Response of whole blood, erythrocyte and plasma vitamin E content to dietary vitamin E intake in the chick

Whole blood, red blood cells (RBC), and plasma vitamin E (VE) levels in chicks fed dietary VE (dl-alpha-tocopheryl acetate, dl-alpha Ta) supplementation in steps of 0.0, 5.0, 10.0, 15.0, 20.0 and 30.0 mg/Kg were determined to examine their usefulness as an index of VE status. The increase in VE level was significant and linear in whole blood (r = 0.90), RBC (r = 0.89) and plasma (r = 0.93) in response to dietary VE intake. There was a close correlation between VE in plasma vs whole blood (r = 0.90), plasma vs RBC (r = 0.91) and whole blood vs RBC (r = 0.95). The plasma VE content was 1.2-1.8 times greater than that of whole blood, and 6.6-12.5 times greater than that of RBC. The plasma total lipids content was not affected by the dietary VE intake, whereas the level of VE in the plasma total lipids was significantly increased with increasing supplementation. Alpha tocopherol was the major isomer (ca 92%) of VE in whole blood, RBC and plasma at hatching. The small proportions of beta-tocopherol (ca 2%), gamma-tocopherol (ca 5%) and alpha-tocotrienol (ca 1%) observed at 1 day of age had decreased or totally disappeared by 7 days of age after feeding the VE-free basal diet. The data showed that in the chick, the whole blood and RBC levels of VE were as sensitive and reliable indexes of dietary VE status as was that of the plasma.     

干旱胁迫下γ-氨基丁酸保护玉米幼苗光合系统的生理响应

为明确干旱胁迫下γ-氨基丁酸(GABA)保护玉米幼苗光合系统的生理响应,以郑单958为试验材料,依据玉米幼苗生长数据,选择1 mmol·L^-1为γ-氨基丁酸(GABA)供试浓度,设置空白对照(CK)、1 mmol·L^-1 GABA(G)、20%聚乙二醇(PEG-6000)模拟干旱胁迫(D)、20%聚乙二醇(PEG-6000)模拟干旱胁迫和1 mmol·L^-1 GABA(DG)4个处理开展玉米水培试验。结果表明:不同浓度GABA能缓解干旱胁迫的抑制作用,使玉米幼苗恢复生长,其中1 mmol·L^-1 GABA效果最好。干旱胁迫下,外源施用1 mmol·L^-1 GABA能显著提高叶片超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)活性,减少丙二醛(MDA)、超氧阴离子($O_{2}^{-}$)和过氧化氢(H₂O₂)积累,降低叶片相对电导率。此外,外源GABA能显著提高干旱胁迫下叶片内可溶性蛋白、可溶性糖和脯氨酸含量,从而提高细胞保水能力。外源施用GABA能显著降低叶片干旱胁迫下初始荧光(F₀),提高暗适应下最大可变荧光(F_v)、最大荧光(F_m)和最大光量子效率(F_v/F_m),从而降低叶片光化学损伤。在干旱胁迫第5天,与D处理相比,DG处理SPAD数值、净光合速率(P_n)、蒸腾速率(T_r)和气孔导度(G_s)分别提高8.25%、7.69%、9.13%和7.38%,胞间CO₂浓度(C_i)降低2.93%。因此,外源GABA能通过降低叶片的氧化损伤和提高细胞保水能力来改善叶片对干旱胁迫的适应能力,从而保护玉米幼苗光合系统。     

Serum-gamma-glutamyltranspeptidase (gamma-GT) and aspartate-aminotransferase (AspAT) activities in adult cattle with chronic Fasciola hepatica infection

Sera from adult cattle with fluke-infected and normal livers were analyzed for γ-GT and AspAT activities. The analyses revealed no significant increases in either γ-GT or AspAT values of the animals with fascioliasis as compared with the non-infected animals. The advanced chronic stage of the infection characterized by hepatic fibrosis, limited injury of parenchyma and low fluke burdens may explain the lack of enzymatic increase.     

两种活性羰基类物质对 ECV304细胞的毒性研究

为探讨乙二醛（GO）和甲基乙二醛（MGO）对上皮细胞的细胞毒性及其机制,分别以 MTT 法检测两种活性羰基化合物（RCS）对 ECV304细胞活力的影响,评价其细胞毒性;DCFH-DA 探针法检测氧化应激;用罗丹明123检测线粒体膜电位及线粒体形态变化。结果表明,GO 和 MGO 对 ECV304细胞的 IC50分别为3．6 mmol／L±0．1 mmol／L 和1．3 mmol／L±0．1 mmol／L;氧化应激水平小幅度增高,线粒体膜电位随发生改变;线粒体形态发生改变。GO 和 MGO 的细胞毒性与氧化应激水平增加相关性较低而与线粒体损伤相关。     

Evidence for estrogen receptor expression during medullary bone formation and resorption in estrogen-treated male Japanese quails (Coturnix coturnix japonica)

The temporal expression of estrogen receptor (ER)-α and ER-β mRNA was examined in male Japanese quails. Femurs of quails receiving 17β-estradiol underwent RTPCR and histochemical analysis 1 to 15 days after treatment. Untreated quails were used as controls (day 0). Between days 0 and 5, cells lining the bone endosteal surface differentiated into osteoblasts, which in turn formed medullary bone. Expression of ER-α was already observed on day 0 and increased slightly during bone formation whereas ER-β was hardly detected throughout this process. After osteoclasts appeared on the medullary bone surface, this type of bone disappeared from the bone marrow cavity (days 7~15). ER-α expression simultaneously decreased slightly and ER-β levels remained very low. These results suggest that estrogen activity mediated by ER-α not only affects medullary bone formation but also bone resorption.     

Effect of transportation and relocation in post-weaning anoestrous primiparous sows

The object of this investigation was to study the clinical and endocrine responses to transportation and relocation in 8 post-weaning anoestrous sows. They had been anoestrous for at least 24 days after weaning before transportation/relocation was performed. Laparoscopy, performed at the beginning of the experiment, revealed that the ovaries contained many follicles (less than or equal to 6 mm in diameter), but no corpora lutea. Blood samples, taken before and after transportation/relocation, showed that LH activity was low at the beginning of the experiment and increased after transportation/relocation in the majority of the sows. Peripheral plasma concentrations of of oestradiol-17 beta increased 1-4 days after transportation/relocation in 6 out of 8 sows which was followed by oestrus and ovulation. Progesterone concentrations were also below the practical detection limit until the end of oestrus. This study has demonstrated that a change in environment by transportation and relocation can induce oestrus by increasing the LH activity in post-weaning anoestrous sows.     

Effect of fractionated weaning on hormonal patterns and weaning to oestrus interval in primiparous sows

The effect of weaning the 4-5 heaviest piglets in the litter on day 33 of lactation and the remainder 2 days later (fractionated weaning) on plasma levels of prolactin, cortisol, oestradiol-17 beta (E2), progesterone (P4) and LH, as well as on the weaning to oestrus interval in primiparous sows was studied. Twelve crossbred sows were grouped into 6 pairs according to farrowing date and litter size. The litter of 1 sow in each pair (F) was weaned in 2 stages, and the other conventionally weaned at 35 days (C). Blood samples were collected via a permanent jugular vein catheter every 3 h from 9 a m to 9 p m daily throughout the experimental period, and intensively at 15 min intervals for 12 h on the day of first and final weaning and for 6 h on the day after each weaning. All sows were slaughtered following their first post-weaning oestrus and the reproductive organs were macroscopically examined. Lactational oestrus was not observed in any of the sows. Sows from 5 out of 6 pairs showed oestrus within 8 days of weaning and post-mortem examination showed normal ovulation. There was a tendency for the F sows to have a shorter weaning to oestrus interval, as compared with the C sows (5 of 6 pairs, 4.8 days v 5.6 days). The plasma levels of prolactin around weaning were not significantly different between the 2 groups. Within 6 h after final weaning, the prolactin concentrations decreased gradually from 7.6 and 8.7 to 1.6 and 1.7 microgram/l in the control and treatment groups, respectively. The plasma levels of cortisol, showing a diurnal rhythm (with the lowest level at 6 and/or 9 p m), did on no occasion differ between the 2 groups. On the day of final weaning, no diurnal rhythm was observed, with cortisol remaining high at 6 and 9 p m. The plasma levels of E2 and P4 were low until final weaning in both groups. After final weaning the E2 levels rose faster in the F sows than in the C sows, to 44.3 and 34.8 pmol/l, respectively, on day 2 (p less than 0.01). No significant differences in levels of plasma LH and the number of LH pulses were observed between the groups. After final weaning the average and base levels of LH and the number of LH pulse(s) increased significantly.     

5alpha-androstenone in fat from boars selected for rate of gain and thickness of back fat, and from boars used in artificial insemination service.

Large variations in the level of 5α-androstenone in fat from different boars have been found. No significant difference in the level of 5α-androstenone was detected in fat from boars selected for high rate of gain and low back fat (HP-line), low rate of gain and high degree of fatness (LP-line) and a control group maintained without deliberate selection (GL-line). In boars used in artificial insemination service relatively high levels of 5α-androstenone in fat were observed, and in these animals a significant (P < 0.05) positive regression of 5α-androstenone level on age was found. Positive but non-significant regression coefficients were found between number of services which the boars had performed and level of 5α-androstenone in fat from the same animals.     

Alpha basic crystallin expression in canine mammary tumors

The aim of this study was to evaluate prognostic and/or diagnostic factors of canine mammary tumors by immunohistochemically analyzing the expression of alpha basic crystallin (αB-c). For this, formalin-fixed, paraffin-embedded blocks of 51 naturally-occurring canine mammary tumors (11 benign and 40 malignant) were used. Tissue from eight normal canine mammary glands were served as a control. Immunohistochemically, in the control mammary tissues, a few luminal epithelial cells were αB-c positive but myoepithelial cells were negative. In benign or simple type malignant tumors, αB-c expression was observed in luminal epithelial cells while the myoepithelial basal cells were negative. In benign or complex type malign tumors, positive staining was predominantly found in the cytoplasm of epithelial cells. Immunoreactivity of αB-c was also observed in neoplastic myoepithelial cells. Statistically, the number of cells immunolabeled with αB-c was found to be significantly different among tissues from normal canine mammary glands, benign lesions, and malignant tumors (p < 0.05). αB-c immunoreactivity was higher in malignant tumors than the control mammary tissues (p < 0.001). Data obtained in the current study revealed a strong association between high expression levels of αB-c and primary mammary gland tumors in canines.     

The effect of detomidine hydrochloride on the electrical activity of uterus in pregnant mares

The effect of detomidine on the electrical activity of the uterus was studied during the last trimester of pregnancy in 6 mares. The effect was observed in 3-5 min after the i.m. injection and it lasted for 50-70 min. 20 and 40 micrograms/kg b.w. doses of detomidine decreased the myometrial electrical activity, whereas 60 micrograms/kg dose did not have any effect on the activity. The results suggested that 20, 40 and 60 micrograms/kg b.w. doses of detomidine can be administered to mares during the last trimester of pregnancy without the risk of abortion induced by increased uterine electrical activity.     

Phytotherapeutic effects of Echinacea purpurea in gamma-irradiated mice

Echinacea (E.) purpurea herb is commonly known as the purple coneflower, red sunflower and rudbeckia. In this paper, we report the curative efficacy of an Echinacea extract in γ-irradiated mice. E. purpurea was given to male mice that were divided into five groups (control, treated, irradiated, treated before irradiation & treated after irradiation) at a dose of 30 mg/kg body weight for 2 weeks before and after irradiation with 3 Gy of γ-rays. The results reflected the detrimental reduction effects of γ-rays on peripheral blood hemoglobin and the levels of red blood cells, differential white blood cells, and bone marrow cells. The thiobarbituric acid-reactive substances (TBARs) level, Superoxide dismutase (SOD) and glutathione peroxidase (GSPx) activities and DNA fragmentation were also investigated. FT-Raman spectroscopy was used to explore the structural changes in liver tissues. Significant changes were observed in the microenvironment of the major constituents, including tyrosine and protein secondary structures. E. purpurea administration significantly ameliorated all estimated parameters. The radio-protection effectiveness was similar to the radio-recovery curativeness in comparison to the control group in most of the tested parameters. The radio-protection efficiency was greater than the radio-recovery in hemoglobin level during the first two weeks, in lymphoid cell count and TBARs level at the fourth week and in SOD activity during the first two weeks, as compared to the levels of these parameters in the control group.     

Clinical and endocrinological studies in primiparous post partum sows. Effects of lactation length and litter size

The object of this investigation was to determine the relationships between clinical findings and hormonal patterns in primiparous sows with different lactation length and litter size during lactation, weaning and to the first oestrus. Seven pairs of primiparous full sib sows were used to determine the effect of lactation length with normal litter size. One sow of each pair was assigned to nurse the piglets for 3 weeks (group A) while the other nusred for 5 weeks (group B). Another 8 primiparous sows (group C) were assigned to nurse 2–4 piglets during a 5-week lactation period. Oestrus detection was performed twice daily and laparoscopic examination every 2 weeks. If the sows did not come in oestrus within 3 weeks after weaning they were slaughtered. Peripheral plasma levels of progesterone, oestradiol-17β and LH were estimated by radioimmunoassays throughout the experimental period.     

Prolonged expression of senescence markers in mice exposed to gamma-irradiation

Although ionizing radiation is known to induce cellular senescence in vitro and in vivo, its long-term in vivo effects are not well defined. In this study, we examined the prolonged expression of senescence markers in mice irradiated with single or fractionated doses. C57BL/6 female mice were exposed to 5 Gy of γ-rays in single or 5, 10, 25 fractions. At 2, 4, and 6 months after irradiation, senescence markers including mitochondrial DNA (mtDNA) common deletion, p21, and senescence-associated β-galactosidase (SA β-gal) were monitored in the lung, liver, and kidney. Increases of mtDNA deletion were detected in the lung, liver, and kidney of irradiated groups. p21 expression and SA β-gal staining were also increased in the irradiated groups compared to the non-irradiated control group. Increases of senescence markers persisted up to 6 months after irradiation. Additionally, the extent of mtDNA deletion and the numbers of SA β-gal positive cells were greater as the number of radiation fractions increased. In conclusion, our results showed that ionizing radiation, especially that delivered in fractions, can cause the persistent upregulation of senescence marker expression in vivo. This should be considered when dealing with chronic normal tissue injuries caused by radiation therapy or radiation accidents.     

Characteristic upregulation of glucose-regulated protein 78 in an early lesion negative for hitherto established cytochemical markers in rat hepatocarcinogenesis

Previously, we reported α(2)-macroglobulin (α(2)M) to be a novel marker characteristic of rat hepatocellular preneoplastic and neoplastic lesions negative for hitherto well-established markers. In the present study, we further examined other candidate markers with specificity for the same type of lesions. Glutathione S-transferase-placental form (GST-P)-negative hepatocellular altered foci (HAF) were generated using a two-stage (initiation and promotion) carcinogenesis protocol with N,N-diethylnitrosamine (DEN) and either Wy-14,643 or clofibrate, two peroxisome proliferators. Microarray analysis using total RNAs isolated from laser-microdissected GST-P-negative HAF (amphophilic cell foci) and adjacent normal tissues was conducted along with immunohistochemistry and real-time RT-PCR. Staining for glucose-regulated protein 78 (GRP78) was detected in GST-P-negative HAF and hepatocellular adenomas, and slightly increased GRP78 mRNA expression was observed in the lesions by real-time RT-PCR analysis. Thus, an early increase of GRP78 expression in hepatocarcinogenesis is likely a feature of the amphophilic subset of HAF.     

Repeat tuberculin skin testing leads to desensitisation in naturally infected tuberculous cattle which is associated with elevated interleukin-10 and decreased interleukin-1 beta responses

The principal surveillance tool used to control bovine tuberculosis in cattle is the removal of animals that provide a positive response to the tuberculin skin-test. In this study we performed a longitudinal investigation of the immunological and diagnostic consequences of repeated short-interval skin-tests in cattle naturally infected with Mycobacterium bovis. Tuberculin skin-test positive cattle were subjected to up to four further intradermal comparative cervical skin-tests at approximately 60-day intervals. A significant progressive reduction in the strength of the skin-test was observed after successive tests. In contrast, the magnitude of interferon-γ (IFN-γ) responses was not influenced by repeat skin-testing either transiently around the time of each skin-test or longitudinally following repeated tests. A significant boost in blood interleukin-10 (IL-10) production was observed within 3 days following each skin-test although the magnitude of this boosted response returned to lower levels by day 10 post-test. The application of a novel multiplex assay to simultaneously measure seven cytokines and chemokines also identified that skin-testing resulted in a significant and progressive reduction in antigen specific interleukin-1β (IL-1β) whilst confirming stable IFN-γ and elevated IL-10 responses in the blood. Therefore, we have demonstrated that in cattle naturally infected with M. bovis, repeat short-interval skin-testing can lead to a progressive reduction in skin-test responsiveness which has potential negative consequences for the detection of infected animals with marginal or inconclusive skin-test responses. The desensitising effect is associated with decreased IL-1β and elevated IL-10 responses, but importantly, does not influence antigen specific IFN-γ responses.     

Implantation of canine umbilical cord blood-derived mesenchymal stem cells mixed with beta-tricalcium phosphate enhances osteogenesis in bone defect model dogs

This study was performed to evaluate the osteogenic effect of allogenic canine umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs) mixed with beta-tricalcium phosphate (β-TCP) in orthotopic implantation. Seven hundred milligrams of β-TCP mixed with 1 × 10⁶ UCB-MSCs diluted with 0.5 ml of saline (group CM) and mixed with the same volume of saline as control (group C) were implanted into a 1.5 cm diaphyseal defect and wrapped with PLGC membrane in the radius of Beagle dogs. Radiographs of the antebrachium were made after surgery. The implants were harvested 12 weeks after implantation and specimens were stained with H&E, toluidine blue and Villanueva-Goldner stains for histological examination and histomorphometric analysis of new bone formation. Additionally, UCB-MSCs were applied to a dog with non-union fracture. Radiographically, continuity between implant and host bone was evident at only one of six interfaces in group C by 12 weeks, but in three of six interfaces in group CM. Radiolucency was found only near the bone end in group C at 12 weeks after implantation, but in the entire graft in group CM. Histologically, bone formation was observed around β-TCP in longitudinal sections of implant in both groups. Histomorphometric analysis revealed significantly increased new bone formation in group CM at 12 weeks after implantation (p < 0.05). When applied to the non-union fracture, fracture healing was identified by 6 weeks after injection of UCB-MSCs. The present study indicates that a mixture of UCB-MSCs and β-TCP is a promising osteogenic material for repairing bone defects.     

乳制品中A1 β-酪蛋白、A2 β-酪蛋白检测方法研究进展

随着消费者对A2 β-酪蛋白的关注度提升,A2 β-酪蛋白相关产品不断更新换代。A2 β-酪蛋白常用检测方法有反相高效液相色谱法（RP-HPLC）、液相色谱–高分辨串联质谱法、毛细管区带电泳法、试剂盒检测法等。本文对β-酪蛋白遗传变体A1 β-酪蛋白和原始形态A2 β-酪蛋白的功能特点、检测方法、定量分析进行综述,说明检测方法的原理和试剂药品的作用机理,加强对A1 β-酪蛋白、A2 β-酪蛋白检测方法的改进与优化,为A2 β-酪蛋白应用于功能性食品、婴幼儿配方食品提供理论依据。     

Oxidant and antioxidant profile of hyperketonemic ewes affected by pregnancy toxemia

Background: Negative energy balance during late pregnancy in ewes is an important cause of hyperketonemia. Ketone bodies can generate superoxide radicals and cause oxidative stress and cellular dysfunction, as noted in cows with subclinical ketosis or in diabetic people. Objective: The aim of this study was to evaluate the role of hyperketonemia in initiating the process of lipid peroxidation. Methods: The study included 10 pregnant ewes (aged 3.5–6 years) with pregnancy toxemia, 10 clinically healthy pregnant ewes, and 10 clinically healthy nonpregnant ewes. Serum concentrations of β‐hydroxybutyrate (BHB), cortisol, and glucose, plasma activities of glutathione peroxidase, superoxide dismutase, and catalase, and plasma concentrations of thiobarbituric acid reactive substances (TBARS), markers of lipid peroxidation, and reduced glutathione were measured. Data from the 3 groups were statistically analyzed and compared. Results: Serum concentrations of BHB, cortisol, and TBARS were significantly higher in ewes with pregnancy toxemia when compared with concentrations in healthy pregnant and nonpregnant groups (P≤.05). In ewes with pregnancy toxemia, a strong positive correlation was found between concentrations of TBARS and BHB (r=.80; P=.002) and between concentrations of BHB and cortisol (r=.76; P=.005). Conclusions: Oxidative stress and lipid peroxidation are involved in the development and complications of pregnancy toxemia. An association between hyperketonemia and the products of lipid peroxidation has also been demonstrated, suggesting that ketosis is a risk factor in the development of lipid peroxidation and oxidative stress in ewes affected by pregnancy toxemia.     

酸奶营养品质提升的加工工艺研究

牛奶中含有丰富的活性营养物质,包括乳铁蛋白、免疫球蛋白、乳过氧化物酶、α-乳白蛋白、β-乳球蛋白等。传统工艺生产的酸奶经过高温杀菌,活性营养物质几乎完全损失。为了避免这一情况,本文推出一种新的酸奶生产工艺,生产出的酸奶可保留大部分活性营养物质。首先将牛奶的乳脂进行分离,乳脂部分进行125 ℃、5 s杀菌,脱脂牛奶部分进行75 ℃、15 s杀菌,将两部分料液混合后加入菌种发酵至酸度75~85°T,破乳后冷却后熟8 h以上。经测定,新工艺酸奶中α-乳白蛋白、β-乳球蛋白、乳铁蛋白、免疫球蛋白、乳过氧化物酶含量分别为966 mg/L、1 963 mg/L、23.4 mg/L、102 mg/L、1 025 U/L,远高于传统工艺酸奶的68 mg/L、200 mg/L、0.7 mg/L、14 mg/L、0 U/L。通过对比新工艺与传统工艺酸奶保质期内的活菌数、黏度、酸度和口感,并无显著差异。