Statistical tools for analyzing water quality data

Water quality data are often collected at different sites over time to improve water quality management. Water quality data usually exhibit the following characteristics: non-normal distribution, presence of outliers, missing values, values below detection limits (censored), and serial dependence. It is essential to apply appropriate statistical methodology when analyzing water quality data to draw valid conclusions and hence provide useful advice in water management. In this chapter, we will provide and demonstrate various statistical tools for analyzing such water quality data, and will also introduce how to use a statistical software R to analyze water quality data by various statistical methods. A dataset collected from the Susquehanna River Basin will be used to demonstrate various statistical methods provided in this chapter. The dataset can be downloaded from website http://www.srbc.net/programs/CBP/nutrientprogram.htm.

猴和人T淋巴细胞表面TRBC受体和E受体的比较研究

1985年贲昆龙等首次发现人和猴T淋巴细胞能与树鼩(Tupaia belangeri)的红细胞(TRBC)形成亲和力极强的玫瑰花结，它与绵羊红细胞(SRBC)玫瑰花结(E花结)明显不同。例如，TRBC经神经氨酸酶处理之后，结花率明显下降，TRBC与T淋巴细胞所形成的玫瑰花结，经45℃保温，仍不受影响，为了进一步探讨TRBC受体和SRBC受体(CD2)，以及与其它T细胞表面分化搞原(CD)的系。我用某些抗人T细胞CD的单克隆抗体(McAb)对人和猴淋巴细胞进行玫瑰花结抑制试验，抗原调变和共调变(Antigenic modulation or co-modulation)实验，并且研究了TRBC受体在其它免疫细胞和某些人类和长臂猿细胞系的分布。结果表明，TRBC与外周血E~+-PBL形成玫瑰花结的百分率为88.8%。而E~--PBL仅为4.16%。TRBC受体存在于所有被试T细胞系(CEM, H33JHJA1, Jurkat, MLA-144, Molt-3, Molt-4, Molt-4 clone-8 和PEER)，但不存在于外周血粒细胞，B细胞，以及B细胞系的绝大多数细胞表面(Daudi, Raji和Reh)。分布于全T细胞的CD3, TCR, CD5, CD6和CD7的相McAb OKT3, T108(F1), T136(F101-15), T149(M-T604)和T152(7G5)均不能调变和共调变TRBC受体。猴和人外周血淋巴细胞与TRBC玫瑰花结的形成，不被T11.1 McAb OKT11所阻断，相反，OKT11显著地阻断猴和人外周血淋巴细胞E玫瑰花结的形成，最大抑制率分别为49.3%和77.7%。在世界各地10个实验室送交第四次国际人白细胞化化抗原讨论会待鉴定的13个CD2 McAb中，除T089 (39C1.5)因抗体量不够未作实验外，对其他12种McAb都进行试验，T081 (x/3)，T082 (GLB-T11.2/1)，T083 (GLB-T11.1/1)，T085 (RPA-2.10)，T1088 (0-275)和T092 (M-T201)对TRBC玫瑰花结和E玫瑰花结都呈现明显的阻断作用，T084 (F110.08)，T091 (AICD2.1)以及已知参数CD2 McAb-T086 (D-66 clonel)和T087 (GT-2)都不阻TRBC玫瑰花结的形成，亦不调变TRBC受体，而对E玫瑰花结则有不同程度阻断效应，并且调变E变体，使E玫瑰花结形成细胞百分率下降。T090 (6F10.3)和T198 (JOR-T2)即不阻TRBC玫瑰花结的形成，也不抑制E玫瑰花结的形成。由此可见，TRBC受体分布于全T细胞，它不同于已知的全T细胞表面分化抗原CD2 (gp50), CD3/TCR复合物，CD5, CD6和CD7。CD2分子不与TRBC玫瑰花结的形成，也不是介导E玫瑰花结的唯一分子。至少有二个或三个以上的蛋白质与E玫瑰花结和TRBC玫瑰花结的形成有关，其中有的分子为E受体和TRBC受体所共有。TRBC受体很有可能包括新的T淋巴细胞分化抗原。

川牛膝多糖的免疫活性及其水解活性片段分离研究

川牛膝多糖（CP）是从传统中药川牛膝（Cyathula officinalis Kuan）中提取的一种活性多糖，现代药理研究表明川牛膝多糖是川牛膝许多生物活性的物质基础。本实验室前期进行了川牛膝多糖的提取、分离、结构鉴定及其部分活性研究，发现川牛膝中多糖含量非常高，在对川牛膝多糖活性的初步研究中也证实了其具有免疫调节作用。我们为了进一步了解其免疫调节活性，并为构效关系的研究奠定基础，对其进行了如下研究： 1． 通过体外毒性检测、淋巴细胞增殖实验、NK细胞杀伤活性和腹腔巨噬细胞吞噬中性红活性测定，发现川牛膝多糖在10～300μg/mL浓度范围内，对细胞无毒性作用；能够促进LPS诱导的B淋巴细胞增殖(P<0.01)、增强NK细胞杀伤活性(P<0.05)和PMΦ吞噬中性红活性(P<0.01)，且随多糖浓度增高而增强；但其对ConA诱导的T淋巴细胞的增殖无促进作用(P>0.05)。 2． 通过正常小鼠体内淋巴细胞转化实验、迟发型变态反应分析、抗体生成细胞检测、碳粒廓清检测、腹腔巨噬细胞吞噬鸡红细胞活性和NK细胞活性测定，发现川牛膝多糖在适应性免疫方面能够促进SRBC免疫小鼠体内的抗体生成细胞的生成（P<0.01）和增强DNFB诱导的DTH（P<0.05），但对ConA诱导的脾淋巴细胞增殖无促进作用（P>0.05）；在固有免疫方面能够提高小鼠碳粒廓清速率（P<0.05），PMΦ吞噬 CRBC 活性（P<0.01）和NK细胞杀伤活性（P<0.05）。同时还发现其对由环磷酰胺(Cy)引起的白细胞数下降具有很好的抑制作用（P<0.01）。 3． 为了获得结构明确、均一的保留活性的川牛膝多糖片段，为其作用机制、构效关系研究提供关键研究材料，我们开展了“保留免疫活性的最小片段”的分离制备的初步研究。建立并优化了川牛膝多糖的酸水解条件，发现在6%的样品浓度，0.025mol/L的硫酸浓度，65℃的水解温度，水解时间为8min的条件下可以得到一系列连续的多糖片段；采用Bio-Gel P2 分子筛柱层析分离得到5个级分，通过体外淋巴细胞增殖实验、NK细胞活性测定、腹腔巨噬细胞吞噬中性红实验发现其中的一个片段仍保留较强的免疫活性，并测得其分子量约为2057Da，为保留免疫活性的最小片段的进一步分离奠定了基础。 Cyathula officinalis Kuan is a commonly-used Traditional Chinese Medicine (TCM) with a wide range of pharmacological activities. Modern pharmacological researches showed the polysaccharide extracted from it (CP) is an important component for many bioactivities of this TCM. In the previous studies, we found CP showed significant immuno-regulative activities. In order to evaluate this activity systematically and lay foundations for revealling its immuno-regulative machanisms and the Structure -Function relationship, we carried out the following research works: 1． The in vitro immunoactivities of CP were evaluated by using normal mice immunocytes with respects to cytotoxicity, lymphocytes proliferation, NK activity and the ability of peritoneal macrophage phagocytizing neutral red. The polysaccharide showed no cytotoxicity below the concentration of 300 μg/mL, and could promote B lymphocytes proliferation (P<0.01), enhance NK activity (P<0.05) and the ability of peritoneal macrophage phagocytizing neutral red (P<0.01) at the concentration of 10-300 μg/mL. The above effects were positively correlated with the concentration of the polysaccharides. But it could not promote T lymphocytes proliferation (P>0.05). 2． The in vivo immunoactivities of CP were observed on normal mice through the following indices: splenic lymphocyte transformation efficiency, delayed-type allergy, antibody-forming cells activity (AFC), rate of carbon clearance, rate of peritoneal macrophage phagocytizing chicken red blood cell (CRBC) and NK activity, and its influence on the decline of the mouse leucocyte count induced by Cy. The polysaccharide at medium-dose enhanced delayed-type allergy （P<0.05）and NK activity（P<0.05） and increased the rate of carbon clearance（P<0.05）, AFC activity（P<0.01） and the rate of peritoneal macrophage phagocytizing CRBC（P<0.01）. The polysaccharides also effectively resisted the decline of the mouse leucocyte count induced by Cy（P<0.01）. However, it couldn’t increase the splenic lymphocyte transformation efficiency（P>0.05）. 3． Attempting to isolate and prepare the minimal fragments retaining activity with identical structure for further studying on immuno-regulative mechanism and Structure-Function relationship, we carried out the study on hydrolysis of CP, isolation of hydrolysed fragments, and the activity evaluation of the isolated fragments. CP with concentration of 6% was hydrolysed at 65℃ for 8 min with sulfuric acid of 0.025 mol/L，then the hydrolysate was separated using Bio-Gel P2 chromatography, 5 portions of fragments were obtained. The immunoactivities of these fragments were evaluated by using normal mice immunocytes with respect to lymphocytes proliferation, NK activity and ability of peritoneal macrophage phagocytizing neutral red. One fragment with relative molecular mass of 2057Da was found retaining immunoactivity.

低氧对大鼠体液免疫的抑制作用

本实验以模拟高原低氧的方法，探讨了低氧对大鼠体液免疫的作用，并与高原鼠免比较，体液免疫以溶血亲和IgG产生为指标。实验结果：与对照组相比，大鼠低氧10d，5km海拔抑制溶血素形成10．3％，7km海拔抑制溶血素形成21．9％；经再次免疫后又低氧10d，5km海拔抑制溶血素形成4．2％，7km海拔抑制溶血素形成4．6％，高原土著动物高原鼠兔（Ochotona curzoniae）则不表现上述的抑制现象；大鼠经SRBC腹腔致敏2d后低氧5d和8d，未表现低氧抑制溶血素形成的作用；大鼠侧脑室给予外源性CRF（1μg／rat），可抑制溶血素形成和IgG产生，溶血素形成抑制8.6％，IgG产生抑制14．0％；7km低氧大鼠侧脑室给予CRF受体阻断剂（α-helicalCRF(9-41)）50μg对低氧抑制溶血素形成一定的阻断作用，可部分阻断低氧抑制IgG产生，使IgG产生的抑制率由24.2%提高到12.1%；大鼠腹腔注射1μgCRF对溶血素形成和IgG产生无影响；与假手术组比，大鼠在摘除双侧肾上腺后，低氧对溶血素形成仍抑制6.6%。因而，本研究认为低氧对初次和再次体液免疫均产生抑制作用，而且抑制作用发生在体液免疫的起始阶段，其抑制作用与中枢CRF升高有关。

Impaired phagocytosis by alveolar macrophages from diabetic rats is related to the deficient coupling of LTs to the Fe gamma R signaling cascade

Diabetic individuals are more susceptible to infections and this seems to be related to impaired phagocyte function. Alveolar macrophages (AMs) are the first barrier to prevent respiratory infections Leukotrienes (LTs) increase AM phagocytic activity via Fc gamma R. In this study, we compared AMs from diabetic and nondiabetic rats for phagocytosis via Fc gamma R and the roles of LTs and insulin Diabetes was induced in male Wistar rats by alloxan (42 mg/kg, i.v); macrophages were obtained by bronchoalveolar lavage and IgG-opsonised sheep red blood cells (IgG-SRBC) were used as targets. LTs were added to the AMs 5 min before the addition of IgG-SRBC. AMs were treated with a LT synthesis inhibitor (zileuton, 10 mu M), or antagonists of the LTB(4) receptor (CP105 696, 10 mu M) cys-LT receptor (MK571, 10 mu M), 30 or 20 min before the addition of IgG-SRBC, respectively. We found that the phagocytosis of IgG-SRBC by AMs from diabetic rats is impaired compared with non-diabetic rats. Treatment with the LT inhibitor/antagonists significantly reduced AM phagocytosis in non-diabetic but not diabetic rats. During the phagocytosis of IgG-SRBC LTB(4) and LTC(4) were produced by AMs from both groups. The addition of exogenous LTB(4) or LTD(4) potentiated phagocytosis similarly in both groups Phagocytosis was followed by the phosphorylation of PKC-delta. ERK and Akt This was reduced by zileuton treatment in AMs from non-diabetic but not diabetic rats The addition of insulin to AMs further increased the phagocytosis by increasing PKC-delta phosphorylation These results suggest that the impaired phagocytosis found in AMs from diabetic rats is related to a deficient coupling of LTs to the Fc gamma R signaling cascade and that insulin has a key role in this coupling An essential role for insulin in Innate immunity is suggested (C) 2010 Elsevier Ltd. All rights reserved.

The effects of testosterone on immune function in quail selected for divergent plasma corticosterone response

The immunocompetence handicap hypothesis (ICHH) suggests that the male sex hormone testosterone has a dual effect; it controls the development and expression of male sexually selected signals, and it suppresses the immune system. Therefore only high quality males are able to fully express secondary sexual traits because only they can tolerate the immunosuppressive qualities of testosterone. A modified version of the ICHH suggests that testosterone causes immunosuppression indirectly by increasing the stress hormone corticosterone (CORT). Lines of Japanese quail (Coturnix japonica) selected for divergent responses in levels of plasma CORT were used to test these hypotheses. Within each CORT response line (as well as in a control stock) we manipulated levels of testosterone in castrated quail by treatment with zero (sham), low or high testosterone implants, before testing the birdsʼ humoral immunity and phytohaemagglutinin (PHA)-induced immune response, as well as body condition. The PHA-induced response was not significantly affected by CORT selected line, testosterone treatment or their interaction. There was, however, a significant effect of CORT line on humoral immunity in that the control birds exhibited the greatest antibody production, but there was no significant effect of testosterone manipulation on humoral immunity. The males in the sham implant treatment group had significantly greater mass than the males in the high testosterone group, suggesting a negative effect of high testosterone on general body condition. We discuss these results in the context of current hypotheses in the field of sexual selection.

The effects of propolis on antibody production by laying hens

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Cell wall fractions from Paracoccidioides brasiliensis induce hypergammaglobulinemia

The antibody response against the antigen sheep red blood cells (SRBC) was investigated in mice pre-treated with formalin-killed Paracoccidioides brasiliensis or with cell wall fractions of the fungus. Pre-treatment with P. brasiliensis, as well as with the F1 fraction and beta-glucan significantly increased the anti-SRBC antibody response in the experimental groups as compared to the control group that received only SRBC. This immunomodulatory effect varied with the different doses employed and with pre-treatment time. We conclude that the cell wall fractions of P. brasiliensis might play an important role in the hypergammaglobulinemia associated with Paracoccidioidomycosis. © 1993 Kluwer Academic Publishers.

Involvement of the major glycoprotein (gp43) of Paracoccidioides brasiliensis in attachment to macrophages

The yeast form of Paracoccidioides brasiliensis, the causative agent of a deep mycosis in humans, is known to be phagocytized by, and to multiply inside, macrophages. In this work we describe the involvement of gp43, a major antigenic protein of P. brasiliensis, in the initial steps of attachment of the fungus to macrophages. Anti-gp43 F(ab) polyclonal fragments were capable of inhibiting phagocytosis in a concentrationdependent manner. Sheep red blood cells sensitized with purified gp43 were more endocytized than SRBC alone, and this process was also inhibited by anti-gp43 F(ab) fragments. Inhibition tests indicated the involvement of fucose and mannose residues in the phagocytosis of the fungus and of SRBC-gp43 by macrophages. Taken together, these results suggest that gp43 may be involved in the adherence and uptake of the fungus by murine peritoneal macrophages, and that this binding may be dependent on monosaccharide residues that are part of the gp43 glycoprotein. © 1998 ISHAM.

Ações antioxidantes e imunológicas do selênio em cães

Pós-graduação em Zootecnia - FMVZ

Diferentes fontes e níveis de selênio sobre a imunidade humoral de frangos de corte

The use of organic trace minerals is getting strength and is an alternative to increase production and improve other characteristics such as humoral immunity. The present study was conducted to evaluate the effect of different levels and sources of selenium (Se) on humoral immunity of broilers. A six-week research was conducted using 1440 one-day-old males broiler chickens. The experimental design was randomized with six experimental diets (A: 0.15mg kg(-1) inorganic (inorg.); B: 0.15mg kg(-1) organic; C: 0.15mg kg(-1) inorg. + organic; D: 0.45mg kg(-1) inorganic; E: 0.45mg kg(-1) organic; F: 0.45mg kg(-1) inorg. + organic) calculated to provide the described amount of Se. Each diet was replicated in six box with 40 birds. A 3x2 factorial arrangement was used and the data were analyzed by ANOVA. The immunity was evaluated by means of the reaction against vaccine of Newcastle disease, and a reaction against sheep red blood cells (SRBC). No significant effects were observed at 5% significance level in NewCastle antibody (P >0.05). However at 14 day-old the source factor had p value at 0.0580, that show a trend of inorganic source in prolong the maternal immunity. No effect was observed in the immune response against SRBC (P >0.05). The results showed a immunologic response against Newcastle vaccine and SRBC, but the treatments was not able to induce a significant difference. The source and the level of Se showed no effect on the response against Newcastle vaccine and SRBC.

The role of SENP1 in B cell development and differentiation

SUMOylation is a highly dynamic and reversible posttranslational protein modification closely related to ubiquitination. SUMOylation regulates a vast array of different cellular functions, such as cell cycle, nuclear transport, DNA damage response, proliferation and transcriptional activation. Several groups have shown in in vitro studies how important SUMOylation is for early B cell development and survival as well as for later plasma cell differentiation. This thesis focuses on the deSUMOylation protease SENP1 and its in vivo effects on B cell development and differentiation. For this a conditional SENP1 knockout mouse model was crossed to the CD19-Cre mouse strain to generate a B cell specific SENP1 knockout mouse.rnIn our conditional SENP1ff CD19-Cre mouse model we observed normal numbers of all B cell subsets in the bone marrow. However in the spleen we observed an impairment of B cell survival, based on a 50% reduction of the follicular B cell compartment, whereas the marginal zone B cell compartment was unchanged. T cell numbers were comparable to control mice. rnFurther, impairments of B cell survival in SENP1ff CD19-Cre mice were analysed after in vivo blocking of IL7R signalling. The αIL7R treatment in mature mice blocked new B cell formation in the bone marrow and increased apoptosis rates could be observed in splenic SENP1 KO B cells. Additionally, a higher turnover rate of B cells was measured by in vivo BrdU incorporation.rnSince it is known that the majority of transcription factors that are important for the maintenance of the germinal centre reaction or for induction of plasma cell development are SUMOylated, the question arose, how defective deSUMOylation will manifest itself in these processes. The majority of in vitro cultured splenic B cells, stimulated to undergo class switch recombination and plasma cell differentiation underwent activation induced cell death. However, the surviving cells increasingly differentiated into IgM expressing plasma cells. Class switch recombination to IgG1 was reduced. These observations stood in line with observation made in in vivo sheep red blood cell immunization experiments, which showed increased amounts of germinal centres and germinal centre B cells, as well as increased amounts of plasma cells differentiation in combination with decreased class switch to IgG1.rnThese results lead to the conclusion that SENP1 KO B cells increasingly undergo apoptosis, however, B cells that survive SENP1 deficiency are more prone to undergo plasma cell differentiation. Further, the precursors of these plasma cells either are not as capable of undergoing class switch recombination or they do switch to IgG1 and succumb to activation induced cell death. One possible explanation for both scenarios could be a defective DNA damage response mechanisms during class switch recombination, caused by impaired deSUMOylation. rn

ADAMTS13 activity in sickle cell disease

Sickle red blood cell (SRBC)-endothelial adhesion plays a central role in sickle cell disease (SCD)-related vaso-occlusion. As unusually large von Willebrand factor (ULVWF) multimers mediate SRBC-endothelial adhesion, we investigated the activity of ADAMTS13, the metalloprotease responsible for cleaving ULVWF multimers, in SCD. ADAMTS13 activity was determined using a quantitative immunoblotting assay. VWF:Ag and VWF:RCo were determined using commercial assays. The high-molecular-weight VWF multimer percentage was determined by employing gel electrophoresis. ADAMTS13 activity was similar among asymptomatic patients (n = 8), patients at presentation with a painful crisis (n = 23), and healthy controls. ADAMTS13/VWF:Ag ratios were lower in patients compared to healthy HbAA controls, with the lowest values at presentation with a painful crisis (P = 0.02). Division of samples in those with VWF:RCo/VWF:Ag ratios < 0.70 and those with ratios >or= 0.70 revealed significantly more samples with ratios >or= 0.70 (P = 0.01) collected during painful crises. ULVWF multimers were detected in 6 patient samples and in 1 control sample. ADAMTS13/VWF:Ag ratios were inversely related to the duration of symptoms at presentation with an acute vaso-occlusive event (r(s)-0.67, P = 0.002). Although SCD is characterized by elevated VWF:Ag levels, no severe ADAMTS13 deficiency was detected in our patients.