Metformin acts to suppress β-hydroxybutyric acid-mediated inflammatory responses through activation of AMPK signaling in bovine hepatocytes

The occurrence of bovine ketosis involves the accumulation of β-hydroxybutyric acid (BHBA), which contributes to the initiation and acceleration of hepatic metabolic stress and inflammation. Metformin has other beneficial effects apart from its medical intervention for diabetes, such as prevention of laminitis and hyper-triglyceridemic. AMPK maintains energy homeostasis and is the intracellular target of metformin action. This study aims to uncover the role of metformin in modulating BHBA-induced inflammatory responses through the activation of AMPK signaling. The hepatocytes were isolated from the liver tissue of mid-lactation multiparous Holstein cows (~160 d postpartum). Treatments were conducted as follows: treated with PBS for 18 h (control); pretreated with PBS for 12 h followed by treatment of 1.2 mM BHBA for 6 h (BHBA); pretreated with 1.5 mM or 3 mM metformin for 12 h followed by the BHBA treatment (1.2 mM) for 6 h (M(1.5)+B; M(3)+B). The inhibitor of AMPK, Compound C, at a concentration of 10 μM, was applied to substantiate the AMPK-dependent responses. RT-qPCR were applied for the mRNA expression while Western-blots and immunofluorescence were conducted for the target proteins expression. Among dose-dependent assays for BHBA, the concentration of BHBA at 1.2 mM activated NF-κB signaling by upregulating the expression of phosphorylated NF-κB and pro-inflammatory cytokines compared with the control cells (P < 0.05). Along with the upregulation of phosphorylated AMPKα and ACCα, metformin at 1.5 and 3 mM inactivated NF-κB signaling components (p65 and IκBα) and the inflammatory genes (TNFA, IL6, IL1B and COX-2) which were activated by BHBA. Additionally, BHBA inhibited cells staining intensity in EdU assay were increased by pretreatment with metformin. The activation of AMPK resulted in the increased gene and protein expression of SIRT1, along with the deacetylation of H3K9 and H3K14. However, the AMPK inhibitor compound C blocked this effect. Compared with BHBA treated cells, the protein expression of COX-2 and IL-1β were decreased by the pretreatment with metformin, and the inhibitory effect of metformin was released by compound C. The bound of NF-κB onto IL1B promoter displayed higher in BHBA group and this was suppressed by pretreatment with metformin (P < 0.05). Altogether, metformin attenuates the BHBA-induced inflammation through the inactivation of NF-κB as a target for AMPK/SIRT1 signaling in bovine hepatocytes.     

Evaluation of the influence of irrigation methods and water quality on sugar beet yield and water use efficiency

Rapid urbanization and industrialization have increased the pressure on limited existing fresh water to meet the growing needs for food production. Two immediate responses to this challenge are the efficient use of irrigation technology and the use of alternative sources of water. Drip irrigation methods may play an important role in efficient use of water but there is still limited information on their use on sugar beet crops in arid countries such as Iran. An experiment was conducted to evaluate the effects of irrigation method and water quality on sugar beet yield, percentage of sugar content and irrigation water use efficiency (IWUE). The irrigation methods investigated were subsurface drip, surface drip and furrow irrigation. The two waters used were treated municipal effluent (EC = 1.52 dS m⁻¹) and fresh water (EC = 0.509 dS m⁻¹). The experiments used a split plot design and were undertaken over two consecutive growing seasons in Southern Iran. Statistical testing indicated that the irrigation method and water quality had a significant effect (at the 1% level) on sugar beet root yield, sugar yield, and IWUE. The highest root yield (79.7 Mg ha⁻¹) was obtained using surface drip irrigation and effluent and the lowest root yield (41.4 Mg ha⁻¹) was obtained using furrow irrigation and fresh water. The highest IWUE in root yield production (9 kg m⁻³) was obtained using surface drip irrigation with effluent and the lowest value (3.8 kg m⁻³) was obtained using furrow irrigation with fresh water. The highest IWUE of 1.26 kg m⁻³ for sugar was obtained using surface drip irrigation. The corresponding efficiency using effluent was 1.14 kg m⁻³. Irrigation with effluent led to an increase in the net sugar yield due to an increase in the sugar beet root yield. However, there was a slight reduction in the percentage sugar content in the plants. This study also showed that soil water and root depth monitoring can be used in irrigation scheduling to avoid water stress. Such monitoring techniques can also save considerable volumes of irrigation water and can increase yield.     

Ovaprim treatment promotes oocyte development and milt fertilization rate in diploid and triploid African catfish (<Emphasis Type="Italic">Clarias gariepinus)</Emphasis>

Triploid fish are increasingly used in aquaculture because they are generally unable to reproduce successfully. Energy is channeled into somatic growth rather than gonadal development, and in the event of escape, the animals are unlikely to breed successfully among themselves or with wild conspecifics. This study tested the ability of recently matured triploid African catfish (Clarias gariepinus) to produce and fertilize eggs with and without ovaprim treatment. Triploid females did not show the increase in ovary size observed in diploid members of the same cohort between 8 and 9 months of age, or the coincident decrease in visceral fat deposits, and this was unaffected by up to 5 weekly i.m. injections of 0.5 ml kg⁻¹ Ovaprim. However, we observed advanced vitellogenin (Vtg) sequestration in oocytes of triploid females, albeit to a lesser degree and with lesser cortical alveoli, compared to oocytes from diploid cohort members. Histological sections revealed a positive trend of oocyte development up to the third weekly ovaprim injection followed by a negative gonadal development in weeks four and five. Milt from triploid males injected 9–12 h earlier with 0.25 ml kg⁻¹ ovaprim i.m. fertilized more diploid eggs than milt from untreated triploid males (30 vs. 20%), but none of the developing embryos of triploid paternity survived to hatch. In contrast, milt of diploid males fertilized 49% of eggs, and 20% of the developing embryos hatched successfully. These rates were improved in ovaprim-injected diploid males to 70% fertilization and 33% hatch. This study demonstrates potential of overcoming non-viability of eggs from triploid female African catfish, and enhancing the ability of triploid milt to fertilize eggs.     

Evaluation of gallbladder volume and contraction index with three-dimensional ultrasonography in healthy dogs

Three-dimensional (3D) ultrasonography has been shown to be an accurate and appropriate tool for measurement of gallbladder volume in humans. Therefore, we applied this novel technique for the first time to study fasting and postprandial gallbladder volume in 10 healthy dogs and compared the results with those of 2-dimensional (2D) ultrasonography. Fasting gallbladder volumes determined by 3D ultrasonography were significantly higher than corresponding volumes determined by 2D ultrasonography (P<0.01). Additionally, gallbladder volumes were significantly decreased in the postprandial state compared with the fasting state using 3D ultrasonography (P<0.001), but 2D ultrasonography showed no significant difference (P=0.189). The Gallbladder contraction index was higher in 3D ultrasonography than 2D ultrasonography; however, it did not reach statistical significance (P=0.25). In conclusion, 3D ultrasonography was able to measure gallbladder volume in healthy dogs in this study. It is suggested that 3D ultrasonography can be used to accurately estimate gallbladder volume and contractility.     

The efficacy and safety of long-term Helicobacter species quadruple therapy in asymptomatic cats with naturally acquired infection

Owing to rising drug-resistant Helicobacter species infections in people and animals, currently therapies are losing their efficacy; therefore, regimens efficacious in the presence of drug resistance are needed. This study assessed the efficacy and safety of a 14-day quadruple Helicobacter species therapy in cats with naturally acquired infection. Thirteen asymptomatic adult stray cats with Helicobacter species infection (identified by analysis of gastric biopsies using polymerase chain reaction and Helicobacter-specific primers) received omeprazole 0.7mg/kg q 8h plus amoxicillin 20mg/kg q 12h, metronidazole 20mg/kg q 12h and clarithromycin 7.5mg/kg q 12h, for 14 days. Second molecular analysis of gastric biopsies revealed persistence of Helicobacter species DNA in four cats that were negative on quantitative urease testing, cytology and histopathology. Our results suggest that antibiotic regimens that are effective against Helicobacter pylori in people cannot eradicate Helicobacter species in cats with naturally acquired infection, although transient suppression may occur.     

White striping prevalence and its effect on meat quality of broiler breast fillets under commercial conditions

White striping (WS), characterized by white striations on the Pectoralis major muscle, is an emerging problem for the broiler industry. Thus, the impact of WS on meat quality has become a current research topic. In this context, a 2‐year follow‐up study, to reveal the prevalence of WS in broiler integrations, was undertaken. Additionally, a laboratory experiment was conducted to determine the oxidative quality during storage and the differences in proximate composition, colour and fatty acid and amino acid profiles of broiler breast fillets scored visually as normal, moderate and severe. In integrations, the WS incidence increased with increasing broiler age and more than 50% of breast fillets obtained from 32‐ to 35‐day‐old broilers and 36‐ to 39‐day‐old broilers had stripes with different scores. Lower protein and higher fat contents were measured in the moderate and severe scored fillets than the normal fillets (p < 0.001). Severe score stripes induced the formation of less redness on the ventral surface (p < 0.05) and a darker colour on the dorsal surface (p < 0.01). The lowest histidine, arginine and tryptophan amounts were determined in severe fillets (p < 0.05). Moderate and severe scored fillets were rich in polyunsaturated fatty acids implying that they were sensitive to lipid oxidation. That suggestion was further proven by the higher TBARS values. Freeze‐thawing induced the formation of carbonyl compounds, but it did not cause differences in sulphydryl groups. Briefly, we suggest that different WS scores may affect the physicochemical and oxidative quality of breast fillets; however, more studies are needed to assert an accurate and explicit judgement.     

森林生态蓄水功能的经济评价(个案研究:伊朗扎格罗斯森林)(英文)

森林生态服务直接或间接地造福人类。本文利用模拟模型和地理信息系统作为工具,分析生态因子对生态系统服务功能的作用,评估了伊朗西部扎格罗斯森林对Bazoft河流域水源保护的经济价值。应用HEC-HMS模型中的曲线数字法对降雨量-径流进行了模拟。该模型需要输入土地覆被、土壤、短期降雨和流出数据。用观测数据并分阶段校对对拟合模型的功效进行了修订。测定了森林在4种设定的土地覆盖方案中保水性和减少地表径流量的作用以及土地用途改变对该区降雨量-径流行为的影响。结果表明:方案一假定该流域全被森林覆盖,总的流出量将最小,降雨初期损失将增加。用重置成本法估算了森林水文功能之保水性经济价值。评估结果表明,每公顷Bazoft河流域的森林可以蓄存84.8立方米水,年产值0.5 US$/m3。因此,每公顷BazoR河流域的森林保水性经济评价值是43US$。说明覆被森林有利于该流域经济发展,该研究有助于决策者选择适当的、经济可行的发展策略。     

Variation for stomatal characteristics and water use efficiency among diploid,tetraploid and hexaploid Iranian wheat landraces

Wheat is one of the most important crops in the world. Its study has involved anatomy, ecology, physiology, molecular biology, biotechnology, and cultivation. Knowledge of the relationship of diploid, tetraploid and hexaploid wheat with stomatal characteristics and water-use efficiency is spare and is subject to argument. The present study was aimed to examine different characteristics of stomata in 28 Iranian landraces belonging to the species Triticum monococcum (2n = 2x = 14), Triticum durum (2n = 4x = 28) and Triticum aestivum (2n = 6x = 42), respectively and investigated a possible relationship between these characteristics and water use efficiency by pot cultivation experiments. The results revealed large variation among landraces as well as between the different species for stomatal frequency and size. The diploid wheat species had the highest stomatal frequency and the lowest stomatal length and width. The hexaploid species had a lower stomatal frequency than the tetraploid species. A highly significant relationship was noted between the value of those traits on the adaxial and abaxial surfaces of the leaf. The ratio of adaxial to abaxial values appeared to be constant for a given species. Both stomatal frequency and size were associated with ploidy level, but also with the nature of the constituting genomes of the species. A highly significant negative association was found among accessions between stomatal frequency and size. As a consequence, the variation for stomatal area per unit leaf area was less than for the other traits. Diploid and hexaploid wheat have significantly higher water use efficiency, compared to tetraploid wheat. No clear association was found, however, between water use efficiency and stomatal characteristics.     

Changes in carbohydrate contents in shoot tips,leaves and roots of strawberry (Fragaria × ananassa Duch.) during flower-bud differentiation

Flowering is an important step in crop production. Under flower-inducing conditions, biochemical or physiological changes can be recognized. Changes in carbohydrates have an important role in flower development in plants; however, carbohydrate changes during flower bud differentiation in strawberry have not been thoroughly investigated. In this study, runner plants potted in 18 cm diameter pots and grown under non-inducing conditions (28 ± 3/22 ± 3 °C day/night; 16 h day length). When the plants were established, half of the plants then were put under inducing conditions (23 ± 3/13 ± 3 °C day/night; 8 h day length). After the induction period of 21 short-day cycles, plants were brought to non-inductive conditions again. In order to evaluate the carbohydrate changes during flower differentiation, shoot tips, leaves and roots were sampled from four replicated plants collected weekly for the period of 7 weeks. Sucrose, glucose and fructose contents were determined by HPLC, and starch by the anthrone method. The results obtained indicated that the most abundant soluble sugar in all organs tested was sucrose. Sucrose in the shoot tips of induced plants at 42, 56 and 70 days after the start of the short-day treatment were significantly higher than corresponding time in non-induced plants. However, the glucose, fructose and starch contents in shoot tips, leaves and roots of non-induced plants in most sampling dates were greater than those of induced plants. In other words, the shoot tips (bud) of induced plants acted as strong ‘utilizing sink’ and preferentially metabolized carbohydrates rather than storing them. It seems that non-structural carbohydrate contents in shoot tips, leaves and roots of strawberry may have an important role in flower-bud differentiation.