Mitogen-activated protein kinase is involved in the changes of intracellular free calcium concentration induced by wound exudate in cultured rat epidermal stem cells

AIM: To observe the effects of harvested wound exudate on intracellular free Ca²⁺ in epidermal stem cells (ESCs) in vitro, and to investigate the relationship between mitogen-activated protein kinases (MAPKs) signal pathways and Ca²⁺ mobilization in this condition.METHODS: Wound exudate was harvested from the 80 full-thickness wounds produced on both sides of the back in 40 adult Wistar rats. ESCs were isolated, purified from neonatal Wistar rats by referring to the formerly records and binding our ideas. When the cultured cells showed up clone growing, they were divided into five groups as follows: group A: control group (no-treatment); group B: only treatment with wound exudate; group C: treatment with wound exudate and PD98059; group D: treatment with wound exudate and SB203580; group E: treatment with wound exudate, PD 98059 and SB203580. Then, the cells were incubated with fluorescence Ca²⁺ dye fluo-3/AM at 37 ℃ for 30 min, and measured by using laser scanning confocal microscope.RESULTS: The results showed that the fluorescent intensity of group B was higher than that in group A. A phenomenon of calcium oscillation was found in group C and group D. Furthermore, a rapid decrease of fluorescent intensity was observed in the cells that were preincubated with PD98059 and SB203580 at the same time. CONCLUSION: Based on above results, we propose that wound exudate can directly induce an increase in intracellular free Ca²⁺concentrations of ESCs. MAPKs signaling pathway has an important function of feedback regulation for free Ca²⁺ mobilization of ESCs in this condition, and also is capable of affecting the biological behaviour of epidermal stem cells.     

Neuropeptide Y induced redistribution of intracellular free calcium in rat cardiomyocytes

AIM: To investigate the effect of neuropeptide Y (NPY) on intracellular free calcium(［Ca²⁺］_i) and Ca²⁺ sarcoplasmic reticulum of cardiomyocytes in rats.METHODS: Cardiomyocytes of neonatal Sprague-Dawley rats were incubated with NPY at concentration of 100 nmol/L for 24 h. Fluorescent indicator Fluo-4 AM was used to detect ［Ca²⁺］_i and Fluo-5N AM was used to detect Ca²⁺ in sarcoplasmic reticulum (SR). Calcium image was recorded by laser scanning confocal microscope. The SR Ca²⁺ load was estimated by caffeine-induced Ca2+ transient (CCT). RESULTS: 24 h after incubation with NPY, compared with control group, the concentration of ［Ca²⁺］_i was significantly elevated (P<0.05), and the concentration of free Ca2+ in SR (［Ca²⁺］_SR) was significantly decreased (P<0.05), and the peak of CCT was attenuated.CONCLUSION: Stimulation with NPY for 24 h causes redistribution of free calcium in rat cardiomyocytes, namely the elevation in ［Ca²⁺］_i and decline in ［Ca²⁺］_SR.     

Potato brown rot incidence and severity under different management and amendment regimes in different soil types

Ralstonia solanacearum race 3 biovar 2, the causative agent of potato brown rot (bacterial wilt), is an economically important disease in tropical, subtropical and temperate regions of the world. In view of previous reports on suppression of the disease by organic amendments, and the expansion of organic agriculture, it was timely to compare the effects of organic and conventional management and various amendments on brown rot development in different soils (type: sand or clay; origin: Egypt or the Netherlands). Brown rot infection was only slightly reduced in organically compared to conventionally managed sandy soils from Egypt, but organic management significantly increased disease incidence and pathogen survival in Dutch sandy and clay soils, which correlated with high DOC contents in the organic Dutch soils. There was no correlation between disease incidence or severity and bacterial diversity in the potato rhizosphere in differently managed soils (as determined by 16S DGGE). NPK fertilization reduced bacterial wilt in conventional Egyptian soils but not in Dutch soils. Cow manure amendment significantly reduced disease incidence in organic Dutch sandy soils, but did not affect the bacterial population. However, cow manure did reduce densities of R. solanacearum in Egyptian sandy soils, most probably by microbial competition as a clear shift in populations was detected with DGGE in these and Dutch sandy soils after manure amendment. Amendment with compost did not have a suppressive effect in any soil type. The absence of a disease suppressive effect of mineral and organic fertilization in Dutch clay soils may be related to the already high availability of inorganic and organic nutrients in these soils. This study shows that the mechanism of disease suppression of soil-borne plant pathogens may vary strongly according to the soil type, especially if quite different types of soil are used.     

Calcium ion promotes successful penetration of powdery mildew fungi into barley cells

To determine whether Ca²⁺ promotes powdery mildew penetration, Ca²⁺-treated barley coleoptiles were inoculated with conidia of pathogenic and nonpathogenic fungi. Penetration efficiency of the pathogenic powdery mildew Blumeria graminis was enhanced by Ca²⁺ treatment, but that of the necrotrophic pathogen Helminthosporium sp. remained unaffected. Similarly, when actin-dependent penetration resistance is suppressed with cytochalasin A, Ca²⁺ treatment specifically enhanced penetration of the nonpathogenic powdery mildew Erysiphe pisi but not that of other nonpathogens. Calmodulin inhibitors suppressed the promotive effect of Ca²⁺ on B. graminis penetration. These results suggest that barley powdery mildew specifically requires Ca²⁺ and calmodulin for penetration.     

酶解法提取海藻酸钠研究

采用正交实验研究了酶解法提取海藻酸钠的工艺及影响因素,并分析了单因素对酶解反应的影响,从而得出最佳酶解反应条件:缓冲液用量55 ml,pH值5.0,温度45 ℃,反应时间18 h,加酶量90 U/g,此时海藻酸钠浸出量可达2.448 7 g.     

外源钙处理对马铃薯块茎重量和数量的影响

以马铃薯品种费乌瑞它(Favorita)为试材,采用营养液无土栽培法,设5个外源钙浓度水平(25,75,150,300和500 mg/L),研究外源钙浓度对马铃薯单株块茎平均重量、数量和块茎内钙浓度的影响。结果表明,随外源钙浓度的升高,马铃薯单株块茎的数量减少。单株块茎的平均重量,以钙水平150,300和500 mg/L增幅较大,显著高于钙水平为25和75 mg/L的处理。单株产量以钙水平25、75、150和300 mg/L的处理显著高于钙水平500 mg/L处理。然而随外源钙浓度的升高,马铃薯商品率明显提高,块茎内钙浓度也相应升高,但块茎内钙浓度与块茎数量呈负相关关系。     

9—羟基芴—9—甲酸和醇酯化反应的无水氯化钙催化剂

无水氯化钙可代替硫酸催化酯化反应，探讨了氯化钙催化９－羟基芴－９－甲酸甲酯的条件，操作简便，反应温和，无腐蚀，减少了污染与产品纯化。     

饲粮锌和钙水平对产蛋鸡生产性能、血液生化指标和组织中锌含量的影响

试验在玉米-豆饼型基础饲粮中(含锌29.2ppm)添加不同水平锌和钙研究了饲粮锌和钙水平对蛋用型鸡产蛋期生产性能、血液生化指标和组织中锌含量的影响。结果表明:1.仅喂未加锌的基础饲粮的试验鸡开产晚(P<0.05),产蛋率低(P<0.05)、蛋重小(P<0.05)、产蛋量少(P<0.01)、饲料利用效率差(P<0.01)、破壳率高(P<0.01)。饲粮加锌1000ppm与加锌40ppm对照相比,生产性能无显著差异。饲粮高钙对产蛋鸡生产性能无不良影响。2.血清碱性磷酸酶活性随饲粮加锌水平的提高而显著提高(P<0.01),但不受饲粮钙水平显著影响(P>0.05)。饲粮加锌1000ppm使血清总蛋白含量(P<0.01)、血清白蛋白和球蛋白含量(P<0.05)显著降低。饲粮高钙使血清总蛋白含量(P<0.01)和白蛋白含量(P<0.05)显著降低,而球蛋白含量不受影响(P>0.05)。3.随饲粮加锌水平的提高,肝脏(P<0.01)、胰脏(P<0.01)和胫骨(P<0.01)中锌含量大幅度升高;血液(P<0.01)、肾脏(P<0.01)和肺脏(P<0.05)中锌含量升高幅度较小;而输卵管(P>0.05)、心脏(P>0.05)和肌肉(P>0.05)中锌含量无显著变化。饲粮高钙使肝脏(P<0.05)和肌肉(P<0.01)中锌含量显著降低,对其它器官无显著影响(P>0.05)。     

钙镁磷肥对元胡种植区酸化土壤改良效果的探究

为了探究钙镁磷肥对元胡种植区土壤酸化的阻控作用,通过连续两年的钙镁磷肥田间定位试验,以上元观镇当地元胡种植户施肥习惯为对照(CK),研究分析了在氮、钾肥用量相同的情况下,施用钙、镁、磷肥750 kg/hm~2、1 500 kg/hm~2和2 250 kg/hm~2,对土壤pH、交换性酸、阳离子交换量(CEC)、交换性盐基离子以及土壤基本肥力水平的影响。通过测算投入、产出效益,结果表明：施用钙镁磷肥能显著提升土壤pH水平,较(CK)处理提升0.27～0.31个单位,T100、T150两个处理能显著降低土壤交换性酸总量和交换性氢含量,对降低交换性铝有一定作用。施用钙镁磷肥较(CK)处理能显著提升阳离子交换量(CEC)以及交换性钙和交换性镁,是阻控土壤酸化的主要作用机制。但当施用量达到2 250 kg/hm~2时,种植元胡的新增纯收益会有所下降,伴随产生土壤中磷素过量的问题,会有造成磷素流失产生农业面源污染的风险。因此在汉中元胡种植区土壤酸化改良中推荐钙镁磷肥施用量750～1 500 kg/hm~2。     

Alcohol diluent provides the optimal formulation for calcium chloride non-surgical sterilization in dogs

Background

Surgical castration is widely used to sterilize male dogs, but has significant impacts on time to perform the operation, recovery of the animals as well as cost, which can limit population control programs. Previous research has shown intratesticular injection of calcium chloride dihydrate (CaCl₂) in saline to be a promising alternative to surgery. However, long-term azoospermia was not maintained at dosages low enough to avoid side effects. In the search for an optimized formulation, the current investigation is the first study on long-term sterilization effects of intratesticular injection of CaCl₂ in either lidocaine solution or alcohol in dogs. CaCl₂ at 20% concentration in lidocaine solution or alcohol was administered via intratesticular injection to groups of 21 dogs each. The treated animals were examined at 2, 6, and 12 months for sperm production, blood levels of testosterone, and side effects; at time zero and 12 months for testicular size and semen volume. The experimentally treated animals were compared to a control group receiving saline injection only.

Results

Testicles of dogs treated with CaCl₂ in either diluent significantly decreased in size. After administration of CaCl₂ in lidocaine solution, sterility was achieved for at least 12 months in 75% of treated dogs. However, optimal long-term contraceptive effectiveness was achieved with CaCl₂ in alcohol, which resulted in azoospermia over the 12-month study period. Testosterone levels significantly decreased following treatment with CaCl₂, and sexual activity disappeared. Although testosterone returned to baseline levels by 12 months for the group treated with CaCl₂ in lidocaine, dogs injected with CaCl₂ in alcohol had a 63.6% drop in testosterone level, which remained at the low end of physiological range throughout the study. No adverse effects were noted.

Conclusions

A single, bilateral intratesticular injection of 20% CaCl₂ in 95% ethanol was a reliable method for induction of sterilization in 18–28 kg male dogs in this study. The approach showed long-term efficacy and reduced sexual behavior. This chemical method of sterilization might provide an effective, efficient alternative to surgical castration that can have positive impacts on dog welfare.