A study of the pathogenesis of Trichostrongylus colubriformis infection in lambs with observations on the contribution of gastrointestinal plasma loss

A study of the pathogenesis of Trichostrongylus colubriformis infection in lambs with observations on the contribution of gastrointestinal plasma loss. International Journal for Parasitology 3: 743–757. Increased effusion into the gut of⁵¹Cr-labelled plasma protein was observed in 20-week-old Merino lambs which showed signs of trichostrongylosis after administration of 100,000 T. colubriformis larvae. In another trial, similarly infected 12-week-old meat type crossbred lambs did not have elevated gastrointestinal plasma loss, or signs of trichostrongylosis, although they generally carried heavier worm burdens than the Merinos. Differences in breed susceptibility were considered a possible explanation. Nine-week-old Merino lambs inoculated with 85,000 T. colubriformis had increased plasma loss into the gut beginning 10–12 days after infection, coinciding with onset of inappetance, hypoproteinaemia and weight loss. Weight loss was considered largely due to reduced feed intake, but infected lambs during the period of protein effusion into the gut lost more weight, or gained less, than pair fed controls. Hypoproteinaemia was related mainly to effusion of plasma into the gut. One infected lamb, and its pair fed control, died, probably due to malnutrition. Slight anaemia, and depressed faecal dry matter per cent were observed in infected lambs.     

Sea trout as a carrier of infectious salmon anaemia virus

Sea trout did not develop symptoms or gross clinical signs of infectious salmon anaemia (ISA) when injected with ascites from salmon, but ISA virus was propagated and haematocrit values dropped. The virus was transmitted from trout to salmon.     

Kinetics of the Hb A to Hb C switch and erythropoietin plasma levels in sheep

The induction of Hb A (alpha 2 beta A2) and Hb C (alpha 2 beta C2) synthesis in three adult sheep has been sequentially analysed, in relation to the reduction of the haematocrit (Ht) and to the changes of erythropoietin (Epo) concentration in plasma. Hb A production is detected in peripheral reticulocytes when the Ht approaches 70% of its initial value in correspondence with the first rise of EPO plasma level, whereas HB C synthesis appears when the Ht is further reduced to about 50%, at an Epo concentration two to three times higher. The assumption that the cell committed to HB C synthesis is close to the erythroid colony-forming unit (CFU-e) progenitor is also discussed.     

鸡贫血病毒VP1和VP1蛋白在家蚕中的联合表达

将鸡贫血病毒vp1和vp2基因分别克隆入转移载体pBacPAK8中,获得重组转移质粒pBac-vp1和pBac-vp2。以上两质粒分别与CunI酶切线性化的亲本病毒Bm-Bacpak6DNA共转染家蚕细胞,通过蓝白斑筛选,纯化得到重组病毒Bm-vp1和Bm-vp2。PCR分析表明vp1和vp2基因已整合进杆状病毒基因组中。将Bm-vp1和Bm-vp2共感染5龄家蚕,通过表达产物免疫SPF鸡产生的抗血清与CAV感染的MDCC－MSB1细胞的间接荧光抗体分析,证明表达产物能诱导鸡产生的抗体,而且能够保护子代鸡免受CAV的攻击。该研究表明,表达VP1和VP2蛋白的重组家蚕杆状病毒（Recombinant BmNP)是很有前途的CAV亚单位疫苗的生产系统。     

用PCR扩增和克隆鸡传染性贫血病毒全基因组DNA

根据已发表的序列,分别在鸡贫血病毒（ＣＡＶ）环形基因组ＤＮＡ（全长２．３ｋｂ）的ＥｃｏＲＩ位点和ＢａｍＨＩ位点的两侧选择适当序列合成两对引物,用ＰＣＲ技术,从斑点杂交检测到病毒核酸的ＣＡＶ感染的ＭＤＣＣＲＰ１细胞基因组ＤＮＡ中,分别扩增出包含ＥｃｏＲＩ和ＢａｍＨＩ分割开的病毒基因组两部分（１．５ｋｂ和０．８ｋｂ）约１．５ｋｂ和约１．２５ｋｂ的两个片段。再将其中相应序列拼接克隆进ｐＵＣ１８载体,获得包含ＣＡＶ全基因组序列ＤＮＡ片段的克隆质粒ｐＣＡＶ２．４。酶切分析表明,该质粒具有预期的ＢａｍＨＩ位点、ＰｓｔＩ位点、ＨｉｎｄⅢ位点,而预期的ＥｃｏＲＩ位点消失。重组质粒插入ＤＮＡ片段的两端序列分析表明,质粒ｐＣＡＶ２．４是包含ＣＡＶ全基因组序列的重组质粒,插入ＤＮＡ片段序列中的ＥｃｏＲＩ位点序列发生了一个碱基突变。     

Validation of the use of phenylhydrazine hydrochloride (PHZ) for experimental manipulation of haematocrit and plasma haemoglobin in birds

The levels of haematocrit (Hct) and plasma haemoglobin (Hb) vary markedly through the annual cycle of birds, as well as among individuals at all life‐stages (embryos, chicks, adults). It is thus surprising that the functional, fitness‐related consequences of this variation are poorly understood. Putative ‘adaptive’ variation in these haematological traits has been associated with varying demands for aerobic capacity and oxygen transport, for example during migration, winter acclimatization, with increasing altitude, or during parental care. It has also been proposed that changes in Hct might reflect ‘costs’ of parental investment, for example during ‘reproductive anaemia’. However, almost all studies to date have been correlative. Here we describe a series of experiments that validate the use of phenylhydrazine hydrochloride (PHZ) for the transient, reversible experimental manipulation of Hct and Hb in birds. A single bolus injection (12.5 μg PHZ/g body weight delivered via intra‐muscular injection) caused a rapid decrease in Hct and plasma Hb within 24 h, from pretreatment values of 50–54% to 40–44% in non‐breeding Zebra Finches Taenipoygia guttata and European Starlings Sturnus vulgaris, and to 35% in breeding female Zebra Finches, changes within the normal physiological range. Hct and Hb returned to pre‐injection levels within 5–10 days of treatment. Changes in plasma Hb paralleled those for Hct. We suggest that PHZ treatment provides a widely applicable technique for use in experimental work to establish relationships between haematological status, aerobic capacity, workload (e.g. migration, parental care, thermoregulation), individual quality (of both adults and chicks) and trade‐offs such as costs of reproduction.     

鸡贫血病毒VP1基因在大肠杆菌中的高效表达及其生物学特性的初步分析

将鸡贫血病毒(CAV)VP1蛋白449个氨基酸中的424个氨基酸(占95%)编码序列分二段,分别克隆入表达性载体pGEX-5X-3,在大肠杆菌以GST融合蛋白的形式获得了高效表达.以表达产物免疫小鼠4次,制备了抗CAV的多克隆抗体.通过对CAV感染的MSB1细胞做间接免疫荧光试验(IFA),结果为阳性.这表明表达物保留了天然VP1相关的抗原性.用此抗CAV抗体和所建立的IFA方法对临床CAV凝似病料成功地进行了实验室诊断.     

?Np73 is capable of inducing apoptosis by co-ordinately activating several BH3-only proteins

Inactivation of p53 is one of the most relevant events in human cancer, since it allows transformed cells to escape their own proliferation control and leave them irresponsive to drugs that aim to damage their DNA. When p53 falls, other members of its family may become targets to attack tumoural cells. p73 has shown capacity to mediate these attacks. However, its N-terminal truncated isoforms have been associated with oncogenesis due to their capacity to act as dominant negatives of p53 and the transactivation (TA) isoforms of p73. We previously found a relationship between the overexpression of N-terminus-truncated p73 isoform (∆Np73) and that of the proapoptotic gene Bcl-2-interacting killer (BIK). In the present report we demonstrate that ∆Np73-α has the capacity to induce apoptosis through the co-ordinated activation of a group of genes harbouring GC-rich elements in their regulatory regions. ∆Np73-α synergizes with specificity protein (Sp1) on these elements but the overall response of these genes probably depends on the additional presence of consensus p53 elements. We explore the domains of ∆Np73-α involved in this transactivation capacity and found divergences with the previously described functions for them. Moreover, we found that the transforming mutation V12 of HRas impairs this transactivation capacity of ∆Np73-α, further supporting the anti-tumoural function of this later. Our data add complexity to the action of p73 on the induction of apoptosis and tumourogenesis, opening new interpretations to the expression profile of p73 isoforms in different human neoplasias.     

Babesia major in Britain: blood-induced infections in splenectomized and intact calves

A strain of Babesia major, originally isolated from field collections of Haemaphysalis punctata in Kent, England was maintained in splenectomized calves by the intravenous inoculation of infected blood. Rapid passage from carrier calves, that had recovered from a tick-induced infection, resulted in a marked increase in virulence; 4 out of 6 calves of the second passage underwent fatal infections and the others suffered severe reactions.Five splenectomized and 5 intact calves of the same breed and age were infected with the same number of infected erythrocytes (RBC). The intact calves reacted mildly with maximum parasite counts ranging from < $\text{1}\text{1000}$ RBC to $\text{5}\text{1000}$ RBC; haemoglobin levels and packed cell volume values, however, fell sharply but recovered swiftly. The group of splenectomized calves exhibited one fatal case, 2 severe reactions and 2 mild infections; maximum parasitaemias varied from $\text{7}\text{1000}$ RBC to $\text{322}\text{1000}$ RBC. Packed cell volumes and haemoglobin concentrations declined to low levels and took several weeks to return to normal.It is concluded that B. major should be regarded as a potential pathogen of British cattle.     

The identification of six risk genes for ovarian cancer platinum response based on global network algorithm and verification analysis

Ovarian cancer is the most lethal gynaecological cancer, and resistance of platinum‐based chemotherapy is the main reason for treatment failure. The aim of the present study was to identify candidate genes involved in ovarian cancer platinum response by analysing genes from homologous recombination and Fanconi anaemia pathways. Associations between these two functional genes were explored in the study, and we performed a random walk algorithm based on reconstructed gene‐gene network, including protein‐protein interaction and co‐expression relations. Following the random walk, all genes were ranked and GSEA analysis showed that the biological functions focused primarily on autophagy, histone modification and gluconeogenesis. Based on three types of seed nodes, the top two genes were utilized as examples. We selected a total of six candidate genes (FANCA, FANCG, POLD1, KDM1A, BLM and BRCA1) for subsequent verification. The validation results of the six candidate genes have significance in three independent ovarian cancer data sets with platinum‐resistant and platinum‐sensitive information. To explore the correlation between biomarkers and clinical prognostic factors, we performed differential analysis and multivariate clinical subgroup analysis for six candidate genes at both mRNA and protein levels. And each of the six candidate genes and their neighbouring genes with a mutation rate greater than 10% were also analysed by network construction and functional enrichment analysis. In the meanwhile, the survival analysis for platinum‐treated patients was performed in the current study. Finally, the RT‐qPCR assay was used to determine the performance of candidate genes in ovarian cancer platinum response. Taken together, this research demonstrated that comprehensive bioinformatics methods could help to understand the molecular mechanism of platinum response and provide new strategies for overcoming platinum resistance in ovarian cancer treatment.