Familial Congenital Methemoglobinemia in Pomeranian Dogs Caused by a Missense Variant in the NADH‐Cytochrome B5 Reductase Gene

Background

In veterinary medicine, congenital methemoglobinemia associated with nicotinamide adenine dinucleotide (NADH)‐cytochrome b5 reductase (b5R) deficiency is rare. It has been reported in several breeds of dogs, but little information is available about its etiology.

Objectives

To analyze the NADH‐cytochrome b5 reductase gene, CYB5R3, in a Pomeranian dog family with methemoglobinemia suspected to be caused by congenital b5R deficiency.

Animals

Three Pomeranian dogs from a family with methemoglobinemia were analyzed. Five healthy beagles and 5 nonrelated Pomeranian dogs without methemoglobinemia were used as controls.

Methods

Methemoglobin concentration, b5R activity, and reduced glutathione (GSH) concentration were measured, and a turbidity index was used to evaluate Heinz body formation. The CYB5R3 genes of the affected dog and healthy dogs were analyzed by direct sequencing.

Results

Methemoglobin concentrations in erythrocytes of the affected dogs were remarkably higher than those of the control dogs. The b5R activity of the affected dogs was notably lower than that of the control dogs. DNA sequencing indicated that this Pomeranian family carried a CYB5R3 gene missense variant (ATC→CTC at codon 194) that resulted in the replacement of isoleucine (Ile) by leucine (Leu).

Conclusions and Clinical Importance

This dog family had familial congenital methemoglobinemia caused by b5R deficiency, which resulted from a nonsynonymous variant in the CYB5R3 gene. This variation (c.580A>C) led to an amino acid substitution (p.Ile194Leu), and Ile194 was located in the proximal region of the NADH‐binding motif. Our data suggested that this variant in the canine CYB5R3 gene would affect function of the b5R in erythrocytes.     

Interaction between the sequence of feeding of hay and concentrate,and boiling of barley on feed intake,the activity of hydrolytic enzymes and fermentation in the hindgut of Arabian mares

The interaction between the sequence of feeding of hay and concentrate and the hydrothermal processing of barley in alleviating concentrate effects on intake, and hindgut fermentation in horses was tested. Six Arabian mares (4–10 years of age, 410 ± 35 kg body weight) were used to evaluate the effects of feeding sequence (FS) and type of barley (TB) on intake, and faecal volatile fatty acids (VFA), activities of α‐amylase (AA: EC 3.2.1.1), carboxymethyl cellulase (CMCase: EC 3.2.1.4), microcrystalline cellulase (MCCase: EC 3.2.1.91) and general filter paper degrading activity (FPD). Mares were offered a ration of air‐dried alfalfa and concentrate (70:30 as‐fed) in four subsequent periods of 14 days including 8 days of adaptation and 6 days of sampling. In each period and each meal, mares received concentrate either 30 min after (HC) or 30 min before (CH) alfalfa hay. Barley was either milled or boiled in water. Rectal samples were grabbed directly from rectum once per period. Mares subjected to CH had higher dry matter intakes than mares under HC regime. The acetate:propionate ratio (A:P ratio) in rectal content was higher with CH than HC. The AA activity was higher under CH than under HC. Mares fed boiled barley had lower rectal concentrations of VFA and propionate and a higher A:P ratio than mares fed milled barley. Furthermore, the rectal content showed a higher MCCase activity but a lower AA activity when mares were fed boiled compared with milled barley. Interactions between FS and TB were observed with respect to CMCase activity, and concentrations of propionate and valerate. In conclusion, the present results suggest that both, feeding concentrate before hay and boiling the barley, might improve the hindgut environment in Arabian mares, and that the two measures were mostly additive and sometimes even synergistic.     

Disaccharide combinations and the expression of enolase3 and plasma membrane Ca2+ ATPase isoform in sturgeon sperm cryopreservation

Acipenser sinensis and Acipenser dabryanus are critically endangered species, so germplasm conservation via cryopreservation of sperm is necessary. Disaccharides can act as membrane‐impermeable cryoprotectants, and enolase3 (ENO3) and plasma membrane Ca²⁺ ATPase isoform (PMCA2) are proteins associated with sperm quality. We considered seven characteristics of sperm quality in cultured brood stock from A. sinensis and A. dabryanus. We tested use of sucrose or trehalose alone and in combination at different concentrations for cryopreservation of A. dabryanus sperm. A low concentration of sucrose plus trehalose (S₁₅T₁₅) was optimal. Mixing of the extender with sucrose, lactose, or trehalose alone or with pairwise mixtures revealed that a mixture of lactose and trehalose (L₁₅T₁₅) gave the best results for both A. sinensis and A. dabryanus. Enolase3 and PMCA2 expression levels were measured in cryopreserved A. sinensis sperm via Western blotting. Relative ENO3 and PMCA2 expression levels were examined, and the relationship between disaccharide composition, sperm quality and protein expression was explored in A. sinensis. The results showed that relative ENO3 and PMCA2 expression levels were the highest at L₁₅T₁₅ in cryopreserved A. sinensis sperm. There were significant positive correlations between ENO3 expression and percentage membrane integrity, and between PMCA2 expression and sperm motility parameters (percentage of motile sperm, curvilinear velocity, straight‐line velocity and average path velocity; p < .05) in cryopreserved A. sinensis sperm. Our results indicate the optimal disaccharide combination and concentrations for cryopreservation of A. sinensis and A. dabryanus sperm and suggest that ENO3 and PMCA2 expression levels could serve as a valuable indicator of sperm quality in A. sinensis.     

南果梨周年干物质与氮磷钾积累动态

【目的】明确南果梨干物质积累特征和氮磷钾养分周年动态积累规律,为南果梨优化施肥量和施肥时期提供依据。【方法】以12年生南果梨树为试材,采用田间采样和树体分解方法,分别于萌芽后10 d(萌芽期)、 30 d(花期)、 65 d(幼果膨大期)、 100 d(果实膨大或新稍停止生长期)、 130 d(果实着色前)、 155 d(果实采收期)、 185 d(采收后)、 210 d(落叶前)8个生育期,选干周和树高一致的3株树,将树体连根从土壤中挖出,分出果实、 叶片、 枝条、 主干、 主根、 侧根、 须根,各器官单独称重,并取200 g左右的鲜样按清水、 洗涤剂、 清水、 1%盐酸、 3次去离子水冲洗、 杀青、 烘干后,电磨粉碎过0.15 mm筛,测定样品中氮、 磷、 钾含量。【结果】1)南果梨周年生育期内,树体干物质当年净积累量为18.4 kg/plant,干物质累积速率出现两次累积高峰,分别是幼果膨大期(0.15 kg/d)和采收期(0.11 kg/d)。2)南果梨树体总氮周年积累量为154.0~301.0 g,新生器官为0~116.2 g,果实膨大期达到最高;多年生器官氮积累量为154.0~194.8 g,落叶前达到最高。3)南果梨树体总磷周年积累量为17.1~37.2 g,果实着色前最高。其中新生器官为13.7 g,果实采收期最高;多年生器官为17.1~24.9 g,果实转色期最高。4)南果梨树体总钾周年积累量为27.9~174 g。新生器官钾为97.3 g,采收期最高;多年生器官钾为27.6~76.6 g,落叶前最高。5)产量大约为17 t/hm2的12年生南果梨从萌芽到落叶前树体当年氮磷钾的单株净累积量分别为146.2、 20.1、 146.1 g,折合1000 kg果实经济产量需吸收氮(N)、 磷(P)、 钾(K)5.4、 0.7、 5.4 kg。【结论】南果梨周年干物质单株积累总量为41.4 kg,当年净积累量为19.7 kg。南果梨干物质积累主要集中在花期到果实膨大期和果实转色到落叶前,分别占47.3% 和47.5%。南果梨从萌芽到落叶前氮、 磷、 钾的单株净累积量分别为146.2、 20.1、 146.1 g,每1000 kg果实经济产量需吸收氮(N)、 磷(P)、 钾(K)5.4、 0.7、 5.4 kg。从开花到果实膨大期和从果实着色到采收后30天对氮吸收分别占总氮累积量的39.0%和49.0%,而磷、 钾的累积从萌芽到开花较快,到果实膨大期磷的累积达67.4%,钾的累积达65.1%,果实膨大期是干物质和氮磷钾积累的关键时期。     

三维CAD技术在机械设计中的应用

现代计算机在技术和性能上越发成熟,再加之网络的普及和信息的智能化处理技术的提高,三维CAD技术本身也正逐步成熟,因此,正确地把握CAD技术的发展趋势对我国机械工程中的设计工作具有深远的意义.     

农五师葡萄钾素动态变化及调节措施研究

通过叶分析研究了不同葡萄品种叶柄全钾5～9月含量动态变化规律.结果表明:葡萄叶柄钾含量在生育期内整体呈波浪式下降趋势,前期下降迅速,中期平缓,进入浆果成熟期后又趋于下降；强势品种在整个生长季节内对钾的吸收大于弱势品种；早熟品种叶柄内钾含量变化趋势比晚熟品种提前.根据新疆农五师葡萄园区土壤有效钾含量分布现状,建议生产中应根据葡萄钾元素动态变化特点,不同团场应结合当地土壤有效钾含量相应补充.     

基于“3S”技术的辽宁省某尾矿库扩容工程生态影响预测与评价

以辽宁省某尾矿库为例,基于GIS技术,利用2010年的TM遥感影像及地形图,预测该尾矿库扩容后的生态影响。分析结果显示:尾矿库扩容工程对生态的影响主要体现在土地利用、植被、生态景观和地形地貌的变化4个方面。     

库车坳陷克深地区现今地应力场及储层裂缝数值模拟

克深三维区目前处于勘探开发的前期,需要明确地应力场及地下储层裂缝的分布规律,指导开发工作,但该区存在地层埋深大、取心困难、成像测井资料缺乏等实际情况,应力场研究及裂缝预测难度大。利用有限元分析方法,以井点现今地应力值为约束进行了克深三维区现今地应力场的数值模拟,同时建立了应力应变与裂缝参数的定量关系模型,利用古今应力场结合法对克深三维区的裂缝参数进行了预测,模拟结果显示构造高部位和断层带是低应力值区及裂缝的有利发育区。裂缝的数值模拟结果与测井解释结果对比表明,预测结果与实钻吻合度较高,裂缝预测效果较好。     

类器官的研究进展及应用前景

类器官（organoids）来源于自组织和自我更新的干细胞，是利用干细胞的自组织特性进行体外3D培养后形成的细胞团，与来源器官密切相关，再现了来源器官的三维细胞结构，并为探索来源器官的发病机制提供了新的模型。类器官系统是由自分泌、旁分泌或邻分泌信号调节下的细胞，或者外源性添加的细胞外基质（extracellular matrix，ECM）底物、小分子和生长因子等衍生而来，这些因素的相互作用创造了一个动态的环境，指导干细胞的自我更新和分化，以及细胞在类器官中的自我组装。诱导多能干细胞（induced pluripotent stem cells，iPSC）重编程方法结合3D类器官工具，使患者来源的类器官作为动物模型和人类临床试验之间的桥梁，是对细胞研究和在体试验的补充。在研究来源器官发育、生物学和病理生理学方面，类器官不仅是一种比传统细胞培养更具生理相关性的体外模型，而且还是再生医学和个性化医学领域中的新模型，有望成为研究营养素、药物、毒物及毒素等的作用机制及药物的筛选、再生医学等领域的重要模型。总之，类器官技术的发展增强了人们对器官和组织生理生化功能的认知。作者对肠、脑、肺脏、肝脏、子宫、卵巢等类器官培养和应用的研究进展进行综述，以期为类器官相关科研及应用提供参考。