Models for Trypanosoma evansi (surra), its control and economic impact on small-hold livestock owners in the Philippines

Simple demographic and infectious disease models of buffaloes and other domestic hosts for animal trypanosomosis (surra) caused by Trypanosoma evansi were developed. The animal models contained deterministic and stochastic elements and were linked to simulate the benefit of control regimes for surra in village domestic animal populations in Mindanao, Philippines. The impact of the disease on host fertility and mortality were key factors in determining the economic losses and net-benefit from the control regimes. If using a high (99%) efficacy drug in surra-moderate to high risk areas, then treating all animals twice each year yielded low prevalence in 2 years; targeted treatment of clinically sick animals, constantly monitored (monthly), required 75% fewer treatments but took longer to reach a low prevalence than treating all animals twice each year. At high drug efficacy both of these treatment strategies increased the benefit over untreated animals by 81%. If drug efficacy declined then the benefit obtained from twice yearly treatment of all animals declined rapidly compared with regular monitoring and targeting treatment to clinically sick animals. The current control regimen applied in the Philippines of annual sero-testing for surra and only treating sero-positive animals provided the lowest net-benefit of all the control options simulated and would not be regarded as effective control. The total net-benefit from effective surra control for a typical village in a moderate/high risk area was 7.9 million pesos per annum (US $158,000). The value added to buffaloes, cattle, horses, goats/sheep and pigs as a result of this control was US $88, $84, $151, $7, $114 per animal/year, respectively.     

Characterization of tubule and monomer derived from VP4 protein of infectious bursal disease virus

The VP4 protein of infectious bursal disease virus (IBDV) is a serine protease that processes the polyprotein for viral assembly. VP4 has been found to associate primarily with type II IBDV tubules that are 24 nm in diameter. In this study, a chimeric VP4, assigned as HS1VP4, was constructed with a VP4-autocleavage site inserted between the N-terminal His-tag and the VP4 sequence. The results showed that the VP4 forms tubules after the self-cleavage of HS1VP4 when expressed in Escherichia coli. Furthermore, a deletion of 28 amino acids at the C-terminus of VP4 resulted in monomers and dimers instead of tubule formation; mutants of S652A and K692A at active site destroyed the activity. The endopeptidase activity of these monomers and dimers was approximately 12.5 times higher than that of VP4 tubules. Additionally, the formation of tubules inhibited VP4 protease activity, as demonstrated through in vitro assays. The production and characterization of monomers or dimers that have greater endopeptidase activity and protease activity than tubules can provide further insight into VP4 tubule assembly and the regulation of VP4 activity in host cells; this insight will facilitate the development of new anti-IBDV strategies.     

Inhibition of human lymphocyte activation by wheat germ agglutinin: a model for saccharide-specific suppressor factors

The suppressive effect of wheat germ agglutinin (WGA) on lectin-stimulated blastogenesis and immunoglobulin production was studied. Addition of WGA at 10 micrograms/ml inhibited phytohemagglutinin (PHA)-, concanavalin-A (Con-A)-, and pokeweed mitogen (PWM)-induced mitogenic responses by 70-80%. PWM-driven immunoglobulin synthesis was suppressed by 45% with WGA. The inhibitory effects of WGA were not due to cell death or to interference with lectin binding at the cell surface. Inhibition was dependent on the presence of WGA in the cell culture during the first 24 hr of mitogen exposure and was observed in cultures of both monocyte-depleted peripheral blood mononuclear cells as well as T-cell-enriched populations. WGA-induced inhibition of blastogenesis was blocked by the addition of N-acetylglucosamine (GluNAc) which prevents WGA binding to the cell surface. WGA was found to mimic the suppressive effect of a soluble immune suppressor supernatant (SISS) derived from Con-A-activated mononuclear cell cultures. PHA responses were inhibited by 80 and 95% with SISS and WGA, respectively. The inhibition by both WGA and SISS was totally reversed with addition of GluNAc. Furthermore, WGA and SISS demonstrated competition for the same cell surface receptor site. WGA may therefore be useful as an in vitro model of a saccharide-specific, biologically relevant, soluble mediator for the investigation of mechanisms of immunologic suppression.     

Immunologic and clinical improvement of progressive coccidioidomycosis following administration of transfer factor

Three patients with progressive coccidioidomycosis were given preparations of transfer factor (TF). Adverse reactions to TF were minimal. Following TF administration two of these patients had prolonged clinical remissions in their coccidioidal disease. Cellular immune responses were sequentially evaluated by coccidioidininduced delayed-type skin tests, lymphocyte blast transformation and macrophage inhibition factor production (MIF). These three patients each exhibited different cellular immune patterns before and after TF administration. Two patients converted their coccidioidin skin tests, and one converted lymphocyte transformation response to coccidioidin. Also, TF apparently favorably affected the MIF response in all three patients.     

牛肝提取物提高仔鸡免疫力的研究

将牛肝提取物注射到第13日龄和第15日龄鸡胚中能显著提高孵出后仔鸡的抗SRBC血清抗体的效价,促进脾和法氏囊淋巴细胞增殖、分化。同时,牛肝提取物与雏鸡法氏囊粗提取液,抗鸡法氏囊病毒抗体均有增强仔鸡抵抗传染性法氏囊病的能力。实验结果表明牛肝提取物中可能含有类似法氏囊素的物质。     

表达HSV－2 gD基因的重组痘苗病毒保护小鼠对抗致死量HSV病霉攻击

利用ＰＣＲ方法对单纯疱疹病毒Ⅱ型糖蛋白Ｄ（ＨＳＶ－２ｇＤ）基因进行了修饰,在其５＇端删去约５００ｂｐ的非编码区,仅保留ＡＴＧ上游７个ｂｐ。将修饰后的ＨＳＶ－２ｇＤ基因插入到带有痘苗病毒天坛株ＴＫ基因区段的痘苗表达质粒ｐＪＳＡ１１７５,置于痘苗病毒Ｐ７．５ｋ早／晚期启动子控制下。将此重组质粒用脂质体Ｌｉｐｏｆｅｃｔｉｎ方法转染已受野型ＴＫ￣＋痘苗病毒天坛株感染的ＴＫ￣－１４３细胞,通过同源重组机制和标志基因ＬａｃＺ产物的蓝斑显色作用,以及ＢｕｄＲ试剂对ＴＫ表型的选择压力,筛选出整合有ＨＳＶ－２ｇＤ基因的重组痘苗病毒。Ｓｏｕｔｈｅｍ杂交表明,ＨＳＶ－２ｇＤ基因已正确地插入痘苗病毒ＴＫ基因区内;间接免疫荧光检测显示,ＨＳＶ－２ｇＤ蛋白已得到有效表达,且主要分布于细胞膜。重组病毒免疫家兔可产生明显的抗ＨＳＶ－２ｇＤ中和抗体。用重组病毒免疫小鼠,３周后可使９４％（１７／１８）的小鼠对抗ＨＳＶ－２的致死量攻击,表明重组病毒具有明显的免疫保护作用。     

Chronic Pseudomonas aeruginosa endobronchitis in rhesus monkeys: II. A histopathologic analysis

We have recently established a rhesus monkey model of chronic Pseudomonas aeruginosa (PA) endobronchitis by bronchoscopic instillation of PA-embedded agar beads. All experimental animals developed chronic neutrophilic endobronchitis similar to chronic PA endobronchitis in cystic fibrosis (CF). Histopathologic studies further confirmed similarities to chronic PA endobronchitis in CF, including marked peribronchial inflammation, epithelial damage, presence of degraded cilia and ciliary abnormalities, appearance of PA bacterial clusters, mucosal hyperplasia, goblet cell hypertrophy/hypersecretion, airway obstruction, alveolar abnormalities, bronchiectasis, and fibrosis.     

犬传染性肝炎病毒在体外细胞质内的发生

通过对犬传染性肝炎病毒（ＩＣＨＶ）在犬肾传代细胞内形态发生及其抗原定位的电镜和免疫胶体金电镜研究,发现ＩＣＨＶ除了在宿主细胞核内发生外,还有一条细胞质内的发生途径。在细胞质内病毒核壳体的装配是以均质致密包涵体和副晶格包涵体为“基地”,这与人们熟知的细胞核内形态发生方式相似。免疫胶体金标记显示,细胞质包涵体中含有大量的ＩＣＨＶ抗原成分,显核壳体在细胞质内装配病毒的结构蛋白来源。此外,在感染的细胞质内还观察到与核内相同的病毒核心样结构。