Chicken PRDX3 is required for proliferation of chicken embryo fibroblast cells.

1. This experiment investigated the influence of chicken PRDX3 on cell proliferation in chick embryo fibroblast cells using PRDX3 knockdown technology.2. A methyl thiazolyl tetrazolium (MTT) assay was performed to assess the effect of chPRDX3 knockdown on fibroblast proliferation. The antioxidant effect was investigated to determine if it directly mediated fibroblast cell proliferation.3. To determine the role of chPRDX3 on cell proliferation, an siRNA mediated knockdown was performed in chick fibroblast cells using an in vitro assay. The proliferation of fibroblast cells transfected with siPRDX3 #3 and siPRDX3 Mix was significantly decreased after 48 h (P < 0.01). In addition, the knockdown of chicken PRDX3 suppressed cell proliferation through an increase in oxidative stress.4. The results demonstrated that chPRDX3 is required for cell proliferation in chicken fibroblast cells. Such findings have important implications for the maintenance of chicken fibroblast cells.

Preparation of alpha-bisabolol and phenylethyl resorcinol/TiO2 hybrid composites for potential applications in cosmetics.

OBJECTIVE: Bifunctional alpha-bisabolol and phenylethyl resorcinol/TiO2 hybrids were prepared to apply in cosmetic fields, particularly in anti-ageing and hyperpigmentation treatment. The synergistic effect of combined antioxidant and UV filtering properties was achieved through functionalization of TiO2 particles with skin-lightening materials such as alpha-bisabolol and phenylethyl resorcinol. METHODS: TiO2 microspheres with a diameter of about 1 µm were synthesized through surfactant-assisted sol-gel method for use as supporting materials in the formation of hybrid composites. Carboxylation treatment was performed for surface modification of the TiO2 surface with carboxyl groups as chemical binders. Esterification reaction between carboxyl groups of carboxylated TiO2 and hydroxyl groups of alpha-bisabolol or phenylethyl resorcinol was performed. The hybrids were characterized using various techniques such as FE-SEM, DLS, EDS, ATR-FTIR, XPS and TGA. For application of prepared TiO2 composites in the field of cosmetics, the anti-radicular antioxidant abilities were evaluated using ABTS and DPPH colorimetric antioxidant assay. RESULTS: Organic/inorganic hybrid composites were successfully formed using esterification reaction between the carboxyl groups at TiO2 surface and the hydroxyl groups of the skin-lightening materials. The results demonstrate that both functionalized microspheres show scavenging ability towards the ABTS(•) and DPPH(•) radicals. Specifically, the phenylethyl resorcinol/TiO2 composites exhibited the highest antioxidant ability among the prepared samples owing to the presence of phenolic groups to scavenge free radicals. CONCLUSION: Using this strategy, it could be possible to prepare not only inorganic UV filter but also hybrid organic/inorganic materials with multifunctions and advantages which would be in a great demand for cosmetic applications.

Incidence and risk factors of poor mobilization in adult autologous peripheral blood stem cell transplantation: a single-centre experience.

BACKGROUND AND OBJECTIVES: Collection of sufficient CD34+ cells for autologous peripheral blood stem cell (PBSC) transplantation is frequently failed in patients with lymphoma or multiple myeloma (MM). We investigated the incidence and the predictive factors for poor mobilization. MATERIALS AND METHODS: A total of 205 adult patients (101 lymphoma and 104 MM) were retrospectively included for identifying the incidence of mobilization failure and the predictive factors for poor mobilization in conventional G-CSF-based mobilization regimen. Another 17 patients who used plerixafor for mobilization were included. RESULTS: Overall, 14·1% of patients (21·8% of patients with lymphoma, 6·7% of patients with MM) were poor mobilizers. Univariate analysis and multivariate analysis revealed an interval from G-CSF administration to PBSC collection exceeding 10 days and peripheral blood mononuclear cells count on the first day of collection were predictive factors for poor mobilization in lymphoma, but not in MM. Among plerixafor-treated patient group, 9 of 11 poor mobilizers who received second-cycle plerixafor mobilization were able to collect higher number of CD34+ cells than that of CD34+ cells during the G-CSF-based first mobilization. All patients who had received initial plerixafor mobilization reached 2·0 × 10(6) CD34+ cells/kg during the four leukaphereses. CONCLUSION: In conventional G-CSF-based mobilization, early PBSC collection after G-CSF administration might enhance CD34+ cell yield. A combination of a new mobilizing agent, plerixafor, would be helpful to harvest sufficient number of CD34+ cells for successful transplantation outcome while reducing the effort of collection procedures in poor mobilizers.

Detection and characterization of enterovirus associated with herpangina and hand, foot, and mouth disease in Seoul, Korea.

BACKGROUND: Human enteroviruses (HEVs) are a major cause of herpangina, HFMD (hand, foot, and mouth disease), and other neurological diseases in Seoul, Korea. METHODS: A total of 56 specimens from hospitalized patients collected from February to December 2009 (37 females and 19 males) in Seoul were tested for HEV from stool, throat swab, and vesicle swab samples taken from patients with herpangina or HFMD using cell culture and RT-PCR in 2009. By the 1D gene, encoding the VP1 capsid protein, seven different HEV genotypes were detected with Coxsackievirus A2, A4, A5, A9, A16 (CA), Coxsackievirus B1 (CB), and Enterovirus 71 (EV71). The most prevalent genotype was CA16 (6, 10.7%), followed by CA2 (4, 7.1%), CA5 (4, 7.1%), EV71 (2, 3.6%), CA4 (1, 1.8%), CA9 (1, 1.8%), and CB1 (1, 1.8%). The 1D gene sequences of two EV71 strains were closely related with one another (98.5% nucleotide similarity) and belonged to the C4 genotype. CONCLUSIONS: It is important to continuously survey the genetic characteristics of EV71 and CA16 from patients, which will provide useful data that aids in our understanding of HFMD infections in Seoul, Korea and may contribute to future control.

PPAR gamma partial agonist, KR-62776, inhibits adipocyte differentiation via activation of ERK.

Indenone KR-62776 acts as an agonist of PPAR gamma without inducing obesity in animal models and cells. X-ray crystallography reveals that the indenone occupies the binding pocket in a different manner than rosiglitazone. 2-Dimensional gel-electrophoresis showed that the expression of 42 proteins was altered more than 2.0-fold between KR-62776- or rosiglitazone-treated adipocyte cells and control cells. Rosiglitazone down-regulated the expression of ERK1/2 and suppressed the phosphorylation of ERK1/2 in these cells. However, the expression of ERK1/2 was up-regulated in KR-62776-treated cells. Phosphorylated ERK1/2, activated by indenone, affects the localization of PPAR gamma, suggesting a mechanism for indenone-inhibition of adipogenesis in 3T3-L1 preadipocyte cells. The preadipocyte cells are treated with ERK1/2 inhibitor PD98059, a large amount of the cells are converted to adipocyte cells. These results support the conclusion that the localization of PPAR gamma is one of the key factors explaining the biological responses of the ligands.

Calreticulin, a calcium-binding molecular chaperone, is required for stress response and fertility in Caenorhabditis elegans.

Calreticulin (CRT), a Ca(2+)-binding protein known to have many cellular functions, including regulation of Ca(2+) homoeostasis and chaperone activity, is essential for heart and brain development during embryogenesis in mice. Here, we report the functional characterization of Caenorhabditis elegans calreticulin (crt-1). A crt-1 null mutant does not result in embryonic lethality but shows temperature-dependent reproduction defects. In C. elegans CRT-1 is expressed in the intestine, pharynx, body-wall muscles, head neurons, coelomocytes, and in sperm. crt-1 males exhibit reduced mating efficiency and defects late in sperm development in addition to defects in oocyte development and/or somatic gonad function in hermaphrodites. Furthermore, crt-1 and itr-1 (inositol triphosphate receptor) together are required for normal behavioral rhythms. crt-1 transcript level is elevated under stress conditions, suggesting that CRT-1 may be important for stress-induced chaperoning function in C. elegans.

Effects of dietary recombinant chlorella supplementation on growth performance, meat quality, blood characteristics, excreta microflora, and nutrient digestibility in broilers.

The use of chlorella as an immune stimulant to enhance nonspecific host defense mechanisms or as an antimicrobial to inhibit bacterial growth has been reported. Thus, the aim of the present study was to clarify the effect of recombinant chlorella supplementation on growth performance, meat quality, and the blood profile, excreta microflora, and nutrient digestibility in broilers. A total of 375 one-day-old ROSS 308 broilers (male and female) were allotted to 5 dietary treatments using 5 cages with 15 chicks per cage. Treatments were: 1) NC, basal diet supplemented with 1.0% E. coli fermented liquor (EFL); 2) PC1, 0.2% EFL with chlorella; 3) PC2, 1.0% EFL with chlorella; 4) T1, 0.2% EFL with chlorella (anti-viral); and 5) T2, 1.0% EFL with chlorella (anti-viral). The broilers in the T2 treatment groups showed higher body weight gain (BGW) by 2.55% (P < 0.01) and lower feed conversion ratio (FCR) by 2.75% (P < 0.05) compared with those fed the control NC treatment group. Moreover, the blood contents of blood urea nitrogen (BUN), creatinine, and IgA in the broilers of the T2 treatment group were significantly increased by 28.12, 23.07, and 29.72%, respectively -more than those found in the broilers of the NC treatment group (P < 0.01). In contrast, the LDL/C in the blood from the animals in the T2 treatment group was significantly decreased by 23.23% - more than that in the blood from the NC broilers (P < 0.05). Based on these results, we suggest that the dietary supplementation of broilers with recombinant chlorella could improve their growth performance, increase the concentration of IgA and apparently metabolizable nitrogen in the blood, and decrease ammonia emissions. Therefore, our findings have important implications for the effect of recombinant chlorella supplementation through increasing the concentration of IgA and the level of metabolizable nitrogen.

Fyn is a redox sensor involved in solar ultraviolet light-induced signal transduction in skin carcinogenesis.

Solar ultraviolet (UV) light is a major etiological factor in skin carcinogenesis, with solar UV-stimulated signal transduction inducing pathological changes and skin damage. The primary sensor of solar UV-induced cellular signaling has not been identified. We use an experimental system of solar simulated light (SSL) to mimic solar UV and we demonstrate that Fyn is a primary redox sensor involved in SSL-induced signal transduction. Reactive oxygen species (ROS) generated by SSL exposure directly oxidize Cys488 of Fyn, resulting in increased Fyn kinase activity. Fyn oxidation was increased in mouse skin after SSL exposure and Fyn-knockout mice formed larger and more tumors compared with Fyn wild-type mice when exposed to SSL for an extended period of time. Murine embryonic fibroblasts (MEFs) lacking Fyn and cells in which Fyn expression was knocked down were resistant to SSL-induced apoptosis. Furthermore, cells expressing mutant Fyn (C448A) were resistant to SSL-induced apoptosis. These findings suggest that Fyn acts as a regulatory nexus between solar UV, ROS and signal transduction during skin carcinogenesis.

Apoptosis-inducing protein, AIP, from parasite-infected fish induces apoptosis in mammalian cells by two different molecular mechanisms.

AIP (apoptosis-inducing protein) is a protein purified and cloned from Chub mackerel infected with the larval nematode, Anisakis simplex, which induces apoptosis in various mammalian cells including human tumor cell lines. AIP has shown structural and functional homology to L-amino acid oxidase (LAO) which oxidizes several L-amino acids including L-lysine and AIP-induced apoptosis has been suggested to be mediated by H2O2 generated by LAO activity of AIP. In this study, we confirmed that recombinant AIP generated enough H2O2 in culture medium to induce rapid apoptosis in cells and this apoptosis was clearly inhibited by co-cultivation with antioxidants such as catalase and N-acetyl-cysteine. Surprisingly, however, we found that AIP still could induce H2O2-independent apoptosis more slowly than H2O2-dependent one in HL-60 cells even in the presence of antioxidants. In addition, the HL-60-derived cell line HP100-1, which is a H2O2-resistant variant, underwent apoptosis on treatment with AIP with a similar delayed time course. The latter apoptosis was completely blocked by addition of L-lysine to the culture medium, which is the best substrate of AIP as LAO, indicating that decreased concentration of L-lysine in the culture medium by AIP-treatment induced apoptosis. We also showed that the both apoptosis by AIP were associated with the release of cytochrome c from mitochondria and activation of caspase-9, and overexpressed Bcl-2 could inhibit both of the AIP-induced apoptosis. These results indicate that AIP induces apoptosis in cells by two distinct mechanisms; one rapid and mediated by H2O2, the other delayed and mediated by deprivation of L-lysine, both of which utilize caspase-9/cytochrome c system.

A novel beta-galactoside-binding lectin in adult rat kidney.

Two carbohydrate-binding proteins with subunit molecular weight of about 17,500 and 16,500, respectively, were isolated from Triton X-100 extracts of rat kidney using a lactose affinity column. They did not require Ca2+ for the carbohydrate-binding nor reducing agents for maintaining their activity. The partial amino acid sequence of the 17.5-kDa protein (rkCBP-17.5), the main component, revealed that this protein is a novel member of a superfamily of beta-galactoside-binding animal lectins. The N-terminal amino acid sequence of the 16.5 kDa component (rkCBP-16.5) indicated that it is a fragment derived from the IgE-binding protein (IgEBP). Monoclonal antibodies to rkCBP-17.5 were prepared and used to examine the distribution of the lectin in various organs of adult rats. Immunoreactive protein with the same molecular weight was found in lung, spleen and liver, in lesser amounts in heart, and in trace amounts in brain and skeletal muscle. rkCBP-17.5 exhibits binding activity to various saccharides with the following order of affinity: N-acetyllactosamine > lactose > D-galactose > methyl alpha-D-galactopyranoside > N-acetyl-D-galactosamine > methyl beta-D-galactopyranoside. It binds to Engelbreth-Holm-Swarm(EHS) tumor laminin and rat plasma fibronectin, but does not bind to human plasma fibronectin.

Interaction of human plasma fibronectin with alpha-elastin.

Because there are contradictory reports about the interaction of plasma fibronectin with elastin, we investigated the interaction in vitro. When human plasma was applied to an alpha-elastin-Sepharose column at 4 degrees C, the column-binding fraction contained fibronectin. When isolated plasma fibronectin was applied to the same column at 4 degrees C, most of the fibronectin bound to the column and was eluted with 1 M KBr. However, the binding affinity of plasma fibronectin to the alpha-elastin-Sepharose column was much weaker at 25 degrees C than at 4 degrees C. The elastin-plasma fibronectin interaction was further confirmed by demonstrating the binding of alpha-elastin to fibronectin on polyvinylidene difluoride membranes using an alpha-elastin specific antibody. The elevation of the surface hydrophobicity of plasma fibronectin at 4 degrees C was observed by hydrophobic chromatography, using alkyl-Sepharose columns. It seems that the binding of plasma fibronectin to alpha-elastin involves hydrophobic interaction, which is affected by temperature and possibly by other factors.

A novel beta-galactoside-binding lectin in cultured murine lymphocytic leukemia cells.

Using affinity chromatography on lactosyl-Sepharose, a beta-galactoside-binding protein of 38 kDa was detected in mouse L1210 lymphocytic leukemia cells. Immunoblotting analysis revealed that it is distinct from any known larger molecular weight galectin. The partial amino acid sequences of the 38 kDa protein indicated that it is a novel member of the galectin family. This 38 kDa lectin is expressed in lymphocytic cell lines but not macrophage-like cell lines.

Preferential cytotoxicity to tumor cells of 3,5-diprenyl-4-hydroxycinnamic acid (artepillin C) isolated from propolis.

A tumoricidal substance was isolated from Brazilian propolis as guided by cytotoxicity assay on HuH 13 (human hepatocellular carcinoma) cell and was characterized to be 3-[4-hydroxy-3,5-bis (3-methyl-2-butenyl) phenyl]-2-propenoic acid (3,5-diprenyl-4-hydroxycinnamic acid (artepillin C)). It exhibited preferential cytotoxicity to tumor cells cultured in vitro. The cytotoxicity observed seemed to be partly attributable to apoptosis-like DNA fragmentation. The compound showed anti-tumor activity more effective than that of 5-fluorouracil to transplantable human tumor cell lines when tested on histoculture drug response assay system.

A cross-sectional analysis of the relationship between daily alcohol consumption and serum adiponectin levels among adults aged 40 years or more in a rural area of Korea.

BACKGROUND/OBJECTIVES: Light-to-moderate alcohol consumption has been proposed to raise serum adiponectin levels, but this view is controversial. There is little information on the effect of heavy drinking. The aim of this study was to examine the relationship between alcohol consumption and serum adiponectin levels in healthy Koreans. SUBJECTS/METHODS: The design of the study was cross-sectional, using data from the Korean Multi-Rural Communities Cohort Study (MRCohort), which is a part of the Korean Genome Epidemiology Study (KoGES). The subjects were 1542 individuals (635 men and 907 women) aged ≥ 40 years who were recruited in Yangpyeong-Gun, Kyunggi province, South Korea, in 2005 and 2006. Daily alcohol consumption was calculated from average frequency of alcohol consumption and the amount of alcohol consumed per occasion using a structured questionnaire and serum adiponectin levels were measured. RESULTS: Although adiponectin levels appeared to be higher in those consuming moderate levels of alcohol than in nondrinkers, the difference was not statistically significant. Heavy drinking (≥ 90.0 g/day) was significantly related to reduced serum adiponectin levels (P=0.003), although the significance of the relationship was reduced after adjusting for potential confounders (P=0.061) such as age, waist/hip ratio, blood pressure, fasting blood glucose, current smoker, higher education, protein intake, vitamin C intake and vitamin E intake in men. The relation seemed to be stronger in individuals consuming alcohol in the form of takju (Korean rice wine; P=0.022). CONCLUSIONS: Heavy alcohol drinking (≥ 90.0 g/day) may be related to lower serum adiponectin levels in Korean men.

Polymeric mercaptosilane-modified platinum electrodes for elimination of interferants in glucose biosensors.

An oxidase-based glucose sensor has been developed that uses a mercaptosilane-modified platinum electrode to achieve selectivity of electrochemical interferants. A platinum-iridium (9:1) wire (0.178 mm o.d., sensing area of 1.12 mm2) is modified with (3-mercaptopropyl)trimethoxysilane. The modified sensors show excellent operational stability for more than 5 days. Glucose oxidase is immobilized on the modified surface (i) by using 3-maleimidopropionic acid as a linker or (ii) by cross-liking with bovine serum albumin using glutaraldehyde. Sensitivities in the range of 9.97 nA/mM glucose are observed when the enzyme is immobilized by method ii. Lower sensitivities (1.13 x 10(-1) nA/mM glucose) are observed when immobilization method i is employed. In terms of linear response range, the sensor enzyme-immobilized by method i is superior to that immobilized by method ii. The linearity is improved upon coating the enzyme layer with polyurethane. The sensor immobilized by method ii and coated with polyurethane exhibits a linear range to 15 mM glucose and excellent selectivity to glucose (0.47 nA/mM) against interferants such as ascorbic acid, uric acid, and acetaminophen.

Spinal cord injuries in children in British Columbia.

Fifteen children who sustained traumatic spinal cord injury in British Columbia over a 13-year period have been reviewed. The aetiology, incidence of spinal fracture, length of hospitalisation and subsequent spinal surgery, and their self-care, transfer and ambulatory abilities, bowel and bladder management, schooling, employment and place of abode have been determined.

Cellulitis due to Escherichia coli in three immunocompromised subjects.

In adults, cellulitis is usually caused by group A streptococci and Staphylococcus aureus. However, in patients with underlying disease, it may be caused by other organisms, such as Acinetobacter, Clostridium septicum, Enterobacter, Haemophilus influenzae, Proteus mirabilis or Escherichia coli. We report three cases of cellulitis of the lower legs where E. coli was the causative bacterial organism. It is important to suspect E. coli as a causative organism if blistering cellulitis occurs, especially in patients with underlying diseases.

Detection of multiple patterns of oscillatory oxygen consumption in single mouse islets of Langerhans.

A novel oxygen microsensor was used to measure oxygen levels in single mouse islets as a function of glucose concentration. Oxygen consumption of individual islets was 5.99 +/- 1.17, 9.21 +/- 2.15, and 12.22 +/- 2.16 pmol/min at 3, 10, and 20 mM glucose, respectively (mean +/- SEM, n = 10). Consumption of oxygen was islet-size dependent as larger islets consumed more oxygen than smaller islets but smaller islets consumed more oxygen per unit volume than larger islets. Elevating glucose levels from 3 to 10 mM induced pronounced fast oscillations in oxygen level (period of 12.1 +/- 1.7 s, n = 6) superimposed on top of large slow oscillations (period of 3.3 +/- 0.6 min, n = 6). The fast oscillations could be completely abolished by treatment with the L-type Ca2+-channel blocker nifedipine (40 microM) with a lesser effect on slow oscillations. Slow oscillations were almost completely dependent upon extracellular Ca2+. The oxygen patterns closely mimic those that have previously been reported for intracellular Ca2+ levels and are suggestive of an important role for Ca2+ in amplifying metabolic oscillations.