Nanogel-based nasal ghrelin vaccine prevents obesity.

Obesity is associated with multiple comorbidities such as cardiovascular diseases and has a huge economic impact on the health-care system. However, the treatment of obesity remains insufficient in terms of efficacy, tolerability, and safety. Here we created a nasal vaccine against obesity for the first time. To avoid the injectable administration-caused pain and skin-related adverse event, we focused on the intranasal route of antigen delivery. We developed a vaccine antigen (ghrelin-PspA (pneumococcal surface protein A)), which is a recombinant fusion protein incorporating ghrelin, a hormone that stimulates food intake and decreases energy expenditure, and PspA, a candidate of pneumococcal vaccine as a carrier protein. Ghrelin-PspA antigen was mixed with cyclic di-GMP adjuvant to enhance the immunogenicity and incorporated within a nanometer-sized hydrogel for the effective antigen delivery. Intranasal immunization with ghrelin-PspA vaccine elicited serum immunoglobulin G antibodies against ghrelin and attenuated body weight gain in diet-induced obesity mice. This obesity-attenuating effect was caused by a decrease in fat accumulation and an increase in energy expenditure that was partially due to an increase in the expression of mitochondrial uncoupling protein 1 in brown adipose tissue. The development of this nasal vaccine provides a new strategy for the prevention and treatment of obesity.

Prototyping the Experimental Setup to Quantify the Tissue Oxygen Consumption Rate and Its Preliminary Test.

In order to establish a reliable and practical method to make a diagnosis on the viability of an amputated extremity, we propose a method to evaluate the oxygen consumption rate. To validate this concept, we prototyped an experimental system with which the oxygen transfer rate into tissue can be assessed by the rate of change of the decrease in dissolved oxygen (DO) concentration within the buffer fluid surrounding the target tissue. The purpose of this study is to examine the feasibility of our prototyped experimental system by comparison between fresh and non-fresh rat skeletal muscles. The results show that the fresher tissue transferred more oxygen to the tissue, which suggests that tissue oxygen consumption is highly related to tissue freshness and can indirectly assess the tissue viability.

Weight change after 20 years of age and the incidence of dyslipidemia: a cohort study of Japanese male workers.

BACKGROUND: While heavier weight is known to increase the incidence of dyslipidemia, limited data are available on the relationship between weight gain and its development. METHODS: A total of 2647 males were categorized into the following four groups according to the difference between their self-reported weight at 20 years of age and their measured weight in 1994-95: a loss of ≥5% (decrease), loss of <5% or gain of <5% (no change), gain of ≥5 to <15% (increase) and gain of ≥15% (sizable increase). They were followed up until their 2002-03 health examination. Using the 'no change' group as reference, the multivariable-adjusted odds ratio (adjusted for age, body mass index at 20 years of age, physical activity, smoking and alcohol intake) and 95% confidence interval (95% CI) for the incidence of dyslipidemia were determined using logistic regression models. RESULTS: A total of 1342 participants developed dyslipidemia during the follow-up period. The 'increase' and 'sizable increase' groups had odds ratios for the incidence of dyslipidemia of 1.97 (95% CI, 1.59-2.45) and 2.68 (2.15-3.34), respectively, demonstrating that there was a significant dose-response association between weight gain since 20 years of age and the incidence of dyslipidemia (P < 0.001 for trend). CONCLUSION: These results suggest that dyslipidemia could be prevented by avoiding weight gain in adulthood.

Liposome chaperon in cell-free membrane protein synthesis: one-step preparation of KcsA-integrated liposomes and electrophysiological analysis by the planar bilayer method.

Chaperoning functions of liposomes were investigated using cell-free membrane protein synthesis. KcsA potassium channel-reconstituted liposomes were prepared directly using cell-free protein synthesis. In the absence of liposomes, all synthesized KcsA protein aggregated. In the presence of liposomes, however, synthesized KcsA spontaneously integrated into the liposome membrane. The KscA-reconstituted liposomes were transferred to the planar bilayer across a small hole in a thin plastic sheet and the channel function of KcsA was examined. The original electrophysiological activities, such as voltage- and pH-dependence, were observed. These results suggested that in cell-free membrane protein synthesis, liposomes act as chaperones, preventing aggregation and assisting in folding and tetrameric formation, thereby allowing full channel activity.

Effective CpG DNA delivery using amphiphilic cycloamylose nanogels.

Unmethylated CpG oligodeoxynucleotides induce inflammatory immune responses through cytokine production and have attracted increasing attention as an immunostimulator. However, there remains a challenging issue of the use of 'native CpG DNA'. In the present study, we prepared cationic nanometer-sized gels (nanogels) consisting of cycloamylose modified with cholesterol and diethylaminoethane to form hydrophobic cross-linking points and to add positively charged groups, respectively. The cationic nanogels and native CpG DNA formed nanometer-sized complexes. Complexes of native CpG DNA with cationic nanogels delivered native CpG DNA to macrophage-like cells and induced cytokine production. In addition, complexes of negative control oligonucleotides with cationic nanogels did not induce cytokine production, and the induction of cytokines using complexes of phosphorothioate-modified CpG with cationic nanogels was lower than that of native CpG DNA. These results suggest that the complex of native CpG DNA with cationic nanogels is a promising strategy for nucleic acid adjuvants.

Nanogel-based pneumococcal surface protein A nasal vaccine induces microRNA-associated Th17 cell responses with neutralizing antibodies against Streptococcus pneumoniae in macaques.

We previously established a nanosized nasal vaccine delivery system by using a cationic cholesteryl group-bearing pullulan nanogel (cCHP nanogel), which is a universal protein-based antigen-delivery vehicle for adjuvant-free nasal vaccination. In the present study, we examined the central nervous system safety and efficacy of nasal vaccination with our developed cCHP nanogel containing pneumococcal surface protein A (PspA-nanogel) against pneumococcal infection in nonhuman primates. When [(18)F]-labeled PspA-nanogel was nasally administered to a rhesus macaque (Macaca mulatta), longer-term retention of PspA was noted in the nasal cavity when compared with administration of PspA alone. Of importance, no deposition of [(18)F]-PspA was seen in the olfactory bulbs or brain. Nasal PspA-nanogel vaccination effectively induced PspA-specific serum IgG with protective activity and mucosal secretory IgA (SIgA) Ab responses in cynomolgus macaques (Macaca fascicularis). Nasal PspA-nanogel-induced immune responses were mediated through T-helper (Th) 2 and Th17 cytokine responses concomitantly with marked increases in the levels of miR-181a and miR-326 in the serum and respiratory tract tissues, respectively, of the macaques. These results demonstrate that nasal PspA-nanogel vaccination is a safe and effective strategy for the development of a nasal vaccine for the prevention of pneumonia in humans.

Enhanced expression of the ß4-galactosyltransferase 2 gene impairs mammalian tumor growth.

Altered N-glycosylation of membrane proteins is associated with malignant transformation of cells. We found that the expression of the ß4-galactosyltransferase 2 (ß4GalT2) gene is decreased markedly during the transformation. Here, we examined whether the tumor growth activity of B16-F10 mouse melanoma cells can be reduced by the enhanced expression of the ß4GalT2 gene. We isolated a clone, B16-ß4GalT2, showing its ß4GalT2 transcript 2.5 times higher than a control clone, B16-mock, by transducing its cDNA, and transplanted them subcutaneously into C57BL/6 mice to examine their tumor growth activity. The results showed that the average size of tumors formed with B16-mock cells is 13.1±0.76 mm, whereas that of tumors formed with B16-ß4GalT2 cells is 5.1±1.13 mm (P<0.01) 2 weeks after transplantation. Immunohistochemical analyses showed that the apoptosis and the suppression of angiogenesis are induced in the tumors upon transduction of the ß4GalT2 gene. To pursue a clinical usefulness of the ß4GalT2 gene for suppressing human tumor growth, we injected adenoviruses carrying the human ß4GalT2 cDNA into HuH-7 human hepatocellular carcinomas developed in severe combined immunodeficient mice, and observed marked growth retardation of the tumors. The enhancement of the ß4GalT2 gene expression in tumors is one of the promising approaches to suppress human tumor growth.

Role of aquaporin-5 in gallbladder carcinoma.

BACKGROUND/PURPOSE: Aquaporins (AQPs) are important in controlling bile formation. However, the exact role in human gallbladder carcinogenesis has not yet been defined. METHODS: AQP-5-expressing gallbladder carcinoma (GBC) cell lines (NOZ) were transfected with anti-AQP-5 small interfering RNA (siRNA). Growth, migration, invasion assay, and drug susceptibility tests were performed. Next, microRNA (miRNA) expression was analyzed by miRNA oligo chip (3D-Gene®). AQP-5 and AQP-5-related miRNA target gene expressions were also analyzed using tissue microarray (TMA) in 44 GBC samples. RESULTS: Treatment with AQP-5 siRNA decreased cell proliferation, migration, and invasion. On the other hand, those cells increased IC50 of gemcitabine. By performing miRNA assays, miR-29b, -200a, and -21 were shown to be highly overexpressed in cells treated with AQP-5 siRNA NOZ. When focusing on miR-21, phosphatase and tensin homolog (PTEN) was found to be a target of miR-21. In the TMA, AQP-5/PTEN coexpression was significantly associated with the depth of invasion and MIB-1 index (p = 0.003, 0.010). Survival of patients with a high AQP-5/PTEN coexpression was longer than that of patients with a low coexpression (p = 0.003). CONCLUSIONS: Our result suggested that miR-21 and PTEN may contribute to the role of AQP-5 in GBC. AQP-5 and PTEN cascades are favorable biomarkers of GBC.

CO2 microbubble contrast enhancement in x-ray angiography.

AIM: To demonstrate that carbon dioxide (CO2) microbubble contrast enhancement depicts blood vessels when used for x-ray examinations. MATERIALS AND METHODS: Microbubbles were generated by cavitation of physiological saline to which CO2 gas had been added using an ejector-type microbubble generator. The input pressure values for CO2 gas and physiological saline that produced a large quantity of CO2 microbubbles were obtained in a phantom. In an animal study, angiography was performed in three swine using three types of contrast: CO2 microbubbles, conventional CO2 gas, and iodinated contrast medium. For CO2 microbubble contrast enhancement, physiological saline, and CO2 gas were supplied at the input pressures calculated in the phantom experiment. Regions of interest were set in the abdominal aorta, external iliac arteries, and background. The difference in digital values between each artery and the background was calculated. RESULTS: The input pressures obtained in the phantom experiment were 0.16 MPa for physiological saline and 0.5 MPa for CO2 gas, with physiological saline input volume being 8.1 ml/s. Three interventional radiologists all evaluated the depictions of all arteries as "present" in the CO2 microbubble contrast enhancement, conventional CO2 contrast enhancement, and iodinated contrast enhancement performed in three swine. Digital values for all vessels with microbubble CO2 contrast enhancement were higher than background values. CONCLUSIONS: In x-ray angiography, blood vessels can be depicted by CO2 microbubble contrast enhancement, in which a large quantity of CO2 microbubbles is generated within blood vessels.

Irsogladine maleate reduces the incidence of fluorouracil-based chemotherapy-induced oral mucositis.

BACKGROUND: Oral mucositis is one of the most common side-effects of 5-fluorouracil (5-FU)-based chemotherapy. The objective of this study was to evaluate the effects of irsogladine maleate (IM) on fluorouracil-induced oral mucositis through a double-blind, placebo controlled trial. PATIENTS AND METHODS: Patients (N = 66) were randomly assigned to receive either placebo or IM (4 mg/day for 14 consecutive days). The incidence and maximum severity of fluorouracil-induced oral mucositis and safety of the irsogladine dosing regimen were evaluated. RESULTS: A cohort of 33 patients received placebo and 33 patients received IM. The incidence of oral mucositis was significantly lower for IM than for placebo (27% versus 73%; P < 0.001 by chi-square test). Specific adverse events considered related to IM were not found. CONCLUSION: IM significantly reduced the incidence and maximum severity of oral mucositis in patients treated with 5-FU-chemotherapy.

Search for the Θ+ pentaquark via the π(-)p→K(-)X reaction at 1.92 GeV/c.

The Θ(+) pentaquark baryon was searched for via the π(-)p→K(-)X reaction with a missing mass resolution of 1.4 MeV/c(2) (FWHM) at the Japan Proton Accelerator Research Complex (J-PARC). π(-) meson beams were incident on the liquid hydrogen target with a beam momentum of 1.92 GeV/c. No peak structure corresponding to the Θ(+) mass was observed. The upper limit of the production cross section averaged over the scattering angle of 2° to 15° in the laboratory frame is obtained to be 0.26 µb/sr in the mass region of 1.51-1.55 GeV/c(2). The upper limit of the Θ(+) decay width is obtained to be 0.72 and 3.1 MeV for J(Θ)(P)=1/2(+) and J(Θ)(P)=1/2(-), respectively, using the effective Lagrangian approach.

Transport in simple networks described by an integrable discrete nonlinear Schrödinger equation.

We elucidate the case in which the Ablowitz-Ladik (AL)-type discrete nonlinear Schrödinger equation (NLSE) on simple networks (e.g., star graphs and tree graphs) becomes completely integrable just as in the case of a simple one-dimensional (1D) discrete chain. The strength of cubic nonlinearity is different from bond to bond, and networks are assumed to have at least two semi-infinite bonds with one of them working as an incoming bond. The present work is a nontrivial extension of our preceding one [Sobirov et al., Phys. Rev. E 81, 066602 (2010)] on the continuum NLSE to the discrete case. We find (1) the solution on each bond is a part of the universal (bond-independent) AL soliton solution on the 1D discrete chain, but it is multiplied by the inverse of the square root of bond-dependent nonlinearity; (2) nonlinearities at individual bonds around each vertex must satisfy a sum rule; and (3) under findings 1 and 2, there exist an infinite number of constants of motion. As a practical issue, with the use of an AL soliton injected through the incoming bond, we obtain transmission probabilities inversely proportional to the strength of nonlinearity on the outgoing bonds.

LBH589, a deacetylase inhibitor, induces apoptosis in adult T-cell leukemia/lymphoma cells via activation of a novel RAIDD-caspase-2 pathway.

Adult T-cell leukemia/lymphoma (ATLL), an aggressive neoplasm etiologically associated with human T-lymphotropic virus type-1 (HTLV-1), is resistant to treatment. In this study, we examined the effects of a new inhibitor of deacetylase enzymes, LBH589, on ATLL cells. LBH589 effectively induced apoptosis in ATLL-related cell lines and primary ATLL cells and reduced the size of tumors inoculated in SCID mice. Analyses, including with a DNA microarray, revealed that neither death receptors nor p53 pathways contributed to the apoptosis. Instead, LBH589 activated an intrinsic pathway through the activation of caspase-2. Furthermore, small interfering RNA experiments targeting caspase-2, caspase-9, RAIDD, p53-induced protein with a death domain (PIDD) and RIPK1 (RIP) indicated that activation of RAIDD is crucial and an event initiating this pathway. In addition, LBH589 caused a marked decrease in levels of factors involved in ATLL cell proliferation and invasion such as CCR4, IL-2R and HTLV-1 HBZ-SI, a spliced form of the HTLV-1 basic zipper factor HBZ. In conclusion, we showed that LBH589 is a strong inducer of apoptosis in ATLL cells and uncovered a novel apoptotic pathway initiated by activation of RAIDD.

Integrable nonlinear Schrödinger equation on simple networks: connection formula at vertices.

We study the case in which the nonlinear Schrödinger equation (NLSE) on simple networks consisting of vertices and bonds has an infinite number of constants of motion and becomes completely integrable just as in the case of a simple one-dimensional (1D) chain. Here the strength of cubic nonlinearity is different from bond to bond, and networks are assumed to have at least two semi-infinite bonds with one of them working as an incoming bond. The connection formula at vertices obtained from norm and energy conservation rules shows (1) the solution on each bond is a part of the universal (bond-independent) soliton solution of the completely integrable NLSE on the 1D chain, but is multiplied by the inverse of square root of bond-dependent nonlinearity; (2) nonlinearities at individual bonds around each vertex must satisfy a sum rule. Under these conditions, we also showed an infinite number of constants of motion. The argument on a branched chain or a primary star graph is generalized to other graphs, i.e., general star graphs, tree graphs, loop graphs and their combinations. As a relevant issue, with use of reflectionless propagation of Zakharov-Shabat's soliton through networks we have obtained the transmission probabilities on the outgoing bonds, which are inversely proportional to the bond-dependent strength of nonlinearity. Numerical evidence is also given to verify the prediction.

The role of stress echocardiography and competing technologies for the diagnostic and prognostic assessment of coronary artery disease.

Coronary artery disease remains the leading cause of death in adults in the United States. Non-invasive cardiac imaging is now central to the diagnosis and management of patients with known or suspected coronary disease. The generally accepted indications for stress testing include confirming a diagnosis of coronary disease, assessing prognosis, preoperative risk stratification, and evaluation of medical therapy. Stress echocardiography and single photon computed tomography are well-established non-invasive techniques for all the previously mentioned indications. These modalities provide a relatively high sensitivity and specificity along with an incremental value over clinical risk factors. Cardiac magnetic resonance imaging (CMRI) and multislice computed tomography are new imaging tools in the evaluation of patients with coronary disease. CMRI offers a comprehensive cardiac evaluation which includes wall motion analysis, myocardial tissue morphology, rest and stress first pass myocardial perfusion as well as systolic ventricular function. It is also considered a first line technique for the diagnosis of certain structural heart disease and chamber volume quantification. Cardiac computed tomography allows non-invasive anatomic imaging of the coronary tree. It has a high clinical utility especially in select intermediate risk patient population. Available tests all have advantages and drawbacks and none can be considered suitable for all patients. The choice of the imaging method should be tailored to each person based on the clinical judgment of the a priori risk of cardiac event, clinical history, and risk factors profile.

Perioperative risk predictors of cardiac outcomes in patients undergoing liver transplantation surgery.

BACKGROUND: Cardiac risk assessment for perioperative outcomes of liver transplantation patients is limited. We examined the outcomes of an older intermediate-cardiac-risk group of patients undergoing liver transplantation surgery. METHODS AND RESULTS: Patients who had liver transplantation surgery between 2001 and 2005 were studied. The 3 outcomes analyzed were nonfatal myocardial infarction, death, and either outcome within the first 30 days after the liver transplantation surgery. Of 403 patients (mean age, 52+/-9 years; 67% male), 106 (26%) were diabetic, 84 (21%) were hypertensive, and 173 (43%) had a history of smoking. There were 48 total events (12%), 25 myocardial infarctions (7%), and 38 deaths (9%) recorded during the perioperative period. From the final multivariate model, history of coronary artery disease, prior stroke, and postoperative sepsis predicted greater risk (P=0.014; odds ratio [OR], 4.0; 95% confidence interval [CI], 1.3 to 11.8; P=0.025; OR, 6.6; 95% CI, 1.3 to 33.8; and P<0.001; OR, 7.5; 95% CI, 3.3 to 17.1, respectively). Use of perioperative beta-blockers was protective (P=0.004; OR, 0.20; 95% CI, 0.1 to 0.6) for combined cardiac outcomes. For the outcome of death on multivariate analysis, postoperative sepsis and increased interventricular septal thickness predicted risk (P<0.001; OR, 8.6; 95% CI, 3.5 to 20.9; and P=0.027; OR, 2.8; 95% CI, 1.1 to 7.2, respectively), whereas the use of perioperative beta-blockers was again protective (P=0.012; OR, 0.07; 95% CI, 0.01 to 0.56). CONCLUSIONS: In our study of cardiac risk assessment for liver transplantation surgery, history of stroke, coronary artery disease, postoperative sepsis, and increased interventricular septal thickness were markers of adverse perioperative cardiac outcomes, whereas use of perioperative beta-blockers was significantly protective.

Antiproteinuric effect of angiotensin-converting enzyme inhibitors and an angiotensin II receptor blocker in patients with lupus nephritis.

Although the effects of angiotensin II receptor blockers (ARBs) on non-diabetic glomerulonephritis have been reported, studies of their effects on collagen-vascular diseases, particularly lupus nephritis, are limited. In this retrospective, observational study, systemic lupus erythematosus (SLE) patients (n = 7) with lupus nephritis and uncontrolled proteinuria were treated with an angiotensin-converting enzyme inhibitor followed by the ARB losartan (25 - 50 mg/day). Urinary protein excretion and renal function were evaluated. After 12 months of losartan, mean urinary protein excretion decreased significantly by 84.8%. Mean systolic and diastolic blood pressures also decreased significantly during the 12 months of losartan treatment, although not in normotensive patients. Complement 4, total complement activity and anti-dsDNA antibody levels, which are indices of SLE activity, and serum creatinine levels, which is an index of renal function, showed no change in response to losartan treatment. A more extensive evaluation of the effects of ARBs in patients with lupus nephritis and poorly controlled proteinuria is required.

[Large cell neuroendocrine carcinoma of the lung].

Large cell neuroendocrine carcinoma (LCNEC) is a neuroendocrine tumor comprising a subgroup of large cell carcinoma and is a type of lung cancer showing a neuroendocrine characteristic similar to that of small cell lung carcinoma In our institution, we started to diagnose LCNEC by immunostaining in 2002, and we herein report 9 patients diagnosed with LCNEC from January 2002 to May 2008. The average patient age was 74.9, male/female ratio was 8/1, and all 9 patients had a smoking history. Pathological stages IA/IB/IIB/IIIA comprised 4/1/2/2, respectively. Peripherally located and lobulated tumors were noted on preoperative computed tomography (CT), and moderate uptake of fluoro-2-deoxy-D-glucose (FDG), which balanced with the size, was recognized on positron emission tomography (PET). All 9 patients underwent surgery and 7 underwent radical surgery. Postoperative adjuvant chemotherapy was performed for 4 patients. Three showed recurrence, and 2 of these 3 died of the primary disease. The remaining 7 patients have survived to date. The possibility of LCNEC must be considered when peripherally located lung cancer with lobulation is noted on CT and shows moderate uptake of FDG for its size on PET, and multimodal treatment is needed if the diagnosis is determined postoperatively.