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Epithelial ovarian cancer (EOC) remains localized within the peritoneal cavity in a large number of patients, lending itself to i.p. approaches of therapy. In the present study, we investigated the effect of replication-selective herpes simplex virus-1 (HSV-1) used as an oncolytic agent against EOC and the use of human teratocarcinoma PA-1 as carrier cells for i.p. therapy. HSV-1716, a replication-competent attenuated strain lacking ICP34.5, caused a direct dose-dependent oncolytic effect on EOC cells in vitro. A single i.p. administration of 5 x 10(6) plaque-forming units resulted in a significant reduction of tumor volume and tumor spread and an increase in survival in a mouse xenograft model. PA-1 cells supported HSV replication in vitro and bound preferentially to human ovarian carcinoma surfaces compared with mesothelial surfaces in vitro and in vivo. In comparison with the administration of HSV-1716 alone, irradiated PA-1 cells, infected at two multiplicities of infection with HSV-1716 and injected i.p. at 5 x 10(6) cells/animal, led to a significant tumor reduction in the two models tested and the significant prolongation of mean survival in one model. Histological evaluation revealed extensive necrosis in tumor areas infected by HSV-1716. Immunohistochemistry against HSV-1 revealed areas of viral infection within tumor nodules, which persisted for several weeks after treatment. Administration of HSV-infected PA-1 carrier cells resulted in larger areas of tumor infected by the virus. Our results indicate that replication-competent attenuated HSV-1 exerts a potent oncolytic effect on EOC, which may be further enhanced by the utilization of a delivery system with carrier cells, based on amplification of the viral load and possibly on preferential binding of carrier cells to tumor surfaces.

A levanbiose-accumulating levanase from Pseudomonas sp. No. 43 was purified to a homogeneous state by (NH4)2SO4 fractionation and by chromatography on DEAE-Toyopearl 650 M and phenyl-Toyopearl 650 M columns. The molecular mass and isoelectric point of the enzyme were estimated to be 36 kDa and 5.7 respectively; the optimal pH and temperature for the enzyme reaction were pH 7.0 and 40 degrees C respectively. The purified enzyme was stable in the pH range 6.0-8.0 at 20 degrees C and stable up to 50 degrees C at pH 7.0. The enzyme's activity was inhibited by MnCl2, CoCl2, AlCl3, EDTA and potassium permanganate. The levanase was specific towards the 2, 6-beta-D-fructosidic linkages of levan and did not hydrolyse other polysaccharides among those examined. The enzyme is an exohydrolase of levan and produced levanbiose as a sole product; the limits of hydrolysis of levans from Zymomonas mobilis and Serratia sp. were 65% and 80% respectively.

To evaluate the distribution pattern of apolipoprotein(a) [Apo(a)] phenotypes in Koreans and the effect of dialysis modality on serum lipoprotein(a) [Lp(a)] concentration according to apo(a) phenotype in patients with end-stage renal disease (ESRD).Cross-sectional study.A university hospital.153 normal controls, 99 hemodialysis (HD) patients and 82 continuous ambulatory peritoneal dialysis (CAPD) patients.Fasting serum Lp(a), lipids, and apo(a) phenotypes were measured.The frequencies of the subjects with apo(a) phenotypes of high-molecular weight only, including S3, S4, or S5 or null type were 95.4% of control, 100% of HD patients, and 95.1% of CAPD patients. The frequent apo(a) phenotypes in Koreans consisted of S4, S4S5, S5, and S5S5 isoforms. Significant difference was found in serum Lp(a) concentration among controls and HD and CAPD patients [median (Interquartile range): 0.05 g/L, (0.01-0.19); 0.19 g/L, (0.10-0.35); 0.63 g/L, (0.28-0.90), p0.001]. Lp(a) levels in CAPD patients were significantly higher than in HD patients for all four common apo(a) isoforms found in Korean subjects. CAPD patients had higher total and LDL cholesterol levels, and higher ApoB levels than HD patients. Significant differences were found in serum albumin levels between controls and HD and CAPD patients (44 +/- 3 g/L, 40 +/- 4 g/L, 32 +/- 7 g/L, respectively, p0.05). There were significant inverse correlations between serum albumin and Lp(a) (r = -0.33, p0.01), total cholesterol (r = -0.31, p0.01), LDL (r = -0.39, p0.01) or ApoB (r = -0.35, p0.01) in ESRD patients. A significant positive correlation was found between serum albumin and ApoA1 (r = 0.24, p0.01).These findings indicate that Koreans have mainly high molecular weight apo(a) phenotypes and serum Lp(a) is elevated in CAPD patients compared to HD patients for common apo(a) phenotypes, which may contribute to the frequent cardiovascular mortality in CAPD patients.

Background Recently, a novel scoring system was developed for the diagnosis of gout without joint fluid analysis (1). The performance of this scoring system in the differential diagnosis between acute gout and septic arthritis has not been validated yet. Objectives This study aimed to evaluate the diagnostic performance of this scoring system in the differential diagnosis between acute gout and septic arthritis in patients with acute monoarthritis. Methods The medical records of 33 patients with acute gout and 27 with septic arthritis who presented as acute monoarthritis and were diagnosed at Chung-Ang University Hospital in Seoul, South Korea and Dongguk University Hospital in Goyang, South Korea from 2007 to 2012 were reviewed. Patients with podagra were excluded. All gout patients were MSU positive and all septic arthritis patients had positive results of bacterial culture. The diagnostic scoring system (1) gives different scores to several clinical criteria as follows: 2 to male sex, 2 to previous patient reported arthritis attack, 0.5 to onset within one day, 1 to joint redness, 2.5 to 1 st MTP involvement, 1.5 to hypertension or at least one cardiovasacular disease and 3.5 to serum uric acid greater than 5.88 mg/dL. The probability of gout is high when the sum of the score is ≥ 8, intermediate when between 4 and 8 and low when ≤ 4. Patients were classified to one of each probability groups according to the scores they got. Results Both patient groups were similar in age (55.6±21 vs. 58.4±20.2 years). However, patients with acute gouty arthritis were more likely to be male (90.9% vs. 59.2%), had shorter duration of onset (1.54±1.22 vs.2.92±2.0 days, p=0.004), lower levels of ESR (38.3±73.7mm/hr p=0.001), CRP(7.8±7.0 vs.13.6±9.9 mg/dL, p=0.01), WBC count (9813±2122 vs. 12502±5060/m3, p=0.021) and synovial fluid WBC count (37295±26998 vs. 63210±41211/m3, p=0.014) and had higher levels of serum uric acid compared to those with septic arthritis (7.88±1.8 vs. 5.13±1.7 mg/dL, p Conclusions This novel scoring system showed a good performance in distinguishing between acute gout and septic arthritis. It may be helpful to the primary care physicians when they have to diagnose gout without joint fluid analysis. References Janssens HJ et al. A disgnostic rule for acute gouty arthritis in primary care without joint fluid analysis Arch Intern Med. 2010;170:1120 Disclosure of Interest None Declared

Background Leucine-rich α2-glycoprotein (LRG) is a plasma protein which contains leucine-rich repeats (LRRs). Though physiological functions of LRG have not been clarified yet, it has been reported that LRG could be a marker of granulocytic differentiation and its expression was up-regulated during neutrophil differentiation. LRG could show a significant increase in some inflammatory conditions, such as ulcerative colitis, in where serum LRG concentrations were well correlated with disease activity, and the expressions of LRG were increased in inflamed colonic tissues. However, the association between plasma LRG level and disease activity in RA patients remains obscure. Objectives This study aimed to investigate whether the plasma LRG level is elevated in patients with RA and its correlation with disease activity and other parameters. Methods Our study included 69 patients with RA and 48 age- and sex- matched healthy controls. Plasma samples were obtained from patients with RA during active and inactive disease status and from controls. We assessed the clinical characteristics and laboratory parameters including erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), and disease activity score 28 (DAS28). Plasma concentrations of tumor necrosis factor-α (TNF-α) and LRG were determined by enzyme-linked immunosorbent assay (ELISA). Results Plasma LRG concentrations were significantly elevated in RA patients compared with healthy control (30.8 ± 14.4 ng/mL vs 22.2 ± 6.1 ng/mL, p γ = 0.671, p γ = 0.612, p γ = 0.601, p p Image/graph Conclusions Patients with RA had higher plasma LRG levels than healthy subjects, and plasma LRG concentrations were well correlated with disease activity measures. Our findings suggest that plasma LRG could play a role in the inflammatory process independently of the TNF- α, and that it may be a novel biomarker for reflecting inflammatory activity in RA patients. References Serada S, Fujimoto M, Terabe F, Lijima H, Shinzaki S, Matsuzaki S, et al. Serum leucine-rich alpha-2 glycoprotein is a disease activity biomarker in ulcerative colitis. Inflamm Bowel Dis 2012;18:2169-79 Shirai R, Hirano F, ohkura N, Ikeda K, Inoue S. Up-regulation of the expression of leucine-rich alpha(2)-glycoprotein in hepatocytes by the mediators of acute-phase response. Biochem Biophys Res Commun 2009;382:776-9 Disclosure of Interest None Declared

Background Chemerin is an adipokine that is linked to adipogenesis and chemotaxis of the innate immune system. It is expressed on macrophage, dendritic cells, and synovial lining and sublining cells. It has been reported that chemerin has both pro-inflammatory and anti-inflammatory roles, and higher level of chemerin was detected in various chronic inflammatory diseases. Recent studies have showed that its expression is increased in the synovium of patients with rheumatoid arthritis (RA), and the chemerin may play an important role in the pathogenesis of RA. However, the association between plasma chemerin level and disease activity in RA patients remains unclear. Objectives This study aims to determine whether plasma chemerin level is elevated in patient with RA and its correlation with disease activity and other parameters. Methods This study includes 69 RA patients and 40 age- and sex- matched healthy controls. Plasma samples were obtained from healthy controls and patients with RA during active and inactive disease status. We assessed the clinical characteristics and laboratory parameters including body mass index (BMI), erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), and disease activity score 28 (DAS28). The plasma level of chemerin and tumor necrosis factor (TNF)-α were determined using enzyme-linked immunosorbent assay (ELISA). Results Plasma chemerin level was significantly elevated in patients with RA than healthy control (8.508±12.437 pg/mL vs 0.370±0.219 pg/mL, p p =0.049). Conclusions Patients with RA showed higher plasma chemerin than that of healthy controls. The adjusted plasma chemerin level according to BMI was well correlated with RA disease activity. These findings suggest that plasma chemerin could play a role in the inflammatory process of RA, and that it may be a useful disease activity marker in RA. References Kaneko K, Miyabe Y, Takayasu A, Fukuda S, Miyabe C, Ebisawa M et al. Chemerin activates fibroblast-like synoviocytes in patients with rheumatoid arthritis. Arthritis Res Ther 2011;13(5):R158 Nakajima H, Nakajima K, Nagano Y, Yamamoto M, Tarutani M, Takahashi M et al. Circulating level of chemerin is upregulated in psoriasis. J Dermatol Sci 2010;60(1):45-7 Weigert J, Obermeier F, Neumeier M, Wanninger J, Filarsky M, Bauer S, et al. Circulating levels of chemerin and adiponectin are higher in ulcerative colitis and chemerin is elevated in Crohn’s disease. Inflamm Bowel Dis 2010;16(4):630-7 Disclosure of Interest None Declared

Background Various psychological distresses in systemic lupus erythematosus (SLE) may be associated with systemic conditions. However, the influence of psychological distresses to the SLE disease activity still remains unclear. Objectives We evaluated the levels of depression, anxiety, anger and fatigue in patients with SLE, compared to those of healthy controls. We investigated the association between the psychological distresses and disease activity and damage in patients with SLE. Methods 105 patients with SLE and 51 healthy controls completed a psychological questionnaire. Psychological distresses were assessed as follows: depression, Center for Epidemiologic Studies Depression Scale (CES-D); anxiety, Hospital Anxiety and Depression Scale (HADS); anger, State Trait Anger Expression Inventory (STAXI); fatigue, the Profile of Mood States fatigue-inertia Scale (POMS-F). Disease activity and damage index were measured by the SLE Disease Activity Index (SLEDAI) and the SLE Collaborating Clinics/American College of Rheumatology (SLICC/ACR), respectively. Results Patients with SLE showed higher levels of depression ( p =0.003), anxiety ( p =0.049), anger ( p =0.021) and fatigue ( p =0.003) than healthy controls. In the patients with SLE, psychological distresses were well correlated with each other (depression and anxiety: γ=0.712, p p p p p p 7.5 mg/day) had greater symptoms in all of the psychological distresses than the group with lower doses of glucocorticoids (prednisolone ≤7.5 mg/day) (depression, p =0.002; anxiety, p =0.027; anger, p =0.007; fatigue, p =0.016, respectively). However, none of the psychological distresses showed significant differences between higher disease activity and lower disease activity groups, and between higher damage and lower damage groups. In a multiple linear analysis, depression was affected by SLE disease activity (β=0.223, p =0.026) and daily prednisolone doses (β=0.228, p =0.029). However, SLE disease activity was not affected by the psychological distresses. Conclusions SLE patients had more psychological distresses, such as depression, anxiety, anger and fatigue than healthy controls. Higher levels of disease activity and higher use of glucocorticoids affect on depression, while the results do not support that psychological distress affects disease activity status. Disclosure of Interest None Declared

Background Triggering receptor expressed on myeloid cells 1 (TREM-1) is an immunoglobulin-like cell surface receptor mainly expressed on neurtophils, monocytes and macrophages. TREM-1 serves as a critical amplifier of inflammatory signaling, and its expression is increased in the synovium of rheumatoid arthritis (RA) patients. It can exist as a soluble form. However, the association between plasma sTREM-1 levels and disease activity in RA patients remains unclear. Objectives In this study, we investigated whether plasma soluble TREM-1 (sTREM-1) levels are elevated in patients with RA and whether its levels are correlated with disease activity and other parameters. Methods This cross-sectional study included 71 patients with RA and 50 age- and sex- matched healthy controls. Plasma samples were obtained from patients with RA during active and inactive disease status and from controls. We assessed clinical characteristics and laboratory parameters including the erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), and disease activity score 28 (DAS28). Their plasma levels of sTREM-1 and tumor necrosis factor (TNF)-α were quantitatively measured by enzyme-linked immunosorbent assay (ELISA). Results Patients with RA had significantly higher plasma sTREM-1 levels than healthy controls (170.10±84.71 pg/mL vs 97.41±40.64 pg/mL, p r =0.329, p =0.005; r =0.241, p =0.043; r =0.314, p r =0.261, p =0.028; r =0.278, p =0.019; r =0.313, p =0.009, respectively). Plasma sTREM-1 levels in patients with a active disease status (DAS28 >3.2) were significantly higher than in those with a low disease status (DAS28 ≤3.2) (208.89±100.14 pg/mL vs 150.29±68.70 pg/mL, p =0.005). Conclusions RA patients had higher plasma sTREM-1 levels than healthy controls, and plasma sTREM-1 levels were correlated with disease activity parameters, suggesting that plasma sTREM-1 could play a role in the inflammatory process associated with TNF-α, and that it may be a useful disease activity marker in RA. References Collins CE, La DT, Yang HT, Massin F, Gibot S, Faure G, et al. Elevated synovial expression of triggering receptor expressed on myeloid cells 1 in patients with septic arthritis or rheumatoid arthritis. Ann Rheum Dis 2009;68;1768-74. Kuai J, Gregory B, Hill A, Pittman DD, Feldman JL, Brown T, et al. TREM-1 expression is increased in the synovium of rheumatoid arthritis patients and induces the expression of pro-inflammatory cytokines. Rheumatology 2009:1352-8. Disclosure of Interest None Declared

Background Hyperhomocysteinemia, which is related with cardiovascular diseases and metabolic syndrome, is regarded as one of the important factors in endothelial cell damage processes. It is well known that gout is associated with metabolic syndrome, and cardiovascular diseases are major causes of mortality in gouty patients. However, there are few reports about the serum homocysteine levels in gouty patients, and their results reveal discrepancy. Objectives In this study, we investigated whether or not serum homocysteine levels are elevated in the patients with chronic gout and which factors are associated with the elevated homocysteine levels. Methods This cross-sectional study included 91 male patients with chronic gout and 97 age-matched healthy male controls. The averages of age were 51.19 ± 15.08 and 51.57 ± 17.01 years old, respectively. Serum homocysteine, uric acid (UA), blood urea nitrogen (BUN), creatinine (Cr) and other laboratory findings were tested for all participants. Serum homocysteine levels were measured by a competitive immunoassay using direct chemiluminescent (Siemens Centaur Immunoassay Systems, USA). The estimated glomurular filtration rate (eGFR) was uptained using modification of diet in renal disease (MDRD) formula, then the stages of chronic kidney disease (CKD) were classified according to eGFR levels as follows; stage 1, more than 90 mL/min/1.73m 2 : stage 2, 60-89 mL/min/1.73m 2 : stage 3, 30-59 mL/min/1.73m 2 : stage 4, 15-29 mL/min/1.73m 2 : stage 5, less than 15 mL/min/1.73m 2 . Results The chronic gout group were not significantly different from the control group in serum uric acid levels (6.15 ± 2.23 mg/dL vs 5.82 ± 1.22 mg/dL, p = 0.224). However, the patients with chronic gout showed much higher serum homocysteine levels than healthy controls (13.96 ± 4.05 μmol/L vs 12.67 ± 3.52 μmol/L, p = 0.021). Serum homocysteine levels showed the positive correlations with serum BUN and Cr levels, and the negative correlation with eGFR ( r = 0.429, p r = 0.435, p r = -0.413, p p β = -0.385, p Conclusions Serum homocysteine levels were higher in the male patients with chronic gout than in the healthy male controls. Hyperhomocysteinemia in gouty patients could be related not with serum uric acid levels, but with decreased renal function. Types of uric acid lowering agents did not affect the serum homocysteine levels. Disclosure of Interest None Declared

Background Chemerin is an adipokine that is linked to adipogenesis and chemotaxis of the innate immune system. It is expressed on macrophage, dendritic cells, and synovial lining and sublining cells. It has been reported that chemerin has both pro-inflammatory and anti-inflammatory roles, and higher level of chemerin was detected in various chronic inflammatory diseases. Recent studies have showed that its expression is increased in the synovium of patients with rheumatoid arthritis (RA), and the chemerin may play an important role in the pathogenesis of RA. However, the association between plasma chemerin level and disease activity in RA patients remains unclear. Objectives This study aims to determine whether plasma chemerin level is elevated in patient with RA and its correlation with disease activity and other parameters. Methods This study includes 71 RA patients and 42 age- and sex- matched healthy controls. Plasma samples were obtained from healthy controls and patients with RA during active and inactive disease status. We assessed the clinical characteristics and laboratory parameters including body mass index (BMI), erythrocyte sedimentation rate, C-reactive protein, and disease activity score 28 (DAS28). The plasma level of chemerin and tumor necrosis factor (TNF)-α were determined using enzyme-linked immunosorbent assay (ELISA). Results Plasma chemerin levels were significantly elevated in patients with RA compared with healthy controls (9.074 ± 13.513 ng/mL vs 0.370 ± 0.219 ng/mL, p < 0.001). In RA patients, plasma chemerin levels showed a negative correlation with BMI (γ= -0.264, p = 0.027). Adjusted plasma chemerin levels according to BMI were significantly higher in the active group (DAS28 ≥ 2.6) than in the inactive group (DAS28 < 2.6) (0.591 ± 0.879 ng/mL/Kg/m2 vs 0.220 ± 0.154 ng/mL/Kg/m2, p = 0.015). Plasma chemerin levels adjusted to BMI were correlated with DAS28 (γ = 0.340, p = 0.004), but not with plasma TNF-α levels. Conclusions Patients with RA showed higher plasma chemerin levels than those of healthy controls. Adjusted plasma chemerin levels according to BMI were well correlated with RA disease activity. These findings suggest that plasma chemerin could play a role in the inflammatory process of RA rather than obesity, and that it may be a useful disease activity marker and a candidate target molecule for therapy in RA patients. Disclosure of Interest None Declared

To provide pressure relief at the high pressure areas of the buttocks, small holes were drilled on the poly-urethane (PU) foam cushion under the ischial tuberosities areas. This modification was done to locally lower the supporting property of die cushion. This approach could be easily implemented in the clinical setting. Measurements of interfacial pressures at die ischial tuberosities on such modified cushions for eight nondisabled subjects showed an average pressure 25% lower than those measured on foam cushions without modification (p < 0.01). The maximum pressure was 30% lower after modification (p < 0.01) The peak pressure gradient also showed a 38% decrease on modified foam compared to those measured on unmodified foam cushions (p < 0.01). Simple hole drilling could potentially provide an effective approach to enhance the pressure-relief characteristics of foam cushions.

To assess the efficacy of the chest radiograph in identifying patients with miliary tuberculosis.Retrospective case-controlled review by three independent blinded chest radiologists.Provincial tuberculosis control center.Population-based sample, including all proved cases of miliary tuberculosis diagnosed in the Province of British Columbia, Canada, between November 1982 and November 1992. One hundred cases of miliary tuberculosis were identified, of which 71 had chest radiographs available for review. Forty-four normal chest radiographs and 20 chest radiographs of patients with localized pulmonary tuberculosis were also included as controls.The primary outcome of measurements was the sensitivity and interobserver variability of the chest radiograph in the diagnosis of miliary tuberculosis. The observers were also asked to describe the pattern and extent of pulmonary abnormalities based on the International Labor Organization (ILO) classification of pneumoconioses.The three independent observers identified 42, 44, and 49 of the 71 cases of miliary tuberculosis, respectively (sensitivity, 59 to 69%). The three observers incorrectly diagnosed miliary tuberculosis in 2, 0, and 2 of the 64 controls, respectively (specificity, 97 to 100%). There was good interobserver agreement (90%, kappa = 0.77). The nodules measured less than 3 mm in diameter in 90% of cases in which miliary tuberculosis was correctly identified. In 10% of cases, the nodules measured greater than 3 mm in diameter. The ILO profusion scores ranged from mild (profusion score 1) in 45% of cases, through moderate (profusion score 2) in 27%, and severe (profusion score 3) in 28%.The chest radiograph allowed identification of 59 to 69% of cases of miliary tuberculosis with a high specificity and good interobserver agreement.

To determine the outcome of areas of ground-glass attenuation and assess disease progression on serial high-resolution CT (HRCT) scans of patients with biopsy specimen-proved usual interstitial pneumonia (UIP) and desquamative interstitial pneumonia (DIP).Twelve patients with biopsy specimen-proved UIP and 11 patients with biopsy specimen-proved DIP who had initial and follow-up HRCT scans (median interval, 10 months) were reviewed. Eleven patients with UIP and 11 with DIP received treatment between the initial and follow-up CT scans. The scans were evaluated for the presence and extent of ground-glass attenuation, irregular linear opacities and honeycombing, and overall extent of parenchymal involvement.On initial CT scans, all 12 patients with UIP had areas of ground-glass attenuation (mean +/- SD extent, 30 +/- 16%) and irregular lines (mean +/- SD extent, 17 +/- 7%) and 10 patients had honeycombing (mean +/- SD extent, 10 +/- 6%). All 11 patients with DIP had areas of ground-glass attenuation on initial HRCT scans (mean +/- SD extent, 51 +/- 26%), 5 patients had irregular linear opacities (mean +/- SD extent, 5 +/- 5%), and 1 patient had honeycombing. Nine of the 12 patients with UIP showed increase in the extent of ground-glass attenuation (n = 6) or progression to irregular lines (n = 2) or honeycombing (n = 4) on follow-up as compared with only 2 patients with DIP who showed progression to irregular lines (n = 1) or honeycombing (n = 1) (p0.01 chi 2 test).In patients with UIP, areas of ground-glass attenuation usually increase in extent or progress to fibrosis despite treatment. Areas of ground-glass attenuation in most patients with DIP remain stable or improve with treatment.

Kang-Rotondo CH, Major S, Chiang TM, Myers LK, Kang ES. Upregulation of nitric oxide synthase in cultured human keratinocytes after ultraviolet B and bradykinin. Photodermatol Photoimmunol Photomed 1996: 12: 57–65. © Munksgaard, 1996. Ultraviolet B (UVB) irradiation of the skin has been reported to upregulate nitric oxide synthase (NOS) activity with enhancement of nitric oxide (NO) formation. Bradykinin, a known stimulator of NO production, is produced in the skin within minutes of UVB irradiation. The combined effect of UVB and bradykinin on NOS was therefore examined in a cultured human keratinocyte (KC) line. Activity was determined in KC homogenates by the recovery of [3H]l-citrulline using labeled l-arginine as the substrate in the presence of mM NADPH. Monoclonal antibodies to specific isoforms of NOS that cross-react with their human counterparts were used to determine the isoform(s) in control, UVB, bradykin treated and UVB and bradykinin treated KC. Human KC express NOS activity which is lowest at confluence and highest during proliferation. UVB increased NOS activity when a set dose of irradiation was administered from 32.2–48.3 mJ/cm2 but was inhibitory after 64.4 and 80.5 mJ/cm2. Thirty min after 10−6 M bradykinin, NOS activity nearly doubled followed by return of activity to control levels at 60 min. Activity after UVB and bradykinin was only slightly higher than that observed with bradykinin alone. Immunochemically, an isoform of Mr 155 kDa was detected in control cells with the antibody for the constitutive brain enzyme, bNOS. Recovery of this isoform increased after UVB treatment as well as after bradykinin which was time dependent. When both stimulants were used, the recovery of the 155 kDa enzyme was markedly enhanced, unlike the enzyme activity findings. These data indicate that the expression of NOS activity under unstimulated conditions in human KC in culture is due to the constitutive NOS found in neuronal tissue, bNOS. The recovery of bNOS increased after UVB and after bradykinin while the combination of both resulted in the synergistic increase in bNOS protein with only a marginal further increase in NOS activity.

Three population groups, 1500 blood donors, 513 antenatal women representing a normal population group and 250 sicklers representing a multiply transfused group were studied to determine the prevalence of hepatitis C viral (HCV) infection in Jamaica. The relationship to liver enzyme levels, hepatitis B infection, syphilis and HIV infection was also investigated. Sera were screened by enzyme-linked immunoassay (EIA) for anti-HCV C100-3 and subsequently tested by a supplementary second generation recombinant immunoblot assay (RIBA). In the blood donors, the prevalence of anti-HCV was low, 0.3%-0.4%, the same level as that reported by several European countries. In the multiply transfused sicklers, the prevalence was more than seven times higher. No HCV infection was detected in the antenatal group. There was little correlation between HCV infection and surrogate markers alanine aminotransferase (ALT) and antibody to hepatitis B core antigen (anti-HBc) and no correlation with sexually transmitted diseases.

Effect of oral administration of crude aqueous neem extract on serum testosterone and other blood constituents was studied in the male Wistar rats for 10 weeks. The neem treatment resulted in significant decreases (p0.01) in total testosterone, total bilirubin and K+ in serum. There were also increases (p0.05) in packed cell volume, mean corpuscular haemoglobin concentration, red blood cell, white blood cell and lymphocyte counts without showing any cytotoxic effects in the body.

In this study, the phase transformation and the surface and interface morphologies of TiSi2 formed on atomically clean Si substrates are investigated. 200Å Ti and 400A Si films on Si(111) have been co-deposited at elevated temperatures (400°C - 800°C) in ultrahigh vacuum. The phase transition of TiSi2 is characterized with using XRD. The results distinguish the formation of the C49 and C54 crystalline titanium silicides. The surface and interface morphologies of titanium silicides have been examined with SEM and TEM. A relatively smooth surface is observed for the C49 phase while a rough surface and interface are observed for the C54 phase. The islanding of the C54 phase becomes severe at high temperature (800°C). Islands of TiSi2 have been observed at temperatures above 700°C but no islands are observed at temperatures below 600°C. For films deposited at 400TC and 500°C, weak XRD peaks corresponding to TiSi were observed and TEM micrographs exhibited small crystalline regions of titanium silicide at the interface.

Subacute intraperitoneal administration of the lipid portion of the unripe ackee arillus, referred to as "ackee oil", resulted in marked neutropenia (p less than 0.001) and increase in platelets (p less than 0.01) without anaemia, in rats. Blood urea, sodium and aspartate aminotransferase levels were significantly decreased but glucose and bilirubin levels were similar to those of controls. The lungs showed areas of petechial haemorrhages and a dose-related perivascular and peribronchial mononuclear cell infiltration. The pulmonary toxicity may be interpreted as a hypersensitive reaction to ackee oil. Further research is in progress on the neutropenic effects of ackee oil.

Whether infection with influenza B virus alters hepatic function was examined in the ferret. Also, the possibility that viral-specific antibodies (Ab) could be produced well before their detection in serum was explored. During the febrile period of influenza, reductions in the serum potassium, anion gap, ammonia, albumin and CPK and elevations of the BUN, creatinine and the GGTP levels occurred. With convalescence, the electrolytes, BUN and creatinine normalized, FFA, SGPT and CPK levels rose and the serum GGTP rose even further. Hepatic fatty acid (FA) oxidation, ornithine transcarbamylase (OTC) and carnitine palmitoyltransferase (CPT) activities were minimally altered and liver ATP and total lipid content remained normal. Following experimental secondary viremia, serum FFA continued to rise, TG decreased and CPK remained elevated while SGPT and GGTP levels normalized. In the liver, FA oxidation and OTC rates remained unchanged but CPT activity was inhibited and the liver content of ATP was significantly reduced. Immune complex (IC) protein recovered from postmicrosomal supernatant fractions by polyethylene glycol precipitation was progressively increased in livers from convalescent and viremic animals. While the amount of IC protein recovered in the spleen also increases during convalescence, this is not the case after viremia when the IC formed seem to be processed largely by the liver. By SDS/PAGE, the major proteins identified in the IC were IgM and other viral proteins. However, the viral proteins could not be validated by immunoblot with Ab produced against purified influenza B hemagglutinin (HA) and neuraminidase (NA) most probably due to phagocytic alterations of glycoprotein immunodeterminants. These findings indicate that during influenza, convalescence and post viremia changes in the concentrations of several serum and liver components occur that reflect hepatic involvement. Also, antiviral Ab, largely IgM, appears to be produced early, complexes with Ag and can be found sequestered in both the liver and spleen at a time when Ab is not detectable in the serum.

The cloning of miniature pigs (Mini-pig) are considered to advance in genetic engineering technology and xenotransplantation. A few researches have recently been reported successfully produce cloned Mini-pigs using somatic cells, however its efficiency is still low. The present study was aimed to successfully produce cloned Mini-pigs derived from bone marrow mesenchymal stem cells (MSCs) by NT, and compared the developmental ability of cloned Mini-pigs between fetal fibroblast (FF) and differentiated MSCs. For the production of the cloned Mini-pig derived from MSCs, MSCs were isolated from a 1 month old of female Mini-pigs (T-type, PWG Micro-pig®, PWG Genetics Korea, Ltd.). MSCs were differentiated into osteocytes, adipocytes, and chondrocytes under controlled conditions and characterized by cell surface antigen profile using specific markers. These differentiated or undifferentiated MSCs, FFs of Mini-pig were transferred into enucleated oocytes of domestic pigs, and 2-cell stage of 100 NT embryos were surgical transferred to the synchronized recipients. Statistical analysis was performed using one-way ANOVA, t-test, Duncan’s and Tukey’s multiple comparisons test by SPSS (SPSS, Inc., Chicago, IL, USA). The developmental potential of NT embryos derived from MSCs (differentiated and undifferentiated), the rates of blastocyst formation was significantly (P < 0.05) higher than NT embryos derived from FFs, however the NT embryos derived from three different types of differentiated MSCs were significantly (P < 0.05) lower than undifferentiated MSCs. In addition, total cell numbers in NT blastocysts derived from MSCs were significantly (P < 0.05) higher than NT blastocysts derived from FFs, but it did not significantly (P < 0.05) differ between differentiated or undifferentiated MSCs. NT embryos derived from MSCs were transferred to 5 domestic pig recipients, and 5 cloned Mini-pigs were obtained from 2 recipients (one stillbirth and 4 viable offspring). All of them were confirmed by the microsatellite analysis (8 markers) of the genomes of cloned offspring, donor MSCs and recipients. Physical and histological studies are in the process for the characterization of a cloned Mini-pig derived from MSCs as animal model. The results demonstrated that, in vitro developmental ability of NT embryos derived from undifferentiated MSCs were a higher than those from differentiated MSCs or FFs. Moreover, multipotent MSC might have a potential for the production of viable cloned Mini-pigs. Therefore, MSCs as a nuclear donor might a key to improving the production of cloned Mini-pig as animal model for xenotransplantation. This work was supported by Grant No. 20070301034040 from Bio-organ, Republic of Korea.

Since the birth of the first cloned sheep was reported, fibroblasts are commonly used as donor cells for nuclear transfer. However, in some species there is a higher incidence of abnormal fetuses, still births, and neonatal deaths as a consequence of reprogramming disorders including DNA methylation and histone acetylation. Cloned embryos with mesenchymal stem cells (MSC) have shown higher developmental ability compared to fibroblast, suggesting that undifferentiated genome can required ease reprogramming. Because MSC are relatively difficult to collect from bone marrow, skin is an alternative source of the donor cells. However, molecular and functional analyses remain uncertain between MSC derived from bone marrow and skin stem cells isolated from ear. The present study compared the expression of early transcription factors (Oct-4, Nanog and Sox-2), and differentiation capability to osteocytes, adipocytes and chondrocytes of MSC isolated from bone marrow and skin-derived putative stem cells from miniature pig. Bone marrow was isolated by Ficoll density gradient method, and skin separated from epidermis and dermis was diced into 2-mm diameter explants, and attached to tissue culture dishes. Cells were then cultured in DMEM/F12 supplemented with 10% FBS, 10 ng mL–1 bFGF, 10 ng mL–1 EGF at 38.5°C, in a humidified atmosphere of 5% CO2 in air. Expression of Oct-4, Nanog, Sox2 was analysed by RT-PCR. Osteogenic and adipogenic differentiation were induced following the protocols described previously (Jin et al. 2007 Int. J. Dev. Biol. 51, 85–90; Mohana Kumar et al. 2007 Mol. Cells 24, 343–350) and compared by histological staining and RT-PCR. Osteocytes were defined by the formation of the mineral nodules of deposition of calcium by Von Kossa staining and differentiations into adipocytes and chondrocytes were identified by oil red O staining of lipid vacuoles and alcian blue of proteoglycan, respectively. Skin-derived MSC were revealed to similar mRNA expression of Oct-4, Nanog, Sox2 compared to bone marrow derived MSC. However, bone marrow derived MSC were higher mRNA expression about of osteocytic genes (osteoclacin and osteonectin), chondrocytic gene (collagen type) and adipocytic genes (aP2) than those of skin-derived MSC. In addition, bone marrow derived MSC were revealed greater deposition of calcium, proteoglycan, and lipid vacuoles than those of skin derived MSC by histological staining. The results of present study suggest that cells isolated from skin have lower potential than MSC isolated from bone marrow. However, skin-derived stem cells have properties of multi-lineage differentiation and can be obtained easily. These stem cells, therefore, can serve as easily accessible and expandable source possessing donor cells for cloning, potential animal model, and clinical applications. This work was supported by Grant No. 20070301034040 from Bio-organ, Republic of Korea.

Although cloned canine offspring have been produced, the efficiency of nuclear transfer (NT) is still far from expectations. The most important factor interfering with the success of NT is the shortage of canine matured oocytes. In the present study, the nuclear-cytoplasmic interaction and in vitro development of porcine oocytes cloned with canine fetal fibroblasts (cFF) and canine mesenchymal stem cells (cMSC) were assessed by examining the developmental rate, embryonic morphology, total cell number, canine genome expression, and in vivo development following the transfer of iNT embryos into the oviducts of surrogate mother. Isolated cMSC from bone marrow were characterized by early transcription factors (Oct3/4, Sox2, and Nanog) and by differentiation into adipogenic, osteogenic, and neuronal cells under controlled in vitro conditions. Porcine cumulus–oocyte complexes (COC) were matured in vitro as described by Kim YS et al. (2005 Mol. Reprod. Dev. 70, 308–313). Porcine NT embryos were used for the control group. Canine MSC showed their characteristic property of attaching to plastic culture flasks and forming a monolayer of spindle-shaped cells. Adherent cells showed moderate expression of Oct3/4, Sox2, and Nanog protein by immunostaining. Following osteogenic cell induction, cMSC transformed into mineralized nodules by von Kossa staining. Oil red O staining revealed that MSC produced lipid droplets after incubation in adipogenic media. Neuronal-like cells with multipolar, round cell bodies organized into a network-like structure were confirmed by immunostaining of neuronal markers (NFM). The tissue-specific mRNA levels were confirmed after differentiation by RT-PCR (osteogenic cell, Osteocalcin; adipogenic cell, PPAR γ; neuronal cell, NFM). The control group blastocysts (porcine oocytes-pMSC) exhibited significantly (P < 0.05) greater development and total cell numbers of than blastocysts of interspecies groups (porcine oocytes-cFF and cMSC). The canine genome GAPDH was detected in cFF, cMSC, and interspecies NT embryos using PCR, whereas there was no detection in porcine oocytes and the negative control. Most of the embryos arrested at the 8- to 16-cell stage and only 5% of embryos developed to blastocyst stage. The interspecies-NT embryos (average 35 embryos/recipient dog) were surgically transferred into the oviducts of 3 recipient dogs. Two of them showed exterior signs of implantation; i.e. a slight thickening of the uterus and small pea-sized lumps. No pregnancies were detected in the recipients after ovariohysterectomy surgery. These results demonstrate the potential of cMSC isolated from bone marrow to differentiate into mesoderm (osteogenic and adipogenic cell) and ectoderm (neuronal cell)-like cells under specific induction conditions. Although interspecies NT embryos could not implant in the canine uterine wall, the results provided valuable information on canine MSC and interspecies NT. This work was supported by Grant No. 20070301034040 from Bio-Organ, Republic of Korea.

Monoclonal gammopathies can either be benign or more commonly malignant. The commonest disease associated with it is multiple myeloma. Over the seven-year period 1984-1990, two hundred and thirty-four monoclonal gammopathies were seen at the University Hospital, Jamaica. Multiple myeloma was diagnosed in one hundred and fifty-six cases (84 males and 72 females). The diagnoses of most of the others were not known as the samples came from other institutions. Of the patients with myeloma, the most common immunoglobulin type was IgG followed by IgA and then pure light chain disease. Only in about half of the cases where urine was analysed was Bence-Jones protein found. The majority of the cases had abnormal total serum protein, albumin and total globulin concentrations. Most of the cases also were in renal failure. Hypercalcaemia, hyperphosphataemia, elevated alkaline phosphatase, gammaglutamyl transferase and aspartate aminotransferase occurred in about one-third of them. These results were not much different from those reported in other countries.

109 patients with imperforate anus were followed up postoperatively for 5 to 28 years. Postoperative continence was evaluated by anorectal manometry, barium enema and clinical symptoms. A comprehensive assessment system, combining the clinical and objective systems was designed which can reflect postoperative anal continence function completely and objectively. It is useful for determining the causes of incontinence and its treatment.

Mesenchymal stem cells (MSCs) isolated from bone marrow have unique self-renewal capability for multilineage differentiation and show promise in the field of medicine for cell therapies and tissue engineering. In the present study, we examined the maintenance of cell populations and the differentiation ability of canine MSCs into mesodermal lineages, oesteoblasts and adipocytes, under different culture conditions. MSCs isolated from bone marrow by Ficoll gradient treatment were cultured in (1) ADMEM supplemented with 10% fetal bovine serum (FBS) (basic medium), (2) basic medium supplemented with 10 ng mL–1 basic fibroblast growth factor (bFGF), (3) basic medium supplemented with 1000 IU mL–1 human leukemia inhibitory factor (hLIF), and (4) basic medium supplemented with 10 ng mL–1 bFGF and 1000 IU LIF. In Experiment 1, under a phasecontrast microscope (40�), morphological changes of MSC populations, including the appearance of more spherical shapes and extending processes arranged into network-like structures, were observed in all media with cytokines added, but not in basic medium. In Experiment 2, MSCs cultured under different conditions at 3–5 passages were induced into osteogenic and adipogenic lineages, and their characteristics were evaluated based on the formation of mineral nodules by alkaline phosphatase-positive cells, von Kossa and alizarin red staining of osteoblasts, and oil red O staining of lipid vacuoles following the protocol described earlier (Jin et al. 2007 Int. J. Dev. Biol. 51, 85–90). Only MSCs cultured in basic medium were successfully differentiated into osteoblasts and adipocytes. In conclusion, this study indicates that supplementation of culture media with cytokines might have a negative effect on the pluripotency maintenance of canine MSCs. Further studies are required to evaluate more characteristics of gene expression in canine MSCs.

Adult stem cells are more desirable than somatic cells for nuclear transfer (NT) because of their easy reprogrammability to resemble the genome of the zygote (Zhu et al. 2004 Biol. Reprod. 70, 1088–1095). Mesenchymal stem cells (MSCs) are a heterogeneous population of uncommitted and lineage-committed cells and have a more flexible potential as donor cells for NT. The aim of this study was to compare the developmental potential of NT embryos using undifferentiated (MSCs) and differentiated cells in the same lineage (osteocyte, adipocyte, and chondrocyte) by assessing the cleavage and blastocyst rates. Fetal fibroblasts were used as NT control. MSCs obtained from the aspirated bone marrow of a neonatal pig were cultured in advanced-DMEM (ADMEM) supplemented with 5% FCS. The differentiation potential was demonstrated by culture of MSCs at passage 3 under the conditions that were favorable for adipogenic, osteogenic, and chondrogenic development (Pittenger et al. 1999 Science 284, 143–147). For NT, cells from passages 3–5 were transferred into the perivitelline space of enucleated MII oocytes that had been in vitro-matured after collection from slaughterhouse-derived ovaries. After fusion with a needle-type electrode, eggs were cultured in 7.5 µg mL−1 cytochalasin B for 3 h, and subsequently cultured in PZM-3 medium for 6 days. Statistical significance was tested using ANOVA with Bonferroni and Duncan tests. The results are presented in Table 1. The rates of cleavage and development to blastocyst stage of NT embryos varied among donor cell sources. Most eggs (92.2 ± 2.7%) cloned with MSCs cleaved, and 47.8% of eggs developed to the blastocyst stage. In contrast, NT eggs using differentiated MSCs—osteocytes, adipocytes, chondrocytes, and controls (fetal fibroblasts)—revealed significantly (P < 0.05) lower cleavage (74.5, 63.4, 74.3, and 66.4%, respectively) and blastocyst development (33.7, 30.1, 36.5, and 25.5%, respectively) rates than those using undifferentiated MSCs. The results demonstrate that the genome of donor cells with different differentiated status supports embryonic development to various degrees, and multipotent MSCs might have a greater potential in producing viable cloned porcine embryos. Table 1.Development of NT embryos with undifferentiated and differentiated cells This work was supported by Grant No. R05-2004-000-10702-0 from KOSEF, Republic of Korea.

FeSiBNb amorphous powder was prepared by gas atomization and subsequently consolidated into bulk amorphous cores by hot pressing after creating an oxide layer on the surface of the powder. FeSiBNb amorphous powders exhibiting a saturation magnetization of 127emu∕g and a coercivity of 0.34 Oe were successfully obtained by gas atomization. Hot pressing enabled consolidation of the FeSiBNb amorphous powder core within the supercooled liquid region. The core prepared from the FeSiBNb amorphous powder covered with an oxide layer of 1μm thickness showed excellent high-frequency characteristics with a stable permeability over 10 MHz and a core loss of 400mW∕cm3 at 50 kHz for Bm=0.1T. The oxide layer formed on the FeSiBNb amorphous powder was effective in improving the frequency dependence of magnetic properties and in reducing the core loss for the FeSiBNb amorphous powder cores.

An improved embedded zerotree wavelet (IEZW) is presented, which can significantly reduce the scanning and symbol redundancy of the existing EZW.

Objectives To define the clinical and laboratory features associated with infective dermatitis (ID) and confirm its association with human T-lymphotrophic virus type I (HTLV-I). Design A case series of patients with ID were compared with patients with atopic dermatitis (AD), which is an important disease in the differential diagnosis of ID. Setting Patients were recruited from dermatology and pediatric clinics at the University Hospital of the West Indies and the Bustamante Children's Hospital, Kingston, Jamaica. Main Outcome Measures Clinical and laboratory features of patients with AD were compared with those of patients with ID. Patients Consecutive patients older than 112years diagnosed as having ID (n=50) and AD (n=35) were enrolled based on clinical findings. Results The mean ages of patients with ID and AD were 6.9 and 7.8 years, respectively. Histologically, both diseases were predominantly chronic dermatitis with propensity for skin colonization withStaphylococcus aureusand β-hemolytic streptococci; however, the distribution of sites of skin involvement differed. Infection with HTLV-I was the most distinguishing feature among patients with ID, with seropositive results in 100%; only 5 (14%) of the 35 patients with AD had results seropositive for HTLV-I. Infective dermatitis was further characterized by dermatopathic lymphadenitis in 16 (67%) of 24 patients with palpable nodes. Anemia, lymphocytosis, and low albumin and elevated serum globulin levels were more prevalent among patients with ID. Significant elevations of IgA, IgD, and IgG levels were observed among patients with ID compared with those with AD. However, both patients with AD and those with ID had levels of IgD and IgE elevated above the normal range. T-cell subsets among patients with ID revealed T-cell activation with a high percentage of HLA-DR antigen positivity, elevated CD4 (2.4 × 109/L) and CD8 (1.4 × 109/L) cell counts, with an increased CD4/CD8 ratio of 1:73. Conclusion Infective dermatitis is a distinct clinical entity associated with HTLV-I, which plays a role in the pathogenesis and immune perturbations observed.