The distribution of copper, vitamin B12 and zinc in the livers of sheep

Six livers from one-year-old sheep were each sampled at 27 different sites and the concentrations of copper, vitamin B12 and zinc determined for each sample. The distribution of these components was uniform throughout the liver, indicating that analysis of a single liver biopsy sample would give an accurate assessment of their hepatic concentration.     

Quantitative ultrasound, magnetic resonance imaging, and histologic image analysis of hepatic iron accumulation in pigeons (Columbia livia).

Iron overload was induced by iron dextran i.v. in clinically healthy adult pigeons, Columbia livia, (n = 8). Hemosiderosis was induced in all treated birds. Two control pigeons received no iron injections. Pigeons did not show clinical signs of iron overload during the 6-wk study. Ultrasound examination of the liver in the pigeons receiving iron dextran was performed on days 0, 13, 28, and 42. No ultrasound images were collected on the control pigeons. Magnetic resonance imaging was performed on days 0, 13, 28, and 42 on all study pigeons and imaging sequences were collected in three different imaging formats: T1, T2, and gradient-recalled echo (GRE). Surgical liver biopsies were performed on pigeons receiving iron dextran on days 2, 16, and 45 (at necropsy). A single liver sample was collected at necropsy from the control birds. Histologic examination, quantitative image analysis, and tissue iron analysis by thin-layer chromatography were performed on each liver sample and compared to the imaging studies. Although hemosiderosis was confirmed histologically in each experimental pigeon, no significant change in pixel intensity of the ultrasound images was seen at any point in the study. Signal intensity, in all magnetic resonance imaging formats, significantly decreased in a linear fashion as the accumulation of iron increased.     

A Comparison of Liver Sampling Techniques in Dogs

Background

The liver sampling technique in dogs that consistently provides samples adequate for accurate histopathologic interpretation is not known.

Hypothesis/Objectives

To compare histopathologic results of liver samples obtained by punch, cup, and 14 gauge needle to large wedge samples collected at necropsy.

Animals

Seventy dogs undergoing necropsy.

Methods

Prospective study. Liver specimens were obtained from the left lateral liver lobe with an 8 mm punch, a 5 mm cup, and a 14 gauge needle. After sample acquisition, two larger tissue samples were collected near the center of the left lateral lobe to be used as a histologic standard for comparison. Histopathologic features and numbers of portal triads in each sample were recorded.

Results

The mean number of portal triads obtained by each sampling method were 2.9 in needle samples, 3.4 in cup samples, 12 in punch samples, and 30.7 in the necropsy samples. The diagnoses in 66% of needle samples, 60% of cup samples, and 69% of punch samples were in agreement with the necropsy samples, and these proportions were not significantly different from each other. The corresponding kappa coefficients were 0.59 for needle biopsies, 0.52 for cup biopsies, and 0.62 for punch biopsies.

Conclusion and Clinical Importance

The histopathologic interpretation of a liver sample in the dog is unlikely to vary if the liver biopsy specimen contains at least 3–12 portal triads. However, in comparison large necropsy samples, the accuracy of all tested methods was relatively low.     

维生素A过量对肉鸡早期骨骼钙磷代谢的影响

试验通过研究维生素A过量对肉鸡早期骨形成生化指标及组织中碱性磷酸酶（ALP）和骨钙素（BGP）基因表达的影响,旨在为深入研究日粮维生素A过量对肉鸡骨骼钙磷代谢产生负面影响的机理提供理论基础。采用完全随机试验设计,将450只1日龄健康爱拔益加（AA）肉鸡（公母各半）称重后随机等分为5个处理组,每个处理6个重复,每个重复15只鸡。各处理组日粮分别在基础日粮中添加3 000、6 000、15 000、30 000和60 000 IU/kg维生素A。试验期28 d,分别于试验第14、28天从每个重复中随机抽取1只鸡,翅静脉采血制备血清后随即宰杀,在冰浴上迅速分离右侧肝脏、十二指肠、空肠、回肠、胫骨组织,-80℃保存备用。试验结果表明,随日粮维生素A添加剂量的升高,肉鸡血清BGP浓度、ALP活性和骨特异性碱性磷酸酶（BAP）活性及胫骨、肝脏及小肠组织中ALP、BGP mRNA相对表达量均呈显著（P<0.05）或趋于显著的一次线性或一元二次（P<0.10）降低,通过回归分析得出：维生素A水平在42 766~47 185 IU/kg时,肉鸡14 d胫骨和肝脏ALP mRNA相对表达量、肝脏BGP mRNA相对表达量,28 d的血清ALP活性及肝脏ALP mRNA相对表达量、胫骨和回肠的BGP mRNA相对表达量最低。综合试验结果,维生素A线性降低了肉鸡早期血清BAP活性;日粮维生素A以剂量依赖模式影响肉鸡早期血清BGP浓度、ALP活性及其在组织中的基因表达,42 766~47 185 IU/kg维生素A对上述指标的降低作用较大,提示日粮维生素A过量可抑制肉鸡早期骨形成相关生化指标的活性,并下调胫骨、肝脏及小肠组织中ALP、BGP mRNA的相对表达量。     

Depletion of gentamicin and its major components from various tissues of turkeys

OBJECTIVE: To determine tissue depletion profiles for gentamicin and its 3 major components (C1, C1a, and C2) in turkeys. ANIMALS: Twenty 10-week-old male turkeys. PROCEDURE: birds were maintained as untreated controls. The remaining birds were treated with gentamicin sulfate at a dosage of 2 mg/kg, IM, once daily for 5 days. Treated birds were euthanatized 45, 60, 75, and 90 days (4 birds at each sample time) after the last dose of gentamicin was administered, and samples of muscle, liver, kidney, and skin and fat were collected. Control birds were euthanatized on day 45. Concentrations of the 3 major components of gentamicin were measured by means of reversed-phase high-performance liquid chromatography. RESULTS: Total gentamicin concentration (ie, sum of the concentrations of the 3 major components) was < 100 microg/kg for all muscle and skin and fat samples by day 45 and all liver samples by day 75. At all sample times, concentration of the gentamicin C1 component was higher than concentrations of the C1a and C2 components in all tissues. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that tissue depletion profiles of the 3 major components of gentamicin differ from each other. Withdrawal time, therefore, may depend on the ratio of the components in the pharmaceutical preparation used.     

Radioimmunoassay of the anabolic agent zeranol. IV. The determination of zeranol concentrations in the edible tissues of cattle implanted with zeranol (Ralgro)

Rapid solvent extraction combined with a radioimmunoassay using a monoclonal antibody raised against a derivative of zeranol has been used to measure the residues of the anabolic agent zeranol in the edible tissues (muscle, liver, kidney and fat) of cattle treated with Ralgro. Calibration curves, both with and without, tissue extracts exhibit good parallelism. Regression analysis for the extraction of zeranol from tissues dosed with standard amounts of zeranol have correlation coefficients of 0.979, 0.991, 0.986 and 0.985 for muscle, liver, kidney and fat, respectively. The limits of decision defined as the mean value + 3 SD for the concentrations apparently observed (noise) in tissues from animals not treated with Ralgro were 278, 121, 373 and 110 ng/kg for muscle, fat, liver and kidney, respectively. In the tissues of 4 cows implanted with Ralgro (36 mg), and sampled 70 days after implanting, the highest concentration of zeranol in each tissue was 232 ng/kg (muscle), 391 ng/kg (liver), 287 ng/kg (kidney) and 293 ng/kg (fat), and residues were detected in all samples of fat (4), 3 kidney samples and 1 liver sample.     

Effect of various dietary fat supplementations on liver lipid and glycogen of high-yielding dairy cows in the peripartal period

In a model experiment, Holstein-Friesian dairy cows were fed on a corn-silage-based diet supplemented with 11.75 MJ NE1 per day of calcium soaps of palm oil fatty acids (CAS) or hydrogenated triglyceride (HTG) or without fat supplementation (control). All diets were fed to the cows over a period from 21 +/- 3 days (d) prior to the expected calving to d 100 +/- 5 postpartum. On d 25 (basal sample) and d 14 prepartum as well as on d 5 and 25 postpartum liver samples were collected by percutaneous biopsy. Total lipid content, fatty acid composition and glycogen of liver tissues were determined. At d 5 postpartum, both control and CAS cows had higher liver lipid (P < 0.05) and lower glycogen (P < 0.05) concentrations than cows in the HTG group. No significant (P < 0.05) differences were detected in liver fat content among the groups at d 14 prepartum or d 25 postpartum. The glycogen concentration slightly decreased in the liver of cows in each treatment group from d 14 prepartum to d 5 postpartum; however, this decrease was more intensive in both the control and CAS groups than in the HTG group. The variations in liver lipid concentrations were accompanied by significant changes in the proportion of C16:0, C16:1n-7, C18:0, C18:1n-9, C18:2n-6 and C20:4n-6 fatty acids in the liver lipids. The results show that HTG supplementation exerted more advantageous effects on liver lipid and glycogen metabolism than did CAS supplementation.     

Isolation and multiresidue detection of macrolide endectocides present in animal matrices.

A simple, rapid, and sensitive method for isolation and detection of macrolide endectocides (moxidectin, doramectin, selamectin, ivermectin, and eprinomectin) in animal sera and liver is described. Fortified sera or homogenized liver samples were treated with sodium chloride followed by organic solvent extraction. No additional steps were required prior to analysis. Separation of analytes and collection of mass information was achieved by liquid chromatography/mass spectrometry with positive atmospheric pressure chemical ionization set in selected ion monitoring mode with each sample analysis complete in 15 minutes. Presence of each compound was confirmed based on 2 separate extracted ion profiles. Detection of avermectins and moxidectin in a range of working standards was achieved at 10, 50, and 100 ppb. Quantitation of these compounds in fortified samples was based on standard calibration curves with R2 > 0.99. Detection limits of 10 ppb for ivermectin, moxidectin, and doramectin, 50 ppb for selamectin, and 100 ppb for eprinomectin were achieved in spiked sera. Recoveries of avermectins and moxidectin in 500 ppb fortified sera fell between 61 and 89% (+/-5.7-15.7). Analysis of fortified liver gave comparable results with recovery of selamectin of 83-91% +/- 18.3. A complete mass spectral fragmentation pattern of selamectin and affordable screening method for 6 macrolide endectocides are reported. Method comparison for salt treatment and solid-phase extraction of fortified samples is discussed.     

The effect of storage conditions on samples for the evaluation of copper status in blesbok (Damaliscus pygargus phillipsi)

Investigaltions to determine the effect of sample storage on the concentration of copper in liver tissue and on the activity of erythrocyte superoxide dismutase were undertaken in preparation for a study of blesbok (Damaliscus pygargus phillipsi) that were suspected to be suffering from copper deficiency. Two liver samples were collected from each of 20 culled blesbok in a manner that simulated the collection of biopsies from the live animal. These samples were stored either in 10% formalin or frozen at -20 degrees C until analysed 4 1/2 months later. The effect of different methods of sample storage on superoxide dismutase activity was determined. Erythrocytes collected from 3 Jersey cows and 5 culled blesbok were washed and divided into 0.5 ml portions, stored at room temperature (approximately 20 degrees C), in a refrigerator (4 degrees C), frozen at -20 degrees C in a freezer, and in liquid nitrogen (-200 degrees C). An analysis of superoxide dismutase activity was undertaken using a commercial assay kit at intervals of 2-4 days until the levels of activity had fallen significantly. The copper concentration in formalin-preserved liver samples was significantly lower than that measured in frozen liver tissue apparently as a result of leaching. The activity of superoxide dismutase in cattle blood was unchanged for 4 days at room temperature but fell appreciably after 2 days at 4 degrees C and -20 degrees C. Enzyme activity remained unchanged for 200 days in erythrocytes stored in liquid nitrogen. Superoxide dismutase activity levels in healthy blesbok were considerably lower than those measured in Jersey cows and remained unaffected for up to 6 days in samples stored at 4 degrees C and 20 degrees C. The level of activity fell significantly thereafter. Samples stored in liquid nitrogen were unchanged after 40 days.     

Histopathologic Variation between Liver Lobes in Dogs

Background

Biopsy of the liver evaluates a small portion of tissue, with inferences made to the entire organ. The method and number of biopsies obtained are tempered by consideration of the risks and benefits. Recommendations often include biopsy of more than one liver lobe, although the consistency of histopathology among lobes in dogs is unknown.

Hypothesis/Objectives

To describe the distribution of histopathologic abnormalities between liver lobes. We hypothesized that discordant results would be evenly distributed among all liver lobes.

Animals

Seventy dogs undergoing necropsy.

Methods

Prospective study. Liver samples were obtained from all lobes. A primary diagnosis was assigned to each liver sample based on the predominant histopathologic abnormality.

Results

In this population of dogs, biopsy of at least 2 liver lobes identified the predominant histologic abnormality in 98.6% of the cases. Ten (14%) of the dogs had ≤3 lobes in agreement and could not be assigned a predominant diagnosis. The same diagnosis was present in 6/6 lobes in 39 (56.5%) dogs, 5/6 lobes in 10 (14.5%) dogs, 4/6 lobes in 10 (14.5%) dogs, 3/6 lobes in 7 (10.1%) dogs, and 2/6 in 3 (4.3%) dogs. The number of discordant results did not differ between the liver lobes.

Conclusion and Clinical Importance

The likelihood of obtaining a sample that is reflective of the predominant histologic abnormality in the liver is increased when multiple liver lobes are biopsied.     

QuEChERS-UPLC-MS/MS快速测定鸡肝中七种磺胺类药物残留

建立了一种通过QuEChERS方法和超高效液相色谱-串联质谱法快速测定鸡肝中七种磺胺类药物残留量的方法。样品以磺胺二甲氧嘧啶-D6为内标,经DisQuE萃取管提取,DisQuE净化管净化,浓缩后经超高效液相色谱分离,串联质谱测定。结果表明在12.5～125 ng/mL范围内各磺胺类药物均呈良好的线性关系,高中低浓度回收率在70%～115%,精密度小于15%。该方法快速、简便,适于鸡肝中七种磺胺类药物的批量测定。     

蓝塘仔猪IGF-1水平与组织IGF-1、GHR基因的表达

32头不同日龄(出生、3、21、35d)蓝塘仔猪,由前腔静脉采血后剖杀,取肝脏、背最长肌样品。用RIA法测血液、组织中IGF 1浓度,用放射受体法(RBA)检测肝脏、肌肉组织中GHR结合活性,用实时荧光定量PCR法检测IGF 1、GHRmRNA的表达水平。结果表明:(1)血液中IGF 1在出生日显著高于其它时期(P<0 05)。肌肉组织中IGF 1含量高于肝脏组织,肌肉组织IGF 1含量在3、21、35日龄时都显著高于出生日(P<0 05)。(2)肝脏细胞膜GHR结合活性在出生日显著高于3、21日龄(P<0 05),肝脏细胞膜GHR结合活性高于肌肉组织。(3)肝脏组织IGF 1、GHRmRNA的表达量均显著高于肌肉组织(P<0 05)。肝脏IGF 1mRNA的表达在出生日、21日龄时显著高于3、35日龄(P<0 05),GHRmRNA的表达在出生日显著高于其它日龄(P<0 05)。肌肉IGF 1、GHRmRNA的表达在出生日均显著高于其它日龄(P<0 05)。     

Comparison of selenium determination in liver samples by atomic absorption spectroscopy and inductively coupled plasma-mass spectrometry.

Selenium (Se) is an essential trace element that is often deficient in the natural diets of domestic animal species. The measurement of Se in whole blood or liver is the most accurate way to assess Se status for diagnostic purposes. This study was conducted to compare hydride generation atomic absorption spectroscopy (HG-AAS) with inductively coupled plasma-mass spectrometry (ICP-MS) for the detection and quantification of Se in liver samples. Sample digestion was accomplished with magnesium nitrate and nitric acid for HG-AAS and ICP-MS, respectively. The ICP-MS detection was optimized for 82Se with yttrium used as the internal standard and resulted in a method detection limit of 0.12 microg/g. Selenium was quantified by both methods in 310 samples from a variety of species that were submitted to the Toxicology Laboratory at New Bolton Center (Kennett Square, PA) for routine diagnostic testing. Paired measurements for each sample were evaluated by a mean difference plot method. Limits of agreement were used to describe the maximum differences likely to occur between the 2 methods. Results suggest that under the specified conditions ICP-MS can be reliably used in place of AAS for quantitation of tissue Se at or below 2 microg/g to differentiate between adequate and deficient liver Se concentrations.     

Evaluation of hemorrhage, sample size, and collateral damage for five hepatic biopsy methods in dogs

OBJECTIVES: To compare the volume of hemorrhage, number of lobules, and portal triads available for histologic evaluation, and resultant collateral damage between 5 hepatic biopsy methods: biopsy punch, biopsy needle, ligature method, laparoscopic biopsy forceps, and ultrasonically activated scalpel (UAS). STUDY DESIGN: Experimental, repeated measures, block. ANIMALS: Twelve adult dogs. METHODS: Biopsies were obtained from the periphery and center of the left lateral liver lobe of each dog using each of 5 biopsy techniques. Hemorrhage was quantified and compared between methods and sites. Biopsy samples were evaluated histologically to characterize collateral damage and determine the number of lobules and portal triads sampled. RESULTS: Regardless of technique, liver biopsy resulted in minimal hemorrhage (<2 mL). For peripheral biopsies, UAS was comparable with the ligature method, but caused significantly less hemorrhage than all other methods, whereas for central biopsies, UAS caused significantly less hemorrhage than other methods. Except for the laparoscopic biopsy forceps, UAS caused significantly more collateral damage than other methods. UAS and ligature biopsy methods yielded specimens that had more portal triads per sample than other methods. Eight of 48 biopsy needle samples were inadequate for histologic evaluation, whereas other methods yielded adequate specimens. CONCLUSIONS: All biopsy methods produced minimal hemorrhage and except for needle biopsy yielded adequate tissue samples for histologic evaluation. CLINICAL RELEVANCE: Use of UAS is a reliable, safe alternative technique for liver biopsy and can be used laparoscopically to obtain large tissue samples.     

Distribution of iron and copper in the liver and spleen of veal calves in relation to the concentration of iron in the diet

The distribution of iron and copper in the liver and spleen of veal calves was determined in a group of calves fattened with a commercial milk replacer supplemented with 5 ppm iron and in a control group fed the unsupplemented milk replacer. No marked variations were detected in the distribution of iron and copper. A liver biopsy is therefore likely to be a representative sample of the whole liver with respect to iron and copper concentration.     

Arginase-1 and P-glycoprotein are downregulated in canine hepatocellular carcinoma

BackgroundHepatocellular carcinoma is the most common primary hepatic malignancy in humans and dogs. Several differentially expressed molecules have been studied and reported in human hepatocellular carcinoma and non-neoplastic liver lesions. However, studies on the features of canine hepatocellular carcinoma are limited, especially related to the differential characteristics of neoplastic and non-neoplastic lesions.ObjectivesThe study''s objective was 1) to examine and evaluate the expression of arginase-1, P-glycoprotein, and cytokeratin 19 in canine liver tissues and 2) to investigate the differential features of hepatocellular carcinomas, liver tissue with non-neoplastic lesions, and paracancerous liver tissues in dogs.MethodsThe expression levels of three markers underwent immunohistochemical analysis in 40 non-neoplastic liver tissues, 32 hepatocellular carcinoma tissues, and 11 paracancerous liver tissues. Scoring of each marker was performed semi-quantitatively.ResultsArginase-1 and P-glycoprotein were significantly downregulated in hepatocellular carcinoma, compared with hepatic tissues with non-neoplastic diseases (p < 0.001). Expression levels of arginase-1 and P-glycoprotein were also significantly lower in hepatocellular carcinoma than in paracancerous liver tissues (arginase-1, p = 0.0195; P-glycoprotein, p = 0.047). Few cytokeratin 19-positive hepatocytes were detected and only in one hepatocellular carcinoma and one cirrhotic liver sample.ConclusionsThe results of this study suggest that downregulation of arginase-1 and P-glycoprotein is a feature of canine hepatocellular carcinoma; thus, those markers are potential candidates for use in differentiating hepatocellular carcinomas from non-neoplastic liver lesions in dogs.     

乙酰甲喹及其主要代谢物在鸡体内的残留消除规律研究

建立了高效液相色谱串联质谱（HPLC—MS／MS）法用于乙酰甲喹及其6种主要代谢物的检测。将65只健康白羽鸡分为两组,10只为空白对照组,其余为试验组。试验组施以20mg／kgb．w．灌胃乙酰甲喹悬浊液,1d2次,连续3d。在给药结束后2、4、6、12、16、24、30、36、48、72、120h分别宰杀5只鸡,采集血液和可食性组织（肌肉、肝脏、肾脏和皮脂）样品。结果表明：乙酰甲喹原药在组织和血浆中迅速消除;乙酰甲喹的代谢物广泛存在于鸡的可食性组织和血浆中;代谢物的残留消除过程较为复杂,代谢物在肌肉和肝脏中残留量较多,在皮脂中残留时间最长。研究结果将有助干榍示7．酷甲喹存鸡体内的砖留消除期,律．