically and phenotypically from the recognized species of the genus Novosphingobium. The isolate that has ginsenoside converting ability therefore represents a novel species, for which the name Novosphingobium ginsenosidimutans sp. nov. is proposed, with the type strain FW-6T (= KACC 16615T = JCM 18202T).

Treatment of fingertip amputation in adults by palmar pocketing of the amputated part.

BACKGROUND: First suggested by Brent in 1979, the pocket principle is an alternative method for patients for whom a microsurgical replantation is not feasible. We report the successful results of a modified palmar pocket method in adults. METHODS: Between 2004 and 2008, we treated 10 patients by nonmicrosurgical replantation using palmar pocketing. All patients were adults who sustained a complete fingertip amputation from the tip to lunula in a digits. In all of these patients, the amputation occurred due to a crush or avulsion-type injury, and a microsurgical replantation was not feasible. We used the palmar pocketing method following a composite graft in these patients and prepared the pocket in the subcutaneous layer of the ipsilateral palm. RESULTS: Of a total of 10 cases, nine had complete survival of the replantation and one had 20% partial necrosis. All of the cases were managed to conserve the fingernails, which led to acceptable cosmetic results. CONCLUSIONS: A composite graft and palmar pocketing in adult cases of fingertip injury constitute a simple, reliable operation for digital amputation extending from the tip to the lunula. These methods had satisfactory results.

Down-regulation of mitochondrial F1F0-ATP synthase in human colon cancer cells with induced 5-fluorouracil resistance.

5-Fluorouracil (5-FU) is widely used for treatment of advanced colorectal cancer. However, it is common for such patients to develop resistance to 5-FU, and this drug resistance becomes a critical problem for chemotherapy. The mechanisms underlying this resistance are largely unknown. To screen for proteins possibly responsible for 5-FU resistance, cells resistant to 5-FU were derived from human colon cancer cell lines and two-dimensional gel electrophoresis-based comparative proteomics was done. Two-dimensional gel electrophoresis data showed there was lower expression of the alpha subunit of mitochondrial F(1)F(0)-ATP synthase (ATP synthase) in 5-FU-resistant cells compared with parent cells. Western blotting showed that expression of other ATP synthase complex subunits was also lower in 5-FU-resistant cell lines and that these resistant cells also showed decreased ATP synthase activity and reduced intracellular ATP content. The ATP synthase inhibitor, oligomycin A, strongly antagonized 5-FU-induced suppression of cell proliferation. When 5-FU sensitivity was compared with ATP synthase activity in six different human colon cancer cell lines, a positive correlation has been found. Furthermore, suppressed ATP synthase d-subunit expression by siRNA transfection increased cell viability in the presence of 5-FU. Bioenergetic dysfunction of mitochondria has been reported as a hallmark of many types of cancers (i.e., down-regulation of ATP synthase beta-subunit expression in liver, kidney, colon, squamous oesophageal, and lung carcinomas, as well as in breast and gastric adenocarcinomas). Our findings show that ATP synthase down-regulation may not only be a bioenergetic signature of colorectal carcinomas but may also lead to cellular events responsible for 5-FU resistance.

Identification of genes with differential expression in acquired drug-resistant gastric cancer cells using high-density oligonucleotide microarrays.

PURPOSE: A major obstacle in chemotherapy is treatment failure due to anticancer drug resistance. The emergence of acquired resistance results from host factors and genetic or epigenetic changes in the cancer cells. The purpose of this study was to identify differentially expressed genes associated with acquisition of resistance in human gastric cancer cells. EXPERIMENTAL DESIGN: We performed global gene expression analysis in the acquired drug-resistant gastric cancer cell lines to the commonly used drugs 5-fluorouracil, doxorubicin, and cisplatin using Affymetrix HG-U133A microarray. The gene expression patterns of 10 chemoresistant gastric cancer cell lines were compared with those of four parent cell lines using fold-change and Wilcoxon's test for data analysis. RESULTS: We identified over 250 genes differentially expressed in 5-fluorouracil-, cisplatin-, or doxorubicin-resistant gastric cancer cell lines. Our expression analysis also identified eight multidrug resistance candidate genes that were associated with resistance to two or more of the tested chemotherapeutic agents. Among these, midkine (MDK), a heparin-binding growth factor, was overexpressed in all drug-resistant cell lines, strongly suggesting that MDK might contribute to multidrug resistance in gastric cancer cells. CONCLUSIONS: Our investigation provides comprehensive gene information associated with acquired resistance to anticancer drugs in gastric cancer cells and a basis for additional functional studies.