Leukemia inhibitory factor (LIF) potentiates antinociception activity and inhibits tolerance induction of opioids.

BACKGROUND: The efficacy of opioids typically decreases after long-term use owing to the development of tolerance. Glial activation and the upregulation of proinflammatory cytokines are related to the induction of tolerance. We investigated the effect of leukemia inhibitory factor (LIF) on morphine analgesia and tolerance. METHODS: LIF concentrations in rat spinal cords were measured by polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA) after morphine administration. LIF distribution was examined using confocal microscopy. To evaluate the effects of LIF on morphine analgesia and tolerance, LIF was intrathecally administered 30 min before morphine injection. The analgesic effect of morphine was evaluated by measuring tail-flick latency. Human LIF concentrations from the cerebrospinal fluid (CSF) of opioid tolerant patients were also determined by specific ELISA. RESULTS: Chronic morphine administration upregulated LIF concentrations in rat spinal cords. Intrathecal injection of LIF potentiated the analgesic action of morphine. Patch clamp recording of spinal cord slices showed that LIF enhanced DAMGO ([D-Ala2, N-MePhe4, Gly-ol]-enkephalin)-induced outward potassium current. The development of tolerance was markedly suppressed by exogenous LIF, whereas neutralizing the endogenously released LIF with anti-LIF antibodies accelerated the tolerance induction. Moreover, LIF concentrations in the CSF of opioid-tolerant patients were higher than those in the opioid-naive controls. CONCLUSIONS: Intrathecal administration of LIF potentiated morphine antinociceptive activity and attenuated the development of morphine tolerance. Upregulation of endogenously released LIF by long-term use of opioids might counterbalance the tolerance induction effects of other proinflammatory cytokines. LIF might be a novel drug candidate for inhibiting opioid tolerance induction.

Association between interleukin-27 polymorphisms and pulmonary tuberculosis.

OBJECTIVE: To investigate the effect of interleukin (IL) 27 -964A/G, 2095T/G, 4603G/A and 4730T/C gene polymorphisms on the development of pulmonary tuberculosis (PTB), radiographic characteristics and severity. DESIGN: Differences in the allele and genotype distributions of the -964A/G, 2095T/G, 4603G/A and 4730T/C polymorphisms between 224 PTB patients and 233 healthy controls, between patients with single- and multi-lobe involvement, and between patients with and without cavitation, were investigated. Serum IL-27 concentration was measured using an enzyme-linked immunosorbent assay. RESULTS: There were no significant differences in the allele or genotype distributions between PTB patients and healthy controls. However, the -964A/A genotype was more prevalent in patients with single-lobe involvement than the -964A/G or -964G/G genotype in patients with multi-lobe involvement (50.0% vs. 31.3%, P = 0.01). There was no difference between patients with and without cavitation (P > 0.05). Serum median IL-27 concentration was significantly higher in patients with single-lobe involvement than in those with multi-lobe involvement (P = 0.03) and in those with -964A/A genotypes than in those with -964A/G or -964G/G genotypes (P = 0.02). CONCLUSIONS: In terms of serum IL-27 levels, the -964 A/A genotype may be associated with a protective role that prevents the intrapulmonary spread of PTB rather than its development.

Hepatitis C virus infection: a risk factor for Parkinson's disease.

Recent studies found that hepatitis C virus (HCV) may invade the central nervous system, and both HCV and Parkinson's disease (PD) have in common the overexpression of inflammatory biomarkers. We analysed data from a community-based integrated screening programme based on a total of 62,276 subjects. We used logistic regression models to investigate association between HCV infection and PD. The neurotoxicity of HCV was evaluated in the midbrain neuron-glia coculture system in rats. The cytokine/chemokine array was performed to measure the differences of amounts of cytokines released from midbrain in the presence and absence of HCV. The crude odds ratios (ORs) for having PD were 0.62 [95% confidence interval (CI), 0.48-0.81] and 1.91 (95% CI, 1.48-2.47) for hepatitis B virus (HBV) and HCV. After controlling for potential confounders, the association between HCV and PD remained statistically significant (adjusted OR = 1.39; 95% CI, 1.07-1.80), but not significantly different between HBV and PD. The HCV induced 60% dopaminergic neuron death in the midbrain neuron-glia coculture system in rats, similar to that of 1-methyl-4-phenylpyridinium (MPP(+) ) but not caused by HBV. This link was further supported by the finding that HCV infection may release the inflammatory cytokines, which may play a role in the pathogenesis of PD. In conclusion, our study demonstrated a significantly positive epidemiological association between HCV infection and PD and corroborated the dopaminergic toxicity of HCV similar to that of MPP(+) .

Quantitative optimization of solid freeform deposition of aqueous hydrogels.

Many soft tissues exhibit complex anatomical geometry that is challenging to replicate for regenerative medicine applications. Solid freeform fabrication (SFF) has emerged as an attractive approach for creating 3D tissues, but a detailed understanding of how specific fabrication parameters affect accuracy and viability has not been established to date. In this study, we evaluate the effects of printing parameters of the Fab@Home 3D printing system on accuracy using alginate, photocrosslinkable polyethylene-glycol diacrylate (PEG-DA) and gelatin as commonly used model hydrogel materials. Print accuracy and resolution along the length, width and height were determined based on quantitative image analysis. The effects of extrusion parameters on cell viability were assessed using porcine aortic valve interstitial cells (PAVIC) as a model cell type. We observed that pressure, pathheight and pathspace all significantly affected print accuracy and resolution. Printing conditions did not affect PAVIC viability within the ranges applied. We predicted that optimal pressure, pathheight and pathspace values would be increased linearly with increasing nozzle diameter, and we confirmed that the predicted values generate accurate 3D geometries while poorly chosen parameters yield inaccurate, unpredictable geometries. This systematic optimization strategy therefore improves the accuracy of 3D printing platforms for biofabrication and tissue engineering applications.

Rapid 3D printing of anatomically accurate and mechanically heterogeneous aortic valve hydrogel scaffolds.

The aortic valve exhibits complex three-dimensional (3D) anatomy and heterogeneity essential for the long-term efficient biomechanical function. These are, however, challenging to mimic in de novo engineered living tissue valve strategies. We present a novel simultaneous 3D printing/photocrosslinking technique for rapidly engineering complex, heterogeneous aortic valve scaffolds. Native anatomic and axisymmetric aortic valve geometries (root wall and tri-leaflets) with 12-22 mm inner diameters (ID) were 3D printed with poly-ethylene glycol-diacrylate (PEG-DA) hydrogels (700 or 8000 MW) supplemented with alginate. 3D printing geometric accuracy was quantified and compared using Micro-CT. Porcine aortic valve interstitial cells (PAVIC) seeded scaffolds were cultured for up to 21 days. Results showed that blended PEG-DA scaffolds could achieve over tenfold range in elastic modulus (5.3±0.9 to 74.6±1.5 kPa). 3D printing times for valve conduits with mechanically contrasting hydrogels were optimized to 14 to 45 min, increasing linearly with conduit diameter. Larger printed valves had greater shape fidelity (93.3±2.6, 85.1±2.0 and 73.3±5.2% for 22, 17 and 12 mm ID porcine valves; 89.1±4.0, 84.1±5.6 and 66.6±5.2% for simplified valves). PAVIC seeded scaffolds maintained near 100% viability over 21 days. These results demonstrate that 3D hydrogel printing with controlled photocrosslinking can rapidly fabricate anatomical heterogeneous valve conduits that support cell engraftment.

Carotid atherosclerosis in patients with untreated chronic obstructive pulmonary disease.

BACKGROUND: Chronic obstructive pulmonary disease (COPD) is associated with systemic inflammation. OBJECTIVE: To evaluate carotid artery intima-media thickness (IMT), high sensitivity C-reactive protein (hsCRP) and their correlation in newly diagnosed untreated patients with COPD. DESIGN: Post-bronchodilator spirometry, carotid artery IMT and blood tests were measured in patients with COPD (COPD group). Age, sex, body mass index, smoking status and smoking amount were compared with matched healthy subjects (non-COPD group). Participants taking medications and/or with a history of hypertension, diabetes mellitus, dyslipidaemia, COPD or cardiovascular disease were excluded. RESULTS: A total of 126 patients (COPD group 42, non-COPD group 84) were enrolled. The IMT and hsCRP of the COPD group were significantly higher than in the non-COPD group (P < 0.05). The decrease in the forced expiratory volume in 1 second/forced vital capacity (FEV(1)/FVC) ratio and FEV(1) was significantly correlated with an increase in the hsCRP and IMT (P < 0.05); there was no correlation between the IMT and hsCRP (P = 0.152). CONCLUSION: In newly diagnosed untreated patients with COPD, the carotid artery IMT and hsCRP were significantly higher than in healthy subjects. These findings suggest that systemic inflammation may play a potential role in preclinical atherosclerosis in COPD.

Purple urine bag syndrome: case report and literature review.

Purple urine bag syndrome (PUBS) is a medical syndrome in which there is purple discoloration of the urine of catheterized patients as well as discoloration of the collecting bag and the associated tubing. This rare condition, which mostly affects women, is generally associated with catheter-associated urinary tract infection, chronic constipation and alkaline urine. PUBS may be caused by sequential chemical reactions involving tryptophan from food in the gastrointestinal tract. The clinical course of PUBS is generally benign, and intensive treatment is not usually needed. We present 3 cases of this unusual and interesting phenomenon and a literature review.

Immune regulatory effects of simvastatin on regulatory T cell-mediated tumour immune tolerance.

Statins are potent inhibitors of hydroxyl-3-methylglutaryl co-enzyme A (HMG-CoA) reductase, and have emerged as potential anti-cancer agents based on preclinical evidence. In particular, compelling evidence suggests that statins have a wide range of immunomodulatory properties. However, little is known about the role of statins in tumour immune tolerance. Tumour immune tolerance involves the production of immunosuppressive molecules, such as interleukin (IL)-10, transforming growth factor (TGF)-beta and indoleamine-2,3-dioxygenase (IDO) by tumours, which induce a regulatory T cell (T(reg)) response. In this study, we investigated the effect of simvastatin on the production of IL-10, TGF-beta and IDO production and the proliferation of T(regs) using several cancer cell lines, and Lewis lung cancer (3LL) cells-inoculated mouse tumour model. Simvastatin treatment resulted in a decrease in the number of cancer cells (3LL, A549 and NCI-H292). The production of the immune regulatory markers IL-10, TGF-beta in 3LL and NCI-H292 cells increased after treatment with simvastatin. The expression of IDO and forkhead box P3 (FoxP3) transcription factor was also increased in the presence of simvastatin. In a murine 3LL model, there were no significant differences in tumour growth rate between untreated and simvastatin-treated mice groups. Therefore, while simvastatin had an anti-proliferative effect, it also exhibited immune tolerance-promoting properties during tumour development. Thus, due to these opposing actions, simvastatin had no net effect on tumour growth.

Steric effects of ions in the charge-related wetting phenomena.

Steric effects of ions on the charge-related wetting phenomena are studied. Along with a general treatment, three specific problems in two-dimensional system are considered: a droplet on an electrode, a droplet on a charged surface, and an electrowetting phenomenon on a dielectric. For computation of wetting tension, the electromechanical approach is adopted with the principle of mechanical force balance for each phase. The modified Poisson-Boltzmann equation, which was originally proposed by Bikerman [Philos. Mag. 33, 384 (1942)], is adopted for the analysis of the steric effects. It is found that the steric hindrance reduces significantly both the osmotic pressure and the electrical stress near the triple contact line. This reduction results in a considerable decrease in the wetting tension when the ratio of the capacitance per unit area of the electrical double layer to that of the dielectric layer is small.

New electrohydrodynamic flow caused by the Onsager effect.

We report a new type of electrohydrodynamic (EHD) flow generated around a circular cylinder and a spherical particle in a dielectric liquid under dc and ac electric fields. The EHD flow is observed for various combinations of dielectric liquids and polar additives. We suggest that the EHD flow is caused by a gradient of electrical conductivity produced by a nonuniform electric field and subsequent generation of free charge in the bulk liquid. Analytical and numerical analyses which are based on the leaky-dielectric model show good agreement with experimental results.

Multiple gastrointestinal stromal tumours presenting as a haemoperitoneum in a patient with Type 1 neurofibromatosis: MDCT findings.

We report multidetector row CT images of multiple gastrointestinal tumours in a patient with Type 1 neurofibromatosis who presented with haemoperitoneum. The use of CT demonstrated multiple, variably sized, ovoid, relatively well-enhancing masses in the intramural or subserosal location of the jejunum. The largest mass that showed a haemorrhagic necrosis caused haemoperitoneum. These imaging features are unique and differ from previous imaging features that have been reported in the radiological literature.

Quantitative cerebrovascular reserve measured by acetazolamide-challenged dynamic CT perfusion in ischemic adult Moyamoya disease: initial experience with angiographic correlation.

BACKGROUND AND PURPOSE: CT perfusion is a much more readily accessible imaging method to assess cerebral hemodynamic status than single-photon emission CT. We prospectively assessed quantitative cerebrovascular reserve by using acetazolamide (ACZ)-challenged CT perfusion for evaluating hemodynamic impairment in ischemic adult Moyamoya disease and compared it with angiographic findings. MATERIALS AND METHODS: Sixteen adult patients with ischemic Moyamoya disease and 12 age-matched normal control subjects underwent both ACZ-challenged CT perfusion and digital subtraction angiography. Normalized baseline hemodynamic parameters and their percent changes (PCs) were calculated in 56 hemispheres. We classified the degrees of distal carotid artery stenosis according to modified Suzuki stage and determined the presence of basal Moyamoya vessels (BMVs). The values of normalized parameters and their PCs were compared with angiographic findings. RESULTS: Normalized baseline mean transit time (MTT) and PC of normalized cerebral blood flow (CBF) were significantly correlated with angiographic stages in all of the vascular territories; however, the correlation coefficient of the normalized baseline MTT was lower than that of the PC of CBF. In the external borderzone and the middle cerebral arterial territory, the hemispheres with extensive BMVs exhibited significantly lower PC values of CBF and significantly higher normalized baseline MTT values than those in hemispheres with diminished BMVs and in normal control subjects. CONCLUSION: Among the hemodynamic parameters measured by ACZ-challenged CT perfusion, the PC of CBF correlated highly significantly with angiographic stage; however, the normalized baseline CT perfusion parameters showed weak or no significant correlation.

Substance P regulates macrophage inflammatory protein 3alpha/chemokine C-C ligand 20 (CCL20) with heme oxygenase-1 in human periodontal ligament cells.

Although substance P (SP), a potent proinflammatory peptide, is involved in inflammation and immune responses, the effect of SP on the expression of macrophage inflammatory protein 3alpha[MIP-3alpha, chemokine C-C ligand 20 (CCL20)] in periodontal ligament (PDL) cells is unknown. Equally enigmatic is the link between SP, the stress protein heme oxygenase-1 (HO-1), and CCL20 production. We investigated whether SP induces the release of chemokine CCL20 from immortalized PDL (IPDL) cells, and further clarify SP-mediated pathways. We also examined the relationship between HO-1 and CCL20 by treating PDL cells with SP. Incubating IPDL cells with SP increased expression of CCL20 mRNA and CCL20 protein in a dose-time-dependent manner. Highly selective p38 and extracellular-regulated kinase 1/2 (ERK1/2) inhibitors abrogated SP-induced expression of CCL20 in IPDL cells. SP is also responsible for initiating phosphorylation of IkappaB, degradation of IkappaB and activation of nuclear factor (NF)-kappaB. SP induced expression of HO-1 in both a concentration- and time-dependent manner, and CCL20 reflected similar patterns. The inductive effects of SP on HO-1 and CCL20 were enhanced by HO-1 inducer hemin and the membrane-permeable guanosine 3',5'-monophosphate (cGMP) analogue 8-bromo-cGMP. Conversely, this pathway was inhibited by the HO-1 inhibitor zinc protoporphyrin IX (ZnPP IX) and the selective inhibitor of guanylate cyclase, 1H-(1,2,4)oxadiazole(4,3-a)quinoxalin-1-one (ODQ). We report herein the pathway that connects SP along with other modulators of neuroimmunoregulation to the induction of HO-1 and the inflammatory mediator macrophage inflammatory protein (MIP)-3alpha/CCL20 in IPDL cells, which play an important role in the development of periodontitis or inflammation during orthodontic tooth movement.

The local prevalence of COPD by post-bronchodilator GOLD criteria in Korea.

OBJECTIVE: Chronic obstructive pulmonary disease (COPD) is a disease characterised by not fully reversible airflow limitation. The Global Initiative for Chronic Obstructive Lung Disease (GOLD) committee decided to diagnose COPD using post-bronchodilator spirometry values. We aimed to examine the prevalence and risk factors of COPD in Ansan, an industrialised city of Korea, by using the post-bronchodilator GOLD criteria. We then investigated the implications of brenchodilation on the prevalence of COPD. DESIGN: A total of 3642 participants in the Korean Health and Genome Study were interviewed about age, income, smoking status and respiratory symptoms and completed pulmonary function tests, including postbronchodilator spirometry. RESULTS: COPD prevalence by post-bronchodilator spirometry was 3.7% (134/3642), which was significantly different from that estimated using pre-bronchodilator criteria (7.7%, 282/3642). Exclusion of subjects with significant bronchodilator response (BDR) significantly lowered the prevalence of COPD to 3.3% (117/3572), compared with including subjects with post-bronchodilatory residual obstruction with significant BDR. Prevalence was associated with old age, smoking history, male sex and respiratory symptoms. CONCLUSION: COPD prevalence by post-bronchodilator GOLD criteria was 3.7%, which was much lower than that of pre-bronchodilator criteria. The bronchodilator reversibility test substantially affects estimations of COPD prevalence.

Mapping quantitative trait loci for yield components and morphological traits in an advanced backcross population between Oryza grandiglumis and the O. sativa japonica cultivar Hwaseongbyeo.

Introgression has been achieved from wild species Oryza grandiglumis (2n = 48, CCDD, Acc. No. 101154) into O. sativa subsp. japonica cv. Hwaseongbyeo as a recurrent parent. An advanced introgression (backcross) line, HG101, produced from a single plant from BC5F3 families resembled Hwaseongbyeo, but it showed differences from Hwaseongbyeo in several traits, including days to heading and culm length. To detect the introgressions, 450 microsatellite markers of known chromosomal position were used for the parental survey. Of the 450 markers, 51 (11.3%) detected O. grandiglumis segments in HG101. To characterize the effects of alien genes introgressed into HG101, an F(2:3) population (150 families) from the cross Hwaseongbyeo/HG101 was developed and evaluated for 13 agronomic traits. Several lines outperformed Hwaseongbyeo in several traits, including days to heading. Genotypes were determined for 150 F2 plants using simple sequence repeat markers. Qualitative trait locus (QTL) analysis was carried out to determine the relationship between marker genotype and the traits evaluated. A total of 39 QTL and 1 gene conferring resistance to blast isolate were identified using single-point analysis. Phenotypic variation associated with each QTL ranged from 4.2 to 30.5%. For 18 (46.2%) of the QTL identified in this study, the O. grandiglumis-derived alleles contributed a desirable agronomic effect despite the overall undesirable characteristics of the wild phenotype. Favorable wild alleles were detected for days to heading, spikelets per panicle, and grain shape traits. Grain shape QTL for grain weight, thickness, and width identified in the F(2:3) lines were further confirmed based on the F4 progeny test. The confirmed locus, tgw2 for grain weight is of particular interest because of its independence from undesirable height and maturity. Several QTL controlling amylose content and grain traits have not been detected in the previous QTL studies between Oryza cultivars, indicating potentially novel alleles from O. grandiglumis. The QTL detected in this study could be a rich source of natural genetic variation underlying the evolution and breeding of rice.

Analysis of differentially expressed transcripts from planthopper-infested wild rice (Oryza minuta).

A subtracted library was constructed from planthopper-infested wild rice (Oryza minuta) by suppression subtractive hybridization in combination with mirror orientation selection. To screen the differentially expressed transcripts in the library, we applied a cDNA microarray containing 960 random clones in a reverse Northern blot analysis using cDNA probes prepared from the mRNAs of control and planthopper-infested samples. On the basis of the signal intensities and expression ratios obtained from experiments performed in triplicate, we selected 383 clones. The elevated expression levels and overall profiles over time were verified by Northern blot analysis. Although Southern blot analysis showed similar copy numbers of the screened genes in O. minuta and O. sativa, it also revealed that the expression profiles had a different pattern. Functional categorization placed the identified transcripts in the categories of subcellular localization, metabolism, and protein fate. The presence of these expressed sequence tags implies that resistance of O. minuta to insect infestation can be achieved not only by an elevated expression of defense-related genes but also by enhanced metabolic activities.

Comparative analysis of 5,211 leaf ESTs of wild rice (Oryza minuta).

The expressed sequence tags (ESTs) presented in this report are the first transcriptomes of wild rice. A cDNA library was constructed from 4-week-old leaf samples of greenhouse-grown Oryza minuta. The 5,211 cDNA clones of O. minuta represent 3,401 unique sequences, consisting of 2,787 singletons and 614 assembled sequences. Database comparisons of the cDNAs in GenBank's non-redundant databases using BLAST revealed that 4,957 of the 5,211 cDNAs (95.1%) showed a high degree of sequence homology to genes from other organisms. Most of the transcripts identified were genes related to metabolism, energy, protein biosynthesis and subcellular localization. The metabolism and energy categories of the O. minuta ESTs showed a considerably higher gene expression level than those of O. sativa ESTs. These data and genes can be utilized in rice breeding.