Two Simple Methods for the Collection of Individual Life Stages of Reniform Nematode,Rotylenchulus reniformis

The sedentary semi-endoparasitic nematode Rotylenchulus reniformis, the reniform nematode, is a serious pest of cotton and soybean in the United States. In recent years, interest in the molecular biology of the interaction between R. reniformis and its plant hosts has increased; however, the unusual life cycle of R. reniformis presents a unique set of challenges to researchers who wish to study the developmental expression of a particular nematode gene or evaluate life stage–specific effects of a specific treatment such as RNA-interference or a potential nematicide. In this report, we describe a simple method to collect R. reniformis juvenile and vermiform adult life stages under in vitro conditions and a second method to collect viable parasitic sedentary females from host plant roots. Rotylenchulus reniformis eggs were hatched over a Baermann funnel and the resultant second-stage juveniles incubated in petri plates containing sterile water at 30°C. Nematode development was monitored through the appearance of fourth-stage juveniles and specific time-points at which each developmental stage predominated were determined. Viable parasitic sedentary females were collected from infected roots using a second method that combined blending, sieving, and sucrose flotation. Rotylenchulus reniformis life stages collected with these methods can be used for nucleic acid or protein extraction or other experimental purposes that rely on life stage–specific data.     

Variabilité et homogénéité des modes de débitage en Chine entre 300 000 et 50 000 ans

The position of lithic industries of Chinese Paleolithic in comparison with those of other regions in the world is always a controversial topic. The typological analysis has made Chinese prehistorians reveal some variability between lithic industries of different regions of China in terms of technical tradition and industrial structure. But this recognition, based upon some morphological characters of stone artifacts, failed to define systematically the modes of production that exist in each site. To answer these problematic some new methods need to be applied. So the lithic materials from 8 Chinese sites dated from 300 ka to 50 ka B.P. were selected and analyzed by means of lithic technological analysis and a new approach for analyzing the structures of cores. This paper presents principally the analytical results of modes of débitage and revealed that the modes of débitage during this period were characterized essentially by system C (“simple débitage”) and E (discoid) and that the concept Levallois was absent during this period in technical systems of lithic production in China. The Chinese lithic industries between 300 ka and 50 ka B.P. saw some homogeneity in the scale of the continent and remarkable difference from those of other regions in the world. So a stable, continuous but independent even isolated evolution in comparison with those of other regions was observed in this period. In addition, despite of apparent homogeneity, some variability exists also between these industries.     

小哺乳动物化石牙齿釉质碳、氧同位素测试方法简介及应用前景

探讨古环境和古气候变化与哺乳动物演化之间的关系是目前古生物学研究领域中的一个热点,而哺乳动物化石牙齿釉质的碳、氧同位素分析是恢复古环境和古气候的一个重要手段。以往的哺乳动物化石牙齿釉质稳定同位素分析多集中在大哺乳动物化石,这主要是受到技术手段的限制,所需的样品量较大所决定的。但最近几年随着激光和离子显微探针技术的应用,对小哺乳动物化石(如啮齿类和兔形类)的牙齿釉质碳、氧同位素的分析和应用日趋成熟和广泛。除了传统的化学处理方法之外,对小哺乳动物化石牙齿釉质碳、氧同位素的分析还有以下三种方法:1)激光剥蚀气相色谱/同位素比值质谱分析;2)直接激光氟化技术;3)离子显微探针技术(SHRIMPII)。这些技术需要的样品量少,对标本的破损小,准确度和精密度高,所以在小哺乳动物化石和一些珍贵标本(如古人类化石)的稳定同位素分析中起到了重要作用。相对于大哺乳动物化石,小哺乳动物化石数量多、演化速度快,更能反映多个层位长时间序列的古环境和气候变化;而且小哺乳动物通常没有长距离迁徙的行为,栖息地局限,所以更能准确反映化石埋藏地点的古环境和气候状况。     

Terpenoid Induction in Sweet Potato Roots by Cyclic-3′, 5′-Adenosine Monophosphate

The bar-headed goose, a specialized high-altitude species, has a capacity for high oxygen uptake from a hypoxic environment. It thus has a higher oxygen affinity than other bird species of lower-altitude environments. Oxygen affinity is determined by molecular structures and genetic mutations of hemoglobin (Hb), which can also influence the coordinating structures and dynamics of oxygen-Hb. To explore the structural differences in Hbs as between high and low altitude species, photolysis dynamic parameters, including quantum yield, enthalpy, and conformational volume changes in carboxy-Hbs (HbCO) for the bar-headed goose and low altitude counterparts (the Chinese goose and chicken) were investigated by the laser pumping-probing technique and photoacoustic calorimetry. Comparing the photolysis results for HbCO of the three species, the enthalpy and conformational volume changes of the bar-headed goose were much smaller than those of the others, although the quantum yields of all three species are similar. To explain the possible mechanisms of these differences, modifications of salt bridges and key residue mutations at the α β subunit interfaces of the proteins are described and discussed briefly.     

基于15N示踪技术的植物-土壤系统氮循环研究进展

同位素示踪技术是指外源添加与生物体内的元素或物质完全共同运行的示踪物,用来指示生物体内某元素或物质变化过程的一种方法。利用氮稳定同位素示踪技术,能从本质上揭示生态学过程发生的机理,从而成为生态学科研工作十分重要的工具之一。对近年来15N示踪技术应用于土壤氮素固定、植物氮素营养和植物-土壤系统氮迁移的研究进展进行了综述,并对稳定氮同位素技术在解决相关生态学难题可能的前景和不足方面进行了展望。     

Instrumented Indentation Investigation on the Viscoelastic Properties of Porcine Cartilage

Articular cartilage lubricates the contact surfaces in human joints and provides a shock-absorbing effect which protects the joint under dynamic loading. However, this shock-absorbing effect is gradually reduced as the result of normal wear, tear and aging-related cartilage loss. Thus, with the increasing average human life expectancy, the issue of joint health has attracted significant interest in recent decades. In developing new materials for the repair or regeneration of damaged articular cartilage, it is essential that the difference in the mechanical properties of healthy and damaged cartilages is well-understood. In the present study, the hardness and Young＇s modulus of damaged and healthy porcine articular cartilage samples are evaluated via a quasi-static nanoindentation technique. A dynamic mechanical analysis method is then applied to determine the viscoelastic properties of the two samples. The results presented in this study provide a useful insight into the mechanical properties of articular cartilage at the mesoscale, and therefore fill an important gap in the literature.     

Heterotopic Renal Autotransplantation in a Porcine Model: A Step-by-Step Protocol

Kidney transplantation is the treatment of choice for patients suffering from end-stage renal disease. It offers better life expectancy and higher quality of life when compared to dialysis. Although the last few decades have seen major improvements in patient outcomes following kidney transplantation, the increasing shortage of available organs represents a severe problem worldwide. To expand the donor pool, marginal kidney grafts recovered from extended criteria donors (ECD) or donated after circulatory death (DCD) are now accepted for transplantation. To further improve the postoperative outcome of these marginal grafts, research must focus on new therapeutic approaches such as alternative preservation techniques, immunomodulation, gene transfer, and stem cell administration.Experimental studies in animal models are the final step before newly developed techniques can be translated into clinical practice. Porcine kidney transplantation is an excellent model of human transplantation and allows investigation of novel approaches. The major advantage of the porcine model is its anatomical and physiological similarity to the human body, which facilitates the rapid translation of new findings to clinical trials. This article offers a surgical step-by-step protocol for an autotransplantation model and highlights key factors to ensure experimental success. Adequate pre- and postoperative housing, attentive anesthesia, and consistent surgical techniques result in favorable postoperative outcomes. Resection of the contralateral native kidney provides the opportunity to assess post-transplant graft function. The placement of venous and urinary catheters and the use of metabolic cages allow further detailed evaluation. For long-term follow-up studies and investigation of alternative graft preservation techniques, autotransplantation models are superior to allotransplantation models, as they avoid the confounding bias posed by rejection and immunosuppressive medication.     

The effect of two cryopreservation methods on human sperm DNA damage

Several methods are currently available for selection when conducting sperm cryopreservation, however, these methods might cause different degrees of damage on sperm DNA. The aim of the this study is to compare the effects of storage at −80 °C (in ultra-low temperature refrigerator) and at −196 °C (in liquid nitrogen) on sperm DNA damage, thus to provide a reference for choosing the right method according to different aims. We randomly collected 28 semen samples from college students of Chongqing city. The samples stored at −80 °C were neat semen samples and the samples stored at −196 °C were mixed with additional cryoprotectants. Each sample was subjected to two freezing-thawing cycles, and the sperm DNA damage levels of fresh and thawed samples were measured by single cell gel electrophoresis (SCGE) and sperm chromatin structure assay (SCSA). Both SCGE and SCSA assays showed cryopreservation induced significant damage to sperm DNA. However, storage at −196 °C lead to more severe damage to sperm DNA than storage at −80 °C measured by SCSA. Sperm DNA damage increased simultaneously with the higher frequency of freezing-thawing cycles. We concluded that storage of neat semen samples at −80 °C had milder damage to sperm DNA than storage at −196 °C mixed with cryoprotectants. To avoid additional sperm DNA damage, repeated freezing and thawing should be prevented.     

Horticultural mineral oil influences Plum pox virus transmission by Myzus persicae

The residual activity of horticultural mineral oil (HMO) on the ability of green peach aphids, Myzus persicae (Sulzer), (GPA) to transmit Plum pox virus (PPV) to peach was measured by infection rates of detached leaves from plants sprayed with either HMO or water as a control that were inoculated using transfer of 25 viruliferous aphids per leaf at 0, 2, 4, 7, 9, 11 and 14 days after treatment (DAT). Persistent effects of HMO residue on the probing and feeding behaviours of GPA were also monitored with the electrical penetration graph (EPG) system. For glasshouse‐grown peach seedlings, the residual activity of HMO reduced PPV infection rates by more than 58% for up to 4 DAT following an initial reduction of approximately 81%. EPG recordings of GPA feeding behaviour showed that HMO significantly delayed first feeding probes and first intracellular punctures by more than 50 min without changing the ensuing stylet penetration behaviour. Applying HMO reduced virus infection rates for up to a week depending on the environmental conditions. EPG monitoring of aphid probing showed that HMO reduced the mean duration and mean number of potential drop (PD) phase feeding occurrences, compared with the water control. A reduction in the PD that has been shown to be related to the transmission of non‐persistently transmitted viruses may partly explain the reduction in PPV infection rates.     

梯棱羊肚菌全基因组SNP/Indel分析及基于Indel标记的遗传连锁图谱构建

基于梯棱羊肚菌Morchella importuna两个子囊孢子培养物的全基因组测序数据,对全基因组范围内的变异位点进行分析。共鉴定到18 438个变异位点,平均每Mbp的变异位点数量为361个;变异位点以单核苷酸多态性SNP为主,共计17 104个,基因组中SNP的频率为335SNPs/Mbp;Indel多态性位点1 334个,以2-10bp的插入缺失为主;73.4% SNP/Indel位于基因间隔区域,外显子区域共检测到3 042个变异位点,占总数的16.50%;对基因功能产生确定影响的移码突变有1 088个,占5.90%,错义突变916个,占比4.97%;不同Scaffold上的SNP/Indel出现频率不同,SNP频率最大的为Scaffold80,平均每Mbp包含2 856个SNP,频率最低为Scaffold60和Scaffold75,分别为16SNPs/Mbp和30SNPs/Mbp;对≥11bp的Indel变异位点进行标记开发和多态性群体分析,成功开发出75对Indel标记。采用原生质体单细胞分离技术,获得了梯棱羊肚菌M04的两个可亲和的同核体菌株M04P01和M04P40,同时采用来自M04子囊果的58个单孢菌株作为作图群体,初步构建了包含75个Indel标记和1个交配型基因的梯棱羊肚菌遗传连锁图谱,共获得12个连锁群,连锁群总长度273.7cM。