Serum immunoglobulins of Clarias batrachus (Cb-Ig) were purified by affinity chromatography using bovine serum albumin as capture ligand. Under reducing conditions in SDS-PAGE, Cb-Ig was composed of ...a heavy (H) chain (68.7kDa) and two light (L) chains (27.4 and 26.3kDa). Purified Cb-Ig was used to produce a monoclonal antibody (MAb) designated E4 MAb that belonged to IgG1 subclass. In Western blotting, this MAb showed binding to H chain of purified Cb-Ig and putative H chains in reduced sera of C. batrachus, Clarias gariepinus and Heteropneustes fossilis. However, no binding was observed with serum protein of Labeo rohita and Channa striata. Cross-reactivity of anti-Cb-Ig MAb was observed with serum of C. batrachus, C. gariepinus and H. fossilis in competitive ELISA. In immunoblotting of non-reduced Cb-Ig with E4 MAb, four bands assumed to be tetrameric, trimeric, dimeric and monomeric form were observed. In flow cytometric analysis of the gated lymphocytes, the number of surface Ig-positive (Ig+) cells in blood, spleen, kidney and thymus of C. batrachus was determined to be 50.1±3.1, 55.1±3.36, 42.4±4.81 and 5.1±0.89%, respectively, using E4 MAb. Ig+ cells were also demonstrated in formalin-fixed paraffin embedded tissue sections of spleen, kidney, thymus and smears of blood mononuclear cells in indirect immunoperoxidase test. The developed MAb was employed to detect pathogen-specific immunoglobulins in the sera of C. batrachus immunized with killed Edwardsiella tarda, by an indirect ELISA. This monoclonal antibody can be useful tool in immunological research and assays.
► Serum immunoglobulins of Clarias batrachus were purified by affinity chromatography. ► A monoclonal antibody (E4 MAb) was produced against purified Ig of C. batrachus. ► E4 MAb showed binding to heavy chain of purified Ig in Western blotting. ► The MAb was used to quantify and demonstrate Ig+ cells in blood and lymphoid organs. ► This MAb could detect pathogen-specific immunoglobulins in serum of immunized fish.
The dorsal hippocampus of the rat normally receives its 5-HT innervation from two homologous groups of cells in the median raphe nucleus via the cingulum bundle-induseum griseum (CB-IG) and the ...fornix-fimbria (FF) (J. Comp. Neurol., 179 (1978) 641-667 and Brain Res. Bull., 10 (1983) 445-451). 5-HT immunoreactive (IR) fibers are distributed in a laminar pattern in the hippocampus. These fibers have large varicosities and are densely distributed in the infragranular layer of dentate gyrus, in the stratum lacunosum-moleculare of the cornu Ammonis and in the area fasciola cinerea (FC). The present study provides evidence that the density of the 5-HT-IR fibers in the dorsal hippocampus is greatly decreased but maintained a similar laminar pattern 3 days after lesioning by microinjection of 4 micrograms of 5,7-dihydroxytryptamine (5,7-DHT) into the CB-IG. An apparently normal density and distribution pattern of the 5-HT-IR fiber is seen by 42 days postlesion. The FC in the hippocampus is among the first regions reinnervated by 5-HT-IR fibers with very dense and large varicosities. The restitution of 5-HT-IR fibers in the dorsal hippocampus after the 5,7-DHT lesion in the CB-IG is accompanied by a marked increase in the number and intensity of 5-HT-IR fibers in the FF. No evidence of a regeneration of 5-HT-IR fibers is seen distal to the injection site in the CB-IG. These observations provide direct evidence for homotypic collateral sprouting in the CNS induced by removal of a single fiber type.
Frequent pathogenetic condition of hemoconcentration in calves results in apparent increase in the parameters of metabolic profile above the standard level. These values, however, do not conform to ...the actual state and often lead to unsubstantiated diagnostic conclusions with pathological deviations. On the basis of experimental study on the dynamics of some parameters of hematological, protein, mineral and electrolytic profile in the calves with experimental hemoconcentration, a correction table and diagram for the calculation of corrected values for some parameters of metabolic profile were elaborated. At hematocrit value increased above the physiologic limit (0.45--0.60) the correction values of parameters in percent are subtracted from the actual value. The correction table was used to correct actual values of those parameters of metabolic profile which during the experimental hemoconcentration were found to have direct relations to the dynamics (Er, Hb, Le, CB, Ig, serum Ca, P and Na). The correction of serum potassium and of some enzymes (AST, ALT, gMT) is not indicated because no relations were determined. Whereas the actual values of the studied parameters reached their maxima (statistically significant differences in Er, Hb, CB, Ig levels) at the culmination of hemoconcentration (6th hour--1st sampling), the differences between the corrected values and initial values were in all parameters insignificant. The correction of actual values of parameters by means of the diagram or table is simple, expeditious and reliable.
Objectives were to (a) propose a mathematical model utilizing the Richards' function to interpret germination kinetics, (b) determine temperature effects on seed germination and seedling development, ...and (c) compare strategies of seedling growth. Four perennials, big bluestem (BB) (Andropogon gerardi Vitman), caucasian bluestem (CB) {Bothriochloa caucasica (Prin.) C.E. Hubb.}, indiangrass (IG) {Sorghastrum nutans (L.) Nash}, and switchgrass (SG) (Panicum virgatum L.), and an annual crabgrass (CG) {Digitaria sanguinalis (L.) Scop.} were used in three germination tests. The Richards' autocatalytic function was used to describe the cumulative germination curve. Prechilling enhanced speed and percentage of germination. Optimum temperatures for germination of prechilled seeds were between 20 and 30 C, while those for nonprechilled seeds were between 12 and 20 C. Time to reach 50% of final germination (GT(,50)) decreased with increasing temperature. True germination rate (%/day) of each species was maximum at GT(,50). Base temperatures for germination of IG, BB, SG, and CB were 8.6, 8.9, 10.3, and 10.9 C, respectively. Arrhenius plots indicated a change in temperature response between 20 and 25 C for germination. In controlled-environmental chambers, seedling growth at 20 C was reduced dramatically when compared with that at 25 and 30 C. Crabgrass developed faster than perennial species, had the shortest phyllochron, and the fastest tillering rate index, leading to rapid increase in leaf number and leaf area. Among perennials, SG had the highest leaf elongation rate (LER) and developed permanent roots faster than other species. Alternatively, CG had a low LER, but had the fastest phyllochron and the highest rate of index of tillering, giving it the largest number of leaves and fastest rate index of leaf area development. Both SG and CB developed more leaf area and accumulated dry matter more rapidly than other perennials. Both IG and BB partitioned a greater percentage of dry matter to roots at early stages, but later developed less mass of permanent roots than SG and CB. Generally, seedling weight at final harvest was ranked from highest to lowest as CG, SG, CB, IG, and BB.
To investigate the enhanced cytotoxicity against leukemia cells of natural Killer (NK) cells from cord blood (CB) after expansion in vitro.
NK cells was expanded on a layer of trophoblast cells with ...irradiated K562-mb15-41BBL cell line for 21 days. The levels of receptors on NK cells were detected by flow cytometry. Cytotoxicity of expanded NK cells against leukemia cells and specific ligand of immunoglobulin like(Ig- liKe)receptors were assessed using 51Cr released assay.
There were no differences of inhibitory receptors expression between fresh NK cells and expanded NK cells CD158a:(16.77±11.65)% vs(14.37±11.12)%, P>0.05; CD158b: (42.48±18.11)% vs (40.92±19.02)%, P>0.05; NKG2A: (70.20±18.43)% vs (78.90±13.69)%, P>0.05, but higher activated receptors expression on expanded NK cells NKp30: (54.10±13.27)% vs (4.14±2.05)%, P<0.05; NKp44: (72.10±17.30)% vs (0.52±1.16)%, P<0.05; NKp46: (80.63±14.01)% vs (44.19±6.19)%, P<0.05; NKG2D: (97.50±2.55)% vs (72.25±14.35)%, P<0.05. Expanded NK cells showed higher cyto
Dynamic changes of T and B lymphocyte subsets, and immunoglobulin (Ig) synthesis in cultured B lymphocytes were studied in accordance with the clinical activity in 29 adult patients with minimal ...change nephrotic syndrome (MCNS) . The subsets (T3, T4, T8, sBγ, sBα, sBμμ, and sBε) were identified by detection of cell surface markers such as OKT3, OKT 4, OKT8, and γ, α, μ and s chain of Ig respectively. Cytoplasmic Ig positive cells (cB) were studied after culture of peripheral lymphocytes for 5 days. In the nephrotic phase (n=10), T lymphocyte subsets remained unchanged, but sB (sBγ, sBα, sBμ, sBε) and spontaneous cB (cBγ, cBα, cBε) were increased. However, cB induced by pokeweed mitogen (PWM a 10μg/ml) did not differ significantly from the control levels, and the PWM stimulation index (St. I. = cB with PWM/cB without PWM) in the IgG producing system was decreased (p<0.005) . In the remitting phase by steroid therapy (n=10), T3, T4 and T4/T8 (helper/suppresser) ratio revealed a significant reduction in an accompa-niment with a tendency of normalization of B lymphocyte subsets, However, spontaneous and PWM induced cB were increased no more than the control value, resulting in a significant reduction of cB/sB ratio. In the remitted phase requiring no steroid therapy (n=9), all of the parameters were normalized except for the PWM St. I. in the IgG producing system, which was decreased (p<0.025) . Autologous plasma of nephrotic phase significantly reduced cB with or without PWM, but remitted plasma did not. These results suggest that in the nephrotic phase of MCNS B lymphocyte subsets are already activated in vivo, but their proliferation or maturation from sB to cB is suppressed.
碩士
國立中興大學
材料科學與工程學系所
99
In this study, the conductive polycarbonate（PC）/ acrylonitrile-butadiene-styrene copolymer（ABS）/ multi-walled carbon nanotube（MWNT）composites have been prepared using melt ...compounding method. The MWNT was modified using free radical reaction of maleic anhydride（MA）to graft on the surface of MWNT. For comparison, PC/ABS/carbon black（CB）composites were also fabricated through the same procedure. The electrical conductivity, mechanical and thermal properties of composites with various weight ratio will be discussed.
Raman spectroscopy shows a strong band at 1580 cm-1（G mode）and a disordered-induced peak at 1355 cm-1（D mode）, which may originate from the defects in the curned graphene sheets. Comparing the ID/IG ratio of the samples, which is 1.20 for MWNT and 1.48 for MA-modified MWNT, results the chemical functionalization increases the degree of disorder. The conductivity of fabricated PC/ABS composite would approach 103 Ω/□ by adding 1.0 wt% MWNT、1.5 wt% MWNT-MA and 12 wt% CB. The smal
