Chemotactic responses of Brugia pahangi infective third-stage larvae to tris(hydroxymethyl)aminomethane-related compounds and amino acids.

Since the exogenous compound tris(hydroxymethyl)aminomethane (Tris) showed a potent chemoattractant activity for Brugia pahangi infective third-stage larvae (L3), it was assumed that, in natural infection to a host, filarial L3 can be expected to recognize an endogenous Tris-related compound. In addition, a few amino acids have been identified as water-soluble attractants for second-stage juveniles of Meloidogyne incognita, a plant parasitic nematode. Therefore, the present study assesses the in vitro chemotactic responses of B. pahangi L3 to Tris-related compounds and amino acids using an agar-plate assay. Among Tris-related compounds, 2-amino-1,3-propanediol (APD) and 2-amino-2-methyl-1,3-propanediol (AMPD) exhibited a potent chemoattractant activity for filarial L3 at a level similar to Tris. Furthermore, arginine (Arg) was identified as a potent attractant for filarial L3 among amino acids. In addition, filarial L3 were attracted to Arg, APD and AMPD in mild alkaline conditions rather than acidic conditions. The chemoattractant activity of the three compounds for filarial L3 was observed in concentrations between 6.3 and 200 mm. This is the first report to demonstrate that Arg, APD and AMPD are potent chemoattractants for B. pahangi L3. Endogenous Arg and APD, in particular, may be involved in the regulation of the chemotactic behaviour of filarial L3 in the infection to a host. The present results will help to elucidate the mechanism of filarial skin-penetrating invasion of a host.

Acute Inflammatory Syndrome Paradoxically Induced by De Novo Purine Inhibitors Synthesis Before Renal Transplantation: A Case Report and Review of the Literature.

BACKGROUND: Mycophenolate mofetil (MMF) and mizoribine (MZR) are increasingly used as immunosuppressive agents for organ transplantation and chronic inflammation. We report a patient with rheumatoid arthritis who had an acute inflammatory syndrome triggered by preoperative immunosuppression therapy with both MMF and MZR. CASE REPORT: A 41-year-old woman with IgA nephropathy was referred to our department for living donor renal transplantation. She had rheumatoid arthritis that was adequately treated with prednisolone 5 mg once a day and salazosulfapyridine 2000 mg once a day. MMF 1000 mg twice a day was started for desensitization therapy. Three days later, the patient developed arthritis in the joints of her left hand and elevated inflammatory markers. On day 7, MMF was switched to MZR 150 mg 3 times a day. However, the symptoms extended to both shoulders and the joints of the right foot; MZR was discontinued. The arthritis and inflammatory markers improved. Two months later, the patient was rechallenged with MMF followed by MZR, resulting in a similar clinical course as previously. Tacrolimus (TAC) 3 mg twice a day and everolimus (EVL) 0.5 mg twice a day were introduced as alternative immunosuppressant therapies. No arthritis occurred. ABO-compatible living donor renal transplantation was successfully performed. The patient received TAC, EVL, prednisolone, rituximab, and basiliximab, and her postoperative course was uneventful without arthritis or rejection. At 9 months postoperatively, the serum creatinine was 0.79 mg/dL. CONCLUSIONS: Acute inflammatory syndrome is an extremely rare complication triggered by preoperative immunosuppression therapy. If antimetabolites cannot be used in immunologically high-risk patients, transplantation becomes very difficult. Clinicians should keep in mind this paradoxical reaction.

Chemoattractant activity of tris-(hydroxymethyl)aminomethane for Brugia pahangi infective third-stage larvae.

Urocanic acid (UCA) is known as a major chemoattractant for Strongyloides stercoralis infective third-stage larvae (L3). Since Brugia pahangi is a skin-penetrating parasitic nematode similar to S. stercoralis, UCA was expected to be a chemoattractant for B. pahangi L3. Thus, the chemoattractant activity of UCA for B. pahangi L3 was assessed. The chemotactic responses of B. pahangi L3 to UCA or acetic acid (CH3COOH) dissolved in amine solutions were assessed using an agar-plate assay. A test solution of 200 mm UCA dissolved in aqueous 270 mm tris(hydroxymethyl)aminomethane (Tris) significantly attracted B. pahangi L3 compared with deionized water (DW), while neither a solution of 200 mm UCA dissolved in aqueous 230 mm ammonia (NH3) nor 290 mm triethylamine (TEA) significantly attracted L3. Similarly, a test solution of 200 mm CH3COOH dissolved with 200 mm Tris significantly attracted L3, but neither a test solution of 200 mm CH3COOH plus 200 mm NH3 nor 200 mm TEA attracted L3. Furthermore, L3 were significantly attracted to 200 mm Tris alone, compared with DW, but avoided 200 mm NH3 and 200 mm TEA. Moreover, the chemoattractant activity of Tris for L3 was observed even at a low concentration of 25 mm, and it was observed in a mild alkaline condition but not in an acidic condition. The present study reveals that Tris is a potential chemoattractant for B. pahangi L3 while UCA is not. This finding will contribute to an understanding of the mechanisms of skin-penetrating infection of filarial L3.

Long-acting formulation leading to severe long-term adverse effects: a case report of fluphenazine and persistent extrapyramidal symptoms.

WHAT IS KNOWN AND OBJECTIVE: Long-acting formulations are an important therapeutic option for non-adherent patients with schizophrenia. There is a commonly held view that management of long-acting formulation-induced side effects is difficult. CASE DESCRIPTION: We present a patient with schizophrenia who developed acute and persistent extrapyramidal symptoms requiring tracheostomy and long-term rehabilitation after long-acting injections of fluphenazine decanoate. Extrapyramidal symptoms improved with declining fluphenazine concentration and antiparkinsonian drug therapy with bromocriptine. WHAT IS NEW AND CONCLUSION: Long-acting formulations may lead to severe persistent adverse effects. For preventing fluphenazine-induced side effects, a possible option might be the antiparkinsonian drug therapy with bromocriptine.

MicroRNA-383 located in frequently deleted chromosomal locus 8p22 regulates CD44 in prostate cancer.

A major genomic alteration in prostate cancer (PCa) is frequent loss of chromosome (chr) 8p with a common region of loss of heterozygosity (LOH) at chr8p22 locus. Genomic studies implicate this locus in the initiation of clinically significant PCa and with progression to metastatic disease. However, the genes within this region have not been fully characterized to date. Here we demonstrate for the first time that a microRNA component of this region-miR-383-is frequently downregulated in prostate cancer, has a critical role in determining tumor-initiating potential and is involved in prostate cancer metastasis via direct regulation of CD44, a ubiquitous marker of PCa tumor-initiating cells (TICs)/stem cells. Expression analyses of miR-383 in PCa clinical tissues established that low miR-383 expression is associated with poor prognosis. Functional data suggest that miR-383 regulates PCa tumor-initiating/stem-like cells via CD44 regulation. Ectopic expression of miR-383 inhibited tumor-initiating capacity of CD44+ PCa cells. Also, 'anti-metastatic' effects of ectopic miR-383 expression were observed in a PCa experimental metastasis model. In view of our results, we propose that frequent loss of miR-383 at chr8p22 region leads to tumor initiation and prostate cancer metastasis. Thus, we have identified a novel finding that associates a long observed genomic alteration to PCa stemness and metastasis. Our data suggest that restoration of miR-383 expression may be an effective therapeutic modality against PCa. Importantly, we identified miR-383 as a novel PCa tissue diagnostic biomarker with a potential that outperforms that of serum PSA.

B-RAF mutation and accumulated gene methylation in aberrant crypt foci (ACF), sessile serrated adenoma/polyp (SSA/P) and cancer in SSA/P.

BACKGROUND: Sessile serrated adenomas/polyps (SSA/Ps) are a putative precursor of colon cancer with microsatellite instability (MSI). However, the developmental mechanism of SSA/P remains unknown. We performed genetic analysis and genome-wide DNA methylation analysis in aberrant crypt foci (ACF), SSA/P, and cancer in SSA/P specimens to show a close association between ACF and the SSA/P-cancer sequence. We also evaluated the prevalence and number of ACF in SSA/P patients. METHODS: ACF in the right-side colon were observed in 36 patients with SSA/Ps alone, 2 with cancers in SSA/P, and 20 normal subjects and biopsied under magnifying endoscopy. B-RAF mutation and MSI were analysed by PCR-restriction fragment length polymorphism (RFLP) and PCR-SSCP, respectively, in 15 ACF, 20 SSA/P, and 2 cancer specimens. DNA methylation array analysis of seven ACF, seven SSA/P, and two cancer in SSA/P specimens was performed using the microarray-based integrated analysis of methylation by isochizomers (MIAMI) method. RESULTS: B-RAF mutations were frequently detected in ACF, SSA/P, and cancer in SSA/P tissues. The number of methylated genes increased significantly in the order of ACF

Intraoperative fluorescence vascular imaging using indocyanine green for assessment of transplanted kidney perfusion.

INTRODUCTION AND OBJECTIVE: Indocyanine green (ICG) emits infrared light with exposure to laser light. When intravenously injected, it binds to plasma proteins and predominantly persists in the vasculature, which is very useful for definition of the vascular network. The HyperEye Medical System (HEMS; Mizuho Ikakogyo Co., LTD, Tokyo, Japan) is a new device able to identify both near-infrared and visible rays "in situ" without needing to dim the operation room lighting. We speculated that intraoperative ICG imaging would be applicable for kidney transplantation, by providing "in situ" determination of successful vascular anastomosis. MATERIALS AND METHODS: Four patients underwent intraoperative ICG imaging following intravenous administration of 1 mL of a solution containing 0.25% ICG. After performing vascular anastomosis, the allograft was examined using the HEMS light source device. Fluorescent signals were transmitted to a digital video processor connected to a television monitor and evaluated in real time. RESULTS: In all 4 patients, intraoperative ICG imaging provided excellent resolution of blood flow at each step in real time, namely, coming from the recipient's artery to the allograft renal artery, circulating throughout the whole grafted kidney, and draining through the allograft renal vein to the recipient's vein. HEMS provides ICG fluorescence image in color, allowing surgeons to clearly discriminate the positional relationship between the target tissue and the surrounding tissue. No complications associated with ICG injection were noted. CONCLUSION: Our preliminary results indicate that HEMS is a feasible and safe ICG imaging system that helps prevent technical failure during vascular anastomosis, and also demonstrates blood supply to the grafted kidney.

In vitro chemotactic responses of Brugia pahangi infective larvae to sodium ions.

In vitro chemotactic responses of infective third-stage larvae (L3) of Brugia pahangi to NaCl, Na2HPO4, KCl, K2HPO4, MgCl2 and CaCl2 were assessed. Compared to deionized water as a control, 200 mm NaCl and 100 mm Na2HPO4 significantly attracted L3 (P < 0.01 and P < 0.01), whereas L3 were likely to avoid 200 mm KCl and 100 mm K2HPO4 (P < 0.05 and P < 0.05). L3 showed no significant tendency to avoid or to be attracted to 200 mm CaCl2 and 200 mm MgCl2. Furthermore, NaCl exhibited a significant chemoattractant activity for L3 at a low concentration of 100 mm.

Nocturnal blood glucose and IGFBP-1 changes in type 1 diabetes: Differences in the dawn phenomenon between insulin regimens.

OBJECTIVE: Insulin-like growth factor binding protein-1 (IGFBP-1) is known to regulate the bioavailability of insulin-like growth factor (IGF) and the levels of IGFBP-1 are increased in the morning in patients with type 1 diabetes mellitus. We investigated the nocturnal fluctuations of glucose, IGFBP-1, and free IGF-1 levels with three insulin regimens. RESEARCH DESIGN AND METHODS: Forty-eight type 1 diabetes patients were divided into three groups according to their basal insulin therapy (continuous subcutaneous insulin infusion [CSII], insulin glargine, NPH insulin). Blood samples were obtained every 2 h between 2 300 h and 0700 h to measure plasma glucose, IGFBP-1 and free IGF-1 levels. RESULTS: The dawn phenomenon was more frequent with NPH (62.1%) than with glargine (16.6%, p<0.05) and CSII (14.3%, p<0.05). In the NPH group, the serum IGFBP-1 levels were markedly increased from 21.0+/-3.6 ng/ml at 2 300 h to 200.3+/-21.8 ng/ml at 0700 h and free IGF-1 levels were inversely decreased; these changes were partially suppressed in the CSII and glargine groups. CONCLUSIONS: The use of insulin regimens that provide sufficient insulin levels in the early morning can suppress the dawn phenomenon, leading to improved glycemic control. The increase in circulating IGFBP-1 in the morning, as a result of waning of insulin action, lowers free IGF-1 levels and may cause insulin resistance.

Unconventional ferroelectric transition in the multiferroic compound TbMnO3 revealed by the absence of an anomaly in c-polarized phonon dispersion.

TbMnO(3) exhibits a spontaneous electric polarization along c concomitantly with a spiral spin ordering modulated along b below T_{C} = 28 K. We have performed inelastic x-ray scattering measurements on a single crystal of TbMnO(3) to clarify whether phonon anomalies related to the ferroelectricity exist. We measured transverse modes, especially the Mn-O-Mn bending mode polarized along c and propagating along b, which we expect is most relevant to the ferroelectricity. However, no anomaly was found in the phonon dispersion below 50 meV across T_{C}. The present result suggests that the mechanism of ferroelectricity in TbMnO(3) is different from that of a conventional displacive-type ferroelectric. The weak coupling between electric polarization and lattice in TbMnO(3) strongly suggests that the ferroelectricity is mainly derived from the spiral spin ordering.

In vitro effects of artesunate on the survival of worm pairs and egg production of Schistosoma mansoni.

The effect of artesunate (ART) on the survival time of adult worm pairs of Schistosoma mansoni and on their egg output during in vitro culture was assessed. ART significantly decreased the survival time of both paired male and female worms at concentrations of 5, 10, 20 and 40 mg l- 1 during in vitro cultivation. An inhibitory effect of ART on the daily egg output of paired female worms during in vitro cultivation was also observed.

Isolation and characterization of microsatellite loci in Ainsliaea faurieana (Asteraceae), an endemic plant species on Yakushima Island, Japan, and cross-species amplification in closely related taxa.

Fourteen microsatellite loci were isolated and characterized for Ainsliaea faurieana, an endemic perennial plant species on Yakushima Island. In an analysis of 37 individuals from Yakushima Island, the observed number of alleles ranged from two to six. The expected and observed heterozygosities were 0.109-0.811 and 0.000-0.27, respectively. The inbreeding coefficient was 0.489-1.000. The interspecific applicability of these microsatellites was evaluated by analysing three closely related taxa from the Ryukyu Islands. All primer pairs for the 14 loci tested successfully amplified in all taxa.

Malignancy following kidney transplantation.

A cohort of 429 patients who received kidney grafts between 1973 and 2007 at our hospital was studied for the incidence and sites of malignancy. Sixty-two malignant diseases developed in 57 of 429 patients (13.3%). The cumulative incidences of malignancy increased markedly in the second and third posttransplantation decades. The overall rates were 1.8% at 5 years, 6.7% at 10 years, 12.5% at 15 years, 17.3% at 20 years, and 25.6% at 25 years. In the second and third posttransplantation decades, patients without malignancy showed significantly superior survival versus than those with cancer (P = .0002). Their survival rates were 83.4% versus 86.9% at 10 years and 63.1% versus 80.3% at 20 years, respectively. Skin cancer, renal cell carcinoma of the native kidney, hepatocellular carcinoma, posttransplantation lymphoproliferative disease, uterine cancer, and colorectal cancer were common in our series. The 5-year survival rates after the treatment of malignancy were better for skin cancer and renal cell carcinoma of the native kidney. Concerning the effects of immunosuppression, the tacrolimus-based group displayed a higher incidence among 3 groups (P = .0044).

Drug efficacy of praziquantel and albendazole in school children in Mwea Division, Central Province, Kenya.

The objective of this research was to assess drug efficacy in school children after mass chemotherapy with praziquantel and albendazole conducted in Mwea Division, Kirinyaga District, Central Kenya in 2004. In total 2300 children aged between 4 and 18 years in five primary schools were selected for the study. Before mass chemotherapy, prevalence of infection was 47.4% for Schistosoma mansoni, 16.7% for Necator americanus, 1.6% for Ascaris lumbricoides, and 0.8% for Trichuris trichiura. Post-treatment stool examination was carried out 8 weeks later, and a total of 1942 stool samples were collected. Prevalence decreased to 8.6% for S. mansoni, 0.2% for N. americunus, 0 for A. lumbricoides, and 0.6% for T. trichiura. Efficacy was good for S. mansoni and N. americanus (92.6% and 95.0%, respectively). Results of the first round of treatment of school-age children in Mwea indicate a good reduction in parasite burden.

Interleukin-6-induced activation of signal transducer and activator of transcription-3 in ruptured rotator cuff tendon.

The aim of this study was to examine interleukin-6 production and the activation of signal transducer and activator of transcription-3 (STAT3) in ruptured rotator cuff tendon. Specimens of ruptured rotator cuff tendons were analysed using real-time reverse transcriptase polymerase chain reaction, Western blotting and immunohistochemistry. Specimens of co-existing inflammatory subacromial synovia were examined for comparison. The level of interleukin-6 messenger RNA was increased in ruptured rotator cuff tendon as well as in subacromial synovium. Western blot analysis showed constitutive production of activated, phosphorylated STAT3 in ruptured rotator cuff tendon and co-existing subacromial synovium. Immunohistochemical examination detected cells producing interleukin-6, interleukin-6 receptor and phosphorylated STAT3 in ruptured rotator cuff tendon, mainly in proliferative vessels and, to a lesser extent, in tendon fibroblasts around the vessels. This study demonstrates that activation of STAT3 induced by interleukin-6 is promoted mainly by proliferative vessels in ruptured rotator cuff tendon.

Delayed macrofilaricidal activity of diethylcarbamazine against Brugia pahangi in Mongolian jirds.

The macrofilaricidal activity of diethylcarbamazine (DEC) was confirmed in jirds infected with Brugia pahangi. Seventy jirds were inoculated subcutaneously with 100 infective larvae. At 20 weeks post-infection, the microfilaraemic jirds were divided into two groups, untreated and treated. For the treated group, 200 mg kg(-1) of DEC was injected intraperitoneally for 5 consecutive days. One, 4, 8, 12, 16 and 27 weeks after the final treatment, 4-7 jirds in each group were sacrificed to measure adult worm burdens. The number of adult worms recovered from treated jirds was comparable to controls at earlier necropsy (1 and 4 weeks post-treatment). However, at late necropsy (8 weeks and later) the recovery rate of adult worms in treated jirds was significantly lower than that in untreated controls, indicating an adultcidal effect of DEC. The present study demonstrates that DEC requires 8 weeks to kill B. pahangi adult worms in jirds and that the Mongolian jird is a useful model for screening antifilarial activity.

The effects of topical application of phytonadione, retinol and vitamins C and E on infraorbital dark circles and wrinkles of the lower eyelids.

BACKGROUND: Infraorbital dark circles and wrinkles of the lower eyelids are a cosmetic problem, especially with age. AIMS: To determine whether a gel containing 2% phytonadione, 0.1% retinol and 0.1% vitamins C and E is effective in reducing dark under-eye circles and wrinkles of the lower eyelids in healthy Japanese adults. PATIENTS/METHODS: Fifty-seven adult Japanese volunteers with dark under-eye circles and wrinkles were enrolled in an open label study. The gel formulation was applied twice daily to the lower eyelid site for 8 weeks. Haemostasis, pigmentation and wrinkles were evaluated by a physician and by the patients themselves, using a digital camera and a visual analogue scale respectively, after 4 and 8 weeks of treatment. RESULTS: Topical application of the gel decreased not only haemostasis but also wrinkles after 8 weeks of treatment. Of 57 patients, 27 (47%) had reductions in haemostasis. Wrinkles were also decreased in some patients. However, pigmentation was not clearly removed by this gel. CONCLUSIONS: Topical application of the gel containing 2% phytonadione, 0.1% retinol, 0.1% vitamin C and 0.1% vitamin E was fairly or moderately effective in reducing dark under-eye circles, especially in cases of haemostasis, over a short treatment period in healthy Japanese adults. This treatment also slightly decreased wrinkles.

Endothelin-1 stimulates cardiomyocyte injury during mitochondrial dysfunction in culture.

To understand the pathophysiological role of endothelin-1 in the failing heart, we constructed a cellular mitochondrial impairment model and demonstrated the effect of endothelin-1. Primary cultured cardiomyocytes from neonatal rats were pretreated with rotenone, a mitochondrial complex I inhibitor, and the cytotoxic effect of endothelin-1 on the cardiomyocytes was demonstrated. Rotenone gradually decreased the pH of the culture medium with incubation time and caused slight cell injury. Endothelin-1 markedly enhanced the effect of rotenone that decreased the pH of the medium and enhanced cellular injury. The enhancement of the decrease in pH and cell injury induced by endothelin-1 was counteracted by the endothelin ET(A) receptor antagonist BQ123 or by maintaining the pH of the medium by the addition of 50 mM HEPES. Endothelin-1 markedly increased the uptake of 2-deoxyglucose and lactic acid production when the cardiomyocytes were pretreated with rotenone. These findings suggest that the stimulation of glucose uptake and anaerobic glycolysis followed by the increase in lactic acid accumulation in cardiomyocytes under the condition of mitochondrial impairment may be involved, at least in part, in the cellular injury by endothelin-1. Moreover, these findings suggest the possibility that the effect of endothelin-1 on myocardium is reversed by the condition of the mitochondria, and endogenous endothelin-1 may deteriorate cardiac failure with mitochondrial dysfunction. This may contribute to clarify the beneficial effect of endothelin receptor blockade in improving heart failures.

A small dose of the immunosuppressive agent FK506 (tacrolimus) protects peripheral nerve from ischemic fiber degeneration.

The immunosuppressant agent FK506 (tacrolimus) has proven to be neuroprotective against brain ischemia, but there are no data on potential neuroprotective effects of FK506 in peripheral nerve ischemia. We examined the potential effects of two doses of FK506 in protecting peripheral nerve from ischemic fiber degeneration. Ischemia in the left sciatic nerve of the rat was produced by injecting 2 x 10(6) microspheres (14 microm) into the left femoral, hypogastric, and superior gluteal arteries in proportions of 47%, 37%, and 17%, respectively. After embolization, FK506 was injected into the left femoral, hypogastric, and superior gluteal arteries in doses of 9.4, 7.4, and 3.4 microg, respectively, for the high-dose group and 4.7, 3.7, and 1.7 microg, respectively, for the low-dose group. The control rats were injected with saline. FK506 treatment resulted in dramatic behavioral improvement in nerve function, in the number of functioning nerve fibers, and in the salvage of a majority of nerve fibers from ischemic fiber degeneration in a dose-dependent fashion. These results suggest that a small dose of FK506 protects peripheral nerve from ischemic fiber degeneration and that it may have potential in the treatment of ischemic neuropathy.