Micro-scaled high-throughput digestion of plant tissue samples for multi-elemental analysis

Background  

Quantitative multi-elemental analysis by inductively coupled plasma (ICP) spectrometry depends on a complete digestion of solid samples. However, fast and thorough sample digestion is a challenging analytical task which constitutes a bottleneck in modern multi-elemental analysis. Additional obstacles may be that sample quantities are limited and elemental concentrations low. In such cases, digestion in small volumes with minimum dilution and contamination is required in order to obtain high accuracy data.     

Safflower petals: A source of gamma linolenic acid

Safflower petals have been shown to have a lot of medicinal and therapeutic values. Indian safflower petal samples were analyzed for the red pigment carthamin, protein and oil contents. The petal oil (4.0–5.8%) was further analyzed for its fatty acids followed by alpha linolenic acid (15–19%) and palmitic acids (14–16%). Gamma linolenic acid, which has a lot of therapeutic value was present to an extent of 2–3%; decanoic and dodecanoic acids (2–5%) were also present.     

添加木质素和生物炭对土壤氮、磷养分及水分损失的影响

研究添加木质素和生物炭对土壤氮、磷养分及水分损失的影响,降低土壤改良剂使用成本,增加造纸黑液中木质素的利用途径,促进木质素在土壤改良中的应用。试验选取生物炭改良剂与木质素作为对比,通过土柱淋溶和静态吸收法,研究不同添加量(质量分数为0,1%,2%,4%)的木质素和生物炭对土壤氮磷养分、水分、脲酶活性以及pH的影响。土壤中添加木质素和生物炭均能减缓pH变化程度;抑制土壤脲酶的活性,且在添加氮肥后的1～20天抑制效果明显,其抑制效果与添加量呈正相关。添加量为1%,2%,4%的木质素和生物炭与对照组相比,铵态氮挥发量分别显著减少8.29%,14.29%,14.86%和3.79%,11.65%,15.26%;全氮淋溶量分别显著减少32.37%,37.70%,42.49%和25.43%,30.70%,39.54%;全磷淋溶量分别显著减少23.68%,40.48%,48.12%和6.97%,22.88%,35.30%;水分淋溶量分别显著减少7.71%,15.82%,9.29%和9.91%,15.00%,16.06%。在本试验中,木质素、生物炭质量分数分别为2%和4%时对降低土壤氮、磷养分和水分损失的效果最佳。因此,可以说明木质素和生物炭在水肥保持上的效果相近,在一定程度上可以替代生物炭改良剂在土壤中的应用。     

Characterization of monoclonal antibodies against avian reovirus S1133 protein sigmaA synthesized in Escherichia coli

Monoclonal antibodies (MAbs) were prepared against avian reovirus S1133 protein sigmaA (esigmaA) synthesized in Escherichia coli. MAbs were characterized and used to develop a diagnostic test. Ten MAbs were selected for competitive binding assay following coupling with horseradish peroxidase. The results indicated that these MAbs delineated two epitopes I and II of esigmaA. An immuno-dot binding assay was used to detect the effect of denaturation on antibody recognition of the epitopes. All MAbs bound to esigmaA in its native form. After denaturation by boiling in SDS and 2-mercaptoethanol, the binding of MAbs recognizing epitope I was fully abolished. However, the reactivity of MAbs recognizing epitope II was not affected. MAbs 31 and 32, recognizing epitopes I and II, respectively, were selected for the cross-reactivity to heterologous reovirus strains. The results suggest that the two epitopes are highly conserved among these virus strains. A MAb capture enzyme-linked immunosorbent assay (ELISA) procedure was developed using MAbs 32 and 31 to detect reovirus protein sigmaA in samples from tendon tissues of infected bird and chicken embryo fibroblast (CEF) cell cultures. Avian reovirus sigmaA antigens in tendon specimens were detected from the inoculated birds as early as 2 days post-inoculation (PI), approximated a peak at 7 days PI, and maintained this until 16 days PI, then decreased gradually. A clear difference in absorbance values between the tendon samples of the avian reovirus- and mock-infected birds is obtained. Positive results were also obtained from avian reovirus-infected CEF and from the tendon tissues of naturally infected broilers. These results indicated that the MAb capture ELISA is a useful methods for the detection of avian reovirus from chickens suspected to have avian reovirus infections.     

Anabolism,catabolism and proteomic analysis in the slow‐twitch muscle of pacu (Piaractus mesopotamicus) submitted to prolonged fasting followed by refeeding

Here, we aimed to study the slow muscle of the fish Piaractus mesopotamicus submitted to 30 days of fasting (D30) followed by 1 day (D31) or 30 days of refeeding (D60). Histological analysis of fibre diameter was performed in D30 and D60. The gene expression of parvalbumin (pvalb), atrogenes (murf1a, murf1b, mafbx) and anabolic genes (igf‐1, mtor) was analysed using RT‐qPCR in D30, D31 and D60. The proteome was obtained by shotgun proteomics at D30 and D60, and the set of differentially expressed proteins was analysed by bioinformatics. In all experiments, the control was regularly fed fish. The histological analysis showed no changes in muscle fibre diameter. The expression of catabolic and anabolic genes was not changed, except for the downregulation of igf‐1 in D30 and of mafbx in D31. The expression of pvalb was not changed in D30 and D60 but was decreased in D31. The proteomic analysis identified 169 proteins in D30 (24 upregulated and 18 downregulated) and 170 proteins in D60 (17 upregulated and 21 downregulated); many of them were related to energetic metabolism and intracellular Ca²⁺ homoeostasis. Overall, our results indicate that the slow‐twitch muscle presented few changes upon prolonged fasting and refeeding condition.     

Immunopathogenic behaviors of canine transmissible venereal tumor in dogs following an immunotherapy using dendritic/tumor cell hybrid

Canine transmissible venereal tumor (CTVT) is a naturally occurring tumor that can be transmitted between dogs via live tumor cell inoculation. It is also a spontaneous self-regression tumor and its behavior is closely related to host immune responses. Since CTVT had been widely used for tumor models in canine cancers, whether this self-regression may overtake the immunity elicited from an exogenous tumor vaccine remains unclear and certainly worthwhile to be investigated. In this study, we used DCs/tumor hybrids as a tumor vaccine to evaluate the CTVT model. We prepared mature allogeneic dendritic cells from bone marrow and then assessed their phenotype (CD80, CD83, CD86, CD1a, CD11c, CD40 and MHC II), antigen uptake and presenting abilities. Fused dendritic cell/CTVT hybrids were then used as a vaccine, administered three times at two-week intervals via subcutaneous injection near the bilateral auxiliary and inguinal lymph nodes. In comparison with unvaccinated dogs (spontaneous regressed group), within a period of 2.5 months, the vaccinations substantially inhibited tumor progression (p<0.05) and accelerated the rate of regression by a mechanism involving amplification of the host tumor-specific adaptive immune responses and NK cytotoxicity (p<0.001). Pathologic examination revealed early massive lymphocyte infiltration resulting in final tumor necrosis. In addition, there are not any detectable effects on routine physical, body temperature or blood chemistry examinations. In conclusion, our data furnishes a reference value showing that CTVT is a model of potential use for the study of immunity elicited by vaccines against tumors, and also enable early-phase evaluation of the dendritic cell/tumor vaccine in terms of raising host immunity.     

Importance of extensional rheological properties on fiber-enriched corn extrudates

Incorporation of insoluble fibers into extruded cereals severely limits their expansion and reduces crispness. The specific objective of this work was to investigate how corn bran and its fractions mixed with cornmeal affect rheology and extrudate expansion. Alkali-treated bran (ATB) and alkaline-soluble bran (ASB) were prepared from unmodified corn bran (UMB). Mixtures of the different corn bran fractions and degermed cornmeal having a 26% (w/w) of total dietary fibers were extruded in a twin screw extruder and expansion of the extrudates was determined. Melt shear rheology of mixtures of cornmeal and different corn bran fractions having a 20% total dietary fiber was determined using a capillary rheometer at 120 °C while the extensional rheology was determined using lubricated squeezing flow. The expansion of extrudates containing ATB was larger than those containing UMB, while extrudates containing ASB showed a greater expansion that was close to that of the control. Addition of UMB to cornmeal increased shear and extensional viscosity significantly as compared to the control. ATB addition increased the shear viscosity of the mixture to a small extent but showed the highest extensional viscosity amongst the samples. Addition of ASB resulted in mixtures having lower shear and extensional viscosities than the control.     

大兴安岭林区森林抚育工程系统输出特性和转化特性的研究

为提高大兴安岭林区森林抚育工程作业质量和工程成效,采用系统工程的分析方法,根据大兴安岭林区生态环境、功能和林木的结构特点,对大兴安岭林区最主要的三种抚育类型(透光抚育、生长伐抚育、生态疏伐抚育)的直接输出特性、长效输出特性和转化特性进行了详细分析。在此基础上,利用运行控制和统计学理论,通过调整输出特性偏移量、输出特性标准偏差和输出特性公差要求,提高工程能力,进而优化输出特性,最终实现提高工程作业质量和工程成效的目的。对输出特性和转化特性的分析研究,为大兴安岭林区森林抚育工程技术要求标准的产出提供基础和支撑。     

Studies on the antioxidant activities of natural vanilla extract and its constituent compounds through in vitro models

Vanilla extract was prepared by extraction of cured vanilla beans with aqueous ethyl alcohol (60%). The extract was profiled by HPLC, wherein major compounds, viz., vanillic acid, 4-hydroxybenzyl alcohol, 4-hydroxy-3-methoxybenzyl alcohol, 4-hydroxybenzaldehyde and vanillin, could be identified and separated. Extract and pure standard compounds were screened for antioxidant activity using beta-carotene-linoleate and DPPH in vitro model systems. At a concentration of 200 ppm, the extract showed 26% and 43% of antioxidant activity by beta-carotene-linoleate and DPPH methods, respectively, in comparison to corresponding values of 93% and 92% for BHA. Interestingly, 4-hydroxy-3-methoxybenzyl alcohol and 4-hydroxybenzyl alcohol exhibited antioxidant activity of 65% and 45% by beta-carotene-linoleate method and 90% and 50% by DPPH methods, respectively. In contrast, pure vanillin exhibited much lower antioxidant activity. The present study points toward the potential use of vanilla extract components as antioxidants for food preservation and in health supplements as nutraceuticals.