Propagation in Human Cells of a Filterable Agent from the ST Feline Sarcoma

Nineteen lines of human fibroblasts were inoculated with a filtrate prepared from the ST feline sarcoma. Seven lines showed morphological alteration and released focus-forming activity for feline cells, 2 lines showed morphological alteration but did not release focus-forming activity, and 11 lines showed no morphological alteration and released no focus-forming agent. Morphologically altered cells appeared enlarged, hyper-refractile, and intensely stained by hematoxylin. They neither assumed a crisscross pattern nor piled up to form a visible focus. Time-lapse cinegraph showed that the morphologically altered cells did not divide and were motile. The fluid from two human fibroblast cultures, inoculated 4 and 14 weeks previously with the ST sarcoma filtrate, induced fibrosarcoma in newborn kittens.     

Phylogenetic Analysis Reveals a High Prevalence of Sporothrix brasiliensis in Feline Sporotrichosis Outbreaks

Sporothrix schenckii, previously assumed to be the sole agent of human and animal sporotrichosis, is in fact a species complex. Recently recognized taxa include S. brasiliensis, S. globosa, S. mexicana, and S. luriei, in addition to S. schenckii sensu stricto. Over the last decades, large epidemics of sporotrichosis occurred in Brazil due to zoonotic transmission, and cats were pointed out as key susceptible hosts. In order to understand the eco-epidemiology of feline sporotrichosis and its role in human sporotrichosis a survey was conducted among symptomatic cats. Prevalence and phylogenetic relationships among feline Sporothrix species were investigated by reconstructing their phylogenetic origin using the calmodulin (CAL) and the translation elongation factor-1 alpha (EF1α) loci in strains originated from Rio de Janeiro (RJ, n = 15), Rio Grande do Sul (RS, n = 10), Paraná (PR, n = 4), São Paulo (SP, n = 3) and Minas Gerais (MG, n = 1). Our results showed that S. brasiliensis is highly prevalent among cats (96.9%) with sporotrichosis, while S. schenckii was identified only once. The genotype of Sporothrix from cats was found identical to S. brasiliensis from human sources confirming that the disease is transmitted by cats. Sporothrix brasiliensis presented low genetic diversity compared to its sister taxon S. schenckii. No evidence of recombination in S. brasiliensis was found by split decomposition or PHI-test analysis, suggesting that S. brasiliensis is a clonal species. Strains recovered in states SP, MG and PR share the genotype of the RJ outbreak, different from the RS clone. The occurrence of separate genotypes among strains indicated that the Brazilian S. brasiliensis epidemic has at least two distinct sources. We suggest that cats represent a major host and the main source of cat and human S. brasiliensis infections in Brazil.     

Accessory Genes Confer a High Replication Rate to Virulent Feline Immunodeficiency Virus

Feline immunodeficiency virus (FIV) is a lentivirus that causes AIDS in domestic cats, similar to human immunodeficiency virus (HIV)/AIDS in humans. The FIV accessory protein Vif abrogates the inhibition of infection by cat APOBEC3 restriction factors. FIV also encodes a multifunctional OrfA accessory protein that has characteristics similar to HIV Tat, Vpu, Vpr, and Nef. To examine the role of vif and orfA accessory genes in FIV replication and pathogenicity, we generated chimeras between two FIV molecular clones with divergent disease potentials: a highly pathogenic isolate that replicates rapidly in vitro and is associated with significant immunopathology in vivo, FIV-C36 (referred to here as high-virulence FIV [HV-FIV]), and a less-pathogenic strain, FIV-PPR (referred to here as low-virulence FIV [LV-FIV]). Using PCR-driven overlap extension, we produced viruses in which vif, orfA, or both genes from virulent HV-FIV replaced equivalent genes in LV-FIV. The generation of these chimeras is more straightforward in FIV than in primate lentiviruses, since FIV accessory gene open reading frames have very little overlap with other genes. All three chimeric viruses exhibited increased replication kinetics in vitro compared to the replication kinetics of LV-FIV. Chimeras containing HV-Vif or Vif/OrfA had replication rates equivalent to those of the virulent HV-FIV parental virus. Furthermore, small interfering RNA knockdown of feline APOBEC3 genes resulted in equalization of replication rates between LV-FIV and LV-FIV encoding HV-FIV Vif. These findings demonstrate that Vif-APOBEC interactions play a key role in controlling the replication and pathogenicity of this immunodeficiency-inducing virus in its native host species and that accessory genes act as mediators of lentiviral strain-specific virulence.     

Breakdown of Meiosis in a Tetraploid Clone from Dichanthium-Bothriochloa Complex

The paper describes the details of meiosis in a tetraploid clone(collection number 260) from Dichanthium-Bothriochloa complex.Meiosis is reported to have broken down very early in prophaseI. Pollen mother cells are reported to be present in the formof plasmodial mass or syncyte with several nuclei in each ofthem. Each syncyte, presumably, gave rise to one pollen grainwithout undergoing division. Consequently, each pollen grainhad one to several nuclei. A brief note on morphology and taxonomy of the collection isalso added. Although the spikelet morphology indicated it tobe a natural hybrid Bothrio-chloa grahamii x B. intermedia,in other morphological characters, it resembled Dichanthiumannulatum. It was concluded, therefore, that it was a productof continuous natural introgression.     

新疆紫草的无性繁殖及其再生植株的遗传稳定性初探

诱导新疆紫草多种外植体脱分化均已成功,嫩芽切段最快,胚轴切段次之。培养期间,2,4－D不可缺少,KT的作用优于6－BA。降低2,4－D浓度利于愈伤组织扩增和再分化。KT或6－BA都可诱导出不定芽,前者还有利于球形胚的形成。适当的昼夜温差（10－28℃）和自然光照有利于壮苗的形成和移栽成活。的疆紫草种子萌发苗根细胞染色体为14条,再生试管苗根细胞染色体为14条的占97.1%。     

Capsid Functions of Inactivated Human Picornaviruses and Feline Calicivirus

The exceptional stability of enteric viruses probably resides in their capsids. The capsid functions of inactivated human picornaviruses and feline calicivirus (FCV) were determined. Viruses were inactivated by UV, hypochlorite, high temperature (72°C), and physiological temperature (37°C), all of which are pertinent to transmission via food and water. Poliovirus (PV) and hepatitis A virus (HAV) are transmissible via water and food, and FCV is the best available surrogate for the Norwalk-like viruses, which are leading causes of food-borne and waterborne disease in the United States. The capsids of all 37°C-inactivated viruses still protected the viral RNA against RNase, even in the presence of proteinase K, which contrasted with findings with viruses inactivated at 72°C. The loss of ability of the virus to attach to homologous cell receptors was universal, regardless of virus type and inactivation method, except for UV-inactivated HAV, and so virus inactivation was almost always accompanied by the loss of virus attachment. Inactivated HAV and FCV were captured by homologous antibodies. However, inactivated PV type 1 (PV-1) was not captured by homologous antibody and 37°C-inactivated PV-1 was only partially captured. The epitopes on the capsids of HAV and FCV are evidently discrete from the receptor attachment sites, unlike those of PV-1. These findings indicate that the primary target of UV, hypochlorite, and 72°C inactivation is the capsid and that the target of thermal inactivation (37°C versus 72°C) is temperature dependent.     

Transformation and Productive Infection of Human Osteosarcoma Cells by a Feline Sarcoma Virus

FELINE sarcoma virus (FSV) transforms human embryo cells in vitro¹; it therefore seemed interesting to determine whether this virus could transform human osteosarcoma cells. Defective Moloney sarcoma virus genome can be rescued from non-producer hamster tumour cells by feline leukaemia virus (FeLV)² and because FSV stocks also contain excess FeLV (ref. 1 and unpublished observations of R. V. G.), it was hoped that human osteosarcoma cells transformed by FSV and co-infected with FeLV might yield a human sarcoma virus.     

Human Angiostrongyliasis Outbreak in Dali, China

Background

Several angiostrongyliasis outbreaks have been reported in recent years but the disease continues to be neglected in public health circles. We describe an outbreak in Dali, southwest China in order to highlight some key problems for the control of this helminth infection.

Methodology/Principal Findings

All available medical records of suspected angiostrongyliasis patients visiting hospitals in Dali in the period 1 October 2007–31 March 2008 were reviewed, and tentative diagnoses of varying strengths were reached according to given sets of criteria. Snails collected from local markets, restaurants and natural habitats were also screened for the presence of Angiostrongylus cantonensis. A total of 33 patients met criteria for infection, and 11 among them were classified as clinically confirmed. An additional eight patients were identified through a surveillance system put in operation in response to the outbreak. The epidemic lasted for 8 months with its peak in February 2008. Of the 33 patients, 97.0% complained of severe headache. 84.8% patients had high eosinophil cell counts either in the peripheral blood or in cerebrospinal fluid (CSF). Three-quarters of the patients were treated with a combination of albendazole and corticosteroids, resulting in significantly improved overall conditions. Twenty-two patients reported the consumption of raw or undercooked snails prior to the onset of the symptoms, and approximately 1.0% of the Pomacea canaliculata snails on sale were found to be infected with A. cantonensis. The snails were also found in certain habitats around Dali but no parasites were detected in these populations.

Conclusions/Significance

The import and sale of infected P. canaliculata is the likely trigger for this angiostrongyliasis outbreak. Awareness of angiostrongyliasis must be raised, and standardized diagnosis and treatment are needed in order to provide clinicians with a guide to address this disease. Health education campaigns could limit the risk, and a hospital-based surveillance system should be established in order to detect future outbreaks.     

Whole-Genome Analysis of a Rare Human Korean G3P Rotavirus Strain Suggests a Complex Evolutionary Origin Potentially Involving Reassortment Events between Feline and Bovine Rotaviruses

A rare human rotavirus, G3P[9] strain RVA/Human-tc/KOR/CAU12-2-51/2013/G3P[9], was isolated from the stool of a 9-year-old female hospitalized with acute watery diarrhea in August 2012 in South Korea using a cell culture system, and its genome was analyzed. The complete genomic constellation of the CAU12-2-51 strain revealed a novel genotype constellation for human rotavirus, G3-P[9]-I2-R2-C2-M2-A3-N2-T3-E3-H3. Phylogenetic analysis revealed that the CAU12-2-51 strain originated from feline- and bovine-like reassortment strains. The genes encoding VP4, VP7, NSP1, NSP3, NSP4, and NSP5 were related to human/feline-like and feline rotavirus strains, whereas the remaining five genes encoding VP1, VP2, VP3, VP6, and NSP2 were related to the human/bovine-like and bovine rotavirus strains. This novel strain was identified for the first time, providing evidence of feline/bovine-to-human transmission of rotavirus. The data presented herein provide information regarding rotavirus diversity and evolution.     

CXCR4 Is Required by a Nonprimate Lentivirus: Heterologous Expression of Feline Immunodeficiency Virus in Human, Rodent, and Feline Cells

A heterologous feline immunodeficiency virus (FIV) expression system permitted high-level expression of FIV proteins and efficient production of infectious FIV in human cells. These results identify the FIV U3 element as the sole restriction to the productive phase of replication in nonfeline cells. Heterologous FIV expression in a variety of human cell lines resulted in profuse syncytial lysis that was FIV env specific, CD4 independent, and restricted to cells that express CXCR4, the coreceptor for T-cell-line-adapted strains of human immunodeficiency virus. Stable expression of human CXCR4 in CXCR4-negative human and rodent cell lines resulted in extensive FIV Env-mediated, CXCR4-dependent cell fusion and infection. In feline cells, stable overexpression of human CXCR4 resulted in increased FIV infectivity and marked syncytium formation during FIV replication or after infection with FIV Env-expressing vectors. The use of CXCR4 is a fundamental feature of lentivirus biology independent of CD4 and a shared cellular link to infection and cytopathicity for distantly related lentiviruses that cause AIDS. Their conserved use implicates chemokine receptors as primordial lentivirus receptors.     

一步法扩增克隆IBDV上海超强毒VP2—4—3基因

分离、纯化了鸡传染性法氏囊病病毒超强毒 (vvIBDV)上海株SH95的病毒核酸dsRNA ,应用随机引物将RNA反转录成cDNA ,以此为模板一步扩增出A片段前体融合蛋白基因即VP2 4 3基因 ,将其克隆入 pGEM T载体 ,并进行序列分析 ,其与超强毒株HK4 6的核苷酸序列的同源性达 98% ,整个基因有 5个氨基酸差异 ,同源性达99.5 1% (10 0 7/ 10 12 )。     

人NDRG2启动子的克隆与活性鉴定

目的:克隆人NDRG2的启动子,并进行启动子的活性鉴定。方法:用Advantage-GC Taq酶,采用PCR方法,以BCA克隆R-998D1为模板,克隆人NDRG2(-1455/ 274)的启动子。分别构建NDRG2(-1131/ 274)、NDRG2(-273/ 274)、NDRG2(-135/ 274)、NDRG2(-79/ 274)和NDRG2(-79/ 57)等不同长度的截短体,并分别亚克隆入pGL3-basic报告基因载体,测序鉴定。分别转染HEK293和HeLa细胞后,运用双荧光报告基因系统进行启动子活性分析,从而判断核心启动子的位置。结果:人NDRG2的启动子克隆成功,并构建了不同长度的截短体,DNA测序结果与报道一致。报告基因分析结果将人NDRG2启动子的核心区域定位于NDRG2(-79/ 57)。结论:随着人NDRG2的启动子的长度逐渐被截短,启动子的基础活性也逐渐降低,可将人NDRG2启动子的核心区域定位于NDRG2(-79/ 57)。     

一步法扩增克隆IBDV上海超强毒VP2-4-3基因

分离、纯化了鸡传染性法氏囊病病毒超强毒 (vvIBDV) 上海株SH95的病毒核酸dsRNA,应用随机引物将RNA反转录成cDNA,以此为模板一步扩增出A片段前体融合蛋白基因即VP2-4-3基因,将其克隆入pGEM-T载体,并进行序列分析,其与超强毒株HK46的核苷酸序列的同源性达98%,整个基因有5个氨基酸差异,同源性达99.51%(1007/1012).     

Feline Preleukemia: An Animal Model of Human Disease

In man, hematologic abnormalities precede the development of acute myeloblastic leukemia in about one-third of individuals. This preleukemic state may represent a stage of adult leukemia wherein small numbers of leukemic cells are present and the normal marrow stem cell compartment has not been seriously compromised. A syndrome resembling human preleukemia occurs in cats infected with feline leukemia virus (FeLV). This disorder is characterized by anemia, leukopenia or thrombocytopenia occurring weeks or months prior to the development of feline acute leukemia. The natural occurrence of this syndrome in this domestic animal population makes it a potential model of human preleukemia. Initial poor results of therapy of human preleukemia presently prohibit one from carrying out controlled trials with chemotherapeutic agents in such a group of patients. Preliminary trials with chemo- and/or immunotherapy may be more easily attempted with FeLV infected preleukemic cats.     

An Outbreak of Human Fascioliasis gigantica in Southwest China

Fascioliasis is a common parasitic disease in livestock in China. However, human fascioliasis is rarely reported in the country. Here we describe an outbreak of human fascioliasis in Yunnan province. We reviewed the complete clinical records of 29 patients and performed an epidemiological investigation on the general human population and animals in the outbreak locality. Our findings support an outbreak due to Fasciola gigantica with a peak in late November, 2011. The most common symptoms were remittent fever, epigastric tenderness, and hepatalgia. Eosinophilia and tunnel-like lesions in ultrasound imaging in the liver were also commonly seen. Significant improvement of patients’ condition was achieved by administration of triclabendazole®. Fasciola spp. were discovered in local cattle (28.6%) and goats (26.0%). Molecular evidence showed a coexistence of F. gigantica and F. hepatica. However, all eggs seen in humans were confirmed to be F. gigantica. Herb (Houttuynia cordata) was most likely the source of infections. Our findings indicate that human fascioliasis is a neglected disease in China. The distribution of triclabendazole®, the only efficacious drug against human fascioliasis, should be promoted.     

Genetic Analysis of a Mouse t Complex Locus That Is Homologous to a Kidney Cdna Clone

A mouse kidney cDNA clone, pMK174, identifies restriction fragment length polymorphisms (RFLPs) that map to two unlinked loci. One, designated D17Rp17, has been mapped near quaking, (qk), on chromosome 17 using three sets of recombinant inbred (RI) strains. A study of several t haplotypes resulted in the identification of t-specific alleles of D17Rp17 that map to the proximal half of the t complex. Neither t-specific nor wild-type D17Rp17 alleles are present in chromosomes carrying either the T Orleans (TtOrl) or the T hairpin tail (Thp) deletions. Comparison with other molecular markers indicates that pMK174 identifies a new proximal t complex locus, Rp17. The second locus identified by pMK174, termed D4Rp18, is tentatively assigned to chromosome 4 by mouse-Chinese hamster somatic cell hybrid analysis.     

Infection and Propagation of Human Rhinovirus C in Human Airway Epithelial Cells

Human rhinovirus species C (HRV-C) was recently discovered using molecular diagnostic techniques and is associated with lower respiratory tract disease, particularly in children. HRV-C cannot be propagated in immortalized cell lines, and currently sinus organ culture is the only system described that is permissive to HRV-C infection ex vivo. However, the utility of organ culture for studying HRV-C biology is limited. Here, we report that a previously described HRV-C derived from an infectious cDNA, HRV-C15, infects and propagates in fully differentiated human airway epithelial cells but not in undifferentiated cells. We demonstrate that this differentiated epithelial cell culture system supports infection and replication of a second virus generated from a cDNA clone, HRV-C11. We show that HRV-C15 virions preferentially bind fully differentiated airway epithelial cells, suggesting that the block to replication in undifferentiated cells is at the step of viral entry. Consistent with previous reports, HRV-C15 utilizes a cellular receptor other than ICAM-1 or LDLR for infection of differentiated epithelial cells. Furthermore, we demonstrate that HRV-C15 replication can be inhibited by an HRV 3C protease inhibitor (rupintrivir) but not an HRV capsid inhibitor previously under clinical development (pleconaril). The HRV-C cell culture system described here provides a powerful tool for studying the biology of HRV-C and the discovery and development of HRV-C inhibitors.