Human Organic Anion Transporters and Human Organic Cation Transporters Mediate Renal Antiviral Transport

The Journal of Pharmacology and Experimental Therapeutics. Mar, 2002  |  Pubmed ID: 11861798

Renal excretion is an important elimination pathway for antiviral agents, such as acyclovir (ACV), ganciclovir (GCV), and zidovudine (AZT). The purpose of this study was to elucidate the molecular mechanisms of renal ACV, GCV, and AZT transport using cells stably expressing human organic anion transporter 1 (hOAT1), hOAT2, hOAT3, and hOAT4, and human organic cation transporter 1 (hOCT1) and hOCT2. Time- and concentration-dependent uptake of ACV and GCV was observed in hOAT1- and hOCT1-expressing cells. In contrast, uptake of valacyclovir, L-valyl ester of ACV, was observed only in hOAT3-expressing cells. On the other hand, AZT uptake was observed in hOAT1-, hOAT2-, hOAT3-, and hOAT4-expressing cells. The Km values of ACV uptake by hOAT1 and hOCT1 were 342.3 and 151.2 microM, respectively, whereas those of GCV uptake by hOAT1 and hOCT1 were 895.5 and 516.2 microM, respectively. On the other hand, the Km values of AZT uptake by hOAT1, hOAT2, hOAT3, and hOAT4 were 45.9, 26.8, 145.1, and 151.8 microM, respectively. In addition, probenecid weakly inhibited the hOAT1-mediated ACV uptake. In conclusion, these results suggest that hOAT1 and hOCT1 mediate renal ACV and GCV transport, whereas hOAT1, hOAT2, hOAT3, and hOAT4 mediate renal AZT transport. In addition, L-valyl ester appears to be important in differential substrate recognition between hOAT1 and hOAT3. hOAT1 may not be the molecule responsible for the drug interaction between ACV and probenecid.

Human Organic Anion Transporters and Human Organic Cation Transporters Mediate Renal Transport of Prostaglandins

The Journal of Pharmacology and Experimental Therapeutics. Apr, 2002  |  Pubmed ID: 11907186

Prostaglandin E(2) (PGE(2)) and prostaglandin F(2 alpha) (PGF(2 alpha)) have been used for the induction of labor and the termination of pregnancy. Renal excretion is shown to be an important pathway for the elimination of PGE(2) and PGF(2 alpha). The purpose of this study was to elucidate the molecular mechanism of renal PGE(2) and PGF(2 alpha) transport using cells stably expressing human organic anion transporter (hOAT) 1, hOAT2, hOAT3, and hOAT4, and human organic cation transporter (hOCT) 1 and hOCT2. A time- and dose-dependent increase in PGE(2) and PGF(2 alpha) uptake was observed in cells expressing hOAT1, hOAT2, hOAT3, hOAT4, hOCT1, and hOCT2. The K(m) values of PGE(2) uptake by hOAT1, hOAT2, hOAT3, hOAT4, hOCT1, and hOCT2 were 970, 713, 345, 154, 657, and 28.9 nM, respectively, whereas those of PGF(2 alpha) uptake by hOAT1, hOAT3, hOAT4, hOCT1, and hOCT2 were 575, 1092, 692, 477, and 334 nM, respectively. PGE(2) and PGF(2 alpha) significantly inhibited organic anion uptake by hOATs and organic cation uptake by hOCTs. In conclusion, considering the localization of these transporters, the results suggest that PGE(2) and PGF(2 alpha) transport in the basolateral membrane of the proximal tubule is mediated by multiple pathways including hOAT1, hOAT2, hOAT3, and hOCT2, whereas that in the apical side is mediated by hOAT4.

Molecular Identification of a Renal Urate Anion Exchanger That Regulates Blood Urate Levels

Nature. May, 2002  |  Pubmed ID: 12024214

Urate, a naturally occurring product of purine metabolism, is a scavenger of biological oxidants implicated in numerous disease processes, as demonstrated by its capacity of neuroprotection. It is present at higher levels in human blood (200 500 microM) than in other mammals, because humans have an effective renal urate reabsorption system, despite their evolutionary loss of hepatic uricase by mutational silencing. The molecular basis for urate handling in the human kidney remains unclear because of difficulties in understanding diverse urate transport systems and species differences. Here we identify the long-hypothesized urate transporter in the human kidney (URAT1, encoded by SLC22A12), a urate anion exchanger regulating blood urate levels and targeted by uricosuric and antiuricosuric agents (which affect excretion of uric acid). Moreover, we provide evidence that patients with idiopathic renal hypouricaemia (lack of blood uric acid) have defects in SLC22A12. Identification of URAT1 should provide insights into the nature of urate homeostasis, as well as lead to the development of better agents against hyperuricaemia, a disadvantage concomitant with human evolution.

Characterization of Methotrexate Transport and Its Drug Interactions with Human Organic Anion Transporters

The Journal of Pharmacology and Experimental Therapeutics. Aug, 2002  |  Pubmed ID: 12130730

Life-threatening drug interactions are known to occur between methotrexate and nonsteroidal anti-inflammatory drugs (NSAIDs), probenecid, and penicillin G. The purpose of this study was to characterize methotrexate transport, as well as to determine the site and the mechanism of drug interactions in the proximal tubule. Mouse proximal tubule cells stably expressing basolateral human organic anion transporters (hOAT1 and hOAT3) and apical hOAT (hOAT4) were established. The K(m) values for hOAT1-, hOAT3-, and hOAT4-mediated methotrexate uptake were 553.8 microM, 21.1 microM, and 17.8 microM, respectively. NSAIDs (salicylate, ibuprofen, ketoprofen, phenylbutazone, piroxicam, and indomethacin), probenecid, and penicillin G dose dependently inhibited methotrexate uptake mediated by hOAT1, hOAT3, and hOAT4. Kinetic analysis of inhibitory effects of these drugs on hOAT3-mediated methotrexate uptake revealed that these inhibitions were competitive. The K(i) values for the effects of salicylate, phenylbutazone, indomethacin, and probenecid on hOAT3-mediated methotrexate uptake were comparable with therapeutically relevant plasma concentrations of unbound drugs. In addition, in the presence of human serum albumin, the K(i) values were comparable with therapeutically relevant total plasma concentrations of drugs. In conclusion, these results suggest that methotrexate is taken up via hOAT3 and hOAT1 at the basolateral side of the proximal tubule and effluxed or taken up at the apical side via hOAT4. In addition, hOAT1, hOAT3, and hOAT4 are the sites of drug interactions between methotrexate and NSAIDs, probenecid, and penicillin G. Furthermore, it was predicted that hOAT3 is the site of drug interactions between methotrexate and salicylate, phenylbutazone, indomethacin, and probenecid in vivo.

Mycobacterium Leprae Infection in Monocyte-derived Dendritic Cells and Its Influence on Antigen-presenting Function

Infection and Immunity. Sep, 2002  |  Pubmed ID: 12183567

Host defense against Mycobacterium leprae infection is chiefly mediated by gamma interferon (IFN-gamma)-secreting cytotoxic T cells. Since which antigen-presenting cell populations act to stimulate these T cells is not fully understood, we addressed the role of monocyte-derived dendritic cells (DCs). The DCs phagocytosed M. leprae and expressed bacterially derived antigens (Ags), such as phenolic glycolipid 1 (PGL-1), in the cytoplasm, as well as on the cell surface. The expression of HLA-ABC and -DR Ags on DCs was down-regulated by M. leprae infection, and that of CD86 was up-regulated, but not as fully as by Mycobacterium bovis BCG infection. Induction of CD83 expression required a large number of M. leprae cells. When a multiplicity of infection of >40 was used, the DCs induced a significant proliferative and IFN-gamma-producing response in autologous T cells. However, these responses were significantly lower than those induced by BCG- or Mycobacterium avium-infected DCs. A CD40-mediated signaling in M. leprae-infected DCs up-regulated the expression of HLA Ags, CD86, and CD83 but did not enhance T-cell-stimulating ability. Therefore, M. leprae-infected DCs are less efficient at inducing T-cell responses. However, when the surface PGL-1 on M. leprae-infected DCs was masked by a monoclonal antibody, the DCs induced enhanced responses in both CD4(+)- and CD8(+)-T-cell subsets. M. leprae is a unique pathogen which remains resistant to DC-mediated T-cell immunity, at least in the early stages of infection.

Urate Transport Via Human PAH Transporter HOAT1 and Its Gene Structure

Kidney International. Jan, 2003  |  Pubmed ID: 12472777

We recently cloned the human organic anion transporter 1 (hOAT1) as a p-aminohippurate (PAH) transporter. Whether urate is transported by the PAH transporter in humans remains unclear. Familial juvenile gouty nephropathy (FJGN) is thought to develop as a result of an abnormality in the urate transporter.

Multiple Mechanisms Involved in the Inhibition of Proinflammatory Cytokine Production from Human Monocytes by N-(p-coumaroyl)serotonin and Its Derivatives

International Immunopharmacology. Feb, 2003  |  Pubmed ID: 12586608

We have reported that N-(p-coumaroyl)serotonin(CS) isolated from safflower oil cake (Carthamus tinctorius L.) inhibits the production of proinflammatory cytokines by endotoxin (LPS)- stimulated human monocytes. In this study, the effects of CS and its three derivatives, N-(trans-cinnamoyl)serotonin (Cin.S), N-(trans-cinnamoyl)tryptamine (Cin.T), and N-(p-coumaroyl)tryptamine (CT) on the production of proinflammatory cytokines were compared. Cin.S possessed radical scavenging activity at a comparable level to CS, while CT and Cin.T exhibited lower activity, suggesting that hydroxyl group in serotonin is essential for the antioxidative activity. CS and CT strongly inhibited the production of proinflammatory cytokines (IL-1alpha, IL-1beta, IL-6, IL-8, and TNF-alpha) from LPS-stimulated human monocytes. However, Cin.S inhibited the production of only IL-1alpha and IL-1beta, and Cin.T inhibited none of these cytokines production. CS and CT markedly inhibited the protein synthesis in monocytes, the inhibitory effect of Cin.S was moderate, and that of Cin.T was quite weak. These results indicate that CS and its derivatives inhibit the production of proinflammatory cytokines through multiple mechanisms.

[Urate Transport in Human Kidney]

Nihon Rinsho. Japanese Journal of Clinical Medicine. Jan, 2003  |  Pubmed ID: 12629703

Studies on Murine Thyroiditis: New Insights from Organ Flow Cytometry

Thyroid : Official Journal of the American Thyroid Association. May, 2003  |  Pubmed ID: 12855008

The pathogenesis of autoimmune diseases is frequently studied in murine models, in which disease outcome is traditionally assessed by light microscopy. To determine whether digital imaging improves reliability of the histopathologic assessment, and whether flow cytometry is applicable directly on the murine thyroid, we studied 395 CBA/J mice 3 weeks after thyroglobulin immunization, and 192 nonimmunized CBA/J mice. Digital imaging significantly improved reliability of the histopathological assessment (r = 0.988, 95% confidence interval: 0.980-0.992, p < 0.0001), and flow cytometry on the murine thyroid could be performed successfully. We also found that normal thyroids contained a higher than expected number of hematopoietic cells in the interstitium. We suggest that digital imaging offers a better means of estimating disease outcome, and that flow cytometry performed at the target organ levels reflects the autoimmune pathogenesis more closely than when performed on peripheral lymphoid organs. These methods should also be applicable to other organ systems targeted by autoimmune attack, such as heart, exocrine, and other endocrine glands.

A Patient with MRSA Infection to Prosthesis of Femoral Head Diagnosed Non-invasively Using Bi-digital O-ring Test: a Clinical Case Report

Acupuncture & Electro-therapeutics Research. 2003  |  Pubmed ID: 12934960

Prosthesis of femoral head is a common surgical procedure, but the diagnosis of infection associated with the prosthesis remains difficult. We diagnosed non-invasively methicillin resistant Staphylococcus aureus (MRSA) infection of prosthesis of femoral head with Bi-Digital O-Ring Test (BDORT). BDORT uses the resonance phenomenon between 2 identical substances, and electromagnetic field principle. The method can non-invasively detect viral & bacterial infection. Accuracy of the BDORT findings was confirmed through bacterial culture & sensitivity test to antibiotics. Patient was successfully treated with operation of evulsion of the prosthesis of femoral head and administration of antibiotics and Cilantro. The drug compatibility was tested with BDORT. BDORT was an effective technique for non-invasively detecting infection of prosthesis and selecting the most effective antibiotics.

Does a Viral Infection Cause Complex Regional Pain Syndrome?

Acupuncture & Electro-therapeutics Research. 2003  |  Pubmed ID: 14998056

In 1990 Omura, Y. reported that Herpes Simplex Virus Type 1 as the major cause of chronic intractable pain and its effective treatment using mixture of EPA & DHA with Selective Drug Uptake Enhancement Method. Subsequently among the other causes of pain, he included Chlamydia Trachomatis, Borrelia Burgdorferi, Mycobacterium Tuberculosis, human Herpes Virus type 6, and Circulatory Disturbances. In order to test possible involvement of viral infection in Complex Regional Pain Syndrome (CRPS), a disease which usually occurs in the extremities, we did a study of 17 patients with CRPS. They were examined for Herpes Simplex Virus (HSV) and Varicella Zoster Virus (VZV) by measuring IgG and IgM antibody titers, and 14 of these patients were also examined for Cytomegalo-Virus (CMV). As a control group 100 healthy Japanese employees at SRL, Inc. were also studied. In CRPS group, HSV IgG was positive in 12 of the 17 patients with an average antibody titer of 90.0 EIA value. VZV IgG was positive in all 17 patients with an average antibody titer of 26.8 EIA value. CMV IgG was positive in all 14 patients with an average antibody titer of 66.6 UA/ml. In control group, HSV IgG was positive in 54 subjects with an average antibody titer of 42.3 EIA value. VZV IgG was positive in 97 subjects with an average antibody titer of 26.2 EIA value. CMV IgG was positive in 82 subjects. There were no significant differences of positive rate of IgG antibody for the three viruses between patient and control groups. Although the difference was not significant, the average antibody titers of HSV in CRPS group were more than twice of those in healthy group. Antibody titers were almost equal in both groups for VZV. Possibly, some people in the control group who had latent virus, were also asymptomatic. In 2000, Takasaki, I. et al. in a separate animal study, inoculated with HSV Type-I the shin of the mouse causing allodynia and hyperalgesia (which are some of the characteristic findings seen in CRPS in humans). Also, VZV, which causes shingles which is sometimes followed by Post-Herpetic Neuralgia (PHN), is in the same family of HSV. As PHN resembles CRPS in symptoms, it is possible that HSV contributes to CRPS. Therefore, virus infection theory is an attractive hypothesis that accounts for many enigmas of CRPS.

Comparison of Current Perception Threshold Between Each Side in Unilateral Complex Regional Pain Syndrome Patients Does Not Measure the Patient's Pain

Hiroshima Journal of Medical Sciences. Mar, 2004  |  Pubmed ID: 15274424

The current perception threshold (CPT) test has been developed as one of the neuroselective sensory nerve conduction threshold tests. The score of the CPT of the affected side subtracted from the score of the CPT of the unaffected side in complex regional pain syndrome (CRPS) is expected to show pain objectively. The purpose of this study is to examine first whether the CPT of the affected side is generally lower than that of the unaffected side, and second, whether the greater score shows the more intense pain. The CPT of each side in 25 patients with unilateral CRPS type I was measured and compared. For the 2000 Hz stimulus, the CPT of the affected side was 2677 +/- 262 microAmp (mean +/- standard error) and the CPT of the unaffected side was 2194 +/- 247 microAmp (p = 0.0149). For the 250 Hz stimulus, the CPT was 876 +/- 117 microAmp and 721 +/- 73 microAmp respectively (p > 0.05). For the 5 Hz stimulus, the CPT was 730 +/- 105 microAmp and 448 +/- 56 microAmp respectively (p = 0.0018). In 2000 Hz, 250 Hz, and 5 Hz stimuli, the CPT of the affected side was higher than that of the unaffected side. This shows that generally the affected side is less sensitive than the unaffected side in terms of current perception. The score of the CPT of the affected side subtracted from the score of the CPT of the unaffected side in CRPS does not measure the patient's pain.

Early Chemokine Expression Induced by Interferon-gamma in a Murine Model of Hashimoto's Thyroiditis

Experimental and Molecular Pathology. Dec, 2004  |  Pubmed ID: 15507231

Chemokines represent a group of small, secreted proteins mainly involved in navigating leukocytes towards site of inflammation. Some chemokines have been implicated in the pathogenesis of autoimmune diseases, which are characterized by an ectopic retention of leukocytes within the target organ, ultimately leading to loss of function. To determine the chemokines profile expressed in the thyroid gland upon chronic exposure to interferon-gamma (IFNgamma), we analyzed C57BL6 transgenic mice that aberrantly express IFNgamma under control of the thyroglobulin promoter. We compared by reverse transcriptase PCR the thyroidal expression of 10 chemokines (CCL1 through 5 and CXCL9 through 13) in thyr-IFNgamma transgenics and wild-type littermates. We found that transgenics exclusively expressed CCL4, CXCL9, and CXCL11, and showed increased expression of CCL5 and CXCL10. This chemokine profile was associated with moderate mononuclear cell infiltration of the thyroid stroma that, however, decreased significantly after 2 months of age and did not organize into lymphoid structures. Our findings indicate that the isolated expression of IFNgamma is capable of recruiting mononuclear cells but they do not progress to full lymphoid transformation of the thyroid.

Expression of Class II Major Histocompatibility Complex Molecules on Thyrocytes Does Not Cause Spontaneous Thyroiditis but Mildly Increases Its Severity After Immunization

Endocrinology. Mar, 2005  |  Pubmed ID: 15591134

Class II major histocompatibility complex (MHC) molecules are classically expressed on antigen-presenting cells of the hematopoietic lineage but have also been described on epithelial cells in association with autoimmunity. In this context, however, it remains debatable whether class II MHC molecules are the initiating event or rather the consequence of the autoimmune attack. In addition, the role of epithelial class II expression once the autoimmune attack has begun is unknown. We generated transgenic mice expressing in the thyroid follicular cells the class II transactivator, the master regulator of all the genes in the class II MHC pathway. The study used a cohort of 245 CBA/J mice (127 wild-type and 118 transgenic), both in basal conditions (n = 63) and at different time points after immunization with mouse thyroglobulin (n = 182). In basal conditions, transgenic mice were similar to wild-type controls and did not develop spontaneous autoimmune thyroiditis, despite the aberrant expression of class II MHC molecules on thyrocytes. After immunization, thyroiditis was 8% more severe in transgenics than controls (95% confidence interval from 1.8-13.4%; P = 0.033), especially during the florid stages of disease. These findings suggest that expression of class II MHC molecules on epithelial cells is not sufficient to initiate autoimmunity but mildly modulates an already established autoimmune attack against the target organ.

Increased Thyroidal Fat and Goitrous Hypothyroidism Induced by Interferon-gamma

International Journal of Experimental Pathology. Apr, 2005  |  Pubmed ID: 15810981

Summary Hashimoto's thyroiditis is associated with a diffuse lymphocytic infiltration of the stroma and a production of several cytokines, such as interferon-gamma (IFN-gamma). We previously reported that transgenic mice expressing IFN-gamma under the control of the thyroglobulin promoter develop primary hypothyroidism. In order to determine the long-term changes induced by IFN-gamma in the thyroid gland, we analysed cross-sectionally 202 mice (96 transgenic mice and 106 controls) of 0-650 days of age. Multiple linear regression analysis showed that, after adjusting for age and sex, thyr-IFN-gamma transgenic mice were 14% (3 g) smaller (P < 0.0001) and had a 5- to 6-fold bigger thyroid (P < 0.0001) than wild-type littermates. Transgenic thyroids showed striking histopathological changes in follicles, thyrocytes and stroma. Follicles were enlarged, irregular and were lined by thickened, granular and oxyphilic thyrocytes. The stroma contained a moderate and diffuse mononuclear infiltrate--mainly composed of macrophages--and, interestingly, a clear increase in the content of fat. These findings indicate that, in addition to hypothyroidism, chronic exposure of the thyroid to IFN-gamma leads also to macrophage infiltration and subsequent adipocyte expansion, suggesting a link between inflammation and fat accumulation.

Interleukin (IL)-12-driven Primary Hypothyroidism: the Contrasting Roles of Two Th1 Cytokines (IL-12 and Interferon-gamma)

Endocrinology. Aug, 2005  |  Pubmed ID: 15860554

IL-12, a prototypic T helper 1 cytokine, has been implicated in the pathogenesis of organ-specific autoimmune diseases, such as Hashimoto's thyroiditis, but reported to give conflicting results in murine models of lymphocytic thyroiditis. To determine the effects of chronic, local production of IL-12 within the thyroid gland, we created transgenic mice that express IL-12 p70 under the transcriptional control of the thyroglobulin promoter. Transgenics developed growth retardation, moderate primary hypothyroidism, and mild lymphocytic infiltration of the thyroid gland. The hypothyroidism was associated with increased mRNA levels of the sodium-iodide symporter, an increase partly due to a direct effect of IL-12 on the thyrocyte. Upon immunization with a suboptimal dose of mouse thyroglobulin, IL-12 transgenic mice developed a lymphocytic thyroiditis that was more frequent and severe than that observed in wild-type littermates. The disease-promoting effect of IL-12 was independent of interferon-gamma, as shown by the similar interferon-gamma levels in transgenics and controls. These findings highlight the contrasting roles of two T helper 1 cytokines and report a novel role of IL-12 on thyroid hormonogenesis.

Nerve Tolerance to High-dose-rate Brachytherapy in Patients with Soft Tissue Sarcoma: a Retrospective Study

BMC Cancer. 2005  |  Pubmed ID: 16026629

Brachytherapy, interstitial tumor bed irradiation, following conservative surgery has been shown to provide excellent local control and limb preservation in patients with soft tissue sarcomas (STS), whereas little is known about the tolerance of peripheral nerves to brachytherapy. In particular, nerve tolerance to high-dose-rate (HDR) brachytherapy has never been properly evaluated. In this study, we examined the efficacy and radiation neurotoxicity of HDR brachytherapy in patients with STS in contact with neurovascular structures.

Frequency of Gouty Arthritis in Patients with End-stage Renal Disease in Japan

Internal Medicine (Tokyo, Japan). Jul, 2005  |  Pubmed ID: 16093591

The purpose of this study was to investigate gouty arthritis in Japanese patients with end-stage renal disease (ESRD).

Efficacy of Neurotropin in Chronic Fatigue Syndrome: a Case Report

Hiroshima Journal of Medical Sciences. Mar, 2006  |  Pubmed ID: 16594551

Chronic fatigue syndrome (CFS) is a disorder that causes general fatigue and chronic widespread pain. A 28-year-old male visited an outpatient department due to general fatigue and pain involving the entire body. He did not suffer from fibromyalgia, but he was diagnosed with CFS. At the initial visit, he complained of lack of concentration, memory decline, frequent urination, insomnia and occasional difficulty of emotional control, as well as general fatigue and pain involving the entire body. Four tablets of Neurotropin per day alone were administered. General fatigue and pain were gradually alleviated one week later. His sleep condition, concentration power, and memory also improved two weeks later. Medication was discontinued from 11 weeks based on the patient's judgment as he felt little general fatigue and pain involving the entire body. Treatment was completed 3 months later. The symptoms disappeared and did not recur five months after the discontinuation of Neurotropin. He was looking for a job without fatigue and pain 8 months later (5 months after the cessation of treatment). The functional mechanisms of Neurotropin in CFS are unknown.

IL-12 Overexpression in Mice As a Model for Sjögren Lung Disease

American Journal of Physiology. Lung Cellular and Molecular Physiology. Oct, 2006  |  Pubmed ID: 16751222

Interleukin-12 (IL-12), a Th1 proinflammatory cytokine, is reported to be increased in Sjögren syndrome. To evaluate the effects of local Th1/Th2 deregulation, we generated a transgenic mouse model that overexpresses IL-12 in the lungs. IL-12 transgenic mice developed bronchial and alveolar abnormalities strikingly similar to those found in the lungs of Sjögren patients. Pathologically, lung abnormalities began at approximately 4 mo of age and were characterized by lymphocytic infiltrates around the bronchi, intraluminal periodic acid Schiff-positive debris, increased cell proliferation in the alveolar region, and increased interstitial and alveolar macrophages. Functionally, these abnormalities translated into decreased mucociliary clearance (P<0.05 vs. wild-type littermates) and increased oxidative stress (P<0.01). The pathological and functional abnormalities were accompanied by significant changes in lung natural killer (NK) cells. The number of NK cells was fourfold higher in IL-12 transgenic than wild-type lungs (20% of all lymphoid cells vs. 5%) during the first month of life. NK cells then decreased within a narrow window of time (from 30 to 50 days of age), reaching a nadir of approximately 2% on day 50, and remained at these low levels thereafter. This new mouse model highlights the role of IL-12 in the initiation of Sjögren syndrome.

Stable Top-up Operation at SPring-8

Journal of Synchrotron Radiation. Sep, 2006  |  Pubmed ID: 16924134

Top-up operation allows SPring-8 to provide highly stable X-ray beams with arbitrary filling patterns. The implementation of top-up operation is described, with a focus on the simultaneous achievement of stability of stored current, beam orbit, purity of an isolated single bunch, and beam injection efficiency. Stored-current fluctuations have been routinely reduced to a level of 10(-3). Stored-beam oscillation on frequent beam injection, which was originally regarded as the most serious problem, has been successfully suppressed to a sufficiently low level that it never perturbs imaging experiments. Current impurities in nominally empty buckets have been reduced to a level of 10(-9) over more than one week of operation, making possible the measurement of time-resolved spectra using high-current bunches. Finally, excellent injection efficiency, higher than 80%, is routinely obtained, even for small undulator gaps, which is critical for preventing radiation damage to insertion-device magnets and to reduce leakage radiation. The process of achieving highly stabilized top-up operation at SPring-8 and its utility for user experiments are described.

Antinociceptive Effect of Linear Polarized 0.6 to 1.6 Microm Irradiation of Lumbar Sympathetic Ganglia in Chronic Constriction Injury Rats

Journal of Rehabilitation Research and Development. Jul-Aug, 2006  |  Pubmed ID: 17123194

Linear polarized near-infrared light created with linear polarized near-infrared light therapy equipment (Super Lizer HA-550, Tokyo Iken Co, Ltd, Tokyo, Japan) has been used for the treatment of various painful disorders in Japan. Irradiation near the stellate ganglion with a Super Lizer (ISGL) is an especially notable therapeutic method used with stellate ganglion block (SGB) or substitutes for SGB. ISGL is a safe, simple, well-tolerated, and effective treatment. We examined the effects of irradiation with a Super Lizer applied to an area near the lumbar sympathetic ganglia on the ligated side in a chronic constriction injury (CCI) model, which is believed to be an animal model of complex regional pain syndrome (CRPS). Rats showing thermal hyperalgesia in a radiant heat test 1 wk postoperatively were used in Experiments 1 and 2: (1) Thermal hyperalgesia of irradiation group (n = 11) was less than that of the control or nonirradiation (n = 11) group at 1, 3, and 8 h after irradiation; however, the effect disappeared 12 h after irradiation. (2) Daily irradiation (n = 16) and 1 wk (n = 14) from 7 days after nerve ligation significantly shortened the interval from thermal hyperalgesia until recovery. Rats showing mechanical hyperalgesia in the von Frey hair test 1 wk postoperatively were used in Experiment 3: 1 wk irradiation beginning 7 days after nerve ligation (n = 9) did not promote the recovery from mechanical hyperalgesia. We speculate that repeated ISGL may be more effective than a single ISGL in alleviating pain in CRPS patients. We cannot explain the discrepancy between the results obtained in Experiments 2 and 3. We believe the results of this study are relevant to the effect of ISGL for patients with upper-limb CRPS: irradiation near the lumbar sympathetic ganglia of the rat is effective for thermal but not mechanical pain in CCI.

"Superior Rectus Paralysis" Due to Inferior Rectus Myxedema

Internal Medicine (Tokyo, Japan). 2006  |  Pubmed ID: 17139131

Autoimmune Thyroid Diseases

Current Opinion in Rheumatology. Jan, 2007  |  Pubmed ID: 17143095

Interesting clinical and basic studies have been published in the field of autoimmune thyroiditis (represented by Graves' disease and Hashimoto's thyroiditis) since January 2005. The review is organized into four main areas: genetics, environment, adaptive immune system, and innate immune system.

Toll-like Receptor-MyD88 and Fc Receptor Pathways of Mast Cells Mediate the Thyroid Dysfunctions Observed During Nonthyroidal Illness

Proceedings of the National Academy of Sciences of the United States of America. Apr, 2007  |  Pubmed ID: 17389381

Bacterial infections and other pathologic conditions induce complex dysfunctions of the hypothalamic-pituitary-thyroid axis, collectively known as nonthyroidal illness (NTI). To explore the pathogenesis of bacterial NTI, we injected Mycobacterium tuberculosis extracts or Escherichia coli LPS in mice lacking key components of the Toll-like receptor or crystallizable fragment (Fc) receptor pathways. In wild-type mice, the bacterial components induced a hypothyroidism characterized by elements of both hypothalamic and thyroidal dysfunction. This NTI hypothyroidism did not develop in mice lacking the MyD88 adaptor or in those with a reduced number of mast cells. The hypothyroid responsiveness to LPS, however, was restored upon reconstitution with mast cells derived from the bone marrow of wild-type donors. In addition to bacterial components, whole immunoglobulins induced NTI hypothyroidism in wild-type mice, but not in those lacking activating Fc receptors or mast cells. The study demonstrates a link between Toll-like and Fc receptor signaling and thyroid gland function, uncovering a role of mast cells in murine NTI.

Tenascin-W Inhibits Proliferation and Differentiation of Preosteoblasts During Endochondral Bone Formation

Biochemical and Biophysical Research Communications. May, 2007  |  Pubmed ID: 17395156

We identified a cDNA encoding mouse Tenascin-W (TN-W) upregulated by bone morphogenetic protein (Bmp)2 in ATDC5 osteo-chondroprogenitors. In adult mice, TN-W was markedly expressed in bone. In mouse embryos, during endochondral bone formation TN-W was localized in perichondrium/periosteum, but not in trabecular and cortical bones. During bone fracture repair, cells in the newly formed perichondrium/periosteum surrounding the cartilaginous callus expressed TN-W. Furthermore, TN-W was detectable in perichondrium/periosteum of Runx2-null and Osterix-null embryos, indicating that TN-W is expressed in preosteoblasts. In CFU-F and -O cells, TN-W had no effect on initiation of osteogenesis of bone marrow cells, and in MC3T3-E1 osteoblastic cells TN-W inhibited cell proliferation and Col1a1 expression. In addition, TN-W suppressed canonical Wnt signaling which stimulates osteoblastic differentiation. Our results indicate that TN-W is a novel marker of preosteoblasts in early stage of osteogenesis, and that TN-W inhibits cell proliferation and differentiation of preosteoblasts mediated by canonical Wnt signaling.

Chemokine Orchestration of Autoimmune Thyroiditis

Thyroid : Official Journal of the American Thyroid Association. Oct, 2007  |  Pubmed ID: 17910527

Chemokines are low-molecular-weight proteins that attract leukocytes and other cell types, via interaction with G protein-coupled receptors. Chemokines control leukocyte migration not only during inflammatory processes, but also throughout ontogeny and differentiation of lymphoid tissues. They have been involved in the pathogenesis of numerous diseases, such as human immunodeficiency virus infection, allergy, atherosclerosis, cancer, and autoimmunity. The number of studies focusing on chemokine biology is expanding exponentially. For example, searching PubMed for the terms "thyroid" and "chemokine" retrieved 1 article in 1980s, 18 articles in 1990s, and 81 articles from 2000 to July 2007. This review will focus on studies analyzing the role of chemokine in autoimmune thyroiditis (Graves' disease and Hashimoto's thyroiditis), performed in both patients and experimental animals. The goal is to emphasize how a better understanding of chemokine biology has advanced our knowledge of the pathogenesis of autoimmune thyroiditis.

Reconstruction of a Severe Maxillofacial Deformity After Tumorectomy and Irradiation Using Distraction Osteogenesis and LeFort I Osteotomy Before Vascularized Bone Graft

The Journal of Craniofacial Surgery. Sep, 2007  |  Pubmed ID: 17912098

We present the successful reconstruction of a large mandibular defect with a severe maxillofacial deformity after malignant tumor resection and irradiation. The patient was a 16-year-old boy with a defect in the left mandible, which extended from the mandibular body to the condylar process and hypoplasia of the maxillozygomatic complex on the left side as a result of ablation and radiotherapy of a grown rhabdomyosarcoma in the left infratemporal fossa at the age of 10. We planned a two-stage reconstruction because of his wide mandibular defect and hypoplasia. LeFort I type osteotomy to correct the maxillary declination was combined with mandibular lengthening to decrease the width of the defect in the first stage. New bone formation was confirmed at the distraction site 4 months after surgery, and the second stage was performed. A free latissimus dorsi myocutaneous flap with a vascularized scapula and rib was transferred to reconstruct the ramus of the mandible, zygomatic arch, and soft tissues. This procedure resulted in satisfactory results. In conclusion, the combination of distraction osteogenesis and microsurgical bone transplantation facilitated the straightforward reconstruction of a three-dimensional deformity with huge bony defects. We think that this combined surgical procedure will become a favorable option in the treatment of severe maxillomandibular deformities with bone defects.

Autoimmune Hypophysitis of SJL Mice: Clinical Insights from a New Animal Model

Endocrinology. Jul, 2008  |  Pubmed ID: 18388197

Autoimmune hypophysitis (AH) is a rare but increasingly recognized disease of the pituitary gland. Its autoantigens are unknown, and the management is difficult because it is often misdiagnosed as a nonsecreting adenoma. By immunizing female SJL/J mice with mouse pituitary extracts, we established a new mouse model of experimental AH. Immunized mice developed severe lymphocytic infiltration in the anterior pituitary that closely mimicked the human pathology. In the early phase of experimental AH, the pituitary enlarged, consistent with the compression symptoms reported by hypophysitis patients at presentation. In the florid phase, adrenal insufficiency and pituitary antibodies developed, in strong correlation with the pituitary pathology. In the late phase, hypothyroidism ensued, and the pituitary gland became atrophic. Using immune sera as probes in a two-dimensional immunoblotting screen followed by mass spectrometry, we identified several proteins that could function as pituitary autoantigens. These findings provide new insights into the pathogenesis of AH, and establish a platform for developing novel diagnostic biomarkers and therapeutics.

Operative Technique to Harvest an Arterial Flap from the Posterolateral Calf Region: How Can We Elevate a Lateral Gastrocnemius Perforating Artery Flap Safely?

Journal of Reconstructive Microsurgery. Jan, 2008  |  Pubmed ID: 18548380

The posterior calf region is a useful donor site for skin or composite flaps including muscle and/or nerves. We reported the first clinical use of the lateral gastrocnemius perforating artery flap including a vascularized sural nerve in 2003. This flap was elevated based on a perforator arising from the lateral head of the gastrocnemius muscle. However, we have since encountered vascular variations in these perforators. We subsequently developed a reliable technique for harvesting this flap in the course of treating 10 patients. Safe flap elevation from the lateral aspect of the posterior calf requires preservation of one of the superficial sural arteries until reliable perforators arising from gastrocnemius muscle lateral head are encountered during dissection. When such perforators are not observed, nutrient vessels such as superficial sural arteries or muscle perforators originating from vessels other than the lateral sural artery must be selected as a flap pedicle.

Revision Total Hip Arthroplasty by Nonmodular Short and Long Cementless Stems

Journal of Orthopaedic Science : Official Journal of the Japanese Orthopaedic Association. Jul, 2008  |  Pubmed ID: 18696192

The choice of appropriate implant for reconstruction during revision total hip arthroplasty (THA) is controversial. We use proximally porous-coated cementless short stems and fully porous-coated cementless long stems depending on the state of bone loss during revision surgery.

Pituitary Autoimmunity: 30 Years Later

Autoimmunity Reviews. Sep, 2008  |  Pubmed ID: 18774118

Pituitary autoimmunity encompasses a spectrum of conditions ranging from histologically proven forms of lymphocytic hypophysitis to the presence of pituitary antibodies in apparently healthy subjects. Hypophysitis is a rare but increasingly recognized disorder that typically presents as a mass in the sella turcica. It mimics clinically and radiologically other non-functioning sellar masses, such as the more common pituitary adenoma. Hypophysitis shows a striking temporal association with pregnancy, and it has been recently described during immunotherapies that block CTLA-4. Several candidate pituitary autoantigens have been described in the last decade, although none has proven useful as a diagnostic tool. This review summarizes the advances made in the field since the publication of the first review on pituitary autoimmunity, and the challenges that await clarification.

Cthrc1 is a Positive Regulator of Osteoblastic Bone Formation

PloS One. 2008  |  Pubmed ID: 18779865

Bone mass is maintained by continuous remodeling through repeated cycles of bone resorption by osteoclasts and bone formation by osteoblasts. This remodeling process is regulated by many systemic and local factors.

[Typical MR Images of Cerebral Amyloid Angiopathy Presented with Cerebral Hemorrhage]

Brain and Nerve = Shinkei Kenkyū No Shinpo. Oct, 2008  |  Pubmed ID: 18975609

Efficacy of Distraction Osteogenesis for Mandibular Reconstruction in Previously Irradiated Areas: Clinical Experiences

The Journal of Craniofacial Surgery. Nov, 2008  |  Pubmed ID: 19098554

The efficacy of distraction osteogenesis in an irradiated area is controversial, although this procedure is now widely used in the field of craniomaxillofacial surgery. We report the clinical results from 4 patients with mandibular defects treated by lengthening of the irradiated mandibles. All patients had a mandibular defect caused by ablation of a malignant tumor. They had undergone radiotherapy at a total dose of 30 to 50 Gy to the surgical site after tumorectomy. Distraction osteogenesis was used as the secondary reconstruction method in 6 sites of the remaining irradiated mandibles and in 1 site of the transferred vascularized scapula after radiotherapy. The transported segment was obtained by corticotomy with an initial gap of 0 to 2 mm, and internal extension plates were used. Distraction was commenced after a latency period of 7 to 10 days and performed at the rate of 0.25 to 1.0 mm/d. The total amount of distraction and consolidation periods ranged from 15 to 25 mm and 120 to 193 days, respectively. In 5 of the 6 sites in the remaining irradiated mandibles, satisfactory bone formation in the distraction gap was observed, although a fracture after new bone formation was observed in 1 site. Fibrous callus formation was observed in 1 irradiated site only, and satisfactory results were obtained in another site of transferred vascularized scapula in the same patient. From these experiences, we believe that distraction may provide a reconstruction option for mandibular defects even under irradiated conditions because the procedure is simple and less invasive.

Osteoblast-targeted Expression of Sfrp4 in Mice Results in Low Bone Mass

Journal of Bone and Mineral Research : the Official Journal of the American Society for Bone and Mineral Research. Feb, 2008  |  Pubmed ID: 17907918

Transgenic mice overexpressing Sfrp4 in osteoblasts were established. These mice exhibited low bone mass caused by a decrease in bone formation.

Using Full-thickness Skin Graft from Amputated Foot Can Provide a Stump with Durable Skin

Journal of Plastic, Reconstructive & Aesthetic Surgery : JPRAS. Dec, 2009  |  Pubmed ID: 19231312

The Cyan Fluorescent Protein (CFP) Transgenic Mouse As a Model for Imaging Pancreatic Exocrine Cells

JOP : Journal of the Pancreas. 2009  |  Pubmed ID: 19287108

The use of fluorescent proteins for in vivo imaging has opened many new areas of research. Among the important advances in the field have been the development of transgenic mice expressing various fluorescent proteins.

Development of the Transgenic Cyan Fluorescent Protein (CFP)-expressing Nude Mouse for "Technicolor" Cancer Imaging

Journal of Cellular Biochemistry. May, 2009  |  Pubmed ID: 19306297

A major goal for in vivo biology is to develop models which can express multiple colors of fluorescent proteins in order to image many processes simultaneously in real time. Towards this goal, the cyan fluorescent protein (CFP) nude mouse was developed by crossing non-transgenic nude mice with the transgenic CK/ECFP mouse in which the beta-actin promoter drives expression of CFP in almost all tissues. In crosses between nu/nu CFP male mice and nu/+ CFP female mice, approximately 50% of the embryos fluoresced blue. In the CFP nude mice, the pancreas and reproductive organs displayed the strongest fluorescent signals of all internal organs which vary in intensity. Orthotopic implantation of XPA-1 human pancreatic cancer cells expressing red fluorescent protein (RFP); or green fluorescent protein (GFP) in the nucleus and RFP in the cytoplasm, was performed in female nude CFP mice. Color-coded fluorescence imaging of these human pancreatic cancer cells implanted into the bright blue fluorescent pancreas of the CFP nude mouse afforded novel insight into the interaction of the pancreatic tumor and the normal pancreas, in particular the strong desmoplastic reaction of the tumor. The naturally enhanced blue fluorescence of the pancreas in the CFP mouse serves as an ideal background for color-coded imaging of the interaction of implanted cancer cells and the host. The CFP nude mouse will provide unique understanding of the critical interplay between the cancer cells and their microenvironment.

Influence of Signal Transducer and Activator of Transcription-1 Signaling on Thyroid Morphology and Function

Endocrinology. Jul, 2009  |  Pubmed ID: 19325004

Interferon (IFN)-gamma has been involved in the pathogenesis of Hashimoto thyroiditis. It is a cytokine released by infiltrating mononuclear cells that mediates its actions mainly through signal transducer and activator of transcription-1 (STAT1) but also through other transcription factors. To dissect the effect of IFN gamma on thyroid morphology and function, we crossed transgenic mice that express IFN gamma specifically in the thyroid gland to mice deficient in STAT1. Lack of STAT1 ameliorated the abnormal thyroid morphology and the primary hypothyroidism typical of IFN gamma transgenic mice but not the suppressed iodine accumulation. Interestingly, lack of STAT1 alone decreased iodine accumulation, seemingly through expression of TGFbeta. These results indicate that STAT1 is required to mediate some but not all of the phenotypic changes induced by IFN gamma and that it also regulates iodine accumulation via TGFbeta signaling.

Monopolar Surface Electromyography: a Better Tool to Assess Motoneuron Excitability Upon Passive Muscle Stretching

The Journal of Physiological Sciences : JPS. May, 2009  |  Pubmed ID: 19340538

Bipolar and monopolar surface electromyography (sEMG) are known procedures to measure the H-reflex. However, signal cancellation is a potential experimental problem of bipolar sEMG. The results of our study show that monopolar sEMG was the more sensitive procedure to differentiate motoneuron excitability at different passive muscle stretching speeds as it overcame signal cancellation.

Regenerative Potentials of the Murine Thyroid in Experimental Autoimmune Thyroiditis: Role of CD24

Endocrinology. Jan, 2009  |  Pubmed ID: 18801910

Hashimoto thyroiditis can be partially reproduced in mice by immunization with thyroglobulin or, more recently, thyroperoxidase. This experimental autoimmune thyroiditis (EAT) model has been extensively characterized during early disease phases (up to d 35 after immunization). By extending the analysis of EAT to 100 d after immunization, we noted a remarkable regenerative capacity of the thyroid and the expression of Oct-4, suggesting in vivo the existence of adult thyroid stem cells. After an almost complete destruction of the follicular architecture, occurring between d 21 and 28, the thyroid was capable of restoring its follicles and reducing the mononuclear infiltration, so that by d 100 after immunization, it regained its normal morphology and function. During this regeneration process, thyrocytes expressed high levels of CD24. We therefore assessed the role of CD24 in thyroid regeneration by inducing EAT in mice lacking CD24. Regeneration was faster in the absence of CD24, likely a consequence of the effect of CD24 on the infiltrating lymphocytes. The study suggests that the EAT model can also be used as a tool to investigate adult thyroid stem cells.

Immunoproteasome Overexpression Underlies the Pathogenesis of Thyroid Oncocytes and Primary Hypothyroidism: Studies in Humans and Mice

PloS One. 2009  |  Pubmed ID: 19924240

Oncocytes of the thyroid gland (Hürthle cells) are found in tumors and autoimmune diseases. They have a unique appearance characterized by abundant granular eosinophilic cytoplasm and hyperchromatic nucleus. Their pathogenesis has remained, thus far, unknown.

Interleukin-12 Induces Salivary Gland Dysfunction in Transgenic Mice, Providing a New Model of Sjögren's Syndrome

Arthritis and Rheumatism. Dec, 2009  |  Pubmed ID: 19950301

Interleukin-12 (IL-12) is a pleiotropic cytokine that is elevated in the affected organs of patients with Sjögren's syndrome (SS). We have previously reported that overexpression of IL-12 in CBA mice leads to mononuclear infiltration of salivary and lacrimal glands, as well as to expansion of bronchial lymphoid tissue and decreased mucociliary clearance. Because xerostomia is one of the most important clinical features in SS patients, our main objective in the current study was to evaluate salivary gland function in IL-12-transgenic mice. Our secondary objective was to further characterize this animal model and to determine if the changes observed in these mice are representative of those observed in patients with SS overall.

Ultra-fast Switching of Light by Absorption Saturation in Vacuum Ultra-violet Region

Optics Express. Dec, 2009  |  Pubmed ID: 20052051

Advances in free electron lasers producing high energy photons [Nat. Photonics 2(9), 555-559 (2008)] are expected to open up a new science of nonlinear optics of high energy photons. Specifically, lasers of photon energy higher than the plasma frequency of a metal can show new interaction features because they can penetrate deeply into metals without strong reflection. Here we show the observation of ultra-fast switching of vacuum ultra-violet (VUV) light caused by saturable absorption of a solid metal target. A strong gating is observed at energy fluences above 6J/cm2 at wavelength of 51 nm with tin metal thin layers. The ratio of the transmission at high intensity to low intensity is typically greater than 100:1. This means we can design new nonlinear photonic devices such as auto-correlator and pulse slicer for the VUV region.

Caffeine-potentiated Chemotherapy for Metastatic Osteosarcoma

Journal of Orthopaedic Science : Official Journal of the Japanese Orthopaedic Association. Sep, 2009  |  Pubmed ID: 19802667

The prognosis for patients with metastatic osteosarcoma is still poor despite the development of effective adjuvant and neoadjuvant chemotherapy regimens. We have developed caffeine-potentiated chemotherapy for treatment of high-grade bone and soft tissue sarcomas based on the ability of caffeine to enhance the cytocidal effects of anticancer drugs. We report results of caffeine-potentiated chemotherapy for patients with osteosarcoma with pulmonary metastases.

Real-time Imaging of Single Cancer-cell Dynamics of Lung Metastasis

Journal of Cellular Biochemistry. Jan, 2010  |  Pubmed ID: 19911396

We have developed a new in vivo mouse model to image single cancer-cell dynamics of metastasis to the lung in real-time. Regulating airflow volume with a novel endotracheal intubation method enabled controlling lung expansion adequate for imaging of the exposed lung surface. Cancer cells expressing green fluorescent protein (GFP) in the nucleus and red fluorescent protein (RFP) in the cytoplasm were injected in the tail vein of the mouse. The right chest wall was then opened in order to image metastases on the lung surface directly. After each observation, the chest wall was sutured and the air was suctioned in order to re-inflate the lung, in order to keep the mice alive. Observations have been carried out for up to 8 h per session and repeated up to six times per mouse thus far. The seeding and arresting of single cancer cells on the lung, accumulation of cancer-cell emboli, cancer-cell viability, and metastatic colony formation were imaged in real-time. This new technology makes it possible to observe real-time monitoring of cancer-cell dynamics of metastasis in the lung and to identify potential metastatic stem cells.

Pregnancy, Postpartum Autoimmune Thyroiditis, and Autoimmune Hypophysitis: Intimate Relationships

Autoimmunity Reviews. Jan, 2010  |  Pubmed ID: 19539059

Autoimmune diseases comprise a group of about 85 heterogeneous conditions that can affect virtually any organ and tissue in the body. Many autoimmune diseases change significantly during pregnancy: some ameliorate, some worsen, and others are unaffected. Two autoimmune diseases present prominently in relation to pregnancy: postpartum autoimmune thyroiditis and autoimmune hypophysitis. This article will review the current state of knowledge of the immunological changes that occur during normal pregnancy, and will explore the striking temporal association with pregnancy observed in thyroiditis and hypophysitis.

A Nonclassical Model of Autoimmune Hypothyroidism

Thyroid : Official Journal of the American Thyroid Association. Jan, 2010  |  Pubmed ID: 20067377

Excess Iodide Decreases Transcription of NIS and VEGF Genes in Rat FRTL-5 Thyroid Cells

Biochemical and Biophysical Research Communications. Mar, 2010  |  Pubmed ID: 20132794

Although it is well known that an excess of iodide suppresses thyroid function and blood flow in vivo, the underlying molecular mechanisms are not fully known. The functional effect of iodide occurs at multiple steps, which include inhibition of sodium/iodide symporter (NIS) expression, transient block of organification, and inhibition of hormonal release. The vascular effect likely involves suppression of the vascular endothelial growth factor (VEGF) gene. In this report, we show that excess iodide coordinately suppresses the expression of the NIS and VEGF genes in FRTL-5 thyroid cells. We also demonstrate that the mechanism of iodide suppression of NIS gene expression is transcriptional, which is synergized by the addition of thyroglobulin. Based on the findings of reporter gene assays and electrophoretic gel mobility shift analysis, we also report two novel DNA binding proteins that responded specifically to iodide and modulated NIS promoter activity. The results suggest that excess iodide affects thyroid vascular function in addition to iodide uptake. This study provides additional insights into the mechanism of action of excess iodide on thyroid function.

Response-time Improved Hydrothermal-method-grown ZnO Scintillator for Soft X-ray Free-electron Laser Timing-observation

The Review of Scientific Instruments. Mar, 2010  |  Pubmed ID: 20370156

For pump and probe experiments in x-ray free-electron laser (XFEL) facilities, accurate timing synchronization between short-wavelength femtosecond pulses from XFELs and short optical pulses from other light sources is required. For this purpose, the response time of a hydrothermal-method-grown ZnO is improved by over one order of magnitude via intentional iron ion doping. The fluorescence rise- and decay-time constants are measured to be less than 10 and 100 ps, respectively. Owing to its intense fluorescence even for single pulse XFEL excitation, the timing jitter of the soft x-ray pulse and timing electronics are evaluated to be less than 70 ps.

UV Light Killing Efficacy of Fluorescent Protein-expressing Cancer Cells in Vitro and in Vivo

Journal of Cellular Biochemistry. Aug, 2010  |  Pubmed ID: 20506255

We investigated the cell-killing efficacy of UV light on cancer cells expressing GFP in the nucleus and RFP in the cytoplasm (dual-color cells). After exposure to various doses of UVA, UVB, or UVC, apoptotic and viable cells were quantitated under fluorescence microscopy using dual-color 143B human osteosarcoma cells, HT-1080 human fibrosarcoma cells, Lewis lung carcinoma (LLC), and XPA-1 human pancreatic cancer cells in vitro. UV-induced cancer cell death was wave-length and dose dependent, as well as cell-line dependent. After UVA exposure, most cells were viable even when the UV dose was increased up to 200 J/m(2). With UVB irradiation, cell death was observed with irradiation at 50 J/m(2). For UVC, as little as 25 J/m(2) UVC irradiation killed approximately 70% of the 143B dual-color cells. This dose of UVB or UVA had almost no effect on the cancer cells. UV-induced cancer cell death varied among the cell lines. Cell death began about 4 h after irradiation and continued until 10 h after irradiation. UVC exposure also suppressed cancer cell growth in nude mice in a model of minimal residual cancer (MRC). No apparent side effects of UVC exposure were observed. This study opens up the possibility of UVC treatment for MRC after surgical resection.

UV/thermally Driven Rewritable Wettability Patterns on TiO2-PDMS Composite Films

ACS Applied Materials & Interfaces. Sep, 2010  |  Pubmed ID: 20712336

Composite films of TiO2 and polydimethylsiloxane (PDMS) are prepared by a sol-gel method, cured with UV irradiation, and then treated in hot water to crystallize the TiO2 in the film. The presence of anatase TiO2 contributes to the photoinduced superhydrophilicity of the film under UV irradiation. Contact angle studies reveal that the TiO2-PDMS composite film recovers its original hydrophobic state. Hydrophobic-superhydrophilic patterns are successfully formed on the films. The wettability patterns can be erased by UV irradiation and thermal treatment. New wettability patterns can be reconstructed, demonstrating that the film exhibits rewritable wettability without the need for organic chemicals.

[Optic Nerve Injury Without Optic Canal Fracture Revealed by MR Imaging with Short Inversion Time Inversion Recovery Sequences--a Case Report]

Brain and Nerve = Shinkei Kenkyū No Shinpo. Aug, 2010  |  Pubmed ID: 20714039

GFP-fluorescence-guided UVC Irradiation Inhibits Melanoma Growth and Angiogenesis in Nude Mice

Anticancer Research. Sep, 2010  |  Pubmed ID: 20944099

Melanoma cell lines that stably express green fluorescent protein (GFP) and nude mice that ubiquitously express red fluorescent protein (RFP) have previously been developed to study tumor-host interaction by color-coded imaging. In the present study, the efficacy of fluorescence-guided ultraviolet C (UVC) irradiation on the growth of murine melanoma expressing GFP in the ear of RFP mice was determined using a non-invasive ear-tumor imaging model developed previously. The GFP-expressing melanoma and RFP-expressing blood vessels from the transgenic mice expressing RFP used as hosts were readily visible using non-invasive imaging. The melanoma was treated under fluorescence guidance with UVC at 650 J/m2/minute for 3 minutes. The ears of the mice were observed before and 24 hours after irradiation with UVC. UVC inhibited melanoma growth and also damaged blood vessels in the tumor. Thus, UVC irradiation has a direct effect on melanoma growth as well as an anti-angiogenesis effect. This color-coded tumor-host model is useful for evaluation of treatment efficacy on melanoma growth and angiogenesis, which are readily discernable with non-invasive color-coded fluorescent protein imaging. These results suggest that fluorescence-guided UVC irradiation is a promising therapeutic strategy for melanoma.

Long-working-distance Fluorescence Microscope with High-numerical-aperture Objectives for Variable-magnification Imaging in Live Mice from Macro- to Subcellular

Journal of Biomedical Optics. Nov-Dec, 2010  |  Pubmed ID: 21198203

We demonstrate the development of a long-working-distance fluorescence microscope with high-numerical-aperture objectives for variable-magnification imaging in live mice from macro- to subcellular. To observe cytoplasmic and nuclear dynamics of cancer cells in the living mouse, 143B human osteosarcoma cells are labeled with green fluorescent protein in the nucleus and red fluorescent protein in the cytoplasm. These dual-color cells are injected by a vascular route in an abdominal skin flap in nude mice. The mice are then imaged with the Olympus MVX10 macroview fluorescence microscope. With the MVX10, the nuclear and cytoplasmic behavior of cancer cells trafficking in blood vessels of live mice is observed. We also image lung metastases in live mice from the macro- to the subcellular level by opening the chest wall and imaging the exposed lung in live mice. Injected splenocytes, expressing cyan fluorescent protein, could also be imaged on the lung of live mice. We demonstrate that the MVX10 microscope offers the possibility of full-range in vivo fluorescence imaging from macro- to subcellular and should enable widespread use of powerful imaging technologies enabled by genetic reporters and other fluorophores.

Fragments of Genomic DNA Released by Injured Cells Activate Innate Immunity and Suppress Endocrine Function in the Thyroid

Endocrinology. Apr, 2011  |  Pubmed ID: 21303947

Activation of innate and acquired immune responses, which can be induced by infection, inflammation, or tissue injury, may impact the development of autoimmunity. Although stimulation of cells by double-stranded DNA (dsDNA) has been shown to activate immune responses, the role of self-genomic DNA fragments released in the context of sterile cellular injury is not well understood. Using cultured thyroid cells, we show that cell injury prompts the release of genomic DNA into the cytosol, which is associated with the production of type I interferons, inflammatory cytokines, and chemokines. Molecules necessary for antigen processing and presentation to lymphocytes are also induced in thyroid cells by injury. dsDNA strongly suppressed the expression of sodium/iodide symporter and radioiodine uptake. To identify molecules responsible for sensing cytosolic dsDNA, we directly identified the cellular proteins that bound a dsDNA Sepharose column by mass spectrometry. Our analysis identified histone H2B, which was previously demonstrated to be an essential factor that mediates the activation of innate immunity induced by dsDNA. Knockdown of histone H2B using specific small interfering RNA abolished cell injury-induced innate immune activation and increased sodium/iodide symporter expression. These results indicate that genomic DNA fragments released by cell injury are recognized by extrachromosomal histone H2B, which results in the activation of genes involved in both innate and acquired immune responses in thyroid cells and suppression of thyroid function. These results suggest that sterile thyroid injury, in the absence of infection, may be sufficient to trigger autoimmune reaction and to induce thyroid dysfunction.

The Bulge Area is the Major Hair Follicle Source of Nestin-expressing Pluripotent Stem Cells Which Can Repair the Spinal Cord Compared to the Dermal Papilla

Cell Cycle (Georgetown, Tex.). Mar, 2011  |  Pubmed ID: 21330787

Nestin has been shown to be expressed in the hair follicle, both in the bulge area (BA) as well as the dermal papilla (DP). Nestin-expressing stem cells of both the BA and DP have been previously shown to be pluripotent and be able to form neurons and other non-follicle cell types. The nestin-expressing pluripotent stem cells from the DP have been termed skin precursor or SKP cells. The objective of the present study was to determine the major source of nestin-expressing pluripotent stem cells in the hair follicle and to compare the ability of the nestin-expressing pluripotent stem cells from the BA and DP to repair spinal cord injury. Transgenic mice in which the nestin promoter drives GFP (ND-GFP) were used in order to observe nestin expression in the BA and DP. Nestin-expressing DP cells were found in early and middle anagen. The BA had nestin expression throughout the hair cycle and to a greater extent than the DP. The cells from both regions had very long processes extending from them as shown by two-photon confocal microscopy. Nestin-expressing stem cells from both areas differentiated into neuronal cells at high frequency in vitro. Both nestin-expressing DP and BA cells differentiated into neuronal and glial cells after transplantation to the injured spinal cord and enhanced injury repair and locomotor recovery within four weeks. Nestin-expressing pluripotent stem cells from both the BA and DP have potential for spinal cord regeneration, with the BA being the greater and more constant source.

Role of Thyroglobulin on Negative Feedback Autoregulation of Thyroid Follicular Function and Growth

The Journal of Endocrinology. May, 2011  |  Pubmed ID: 21378092

Thyroid function is tightly regulated by TSH. Although individual follicles are exposed to the same blood supply of TSH and express relatively homogenous levels of the TSH receptor, the function of individual follicles is variable. It was shown that thyroglobulin (Tg), stored in the follicular lumen, is a potent negative feedback regulator of follicular function. Thus, physiological concentrations of Tg significantly suppress thyroid-specific gene expression and antagonize the TSH-mediated stimulation that induces expression of thyroid-specific genes. Tg coordinately regulates both basal and apical iodide transporters in thyroid follicular cells. Recently, it was also reported that Tg could induce thyroid cell growth in the absence of TSH. These results indicate that Tg is an essential autocrine regulator of physiological thyroid follicular function that counteracts the effects of TSH.

Functional Outcomes After Total Scapulectomy for Malignant Bone or Soft Tissue Tumors in the Shoulder Girdle

International Journal of Clinical Oncology / Japan Society of Clinical Oncology. Oct, 2011  |  Pubmed ID: 21480004

The shoulder girdle is a common site for malignant bone and soft tissue tumors. Total scapulectomy represents an attractive alternative to amputation when the whole scapula is invaded with tumor and the neurovascular bundle can be preserved during tumor resection. The purpose of this study was to investigate functional outcomes after total scapulectomy.

Inhibition of Fas Ligand in NOD Mice Unmasks a Protective Role for IL-10 Against Insulitis Development

The American Journal of Pathology. Aug, 2011  |  Pubmed ID: 21718680

Type 1 diabetes mellitus (T1D) is an autoimmune disease caused by the destruction of pancreatic insulin-producing β cells by autoreactive T cells early in life. Despite daily insulin injections, patients typically develop cardiovascular and other complications; and intensive efforts are being directed toward identifying therapeutic targets to prevent the disease without directly impinging on the host defense. Fas ligand (FasL) is one potential target. Fas-FasL interactions primarily regulate T-cell homeostasis, not activation. Nevertheless, spontaneous gene mutation of Fas (called lpr mutation) or FasL (called the gld mutation) prevents autoimmune diabetes in nonobese diabetic (NOD) mice, the widely used model for T1D. Furthermore, although homozygous gld mutations cause age-dependent lymphoproliferation, limiting the gld mutation to one allele (NOD-gld/+) or treating NOD-wild-type mice with FasL-neutralizing monoclonal antibody completely prevents the disease development without causing lymphoproliferation or immune suppression. Herein, we show that the heterozygous gld mutation inhibits the accumulation of diabetogenic T cells in the pancreas, without interfering with their proliferation and expansion in the draining pancreatic lymph nodes. Pancreata from NOD-gld/+ mice contained B cells that expressed CD5 and produced IL-10, which was critical for maintenance of the disease resistance because its neutralization with an IL-10 receptor-blocking monoclonal antibody allowed accumulation of CD4 T cells in the pancreas and led to insulitis development. The results provide novel insights into the pathogenesis of T1D that could have important therapeutic implications.

Activity of Bone Morphogenetic Protein-7 After Treatment at Various Temperatures: Freezing Vs. Pasteurization Vs. Allograft

Cryobiology. Dec, 2011  |  Pubmed ID: 21963423

Insufficient bone union is the occasional complication of biomechanical reconstruction after malignant bone tumor resection using temperature treated tumor bearing bone; freezing, pasteurization, and autoclaving. Since bone morphogenetic protein (BMP) plays an important role in bone formation, we assessed the amount and activity of BMP preserved after several temperature treatments, including -196 and -73°C for 20 min, 60 and 100°C for 30 min, 60°C for 10h following -80°C for 12h as an allograft model, and 4°C as the control. The material extracted from the human femoral bone was treated, and the amount of BMP-7 was analyzed using an enzyme-linked immunosorbent assay. Then, the activity of recombinant human BMP-7 after the treatment was assessed using a bioassay with NIH3T3 cells and immunoblotting analysis to measure the amount of phospho-Smad, one of the signaling substrates that reflect the intracellular reaction of BMPs. Both experiments revealed that BMP-7 was significantly better preserved in the hypothermia groups. The percentages of the amount of BMP-7 in which the control group was set at 100% were 114%, 108%, 70%, 49%, and 53% in the -196, -73, 60, 100°C, and the allograft-model group, respectively. The percentages of the amount of phospho-Smad were 89%, 87%, 24%, 4.9%, and 14% in the -196, -73, 60, 100°C, and the allograft-model group, respectively. These results suggested that freezing possibly preserves osteoinductive ability than hyperthermia treatment.

Comparison of Cancer-cell Seeding, Viability and Deformation in the Lung, Muscle and Liver, Visualized by Subcellular Real-time Imaging in the Live Mouse

Anticancer Research. Nov, 2011  |  Pubmed ID: 22110185

The comparison of cancer cell seeding, deformation and viability in the lung, muscle and liver of nude mice in real-time is reported here. The mice were intubated to support ventilation with positive end-respiratory pressure (PEEP) for imaging on the lung. Human fibrosarcoma cells with green fluorescent protein (GFP) in the nucleus and red fluorescent protein (RFP) in the cytoplasm (dual-color HT-1080 cells) were injected into the tail vein for lung imaging, the portal vein for liver imaging or the abdominal aorta for muscle imaging which was performed with an Olympus OV100 Small Animal Imaging System. The length of the cytoplasm and nuclei in 20 seeded cancer cells were measured. A large number of cells initially arrested in the lung capillaries and many cells formed aggregates. The cell number decreased rapidly at 6 and 24 h. There was no significant difference in cancer cell survival when immunocompetent C57BL/6 mice were used in place of the nude mice, suggesting that T cell reaction is not very important in the first 24 h after seeding of cancer cells in the lung. In the lung and liver, little cancer cell deformation occurred. In contrast in the muscle, the cytoplasm and nuclei of the seeded cells were highly deformed and many fragmented cells were observed. The rate of cancer cell death was highest in the lung and lowest in the muscle. In each organ, single disseminated cells tended to die earlier than aggregated cells. The results of this study suggest that the early steps of metastasis are different in the lung, liver and muscle.

Anterior Corpectomy With Fusion in Combination With an Anterior Cervical Plate in the Management of Ossification of the Posterior Longitudinal Ligament

Journal of Spinal Disorders & Techniques. Nov, 2011  |  Pubmed ID: 22124427

STUDY DESIGN: A retrospective study. OBJECTIVE: The aim of this study is to evaluate the safety and efficacy of an anterior cervical plate (ACP) used in combination with anterior corpectomy with fusion (ACF) for cervical ossification of the posterior longitudinal ligament (OPLL). SUMMARY OF BACKGROUND DATA: Consensus is lacking about the most suitable method to treat cervical myelopathy caused by OPLL. The decision to perform an ACF to treat multilevel myelopathy is controversial because of the potential for problems in the grafted bone. METHODS: We evaluated the surgical outcome of ACF combined with insertion of an ACP for treating cervical myelopathy caused by OPLL. The study group comprised 68 patients who were treated from 2006 to 2009 and followed for an average of 29.6 months. We retrospectively reviewed the information in the patients' charts and radiographs. RESULTS: No dislodgement of the grafted bone or implant was observed, and no patient developed infection, esophageal or tracheal lacerations, or rupture. Radiographs showed no evidence of nonunion. The mean preoperative and the final follow-up C2 to C7 lordotic angles were 6.2±9.5 degrees and 9.4±7.6 degrees, respectively. The preoperative and the final follow-up lordotic angles of the fusion area were 2.0±8.1 degrees and 5.9±6.4 degrees, respectively. The average change in fusion area length was a 1.2 mm increase from before to after the operation and a 1.8 mm decrease from after the operation to the final follow-up. The average recovery rate of the Japanese Orthopaedic Association score was 63.0%. The surgical outcome of ACF with an ACP is satisfactory. CONCLUSIONS: Insertion of an ACP is a good solution for preventing problems with the grafted bone after ACF. Our study suggests that the indications for an anterior-only procedure for the management of cervical OPLL can be expanded.

Pituitary and Systemic Autoimmunity in a Case of Intrasellar Germinoma

Pituitary. Dec, 2011  |  Pubmed ID: 19466616

Germinomas arising in the sella turcica are difficult to differentiate from autoimmune hypophysitis because of similar clinical and pathological features. This differentiation, nevertheless, is critical for patient care due to different treatments of the two diseases. We report the case of an 11-year-old girl who presented with diabetes insipidus and growth retardation, and was found to have an intra- and supra-sellar mass. Initial examination of the pituitary biopsy showed diffuse lymphocytic infiltration of the adenohypophysis and absent placental alkaline phosphatase expression, leading to a diagnosis of hypophysitis and glucocorticoid treatment. Because of the lack of clinical and radiological response, the pituitary specimen was re-examined, revealing this time the presence of scattered c-kit and Oct4 positive germinoma cells. The revised diagnosis prompted the initiation of radiotherapy, which induced disappearance of the pituitary mass. Immunological studies showed that the patient's serum recognized antigens expressed by the patient's own germinoma cells, as well as pituitary antigens like growth hormone and systemic antigens like the Sjögren syndrome antigen B and alpha-enolase. The study first reports the presence of pituitary and systemic antibodies in a patient with intrasellar germinoma, and reminds us that diffuse lymphocytic infiltration of the pituitary gland and pituitary antibodies does not always indicate a diagnosis of autoimmune hypophysitis.

Color-coded Real-time Subcellular Fluorescence Imaging of the Interaction Between Cancer and Host Cells in Live Mice

Anticancer Research. Jan, 2012  |  Pubmed ID: 22213286

Stromal cells are essential for tumor growth. Stromal cells interact with cancer cells during tumor growth and progression. We report here the development of a tri-color imageable mouse model to visualize the interaction between host cells and cancer cells. To observe subcellular cancer cell dynamics in vivo, HT-1080 human fibrosarcoma cells were labeled in the nucleus with histone H2B-green fluorescent protein (GFP) and with retroviral red fluorescent protein (RFP) in the cytoplasm. HT-1080-GFP-RFP cells were sprinkled over a skin-flap in transgenic GFP immunocompetent mice. After 24 h, the mice were imaged with an Olympus IV100 laser scanning microscope. HT-1080-GFP-RFP cells were visualized surrounded by host-derived lymphocytes and macrophages both expressing GFP. It was possible to observe host GFP macrophages contacting, engulfing, and digesting dual-color HT-1080-GFP-RFP cells in real time. The dual-color cancer cells were readily visible after being engulfed in the GFP macrophages. Other cancer cells were visualized being killed by lymphocytes. The results of this study show that differentially labeling cells with spectrally-distinct fluorescent protein can allow subcellular-resolution imaging of cell-cell interactions between host and cancer cells.

Inhibition and Eradication of Human Glioma with Tumor-targeting Salmonella Typhimurium in an Orthotopic Nude-mouse Model

Cell Cycle (Georgetown, Tex.). Feb, 2012  |  Pubmed ID: 22274398

Malignant glioma tumors are the most common primary central nervous system tumors. Despite the multidisciplinary approach to treatment, prognosis remains poor. In this study, we demonstrated that the Salmonella typhimurium A1-R tumor-targeting strain can inhibit and eradicate human glioma in an orthotopic nude-mouse model. S. typhimurium A1-R was administered by injection through a craniotomy open-window or intravenously in nude mice. To establish the model, 2 x 105 U87-RFP human glioma cells were injected stereotactically into the mouse brain through the craniotomy open window. Two weeks after glioma-cell implantation, mice were treated with S. typhimurium A1-R [2 x 10 ( 7) CFU/200 μl intravenous injection (i.v.) or 1 x 10 ( 6) CFU/1 μl intracranial injection (i.c.)] once a week for 3 weeks. Brain tumors were observed by fluorescence imaging through the craniotomy open window over time. S. typhimurium A1-R, administered i.c., inhibited brain tumor growth 7.6-fold compared with untreated mice (p = 0.009) and improved survival 73% (p = 0.001). Two of ten mice appeared to have their tumors eradicated. Intravenous administration of S. typhimurium A1-R was not effective. The craniotomy open window enabled observation of tumor growth in the brain in real time in both treated and untreated mice. The results of the present study demonstrate that bacterial therapy of brain cancer is a novel, effective and safe treatment strategy in a highly treatment-resistance cancer.

Imaging the Inhibition of Anti-β1 Integrin Antibody on Lung Seeding of Single Cells in Live Mice

International Journal of Cancer. Journal International Du Cancer. Feb, 2012  |  Pubmed ID: 22323248

Integrins play a role in tumor growth and metastasis. However, the effect of integrin inhibition has not been visualized on single cancer cells in vivo. In this study, we used a powerful subcellular in vivo imaging model to demonstrate how an anti-integrin antibody affects seeding and growth of osteosarcoma cells on the lung. The 143B human osteosarcoma cell line expressing red fluorescent protein (RFP) in the cytoplasm and green fluorescent protein (GFP) in the nucleus was established. Such double-labeled cells enable imaging of apoptosis and mitosis and other nuclear-cytoplasmic dynamics. Using the double-labeled osteosarcoma cells, single cancer-cell seeding in the lung after i.v. injection of osteosarcoma cells was imaged. The anti-β1 integrin monoclonal antibody, AIIB2, greatly inhibited the seeding of cancer cells on the lung while a control antibody had no effect. To image the efficacy of the anti-integrin antibody on spontaneous metastasis, mice with orthotopically-growing 143B-RFP cells in the tibia were also treated with AIIB2 or control anti-rat IgG1 antibody. After 3 weeks treatment, mice were sacrificed and primary tumors and lung metastases were evaluated with fluorescence imaging. AIIB2 significantly inhibited spontaneous lung metastasis but not primary tumor growth, possibly due to inhibition of lung seeding of the cancer cells as imaged in the experimental metastasis study. AIIB2 treatment also increased survival of mice with orthotopically-growing 143B-RFP. © 2012 Wiley-Liss, Inc.

Prestroke Anticoagulation and Paroxysmal Type Are Correlated with Favorable Outcome in Ischemic Stroke Patients with Atrial Fibrillation

Journal of Stroke and Cerebrovascular Diseases : the Official Journal of National Stroke Association. Jan, 2012  |  Pubmed ID: 20833081

Paroxysmal atrial fibrillation (AF), which often precedes permanent AF, is reported to be a risk factor for milder ischemic stroke. We assessed whether the type of AF and prestroke treatment with an anticoagulant were associated with physical disabilities in patients with AF-related acute ischemic stroke. We identified 162 consecutive acute ischemic stroke patients with AF who were admitted to our hospital over a 3-year period. Disability was measured using the modified Rankin Scale (mRS) at the time of discharge and was categorized according to favorable clinical outcome (mRS score 0-2). Of the 162 patients, 71 (43.8%) had paroxysmal AF and 91 had permanent AF. Fifty-six patients (34.6%) had been treated with a prophylactic anticoagulant. A total of 103 patients (63.6%) had a favorable outcome. Multivariate logistic analysis revealed that paroxysmal AF (odds ratio [OR], 1.58; P = .0187), prestroke anticoagulation treatment (OR, 1.95; P = .0019), and noncardiogenic embolism (OR, 2.20; P = .0073) were independent factors associated with a favorable clinical outcome. Our data indicate that paroxysmal AF and prestroke anticoagulation treatment are independently associated with favorable clinical outcome at the time of hospital discharge in patients with AF.