Sphingobium yanoikuyae Bacteremia, Japan.

We report a case of Sphingobium yanoikuyae bacteremia in an 89-year-old patient in Japan. No standard antimicrobial regimen has been established for S. yanoikuyae infections. However, ceftriaxone and ceftazidime treatments were effective in this case. Increased antimicrobial susceptibility data are needed to establish appropriate treatments for S. yanoikuyae.

Polymethylmethacrylate Membrane Dialyzer: Historic but Modern.

Polymethylmethacrylate (PMMA) hollow fiber membranes are one of the synthetic polymer hollow fiber membranes used to the hollow fiber artificial kidney. A PMMA hollow fiber membrane (PMMA membrane) has unique properties including the uniform structure and the adsorption property. Hemodialyzers using PMMA membranes, Filtryzer®, were approved in Japan in 1977 and have been used worldwide for over 40 years and so is a historical hemodialyzer.Various types in Filtryzer® having different pore sizes are developed and used in the clinical field. Filtryzer® B3 is a low-flux dialyzer. Filtryzer® BK has three types having different pore sizes, and above all, BK-F has the largest pores in the Filtryzer® series. Filtryzer® BG has a more uniform membrane structure by using weak anionic polymers compared with the earlier Filtryzer® series to remove ß2-MG more. Filtryzer® NF is the latest Filtryzer® series and was developed as a dialyzer having improved antithrombogenicity compared with previous models and having protein adsorption property as the same with them. There have been many reports concerning Filtryzer® including improvement of patients' symptoms such as pruritus and nutrition on the advantages for dialysis patients. Although PMMA membranes are historic dialysis tools used for over 40 years, they are also modern dialysis membranes that have been updated to respond to dialysis therapy at those time.

Proxy evaluation of dignity expectations and satisfaction of older patients with dementia by family members and nurses.

AIM: Develop a proxy evaluation questionnaire for patients' family members and nurses to evaluate dignity expectations and satisfaction of patients with dementia. DESIGN: Cross-sectional study. METHOD: A proxy questionnaire draft was prepared with 30 items on expectations for dignity and 23 items on satisfaction with dignity. And administered to three paired groups: 81 older patients with intact cognitive function, 75 family members, and 77 nurses. RESULTS: 18 of 30 items and 21 of 23 were correlated between patients and their family members' responses on dignity expectations and satisfaction, respectively. When limited to nurses with clinical experience of ≥20 years, there were significant correlations between patients' and nurses' responses (p < .05). Exploratory factor analysis of patient's responses to significantly correlated items extracted 3 factors with 13 items of expectations for dignity but no factors of satisfaction with dignity. Using a questionnaire provides insight for proxy evaluation of expectations for dignity.

Cellulitis with persistent bacteremia caused by Campylobacter lari in a patient with mantle-cell lymphoma.

Campylobacter lari is an organism occasionally isolated in humans but rarely causes bacteremia. We report the first case of cellulitis with bacteremia due to C. lari in a patient undergoing mantle-cell lymphoma. A 51-year-old man presented with a two-month history of fever and bilateral leg pain and redness. Despite oral ciprofloxacin administration, his symptoms had not improved. The blood culture sample in the anaerobic bottle yielded positive results and C. lari was identified by mass spectrometry. The bacteremia did not initially respond to oral azithromycin but responded to intravenous meropenem and amikacin for five days followed by oral minocycline. This report indicates that C. lari bacteremia may be treated with oral minocycline following short-term intravenous antimicrobial therapy even among patients undergoing hematological malignancies.

Preliminary evidence that different mechanisms underlie the anger superiority effect in children with and without Autism Spectrum Disorders.

Previous studies have demonstrated that angry faces capture humans' attention more rapidly than emotionally positive faces. This phenomenon is referred to as the anger superiority effect (ASE). Despite atypical emotional processing, adults and children with Autism Spectrum Disorders (ASD) have been reported to show ASE as well as typically developed (TD) individuals. So far, however, few studies have clarified whether or not the mechanisms underlying ASE are the same for both TD and ASD individuals. Here, we tested how TD and ASD children process schematic emotional faces during detection by employing a recognition task in combination with a face-in-the-crowd task. Results of the face-in-the-crowd task revealed the prevalence of ASE both in TD and ASD children. However, the results of the recognition task revealed group differences: In TD children, detection of angry faces required more configural face processing and disrupted the processing of local features. In ASD children, on the other hand, it required more feature-based processing rather than configural processing. Despite the small sample sizes, these findings provide preliminary evidence that children with ASD, in contrast to TD children, show quick detection of angry faces by extracting local features in faces.

Characterization of the intronic VNTR polymorphisms found in a paralog of chicken serotonin transporter gene.

Polymorphisms in the neurotransmitter-related genes can be a major source of behavioral variations. We searched for polymorphic sites in chicken neurotransmitter-related genes and identified two variable number of tandem repeat (VNTR) loci encompassing the paralog of chicken serotonin transporter gene (5-HTT). Both intronic VNTR were highly polymorphic across chicken breeds and the other Galliformes species, even though predominant alleles were considerably different among breeds. One VNTR locus contained sequences complementary to a conserved motif of CCCTC-binding factor (CTCF) within each repetitive unit, indicating that transcription of chicken 5-HTT paralog may be regulated by the CTCF protein. It is of great interest to contrast these results with previous knowledge on the human 5-HTT that also has CTCF binding sites in the repetitive units of intronic VNTR. Additionally, we measured the degree of impulsiveness in domestic chicks for their preference of immediate/small to large/delayed rewards. A significant difference in the impulsiveness score was detected between two chicken breeds (White Leghorn vs. Boris Brown; P < 0.01), as well as between White Leghorn chicks with different 5-HTT genotypes. These findings imply the possibility that 5-HTT VNTR genotypes may have some impact on chicks' impulsive choice by modifying the serotonergic neurotransmission.

Involvement of the parasympathetic nervous system in the initiation of regeneration of pancreatic ß-cells.

The mechanism that initiates regeneration of pancreatic ß-cells is not clear at present. The vagal nerve is implicated in the regulation of gastrointestinal functions, glucose metabolism and proliferation of pancreatic ß-cells under physiological conditions. To elucidate the triggering mechanism of the regeneration of pancreatic ß-cells, we examined the involvement of the vagal nerve. To this end, we employed a rat pancreatic duct ligation (DL) model, in which profound ß-cell neogenesis and ß-cell proliferation were observed within a week. We administered atropine to block the vagal nerve. Administration of atropine inhibited proliferation of ß-cells in both islets and islet-like cell clusters (ICC), without affecting ductal cell proliferation in the ligated pancreas. The numbers of PDX-1 and MafB-positive cells in or attaching to the ducts were significantly reduced by atropine. MafB/glucagon and MafB/insulin double-positive cells were also decreased by atropine. Finally, atropine reduced the number of MafA-positive ductal cells, all of which were positive for insulin, by 50% on day 5. These results strongly suggest that the vagal nerve is involved in ß-cell proliferation, induction of endocrine progenitors and neogenesis of α- and ß-cells.

[Arbekacin sulfate concentrations in peripheral lymph and in serum after intravenous injection: report of four cases].

Antibiotic levels in serum are commonly used to guide antibiotic therapy. The antibiotic levels in peripheral lymph are a more accurate reflection of the efficacy of antibiotic penetration into the tissues of patients with complicated skin and soft-tissue infections. The pharmacokinetics of arbekacin sulfate (ABK) in peripheral lymph after systemic administration has not been studied. Four patients (cases 1-4) with skin and soft-tissue infections (average age 74.3 years, range 54 to 85) received 200 mg of ABK intravenously once a day either by slow bolus (5min.) or by slow infusion (60 min.). The serum concentrations collected 60min. after the start ofABK infusion (C60) and the peripheral lymph concentrations of ABK were measured. 55 min. after initiation of slow 5-min. bolus (case 1), C60 was 32.5l microg/mL. The daily average concentration of ABK in peripheral lymph after slow bolus (case 1) was 14.84 microg/mL. The ratio peripheral lymph on daily average/C60 was 0.46. Patients (cases 2, 3 and 4) had been intravenously administered ABK at an infusion time of 60 min. C60 (cases 2, 3 and 4) were 14.10, 11.48 and 8.26 microg/mL, respectively. The daily average concentration of ABK in peripheral lymph after slow infusion (case 2) was 7.80 microg/mL. The average concentrations of ABK in peripheral lymph during the third eight hours since slow infusion (cases 3 and 4) were 0.72 and 2.23 microg/mL. The ratio peripheral lymph/C60 was 0.55, 0.06 and 0.27, respectively. An increase in the serum peak concentration of ABK may lead to considerable elevation of the concentration of ABK in peripheral lymph.

Essential roles of clinical nurse instructors in Japan: a Delphi study.

In Japan, the clinical nurse instructor is a staff nurse who teaches in clinical practicums. However, there is no consensus on the essential roles that clinical nurse instructors are expected to perform. We conducted a three-round Delphi survey to clarify the essential roles of the clinical nurse instructor in clinical practicums in undergraduate nursing education. The participants were an expert panel of 48 professionals in nursing education and clinical practicums, who rated the importance of 58 role items that were established through a literature review and pilot survey. Thirty one of these items were identified as essential roles, based on agreement of 80% or more of respondents. Further investigation revealed nine of the 31 items to be core roles, defined as the minimum essential roles that must be performed by clinical nurse instructors, however busy they become. The nine core roles are related to proper preparation for the clinical practicum, patient safety, and coordination with the nursing school faculty. It is important for the nursing school faculty to support and work in cooperation with clinical nurse instructors to help them fulfill these roles.

Conophylline suppresses pancreatic stellate cells and improves islet fibrosis in Goto-Kakizaki rats.

Activin A is a differentiation factor for ß-cells and is effective to promote ß-cell neogenesis. Activin A is also an autocrine activator of pancreatic stellate cells, which play a critical role in fibrogenesis of the pancreas. Conophylline (CnP) is a natural compound, which reproduces the effect of activin on ß-cell differentiation and promotes ß-cell neogenesis when administered in vivo. However, its effect on stellate cells is not known. We therefore investigated the effect of CnP on stellate cells both in vitro and in vivo. Unlike activin A, CnP inhibited activation of cultured stellate cells and reduced the production of collagen. We then analyzed the involvement of stellate cells in islet fibrosis in Goto-Kakizaki (GK) rats, a model of type 2 diabetes mellitus. In pancreatic sections obtained from 6-wk-old GK rats, CD68-positive macrophages and glial fibrillary acidic protein- and α-smooth muscle actin-positive stellate cells infiltrated into islets. Later, the number of macrophages was increased, and the α-smooth muscle actin staining of stellate cells became stronger, indicating the involvement of stellate cells in islet fibrosis in GK rats. When CnP was administered orally for 4 wk, starting from 6 wk of age, invasion of stellate cells and macrophages was markedly reduced and islet fibrosis was significantly improved. The insulin content was twice as high in CnP-treated rats. These results indicate that CnP exerts antifibrotic actions both in vitro and in vivo and improves islet fibrosis in Goto-Kakizaki rats.

Extracellular matrix modulates insulin production during differentiation of AR42J cells: functional role of Pax6 transcription factor.

Extracellular matrix (ECM) modulates differentiation of pancreatic ß-cells during development. However, the mechanism by which ECM proteins modulate differentiation is not totally clear. We investigated the effect of ECM proteins on differentiation ß-cells in vitro. We investigated the effect of basement membrane ECM on differentiation of AR42J cells and rat ductal cells. First, we examined the effect of reconstituted basement membrane, Matrigel on differentiation of AR42J cells induced by activin and betacellulin. Matrigel augmented insulin production and increased the expression of GLUT2, SUR1, and glucokinase. Among various transcription factors investigated, Matrigel markedly upregulated the expression of Pax6. When Pax6 was overexpressed in cells treated with activin and betacellulin, the expression of insulin was upregulated. Conversely, knockdown of Pax6 significantly reduced the insulin expression in cells cultured on Matrigel. The effects of Matrigel on insulin-production and induction of Pax6 were reproduced partially by laminin-1, a major component of Matrigel, and inhibited by anti-integrin-ß1 antibody. Matrigel also enhanced the activation of p38 mitogen-activated kinase induced by activin and betacellulin, which was inhibited by anti-ß1 antibody. Finally, the effect of Matrigel on differentiation was reproduced in rat cultured ductal cells, and Matrigel also increased the expression of Pax6. These results indicate that basement membrane ECM augments differentiation of pancreatic progenitor cells to insulin-secreting cells by upregulating the expression of Pax6. .

Identification of differentially expressed genes during proliferative response of the liver induced by follistatin.

The liver mass is controlled strictly and maintained constant in normal and pathological situations. An exception is observed after an administration of follistatin, which induces proliferation in intact liver. In the present study, we identified genes differentially expressed in proliferating liver caused by overexpression of follistatin-288. Adenovirus vector encoding follistatin-288 (Ad-FS) or green fluorescent protein was injected intraperitoneally in rats. Changes in the liver weight, expression of follistatin and nuclear bromodeoxyuridine labeling were measured. Samples taken on day 5 and day 7 were used to prepare RNA for microarray analysis. The expression of the genes was confirmed by quantitative reverse transcriptase PCR. After the injection of Ad-FS follistatin mRNA peaked on day 3, which was followed by progressive increase in the protein expression. A peak in bromodeoxyuridine labeling was observed on day 7. Microarray data from day 5 and day 7 samples showed that follistatin modified the expression of 907 genes, of which 575 were overexpressed and 332 were downregulated taking into consideration a two fold change reference compared to control rats. In particular, significant increases and time related changes in gene expression after the Ad-FS injection were found in nine genes including growth differentiation factor 15 and fibroblast growth factor 21. This study confirmed that follistatin induced proliferation in intact liver, and identified candidate genes involved in follistatin-induced liver cell growth.

Expression of the TRPM8-immunoreactivity in dorsal root ganglion neurons innervating the rat urinary bladder.

The neurochemical phenotypes of the transient receptor potential melastatin-8 (TRPM8)-immunoreactive afferent neurons innervating the rat urinary bladder were examined by using a highly sensitive tyramide signal amplification method, combined with wheat-germ agglutinin-horseradish peroxidase (WGA-HRP) retrograde tracing. TRPM8-immunoreactivity was detected in a small proportion of the WGA-HRP-labeled bladder afferent neurons in the dorsal root ganglia of the Th13-L1 (1.14%) and the L6-S1 (1.27%), and these neurons were small in size (<600 microm(2)). The 82.6+/-3.8% of the TRPM8-immunoreactive bladder afferent neurons and 80.9+/-1.5% of the total population of the TRPM8-immunoreactive afferent neurons in the observed dorsal root ganglia expressed NF200. On the other hand, the proportions of the co-expression of TRPM8 and nociceptive markers such as calcitonin gene-related peptide (CGRP), transient receptor potential vanilloid-1 (TRPV1), and isolectin B4 (IB4) in the bladder afferent neurons (81.5+/-5.2% for CGRP, 36.1+/-4.0% for TRPV1, and 15.8+/-5.5% for IB4) were higher in comparison to those in the total population of the TRPM8-immunoreactive afferent neurons (21.9+/-2.4% for CGRP, 16.6+/-1.7% for TRPV1, and 5.4+/-0.5% for IB4), although no significant difference existed for IB4. Our results suggest that the TRPM8-expressing bladder afferents should be classified as Adelta-fibers and C-fibers, while some of these afferents may be involved in nociceptive sensations.

Administration of conophylline and betacellulin-delta4 increases the beta-cell mass in neonatal streptozotocin-treated rats.

The present study was conducted to examine the effect of administration of conophylline (CnP) and betacellulindelta4 (BTCdelta4) on the beta-cell mass in neonatal streptozotocin-treated rats (neonatal STZ rats). STZ (100 microg/g) was injected into neonatal rats, and then CnP (2 microg/g) and/or BTCdelta4 (200 pmol/g) were administered to neonatal STZ rats for 1 week. The plasma glucose concentration was monitored, and an intraperitoneal glucose tolerance test (ipGTT) was performed on day 8 and at 8 weeks after the STZ injection. In neonatal STZ rats treated with control solution (S group), the plasma glucose concentration increased for several days after the STZ injection, returned to nearly normal levels, and then increased gradually after six weeks of age. Eight weeks after the STZ-injection, the plasma glucose concentration was increased significantly compared to that of normal rats. The glucose response to ipGTT was significantly reduced in neonatal STZ rats treated with CnP (CnP group), BTCdelta4 (delta4 group) and CnP+BTCdelta4 (CnP+delta4 group). The beta-cell mass and the insulin content of the pancreas were significantly increased in the CnP group and delta4 group. The effect of CnP+delta4 was greater than that of CnP alone or BTCdelta4 alone. CnP+BTCdelta4 significantly increased the number of PDX-1-positive ductal cells and the number of insulin/BrdU double-positive ductal cells. These results indicate the efficacy of CnP and BTCdelta4 in increasing the beta-cells mass of neonatal STZ-treated rats.

Sweet taste receptor expressed in pancreatic beta-cells activates the calcium and cyclic AMP signaling systems and stimulates insulin secretion.

BACKGROUND: Sweet taste receptor is expressed in the taste buds and enteroendocrine cells acting as a sugar sensor. We investigated the expression and function of the sweet taste receptor in MIN6 cells and mouse islets. METHODOLOGY/PRINCIPAL FINDINGS: The expression of the sweet taste receptor was determined by RT-PCR and immunohistochemistry. Changes in cytoplasmic Ca(2+) ([Ca(2+)](c)) and cAMP ([cAMP](c)) were monitored in MIN6 cells using fura-2 and Epac1-camps. Activation of protein kinase C was monitored by measuring translocation of MARCKS-GFP. Insulin was measured by radioimmunoassay. mRNA for T1R2, T1R3, and gustducin was expressed in MIN6 cells. In these cells, artificial sweeteners such as sucralose, succharin, and acesulfame-K increased insulin secretion and augmented secretion induced by glucose. Sucralose increased biphasic increase in [Ca(2+)](c). The second sustained phase was blocked by removal of extracellular calcium and addition of nifedipine. An inhibitor of inositol(1, 4, 5)-trisphophate receptor, 2-aminoethoxydiphenyl borate, blocked both phases of [Ca(2+)](c) response. The effect of sucralose on [Ca(2+)](c) was inhibited by gurmarin, an inhibitor of the sweet taste receptor, but not affected by a G(q) inhibitor. Sucralose also induced sustained elevation of [cAMP](c), which was only partially inhibited by removal of extracellular calcium and nifedipine. Finally, mouse islets expressed T1R2 and T1R3, and artificial sweeteners stimulated insulin secretion. CONCLUSIONS: Sweet taste receptor is expressed in beta-cells, and activation of this receptor induces insulin secretion by Ca(2+) and cAMP-dependent mechanisms.

A betacellulin mutant promotes differentiation of pancreatic acinar AR42J cells into insulin-producing cells with low affinity of binding to ErbB1.

Betacellulin (BTC) is one of the members of the epidermal growth factor (EGF) ligand family of ErbB receptor tyrosine kinases. It is a differentiation factor as well as a potent mitogen. BTC promotes the differentiation of pancreatic acinar-derived AR42J cells into insulin-producing cells. It independently and preferentially binds to two type I tyrosine kinase receptors, the EGF receptor (ErbB1) and ErbB4. However, the physiochemical characteristics of BTC that are responsible for its preferential binding to these two receptors have not been fully defined. In this study, to investigate the essential amino acid residues of BTC for binding to the two receptors, we introduced point mutations into the EGF domain of BTC employing error-prone PCR. The receptor binding abilities of 190 mutants expressed in Escherichia coli were assessed by enzyme immunoassay. Replacement of the glutamic acid residue at position 88 with a lysine residue in BTC was found to produce a significant loss of affinity for binding to ErbB1, while the affinity of binding to ErbB4 was unchanged. In addition, the mutant of BTC-E/88/K showed less growth-promoting activity on BALB/c 3T3 cells compared with that of the wild-type BTC protein. Interestingly, the BTC mutant protein promoted differentiation of pancreatic acinar AR42J cells at a high frequency into insulin-producing cells compared with AR42J cells that were treated with wild-type BTC protein. These results indicate the possibility of designing BTC mutants, which have an activity of inducing differentiation only, without facilitating growth promotion.

A simple method to induce differentiation of murine bone marrow mesenchymal cells to insulin-producing cells using conophylline and betacellulin-delta4.

The present study was conducted to establish a method to induce differentiation of bone marrow (MB)-derived mesenchymal cells into insulin-producing cells. When mouse BM-derived mesenchymal cells were cultured for 60 days in medium containing 10% fetal calf serum and 25 mM glucose, they expressed insulin. Addition of activin A and betacellulin (BTC) accelerated differentiation, and immunoreactive insulin was detected 14 days after the treatment. Insulin-containing secretory granules were observed in differentiated cells by electron microscopy. Treatment of BM-derived mesenchymal cells with conophylline (CnP) and BTC-delta4 further accelerated differentiation, and mRNA for insulin was detected 5 to 7 days after the treatment. Mesencymal cells treated with CnP and BTC-delta4 responded to a high concentration of glucose and secreted mature insulin. When these cells were transplanted into streptozotocin-treated mice, they markedly reduced the plasma glucose concentration, and the effect continued for at least 4 weeks. These results indicate an efficacy of the combination of CnP and BTC-delta4 in inducing differentiation of BM-derived mesenchymal cells into insulin-producing cells.

Reversal of streptozotocin-induced hyperglycemia by continuous supply of betacellulin in mice.

Previous studies have shown the efficacy of betacellulin (BTC) to promote beta-cell regeneration. Because of its short half-life, however, the effect of BTC may have been underestimated. This study was conducted to assess the effect of continuous administration of BTC on beta-cell regeneration. Adenovirus vectors encoding proBTC (Ad-proBTC) and mature BTC (Ad-mBTC) were prepared, and the efficacy of secretion of BTC was compared in AML12 hepatocytes. When AML12 cells were infected with Ad-proBTC or Ad-mBTC, cells infected with Ad-mBTC secreted considerably larger amount of BTC. We then infused Ad-mBTC into the mouse tail vein. Expression of BTC was detected in the liver for at least 21 days, and serum BTC was maintained at approximately 1 ng/ml for 7 days. When Ad-mBTC was infused immediately after administration of STZ (170 mg/kg), elevation of the plasma glucose induced by STZ was markedly inhibited, and the plasma glucose concentration remained at less than 200 mg/dl for 21 days. The insulin content and the beta-cell mass were significantly increased in Ad-mBTC-infused mice. These results indicate that continuous administration of BTC is quite effective in promoting regeneration of beta-cells.

Percentages of positive cores, cancer length and Gleason grade 4/5 cancer in systematic sextant biopsy are all predictive of adverse pathology and biochemical failure after radical prostatectomy.

AIM: We investigated whether the quantitative parameters of systematic sextant biopsies were predictive of either adverse pathological findings or disease recurrence after radical prostatectomy (RP). METHODS: We retrospectively evaluated a total of 117 men with untreated prostate cancer whose needle biopsies were matched with RP specimens. The pretreatment parameters of the serum prostate-specific antigen (PSA), the PSA density, the percentage of positive biopsy cores, the percentage of cancer length and the percentage of Gleason grade 4/5 cancer in the biopsy were determined and compared with the pathological features of prostate cancer in RP specimens. These pretreatment parameters and pathological factors in the RP specimens, including the cancer volume, the percentage of Gleason grade 4/5 cancer, the positive surgical margin and the seminal vesicle invasion were evaluated for their ability to predict the disease recurrence. RESULTS: The percentages of positive biopsy cores, the Gleason grade 4/5 cancer in the biopsy and the cancer length in the biopsy had a weak correlation with the cancer volume in RP specimens (r = 0.373, 0.345, 0.408, respectively). All quantitative biopsy parameters were strongly predictive of the non-organ-confined status, the positive surgical margin and the seminal vesicle invasion in the logistic regression analysis. The percentage of positive biopsy cores and the percentage of Gleason grade 4/5 cancer in the biopsy predicted biochemical failure after RP. CONCLUSION: These results indicate that quantitative biopsy parameters are independent predictors of the adverse pathology of prostate cancers and disease recurrence after RP.