Toxoplasma gondii triggers secretion of interleukin-12 but low level of interleukin-10 from the THP-1 human monocytic cell line

Previous studies using both in vitro and in vivo mouse models have demonstrated that a subtle balance between pro- and anti-inflammatory cytokines, among which interleukin-12 (IL-12) and interleukin-10 (IL-10), respectively is crucial to control Toxoplasma infection. However, the few studies performed with human cell lines highlighted important host-related differences in the immune response to Toxoplasma gondii. The goal of our work was thus to study the production of both IL-12 and IL-10 by the THP-1 human monocytic cell line in response to Toxoplasma. We demonstrated that infection by live parasites (RH strain) triggers secretion of IL-12, but low level of IL-10. IL-12 secretion appeared within 8 h, up to 48 h. We also showed that infection by live parasites is not mandatory since heat-killed parasites, crude tachyzoite lysate as well as excreted/secreted antigens induced significant, yet reduced production of IL-12.     

Indo-Fijian Children’s BMI

Health research has shown that overweight and obesity in children and adults are becoming significant public health problems in the developing world. Evidence suggests that this phenomenon is more marked in urban than rural areas and may be associated with modernization. However, the underlying reasons for this nutrition transition remain unclear. Dietary shifts, often in conjunction with income and time constraints in urban environments, may entail a greater reliance on more convenient sugar and fat-dense food. Also, the necessity of labor-intensive agricultural work to meet rural subsistence needs is supplanted in urban environments by sedentary work. This paper extends the application of human behavioral ecology theory into the realm of international development and policy by applying Kaplan’s embodied capital theory to explore differences in food habits and nutritional status of Indo-Fijian children within the context of urbanization. Urban high-embodied-capital women demonstrate higher rates of wage-earning employment than urban low-embodied-capital or rural women. Findings indicate that urban high-embodied-capital households spend significantly more on food purchases, purchase a greater proportion of processed foods, and have children with higher body mass indexes (BMI) than do urban low-embodied-capital or rural households. This suggests that urban high-embodied-capital mothers, who tend to be employed, may be making trade-offs between income and food choices.

Purification and some properties of galectin-1 derived from water buffalo (Bubalus bubalis) brain

An increasing number of galectins have been found in various animal species, the most abundant of which is galectin-1. The purpose of the present study was to purify and characterize galectin-1 from buffalo brain. We purified the galectin using a combination of ammonium sulphate fractionation and affinity chromatography and the homogeneity was determined by both native polyacrylamide gel electrophoresis (PAGE) and denaturing SDS-PAGE. The molecular weight of the galectin as determined by SDS-PAGE under reducing conditions and by gel filtration column under native conditions was 13.8 and 24.5 kDa, respectively, suggesting a dimeric form of galectin. The most potent inhibitor of the galectin activity was lactose, giving complete inhibition of hemagglutination at 0.8 mM. Galectin showed higher specificity towards human blood group A. Free thiol groups were estimated at a molar ratio of 2.9. The effects of alkylating reagents (iodoacetate and iodoacetamide) on saccharide binding of the galectin were studied. Both alkylating reagents significantly inactivated the activity of the galectin within 20 min. The temperature and pH stability of the galectin were determined. Our findings based on physico-chemical properties, carbohydrate and blood group specificities of the galectin may have future implications in biological and clinical applications.     

Anti-proliferation and apoptosis induced by a novel intestinal metabolite of ginseng saponin in human hepatocellular carcinoma cells

20-O-(beta-D-glucopyranosyl)-20(S)-protopanaxadiol (IH-901), a novel intestinal bacterial metabolite of ginseng protopanaxadiol saponins, is reported to induce apoptosis in a variety of cancer cells. We purified the compound and measured its in vitro anti-tumor activity. IH-901 inhibited cell growth of human hepatocellular carcinoma SMMC7721 cells in a dose- and time-dependent manner. We also found that IH-901 induced apoptotic cell death concurrent with cell cycle arrest in G0-G1 phase in SMMC7721 cells. At the molecular level, we show that IH-901 upregulates cytochrome c, p53, and Bax expression, and downregulates pro-caspase-3 and pro-caspase-9 expressions in a dose-dependent manner, while the levels of Bcl-2 and Bcl-X(L) were unchanged in IH-901-treated SMMC7721 cells. These results provide significant insight into the anticarcinogenic action of IH-901.     

Effect of neurotrophins on differentiation, calcification and proliferation in cultures of human pulp cells

Neurotrophins (NTs) are expressed during tooth development. However, little is known about a role of NTs in differentiation of pulp cells into mineralizing cells. In this study, mRNA expressions of hard tissue-related proteins, calcification and proliferation are examined in cultures of human pulp (HP) cells. Nerve growth factor (NGF), brain derived neurotrophic factor (BDNF), neurotrophin (NT)-3 and NT-4/5 increased the mRNA levels of dentin sialophsphoprotein, alkaline phosphatase, osteopontin, type I collagen and bone morphogenetic protein-2 and mineral deposition in cultures of HP cells. The increased levels and manners varied, depending on the concentrations of NTs and hard-tissue related protein tested. On the other hand, only NGF significantly stimulated DNA synthesis in cultures of HP cells. These findings suggest that NTs characteristically regulate hard-tissue related protein expression, calcification and proliferation in pulp cells. NTs may accelerate pulp cell differentiation.     

Different evolutionary fates of recently integrated human and chimpanzee LINE-1 retrotransposons

The long interspersed element-1 (LINE-1 or L1) is a highly successful retrotransposon in mammals. L1 elements have continued to actively propagate subsequent to the human–chimpanzee divergence,  6 million years ago, resulting in species-specific inserts. Here, we report a detailed characterization of chimpanzee-specific L1 subfamily diversity and a comparison with their human-specific counterparts. Our results indicate that L1 elements have experienced different evolutionary fates in humans and chimpanzees within the past  6 million years. Although the species-specific L1 copy numbers are on the same order in both species (1200–2000 copies), the number of retrotransposition-competent elements appears to be much higher in the human genome than in the chimpanzee genome. Also, while human L1 subfamilies belong to the same lineage, we identified two lineages of recently integrated L1 subfamilies in the chimpanzee genome. The two lineages seem to have coexisted for several million years, but only one shows evidence of expansion within the past three million years. These differential evolutionary paths may be the result of random variation, or the product of competition between L1 subfamily lineages. Our results suggest that the coexistence of several L1 subfamily lineages within a species may be resolved in a very short evolutionary period of time, perhaps in just a few million years. Therefore, the chimpanzee genome constitutes an excellent model in which to analyze the evolutionary dynamics of L1 retrotransposons.     

Breeding success and population trends in Adélie penguins in areas with low and high levels of human disturbance

The breeding performance and population trends of Adélie penguins (Pygoscelis adeliae) was studied at Esperanza/Hope Bay, Antarctic Peninsula, by comparing an area with low levels of human disturbance (LLD) and an area with high levels of human disturbance (HLD), close to an Argentine research station. From 1995/1996 to 2004/2005 (except for 1999/2000 and 2003/2004), the following population parameters were measured in both areas: (1) the number of breeding pairs, (2) the number of chicks in creches and (3) the number of chicks produced by breeding pairs. Counts were made for 26 breeding groups situated in the LLD area and 63 breeding groups located in the HLD area. The number of chicks per breeding pair was obtaobtained by following 100 marked nests in each area. All parameters were measured as described in the CCAMLR Monitoring Program protocols. The magnitude and direction (increasing or decreasing) of the changes in breeding population size and the number of chicks creched were similar in both areas. Overall, the number of breeding pairs decreased from 4,744 to 2,968 (37.4%) in the LLD area, and from 8,744 to 5,378 (38.6%) in the HLD area. The number of chicks fledged increased from 3,808 to 4,065 (6.7%) in the LLD area, and decreased from 6,991 to 6,712 (4%) in the HLD area. Breeding success (chicks fledged per marked nest) did not differ significantly between areas for most of the seasons compared. In 1996/1997, breeding success was significantly higher in the HLD area. Our data suggest that environmental influences currently exert greater effects than human disturbance on the penguin population at Esperanza Bay.     

Non-fucosylated therapeutic antibodies: the next generation of therapeutic antibodies

Therapeutic antibody IgG1 has two N-linked oligosaccharide chains bound to the Fc region. The oligosaccharides are of the complex biantennary type, composed of a trimannosyl core structure with the presence or absence of core fucose, bisecting N-acetylglucosamine (GlcNAc), galactose, and terminal sialic acid, which gives rise to structural heterogeneity. Both human serum IgG and therapeutic antibodies are well known to be heavily fucosylated. Recently, antibody-dependent cellular cytotoxicity (ADCC), a lytic attack on antibody-targeted cells, has been found to be one of the critical effector functions responsible for the clinical efficacy of therapeutic antibodies such as anti-CD20 IgG1 rituximab (Rituxan^®) and anti-Her2/neu IgG1 trastuzumab (Herceptin^®). ADCC is triggered upon the binding of lymphocyte receptors (FcγRs) to the antibody Fc region. The activity is dependent on the amount of fucose attached to the innermost GlcNAc of N-linked Fc oligosaccharide via an α-1,6-linkage, and is dramatically enhanced by a reduction in fucose. Non-fucosylated therapeutic antibodies show more potent efficacy than their fucosylated counterparts both in vitro and in vivo, and are not likely to be immunogenic because their carbohydrate structures are a normal component of natural human serum IgG. Thus, the application of non-fucosylated antibodies is expected to be a powerful and elegant approach to the design of the next generation therapeutic antibodies with improved efficacy. In this review, we discuss the importance of the oligosaccharides attached to the Fc region of therapeutic antibodies, especially regarding the inhibitory effect of fucosylated therapeutic antibodies on the efficacy of non-fucosylated counterparts in one medical agent. The impact of completely non-fucosylated therapeutic antibodies on therapeutic fields will be also discussed.     

In situ observation of a cell adhesion and metabolism using surface infrared spectroscopy

In this study, we report on an in situ monitoring system of living cultured cells using infrared absorption spectroscopy in the geometry of multiple internal reflections (MIR-IRAS). In order to observe living cultured cells, the temperature in the sample chamber of a FT-IR spectrometer was maintained at 37 °C and a humidified gas mixture containing 5% CO₂ was introduced into the sample chamber. Human breast cell line MCF-7 cultured on Si MIR prisms were placed in the sample chamber and infrared spectra of MCF-7 cells were collected for 5 h. It was found that the adhesion and metabolism of MCF-7 cells could be monitored by the absorption intensity of amide-II protein band (1,545 cm⁻¹) and also by the absorption intensities of CH _x bands (2,700–3,100 cm⁻¹). These results suggest that our system is useful for a nondestructive and non-label monitoring of cell viability. Our method based on infrared absorption spectroscopy has a potential for bioscreening application.     

Recent advances in the generation of human monoclonal antibody

The use of monoclonal antibodies (mAbs) has now gained a niche as an epochal breakthrough in medicine. Engineered antibodies (Abs) currently account for over 30% of biopharmaceuticals in clinical trials. Several methods to generate human mAbs have evolved, such as (1) immortalization of antigen-specific human B cell hybridoma technology, (2) generation of chimeric and humanized antibody (Ab) from mouse Ab by genetic engineering, (3) acquisition of antigen-specific human B cells by the phage display method, and (4) development of transgenic mice for producing human mAbs. Besides these technologies, we have independently developed a method to generate human mAbs by combining the method of in vitro immunization using peripheral blood mononuclear cells and the phage display method. In this paper, we review the developments in these technologies for generating human mAbs.