Cesium radioactivity in peripheral blood is linearly correlated to that in skeletal muscle: Analyses of cattle within the evacuation zone of the Fukushima Daiichi Nuclear Power Plant

The accident at the Fukushima Daiichi Nuclear Power Plant (FNPP) released a large amount of radioactive substances into the environment. Furthermore, beef contaminated with radioactive cesium above the 500 Bq/kg safety standard was circulated in the food chain in 2011. Japanese consumers remain concerned about the safety of radioactively contaminated food. In our previous study, we detected a linear correlation between radioactive cesium (¹³⁷Cs) activity in blood and muscle around 500 to 2500 Bq/kg in cattle. However, it was unclear whether the correlation was maintained at a lower radioactivity close to the current safety standard of 100 Bq/kg. In this study, we evaluated 17 cattle in the FNPP evacuation zone that had a ¹³⁷Cs blood level less than 10 Bq/kg. The results showed a linear correlation between blood ¹³⁷Cs and muscle ¹³⁷Cs (Y = 28.0X, R² = 0.590) at low radioactivity concentration, indicating that cesium radioactivity in the muscle can be estimated from blood radioactivity. This technique would be useful in detecting high‐risk cattle before they enter the market, and will contribute to food safety.     

Estimation of concentration of radionuclides in skeletal muscle from blood,based on the data from abandoned animals in Fukushima

The damage caused by the earthquake on 11 March, 2011 resulted in a serious nuclear accident in Japan. Due to the damage to the Fukushima Daiichi Nuclear Power Plant (FNPP), large amounts of radioactive substances were released into the environment. In particular, one of the largest safety concerns is radioactive cesium (¹³⁴Cs and ¹³⁷Cs). Due to the FNPP nuclear accident, a 20 km area was restricted from human activity, and various types of domestic animals were left in the zone. We collected the organs and tissues from sacrificed animals to obtain scientific data to evaluate the internal deposition of radioactive compounds. At first, we found there is a strong correlation between blood ¹³⁷Cs and organ ¹³⁷Cs with data from 44 cattle, indicating that skeletal muscle is the target organ of deposition of radioactive cesium. Second, we analyzed the relationship between blood ¹³⁷Cs and muscle ¹³⁷Cs within relatively lower radioactive concentration, suggesting that estimation of concentration of ¹³⁷Cs is possible from blood concentration of ¹³⁷Cs. Finally, we developed computer software to estimate the muscle ¹³⁷Cs concentration from blood samples. Our study contributes to the food safety of livestock products.     

Radioactive cesium and potassium in cattle living in the ‘zone in preparation for the lifting of the evacuation order’ of the Fukushima nuclear accident

To contribute to the reconstruction of livestock industry in Fukushima, radioactive cesium (¹³⁴Cs, ¹³⁷Cs) and potassium (⁴°K) were measured in various tissues of beef cattle living in an area where the evacuation order will be lifted in the near future. Radioactive cesium concentration was less than 100 Bq/kg in most of the samples. Skeletal muscles and kidney had the highest concentrations of radioactive cesium, whereas the liver was lowest among samples, excepting blood. Radioactive cesium concentration in the sirloin, tenderloin and top round was significantly higher than that in the neck muscle. Radioactive cesium concentration in the urine was not correlated with that in the blood, but the relationship became proportional when corrected with urinary ⁴°K. Distribution of ⁴°K was similar but not identical to that of radioactive cesium. These results suggest that it will be possible to resume livestock production in this area after the decontamination measures are completed and the evacuation order is lifted. Contamination level of living cattle can be estimated not only by blood samples but also by urine samples. If 50–100 Bq/kg of cesium is detected in the neck muscle at a slaughterhouse, the cattle should be reinspected using a sample from different muscles.     

A method for estimating radioactive cesium concentrations in cattle blood using urine samples

In the region contaminated by the Fukushima nuclear accident, radioactive contamination of live cattle should be checked before slaughter. In this study, we establish a precise method for estimating radioactive cesium concentrations in cattle blood using urine samples. Blood and urine samples were collected from a total of 71 cattle on two farms in the ‘difficult‐to‐return zone’. Urine ¹³⁷Cs, specific gravity, electrical conductivity, pH , sodium, potassium, calcium, and creatinine were measured and various estimation methods for blood ¹³⁷Cs were tested. The average error rate of the estimation was 54.2% without correction. Correcting for urine creatinine, specific gravity, electrical conductivity, or potassium improved the precision of the estimation. Correcting for specific gravity using the following formula gave the most precise estimate (average error rate = 16.9%): [blood ¹³⁷Cs] = [urinary ¹³⁷Cs]/([specific gravity] ? 1)/329. Urine samples are faster to measure than blood samples because urine can be obtained in larger quantities and has a higher ¹³⁷Cs concentration than blood. These advantages of urine and the estimation precision demonstrated in our study, indicate that estimation of blood ¹³⁷Cs using urine samples is a practical means of monitoring radioactive contamination in live cattle.     

Expression Patterns of Endothelial and Inducible Nitric Oxide Isoforms in the Porcine Umbilical Cord

Stable fetal–placental blood pressure and flow are extremely important in fetal growth and development. Uncontrolled and long-standing increased or decreased vascular blood pressure in the umbilical cord (UC) affects hyperaemia or ischaemia and consequently causes fetal death. Nitric oxide (NO) is one of the most active factors controlling blood flow through relaxation of the vascular smooth muscle. In this study, we investigated endothelial (eNOS) and inducible (iNOS) nitric oxide synthase expression and NADPH-diaphorase activity (NADPH-d) in the porcine UC at various stages of pregnancy. The UCs were collected from pigs on days 40, 60, 75 and 90 of pregnancy and postpartum. Western blot analysis as well as immunohistchemical staining revealed protein presence for eNOS and iNOS in the UC of the pig. The eNOS expression was maintained at a significantly higher level in all analysed days of pregnancy when compared with postnatal stage. Additionally, a significant protein increase for eNOS was observed in a periplacental part of UC on day 90. There were no obvious differences in iNOS protein level in UC samples derived from different stages of pregnancy. NADPH-diaphorase histochemical activity was correlated with NOS immunoreactivity during all analysed days of pregnancy. These results suggest that NOS isoforms are responsible for regulation of blood circulation in UC and immune responses.     

Nucleotide sequence of myostatin gene and its developmental expression in skeletal muscles of Japanese Black cattle

Myostatin, a member of the transforming growth factor‐β superfamily, is a well known negative regulator of skeletal muscle growth. In the present study, the 6660 bp nucleotide sequence of the myostatin gene in Japanese Black cattle (JBC), including the entire coding region of 1128 bp, was determined. The amino acid sequence deduced from the nucleotide sequence of JBC was well conserved with its sequence of other cattle, although it was found that an Α→G transition at nucleotide position 641 results in the substitution of asparagine by serine at amino acid position 214. In order to examine the expression pattern of the myostatin gene in the skeletal muscles of JBC, its expression in three skeletal muscles, Semitendinosus (ST) muscle, Biceps femoris muscle and Longissimus lumborum muscle, of fetal and calf stages was analyzed by real time polymerase chain reaction. The highest level of the myostatin expression was observed in the fetal stage. In calf stages the highest expression was observed in ST muscle compared with the other two muscles. These results suggest that a higher expression of myostatin gene, especially in the fetal stage and in ST muscle during calf stages, is involved in the arrest in skeletal muscle growth and that its functional domains and genomic structure in JBC are well conserved with those in other mammals.     

Comparison of urine and blood as a convenient and practical sample for estimating the contamination level of live cattle with radioactive cesium

Blood and urine were compared to clarify which is a better sample for estimating the concentration of radioactive cesium in muscles of live cattle. The most probable concentration of ¹³⁷Cs in muscles was 21.0 times that in blood, and the error of this estimation was 28%. The concentration of ¹³⁷Cs in blood was estimated accurately using urine samples. The estimation error did not increase so much (33%), even when the concentration of ¹³⁷Cs in muscles was estimated using urine samples. On the other hand, the maximum volume of blood that can be collected with one syringe is 50 ml, whereas it is easy to collect 1,000 ml of urine. It took at least 360 min to confirm that a cow meets the legal standard by inspecting 50 ml of blood. However, with 1,000 ml of urine, a 20 min measurement time was sufficient for this purpose. This difference in the required measurement time is critically important for practical use. In addition, urine can be collected by farmers themselves, whereas the blood collection requires a veterinarian. Therefore, urine is a more convenient and practical sample for estimating the contamination level of live cattle with radioactive cesium.     

生肌调节因子在绵羊不同胎儿期心肌和骨骼肌中的表达

本试验旨在探究生肌调节因子（myogenic regulatory factors，MRFs）在绵羊不同胎儿发育时期心肌和骨骼肌组织中的表达情况。选取妊娠60（E60）、70（E70）、80（E80）和90 d（E90）4个不同时期的滩羊胎儿作为研究对象，采集骨骼肌和心肌组织，通过HE和油红染色观察心肌和骨骼肌在胎儿发育过程中发生的组织形态学变化；通过实时荧光定量PCR研究绵羊胎儿发育过程中MRFs在心肌和骨骼肌组织中的表达规律以及同一胎儿期MRFs在心肌和骨骼肌中的表达差异。HE染色结果显示，心肌组织和骨骼肌组织的微观结构差异明显，心肌肌纤维密度大，纵横交错成网状，和骨骼肌相比发育的更加成熟；骨骼肌组织肌纤维密度小，间隙大，E60~E90肌纤维数量逐渐增加，肌纤维束结构清晰分明。油红染色结果显示，心肌和骨骼肌组织在4个时期均未出现被染成红色的脂滴，表明E60~E90两个组织均没有分化出脂肪。实时荧光定量PCR结果显示，E60~E90胎儿心肌组织中MYOG、Myf5和Myf6基因表达量持续降低，但是没有MYOD基因表达；E60~E90骨骼肌组织中MYOD、MYOG和Myf6基因表达量持续上升，其中MYOG基因表达水平最高，Myf5基因的表达发生周期性波动。以上结果表明，E60~E90胎儿骨骼肌正处于快速发育阶段，而心肌组织此时已大致成型。因此，骨骼肌组织中MRFs的表达量远远高于心肌组织，且MYOG基因对于维持骨骼肌的快速发育发挥了重要作用。     

Expression of Myogenic Regulatory Factors in Myocardium and Skeletal Muscle at Different Fetal Stages

The topic of this research was to investigate the expression pattern of myogenic regulatory factors (MRFs) in the myocardium and skeletal muscle of sheep at different fetal stages,the fetuses of Tan sheep at 60 (E60),70 (E70),80 (E80) and 90 days (E90) of gestation were selected as the research objects,and the myocardium and skeletal muscle were collected.The histogenesis of myocardium and skeletal muscle during fetal development were observed by HE and oil red staining.The expression patterns of MRFs in myocardium and skeletal muscle during fetal development and the expression differences of MRFs between myocardium and skeletal muscle at the same stage were tested by Real-time quantitative PCR.The results of HE staining showed that the microstructure of myocardium and skeletal muscle was very different,myocardial fibers crisscross into a network with high density,which were more mature;the density of skeletal muscle fibers were low with large gaps,the number of fibers from E60 to E90 increased gradually,and the structure fiber bundles was very clear.The results of oil red staining showed that there were no fat droplets stained red in myocardium and skeletal muscle tissues,which means that these two tissues form E60 to E90 did not have fat differentiation.The results of Real-time quantitative PCR showed that the expression of MYOG,Myf5 and Myf6 genes in myocardium decreased continuously from E60 to E90 day,but there was no MYOD gene expression.The expression of MYOD,MYOG and Myf6 genes in skeletal muscle from E60 to E90 increased gradually,among which the level of MYOG gene was the highest,and the expression of Myf5 gene fluctuated periodically.The above results indicated that the skeletal muscle was at the stage of rapid development from E60 to E90,however,the myocardial tissue had been roughly formed,which led to the expression of MRFs in skeletal muscle tissue was much higher than that in myocardial tissue,and MYOG gene played an important role in maintaining the rapid development of skeletal muscle.     

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.     

The effect of cow''s dietary copper intake, sire breed, age on her copper status and that of her fetus in the first ninety days of gestation.

The liver copper concentration of the bovine fetus and the hepatic and plasma copper concentrations of its dam during the first trimester of pregnancy were studied. The effect of the dam's dietary copper intake, age, sire breed and stage of pregnancy on her copper status and that of her fetus was also investigated. Simmental sired heifers had the lowest plasma copper concentration. The younger cows (two, three and four years) had significantly lower liver copper concentration than the older cows (six and seven years). The bovine fetal liver copper concentration was approximately 3.1 mmol/g DM and was not affected by dam's age, breed, stage of pregnancy, or copper status.     

牦牛NGF基因克隆及其在生殖器官中的表达定位

旨在探究NGF基因的生物学特性及其在母牦牛生殖器官中的表达特性.本研究收集黄体期母牦牛的心、肝、脾、肺、肾以及胎牛期、卵泡期、黄体期、妊娠期母牦牛的卵巢、子宫和输卵管(n=3),利用RT-PCR克隆牦牛NGF基因,并对其序列进行生物信息学分析,利用RT-qPCR技术分析NGF基因的组织表达特性,利用免疫组化技术(IHC...     

Serum pharmacokinetics of oxytetracycline in sheep and calves and tissue residues in sheep following a single intramuscular injection of a long-acting preparation

The pharmacokinetics of a long‐acting oxytetracycline (OTC) formulation (Liquamycin® LA‐200®) injected intramuscularly (i.m.) at a dose of 20 mg/kg were determined in four calves and 24 sheep to determine if the approved label dose for cattle provided a similar serum time/concentration profile in sheep. The AUC for the calves was 168±14.6 (μg ? h/mL) and was significantly less than the AUC for sheep (209±43 μg ? h/mL). Using the standard two‐stage approach and a one‐compartment model, the mean Cmax for the calves was 5.2±0.8 μg/mL, and for the sheep was 6.1±1.3 μg/mL. The mean terminal phase rate constants were 0.031 and 0.033 h, and the Vdss were 3.3 and 3.08 L/kg for the calves and sheep respectively. Analysis of the data using the standard two‐stage approach, the naive pooled‐data approach and a population model gave very similar results for both the cattle and sheep data. Sheep tissue residues of OTC in serum, liver, kidney, fat, muscle and injection site were measured at 1, 2, 3, 5, 7 and 14 days after a single i.m. injection of 20 mg/kg OTC. Half‐lives of OTC residues in the tissues were 38.6, 33.4, 28.6, 25.4, 21.3, and 19.9 h for injection site, kidney, muscle, liver, mesenteric fat and renal fat, respectively. The ratio of tissue to serum concentration was fairly consistent at all slaughter times, except for the fat and injection sites. The mean ratios were 1.72, 4.19, 0.11, 0.061, 0.84 and 827 for the liver, kidney, renal fat, mesenteric fat, muscle and injection sites, respectively. The tissue concentrations of OTC residues were below the established cattle tolerances for OTC in liver (6 p.p.m.), muscle (2 p.p.m.) and kidney (12 p.p.m.) by 48 h, and in injection site muscle by 14 days after the single i.m. injection of 20 mg/kg.     

中国西门塔尔牛MC3R和MC4R基因组织表达谱及其在脂肪组织中表达水平与背膘厚度的相关性研究

为探索调控肉牛脂肪沉积的关键基因表达规律,筛选影响肉牛肥育性状的候选基因,本研究采用137头18月龄肉用中国西门塔尔公牛,屠宰并采集组织样品,根据背膘厚度胴体测定值分成高、低2组,每组各选择15个个体,利用荧光实时定量技术对MC3R和MC4R基因在组织中的表达特征及其在不同个体脂肪组织中的表达水平进行分析.结果表明,M...     

Lactate dehydrogenase and its isoenzymes in the tissues and sera of clinically normal dogs

The total activity of lactate dehydrogenase (LDH) and the percentage distribution of its isoenzymes in the tissues and sera of clinically normal adult dogs are presented. Total LDH activity was greatest in skeletal muscle followed by heart muscle, kidney, small intestinal mucosa, liver, lung, pancreas and bone. Each tissue had a unique isoenzyme pattern and the proportions of the isoenzymes in serum suggested that liver is the source of normal serum LDH. The tissue isoenzyme patterns were similar to those obtained by other authors in human beings, horses, cattle, sheep and cats although in liver, differences between ruminants and monogastric animals including dogs were evident. The data presented provide a basis for the interpretation of serum LDH isoenzyme patterns in canine disease.     

Distribution of radioactive cesium and stable cesium in cattle kept on a highly contaminated area of Fukushima nuclear accident

Radioactivity inspection of slaughtered cattle is generally conducted using a portion of the neck muscle; however, there is limited information about the distribution of radioactive cesium in cattle. In this study, therefore, we measured not only radioactive cesium but also stable cesium in various tissues of 19 cattle that had been kept in the area highly contaminated by the Fukushima nuclear accident. Skeletal muscles showed approximately 1.5‐3.0 times higher concentration of radioactive cesium than internal organs. Radioactive cesium concentration in the tenderloin and top round was about 1.2 times as high as that in the neck muscle. The kidney showed the highest concentration of radioactive cesium among internal organs, whereas the liver was lowest. Radioactive cesium concentration in the blood was about 8% of that in the neck muscle. Characteristics of stable cesium distribution were almost the same as those of radioactive cesium. Correlation coefficient between radioactive cesium and stable cesium in tissues of individual cattle was 0.981 ± 0.012. When a suspicious level near 100 Bq/kg is detected in the neck of slaughtered cattle, re‐inspection should be conducted using a different region of muscle, for example top round, to prevent marketing of beef that violates the Food Sanitation Act.     

Development of muscle spindles in the human tongue

The presence, location and morphology of muscle spindles in the tongue of human fetuses and newborns was examined in frontal and sagittal serial sections of the organ or of the whole lower half of the mouth. While in the fetal material of 105, 155 and 170 mrn. C. R. L. there are only very few and small spindles in some extrinsic muscles of the tongue and in the Mm. mylohyoideus and geniohyoideus, the specimens of 260, 310 and 390mm. C. R. L. showed them in the intrinsic muscles as well as in the extrinsic ones, excludicg only the region of the tip of the tongue. In the latter group, the size of the whole spindles is bigger than in the small fetuses, and their number increases towards the root and the midline area of the tongue. In the newborns, there is the same situation in general, with an especially high number of spindles in the Mm. transversus linguae and genioglossus. The size of the intrafusal muscle fibers was of 6.33 ± 0.72µm. in the newborns, and of 4.93 ± 1.16 pm. in the fetal material; the sheat (capsule) is thinner and the periaxi.11 space is smaller in the minor specimens. As it was possible to find spindles in the muscles of the floor of the mouth in a fetus of the 4th lunar month, there seems to be evidence of the earlier appearance of sum proprioceptors in this region than in other parts of the body, an observation which might implicate some functional significance.     

Serum biochemical values and mineral element contents of tissues in yaks

Serum biochemical values and the wet weight to dry weight ratios of tissues were determined in yaks in Shandan county of Gansu province. The liver, kidney, heart and muscle contents of seven elements in yaks were also analysed. Most serum biochemical values were similar to those of cattle, camels and sheep, but the calcium concentration was considerably above the normal range for other ruminants. The liver contained the highest concentrations of copper, manganese and cobalt and the kidney of selenium, iron and calcium.Abbreviations Alb albumin - ALT alanine aminotransferase - AST aspartate aminotransferase - BUN blood urea nitrogen - Chol cholesterol - Glob globulin - -GT -glutamyl transpeptidase - IP inorganic phosphorus - TP total protein     

Activity and properties of adenosine triphosphatase in erythrocytes and various tissues (brain, liver, kidney, small intestine mucosa) of cattle]

In 15 samples of haemolysate of bovine erythrocytes, the splitting of phosphate from adenosine triphosphate average 158 +/- 63 X 10(-3) muMol/min/g haemolysate haemoglobin. Estimation of total adenosine triphosphatase in homogenates of various organs from cattle showed that spleen, liver, kidney and brain possessed high activity, while the activity was moderate in lung, myocardium and skeletal muscle, and low in endometrial mucosa and spinal cord. There was a relatively large proportion of Na-K-adenosine triphosphatase in brain and kidney. In various organs the activity of the enzyme was dependent upon the concentrations of Mg, Na, K and Ca. The inhibition of adenosine triphosphatase in various tissues by ouabain was studied. The optimum pH for the enzyme lay in the weakly alkaline region.     

Fructose utilization for nucleic acid synthesis in the fetal pig

Eight fetal pigs, in utero, were injected ip with 20 microCi/fetus [U14C]-fructose between d 55 and 65 pregnancy. The isotope was allowed to equilibrate between blood and tissues within injected fetuses for a period of 240 min. Fetal pigs were then sacrificed and nucleic acids were extracted from cold tissue homogenates of skeletal muscle and liver. Nuclide disintegrations per minute recovered in extracted DNA and RNA were used to calculate incorporation of labeled C from fructose. The recovery of labeled C per mmol of nucleic acids from skeletal muscle was greater (P less than .05) than that from liver. Relative incorporation of labeled C into skeletal muscle RNA (395.9 pmol/mmol) was greater (P less than .05) than for DNA (189.5 pmol/mmol). The same trend was observed for liver RNA (78.0 pmol/mmol) and DNA (55.6 pmol/mmol), but differences were nonsignificant. These data suggest that at least part of the high concentration of endogenous fructose measured in fetal pigs in utero is involved in synthesis of nucleic acids, thereby providing substrate for anabolic functions necessary for fetal growth and development.