DETEKSI GLUKOSA DALAM URIN ORANGUTAN SUMATERA (PONGO ABELII) MENGGUNAKAN STRIPTEST SEMIKUANTITATIF DI TAMAN HEWAN PEMATANG SIANTAR

Penelitian ini bertujuan mengetahui ada tidaknya glukosa dalam uron orangutan sumatera (Pongo abelii) sebagai penunjang diagnosa di Taman Hewan Pematang Siantar, Sumatera Utara. Pengoleksian urin terhadap 4 ekor orangutan sumatera di dalam kandang yang dilakukan pada pagi hari yaitu saat orangutan bangun tidur atau sebelum pemberian pakan orangutan. Pengulangan uji dilakukan 3 kali selama 10 hari pada bulan Januari 2015. Setelah pengoleksian urin kemudian dilakukan pemeriksaan dengan cara mencelupkan stripstest pada 5-10 ml urin selama 30 detik. Analisis data menggunakan metode deskriptif kualitatif dengan hasil bersifat semikuantitatif melalui pembacaan nilai glukosa pada stripstest yang memiliki skala perubahan warna yaitu : negatif, positif 1 (100 mg/dL), positif 2 (250 mg/dL), positif 3 (500 mg/dL), dan positif 4 (1000 mg/dL). Hasil penelitian menunjukkan bahwa dari 4 sampel urin orangutan sumatera tidak terdeteksi adanya glukosa dalam urin

Data from: Feeding preference as a main determinant of microscale patchiness among terrestrial nematodes

Soil biota are responsible for essential ecosystem services such as carbon storage, nutrient cycling and water retention. However, assessment of the condition of soil biota is hampered by an overwhelming level of diversity. With representatives in all trophic levels of the food web, nematode communities can be used as bio-indicators. Accurate assessment of nematode assemblages requires insight in the distribution of specimens with distinct food preferences. With the availability of taxon-specific quantitative-PCR assays, distribution patterns of multiple nematode groups can be investigated simultaneously. Here, microscale patchiness of 45 nematode taxa was studied on 12 sampling sites (each with four adjacent microplots) located on arable fields or semi-natural grasslands (‘system’), and on marine-, river clay or sandy soils (‘soil type’). From each microplot five composite samples were collected. Contrary to our expectations, an increase of the number of cores per composite sample did not result in more accurate measurements, and apparently the levels of microscale patchiness of the taxa are low compared to what has been reported for oligophagous plant-parasites. System and soil type did not affect microscale distribution. To investigate the level of patchiness in more detail, detection probability (DP) and variability of abundances were calculated. Common and widespread bacterivorous and fungivorous taxa had DP ≥ 90%, confirming low level of microscale patchiness. With DPs of 40-70%, predators and most omnivores showed degrees of local clustering. An overview of mean variabilities of abundances is presented that offers insight in how feeding preferences impact the microscale distribution both between and within trophic groups

Transient Expression of Secretory IgA In Planta is Optimal Using a Multi-Gene Vector and may be Further Enhanced by Improving Joining Chain Incorporation

Secretory IgA (sIgA) is a crucial antibody in host defence at mucosal surfaces. It is a promising antibody isotype in a variety of therapeutic settings such as passive vaccination and treatment of inflammatory disorders. However, heterologous production of this heteromultimeric protein complex is still suboptimal. The challenge is the coordinate expression of the four required polypeptides; the alpha heavy chain, the light chain, the joining chain and part of the polymeric-Ig-receptor called the secretory component, in a 4:4:1:1 ratio. We evaluated the transient expression of three sIgAκ variants, harbouring the heavy chain isotype α1, α2m1 or α2m2, of the clinical antibody Ustekinumab in planta. Ustekinumab is directed against the p40 subunit that is shared by the pro-inflammatory cytokines interleukin (IL)-12 and IL-23. A sIgA variant of this antibody may enable localized treatment of inflammatory bowel disease. Of the three different sIgA variants we obtained the highest yield with sIgA1κ reaching up to 373 μg sIgA/ mg total soluble protein. The use of a multi-cassette vector containing all four expression cassettes was most efficient. However, not the expression strategy, but the incorporation of the joining chain turned out to be the limiting step for sIgA production. Our data demonstrate that transient expression in planta is suitable for the economic production of heteromultimeric protein complexes such as sIgA

A stable monomeric form of human IL-10 has impaired activity.

<p>Bone marrow-derived macrophages, dendritic cells and mast cells from wild-type mice were tested for their response to human IL-10 and a stable monomeric form of IL-10 (IL-10m). Cells pre-treated with IL-10 were stimulated with 100 ng/ml LPS and TNF-α expression was determined to asses anti-inflammatory properties of IL-10 in macrophages (A) and dendritic cells (B) (<i>n</i> = 3, error bars indicate standard error). Mast cells were cultured for 48 hours in the presence of IL-10, where after cell viability was determined (C) (<i>n</i> = 3, error bars indicate standard error). <i>P</i><0.01 at all concentrations in all three assays.</p

The extracellular domain of IL-10R2 is not sufficient to maintain IL-10 activity.

<p>CHO-K1 cells were co-transfected with combinations of the expression vectors for IL-10R1, IL-10R2, IL-10R2^Δ230–330 (extracellular domain), IL-10R2^Δ1–190 (intracellular domain) or an empty vector (mcs) and cultured for 24 hours. Surface expression of IL-10R1 or IL-10R2 was analysed by flow cytometry and mean fluorescent intensity is plotted (<i>n</i> = 6, error bars represent standard error) (A). Phosphorylation of tyrosine 705 (Y705) of STAT3 by IL-10 (100 ng/ml) in CHO-K1 cells upon transient transfection with IL-10 receptors was analysed by western blot (B).</p

The intracellular domain of IL-10R2 mediates conformational changes in IL-10R1.

<p>CHO-K1 cells were co-transfected with combinations of the expression vectors for IL-10R1, IL-10R2, IL-10R2^Δ230–330, IL-10R2^Δ1–190 or an empty vector (mcs) and cultured for 24 hours. Cells were surface stained with the 1B1.3a anti-mouse IL-10R1 monoclonal antibody and analysed by flow cytometry (A). Mean fluorescent intensity for IL-10R1 binding is plotted in a dose-dependent manner (<i>n</i> = 6, error bars represent standard error). IL-10 was cross-linked to the surface of transfected cells and surface-bound IL-10 was detected by flow cytometry (B). Mean fluorescent intensity for IL-10 binding is plotted (<i>n</i> = 3, error bars represent standard error). Asterisk(s) indicate significant differences as determined by a Welch’s <i>t</i>-test (*<i>P</i><0.05; **<i>P</i><0.01).</p

Tyk2 affects early responses to IL-10.

<p>Bone marrow-derived macrophages and dendritic cells from Tyk2^-/- mice were investigated in more detail for their signaling. Phosphorylation of tyrosine 705 (Y705) of STAT3 by IL-10 (0, 1, 10 and 100 ng/ml) was analysed in wild-type and Tyk2^-/- macrophages and dendritic cells by western blot (A and B respectively). Relative up-regulation of SOCS3 mRNA expression by IL-10 was analysed by quantitative PCR in both macrophages and dendritic cells (C). Fold induction of SOCS3 expression was calculated using the 2^ΔCt method using HPRT as a reference gene. Macrophages and dendritic cells from wild-type and Tyk2^-/- transgenic mice were pre-treated with IL-10 and were stimulated with 100 ng/ml LPS and the inhibition of TNF-α expression was determined at 2 and 24 hours (D). Asterisk(s) indicate significant differences as determined by a Welch’s <i>t</i>-test (*<i>P</i><0.05; **<i>P</i><0.01).</p

Tumor Necrosis Factor and Schistosoma mansoni egg antigen omega-1 shape distinct aspects of the early egg-induced granulomatous response.

Infections by schistosomes result in granulomatous lesions around parasite eggs entrapped within the host tissues. The host and parasite determinants of the Schistosoma mansoni egg-induced granulomatous response are areas of active investigation. Some studies in mice implicate Tumor Necrosis Factor (TNF) produced in response to the infection whereas others fail to find a role for it. In addition, in the mouse model, the S. mansoni secreted egg antigen omega-1 is found to induce granulomas but the underlying mechanism remains unknown. We have recently developed the zebrafish larva as a model to study macrophage recruitment and granuloma formation in response to Schistosoma mansoni eggs. Here we use this model to investigate the mechanisms by which TNF and omega-1 shape the early granulomatous response. We find that TNF, specifically signaling through TNF receptor 1, is not required for macrophage recruitment to the egg and granuloma initiation but does mediate granuloma enlargement. In contrast, omega-1 mediates initial macrophage recruitment, with this chemotactic activity being dependent on its RNase activity. Our findings further the understanding of the role of these host- and parasite-derived factors and show that they impact distinct facets of the granulomatous response to the schistosome egg

IL-10R2 mediated signaling via Tyk2 plays a limited role in IL-10 activity.

<p>Bone marrow-derived macrophages, dendritic cells and mast cells from wild-type, IL-10R1^-/-, IL-10R2^-/- and Tyk2^-/- mice were tested for their response to IL-10. Macrophages and dendritic cells from wild-type and IL-10R^-/- mice were pre-treated with IL-10 and subsequently stimulated with 100 ng/ml LPS. The percentage of inhibition of TNF-α expression of macrophages and dendritic cells was determined after overnight incubation (A and B, respectively) (<i>n</i> = 3, error bars indicate standard error). Similarly, macrophages and dendritic cells from Tyk2^-/- mice were tested for their response to IL-10 (D and E, respectively) (<i>n</i> = 4, error bars indicate standard error). Mast cells from wild-type and transgenic mice were cultured for 48 hours in the presence of IL-10 and cell viability was determined (C and F) (<i>n</i> = 3 for IL-10R^-/- mice and <i>n</i> = 4 for Tyk2^-/- mice, error bars indicate standard error). Asterisk(s) indicate significant differences as determined by a Welch’s <i>t</i>-test (*<i>P</i><0.05; **<i>P</i><0.01).</p

The differential response of bone marrow-derived cells to IL-10.

<p>Bone marrow-derived macrophages, dendritic cells and mast cells were analysed by flow cytometry for the expression of cellular markers CD11b & F4/80 (A), CD11c & MHC-II (D) and FcεRI & c-kit (G), respectively. Bone marrow-derived cells were tested for their response to IL-10 by measuring suppression of TNF-α expression by macrophages (B) and dendritic cells (E) or proliferative/anti-apoptotic ability in mast cells (H) (<i>n</i> = 4, error bars indicate standard error). Dose dependent tyrosine phosphorylation of STAT transcription factors by IL-10 (0, 1, 10 or 100 ng/ml) was analysed by western blot in macrophages (C), dendritic cells (F) and mast cells (I).</p