Zika Virus Growth in Human Kidney Cells Is Restricted by an Elevated Glucose Level

Mosquito-borne Zika virus (ZIKV) became a real threat to human health due to the lack of vaccine and effective antiviral treatment. The virus has recently been responsible for a global outbreak leading to millions of infected cases. ZIKV complications were highlighted in adults with Guillain–Barré syndrome and in newborns with increasing numbers of congenital disorders ranging from mild developmental delays to fatal conditions. The ability of ZIKV to establish a long-term infection in diverse organs including the kidneys has been recently documented but the consequences of such a viral infection are still debated. Our study aimed to determine whether the efficiency of ZIKV growth in kidney cells relates to glucose concentration. Human kidney HK-2 cells were infected with different ZIKV strains in presence of normal and high glucose concentrations. Virological assays showed a decrease in viral replication without modifying entry steps (viral binding, internalization, fusion) under high glucose conditions. This decrease replication was associated with a lower virus progeny and increased cell viability when compared to ZIKV-infected HK-2 cells in normal glucose concentration. In conclusion, we showed for the first time that an elevated glucose level influences ZIKV replication level with an effect on kidney cell survival.     

Sofosbuvir Selects for Drug-Resistant Amino Acid Variants in the Zika Virus RNA-Dependent RNA-Polymerase Complex In Vitro

The nucleotide analog sofosbuvir, licensed for the treatment of hepatitis C, recently revealed activity against the Zika virus (ZIKV) in vitro and in animal models. However, the ZIKV genetic barrier to sofosbuvir has not yet been characterized. In this study, in vitro selection experiments were performed in infected human hepatoma cell lines. Increasing drug pressure significantly delayed viral breakthrough (p = 0.029). A double mutant in the NS5 gene (V360L/V607I) emerged in 3 independent experiments at 40–80 µM sofosbuvir resulting in a 3.9 ± 0.9-fold half- maximal inhibitory concentration (IC₅₀) shift with respect to the wild type (WT) virus. A triple mutant (C269Y/V360L/V607I), detected in one experiment at 80 µM, conferred a 6.8-fold IC₅₀ shift with respect to the WT. Molecular dynamics simulations confirmed that the double mutant V360L/V607I impacts the binding mode of sofosbuvir, supporting its role in sofosbuvir resistance. Due to the distance from the catalytic site and to the lack of reliable structural data, the contribution of C269Y was not investigated in silico. By a combination of sequence analysis, phenotypic susceptibility testing, and molecular modeling, we characterized a double ZIKV NS5 mutant with decreased sofosbuvir susceptibility. These data add important information to the profile of sofosbuvir as a possible lead for anti-ZIKV drug development.     

Ionic Strength Effects on Heat-induced Gelation of Myofibrils and Myosin from Fast- and Slow-twitch Rabbit Muscles

The effects of ionic strength on myofibrils and myosin from rabbit fast-twitch Psoas major (PM) and slow-twitch Semimembranosus proprius (SMp) muscles before and after heating were studied by electron microscopy and thermal scanning rheometry. The direct suspension of proteins in low ionic strength (0.2M KCl; pH 6.0) led to very weak gels, whereas a gradual lowering of the ionic strength (by dialysis against 0.2M KCl; pH 6.0) of 0.6M KCl protein solutions induced strand-type networks at low temperature and strong heat-induced gels. As shown by transmission and scanning electron micrographs, in low ionic strength, SMp myosins formed shorter filaments before heating and thinner and shorter structures in heat-induced gels, as well as a lower gel porosity than PM myosins.     

Human DDX3X Unwinds Japanese Encephalitis and Zika Viral 5′ Terminal Regions

Flavivirus genus includes many deadly viruses such as the Japanese encephalitis virus (JEV) and Zika virus (ZIKV). The 5′ terminal regions (TR) of flaviviruses interact with human proteins and such interactions are critical for viral replication. One of the human proteins identified to interact with the 5′ TR of JEV is the DEAD-box helicase, DDX3X. In this study, we in vitro transcribed the 5′ TR of JEV and demonstrated its direct interaction with recombinant DDX3X (K_d of 1.66 ± 0.21 µM) using microscale thermophoresis (MST). Due to the proposed structural similarities of 5′ and 3′ TRs of flaviviruses, we investigated if the ZIKV 5′ TR could also interact with human DDX3X. Our MST studies suggested that DDX3X recognizes ZIKV 5′ TR with a K_d of 7.05 ± 0.75 µM. Next, we performed helicase assays that suggested that the binding of DDX3X leads to the unwinding of JEV and ZIKV 5′ TRs. Overall, our data indicate, for the first time, that DDX3X can directly bind and unwind in vitro transcribed flaviviral TRs. In summary, our work indicates that DDX3X could be further explored as a therapeutic target to inhibit Flaviviral replication     

Misidentification by wild rabbits,Oryctolagus cuniculus,of group members carrying the odor of foreign inguinal gland secretion

To demonstrate that the inguinal gland odor conveys information on the identity of individuals, 60 male, wild-type rabbits,Oryctolagus cuniculus, living in groups of three in outdoor pens, were used in 120 tests. Members of the groups smeared with the inguinal gland secretion from unfamiliar rabbits were attacked by their penmates. In contrast the odors of chin gland secretions and urine of strange rabbits and of a commercial perfume did not cause serious disruption to the social relationships within the groups.     

高强度聚焦超声损伤兔肌肉组织的超声影像学变化的初步研究

探讨高强度聚焦超声损伤兔肌肉组织的超声图像变化。采用高强度聚焦超声经皮照射20只新西兰大白兔双侧后肢肌肉,其中18只于照射前1d、照射后10min、照射后1d,3d,7d,14d,21d,28d进行二维超声、彩色多普勒和能量多普勒超声检查,并于照射后28天处死后测量凝固灶大小。另2只分别于照射后1d及照射后50d解剖,行病理学检查。(1)HIFU照射后凝固灶外周可见强回声带至低回声带的演变过程。(2)HIFU照射后第1d,测量的凝固灶体积最大(1476.59±308.64mm3),第3d～第21d,凝固灶体积逐渐缩小(612.47±127.98mm3)。第28d凝固灶体积最小(343.29±54.79mm3)。超声检查在兔肌肉组织HIFU照射后的随访中发挥了重要的作用,为监测HIFU治疗人体软组织肿瘤超声图像的变化提供了实验依据。     

Partial replacement of sodium chloride with potassium chloride in marinated rabbit meat

This study was aimed at evaluating the consequences of sodium chloride reduction by potassium chloride up to 50% on technological, sensorial and microbiological traits of marinated rabbit meat. In total, 226 rabbit loin meat samples were obtained and subjected to vacuum tumbling using solutions with different NaCl/KCl ratios. Replacing of sodium chloride up to 30% by potassium chloride did not change microbiological traits (total aerobic mesophilic and lactic acid bacteria maximum cell loads), sensorial acceptability (perceived saltiness and overall liking) and technological traits (pH, colour, texture, cooking loss and yield). Otherwise, reduction in sodium chloride to 50% significantly decreased perceived saltiness (4.15 vs. 4.73; P < 0.05) and reduced microbial shelf life by 1 day when compared to control, even if there was still no effect on technological traits. In conclusion, it is feasible imparting an added value for processed rabbit meat products by reduction in sodium content that could increase market interest.     

Preparation,characterisation and structure of rabbit (Hyla rabbit) skin gelatine

Rabbit skin was used as a raw material to prepare gelatine. The yield of extracted rabbit skin gelatine was 8.49 ± 0.91% based on a wet weight basis. The bloom strength of 6.67% rabbit gelatine was 457.57 ± 3.302 g, and its gelling and melting temperatures were 27.8 and 34.4 °C, respectively. Sodium dodecyl sulphate‐polyacrylamide gel electrophoresis analysis demonstrated that the content of the subunit components (α chain, β and γ components) of the rabbit skin gelatine achieved 97.915%. Amino acid composition analysis indicated that the total amino acid contents (proline plus hydroxyproline) in the rabbit gelatine were 222 residues per 1000 residues. The high content of the subunit components and amino acids could contribute to the gelatine's suitable gel properties. Rabbit gelatine had nearly all of the characteristic absorption peaks of collagen from rabbit skin, and the loss of the triple helical structure and damage to part of the hydrogen bonds occurred during the conversion of collagen to gelatine.     

季铵型改性壳聚糖的制备及其在兔毛织物抗菌整理中的应用

以三乙胺和环氧氯丙烷为原料,合成醚化剂3-氯-2-羟丙基三乙基氯化铵。以甲壳素为原料通过醚化反应合成了季铵型甲壳素（CCT）,再经脱乙酰得到季铵型阳离子改性壳聚糖（CCTS）。采用红外光谱（FTIR）、核磁碳谱（¹³C NMR）对其化学结构进行了表征,运用黏度法和分光光度法测定了黏均分子量和溶解性等理化性能。采用最小抑菌法对CCTS的抗菌活性进行了测定,得到其对大肠杆菌的最低有效抑菌浓度（MIC）为0.2g/L,优于天然壳聚糖的MIC值。以柠檬酸为交联剂、次磷酸钠为催化剂,用CCTS对兔毛织物进行抑菌整理,考察织物经整理的抑菌效果和耐洗性。经5次洗涤后结果表明,CCTS对大肠杆菌的抑菌率达99.9%以上,抑菌率高于CCT和天然壳聚糖,是一种针对动物毛织物良好的天然高分子长效抑菌整理剂。