Nerve growth factor (NGF) and NGF mRNA change in rat uterus during pregnancy

During pregnancy, the uterus undergoes a profound sympathetic denervation. To explore whether this is associated with changes in neurotrophic factors, we assayed nerve growth factor (NGF) and NGF mRNA in the uterus of non-pregnant and pregnant rats. In the uterine horn, the concentration of NGF and its mRNA decreased during middle and late pregnancy. However, when values were corrected for the increase of uterine weight and total RNA yield during pregnancy, NGF content and mRNA per horn increased during middle and late pregnancy. Similar, but less pronounced, changes were observed in the cervix. By seven days postpartum, both parameters returned to near normal.     

Isolation and characterization of a new rubella variant with DNA polymerase activity

Summary A rubella variant (HPV-RV) was isolated from high passage rubella virus preparations propagated at 37° C in baby hamster kidney BHK21/WI-2 cells. HPV-RV formed clear plaques in HeLa cells and primary cells of Rhesus monkey kidney although wild type rubella virus did not produce plaques in these cells. Cells persistently infected with rubella virus were insensitive to infection by HPV-RV at both 34° and 39.5° C. HPV-RV agglutinated one day old chick, human group O and sheep erythrocytes. This hemagglutinating activity was inhibited by anti-BHK latent virus serum but not by anti-rubella virus serum. The plaque forming ability of HPV-RV was neutralized by anti-BHK latent virus serum although the same antiserum did not affect the plaque forming ability of wild type rubella virus. Furthermore, HPV-RV was found to have the complement fixation antigen of rubella virus and DNA polymerase activity.From these findings, it is concluded that HPV-RV is a hybrid between rubella virus and a latent virus of BHK21/WI-2 cells.With 6 Figures     

Isolation and characterization of a new Vesivirus from rabbits

This report describes the isolation, cDNA cloning, complete genome nucleotide sequence, and partial characterization of a new cultivable calicivirus isolated from juvenile feeder European rabbits (Oryctolagus cuniculus) showing symptoms of diarrhea. Absence of neutralization by type-specific neutralizing antibodies for 40 caliciviruses and phylogenetic sequence comparisons of the open reading frame 1-encoded polyprotein with those of other caliciviruses demonstrate that this new calicivirus is a putative novel member of the Vesivirus genus which is closely related to the marine calicivirus subgroup. According to its putative classification, this new virus has been named rabbit vesivirus.     

Isolation and characterization of rat carcinoembryonic antigen

A rat tumor-associated antigen with properties similar to those of human carcinoembryonic antigen (CEA) has been detected with rabbit immune sera in extracts of transplantable rat colonic adenocarcinoma, RCA-1. This antigen, termed rat CEA, was also detectable by a monkey antihuman CEA serum and a rat monoclonal antibody to rat CEA. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of perchloric acid extracts of RCA-1 tumor, followed by immunoblotting with the above-mentioned anti-CEA reagents, revealed that rat CEA activity resided in components with a molecular weight of approximately 350 kD. The glycoprotein nature of these components was indicated by positive staining with periodic acid-Schiff. Sephadex G-200 chromatography, as well as Sepharose 4B chromatography with and without sodium dodecyl sulfate indicated that the 350-kD components existed in the extracts as molecular aggregates. The 350-kD material, which had been purified by an affinity column containing rat monoclonal antibodies to rat CEA, reacted with the rabbit and monkey anti-CEA sera. This provided strong evidence that serological activity of rat CEA was confined to the 350-kD components.     

Noopept stimulates the expression of NGF and BDNF in rat hippocampus

Naphtho[2,3-f]indole-5,10-dione aminoalkyl derivatives in cytotoxic concentrations inhibit topoisomerase I, which is an important factor of antitumor activity of compounds of this chemical class. The degree of topoisomerase I inhibition with naphtho[2,3-f]indole-5,10-dione derivatives depends on the structure and position of active (aminoalkyl) groups. The mechanism of topoisomerase I inhibition with aminoalkylnaphtho[2,3-f]indole-5,10-diones differs from specific blocking of the catalytic activity of the enzyme and depends on interactions of these compounds with DNA. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 145, No. 3, pp. 304–308, March, 2008     

神经生长因子及降钙素基因相关肽对局灶性脑缺血再灌注大鼠脑环磷酸腺苷反应元件结合蛋白mRNA表达的影响

目的:探讨外源性神经生长因子(NGF)及降钙素基因相关肽(CGRP)对局灶性脑缺血再灌注大鼠海马和顶叶皮质环磷酸腺苷反应元件结合蛋白(CREB)mRNA表达的影响。方法:用线栓法制作大鼠局灶性脑缺血再灌注模型,应用原位杂交和图像分析技术检测大鼠缺血侧海马CA1区和顶叶皮质CREB mRNA的表达。结果:缺血再灌注组右侧海马CA1区和顶叶皮质神经元CREB mRNA表达比假手术组减少,NGF组及CGRP组的CREB mRNA表达均高于缺血再灌注组,NGF与CGRP联合应用时CREB mRNA的表达分别高于NGF组和CGRP组。结论:NGF及CGRP上调局灶性脑缺血再灌注大鼠海马和顶叶皮质CREB mRNA的表达,NGF与CGRP联合应用则作用更强,NGF和CGRP对缺血神经元的保护作用可能是通过激活CREB基因转录与翻译,从而启动一系列信号通路来实现。     

链脲佐菌素诱导的糖尿病大鼠外周血白细胞iNOS-mRNA表达的变化

目的：研究链脲佐菌素（STZ）诱导的糖尿病大鼠胰岛β细胞的功能与外周血白细胞iNOS-mRNA表达的关系。方法：Wistar大鼠随机分为对照10只,实验15只。测定血糖、血浆胰岛素;用原位杂交方法检测外周血白细胞、肝、肺等组织中iNOS-mRNA表达,观察白细胞iNOS-mRNA阳性细胞率。结果：用STZ腹腔注射损伤胰岛β细胞并导致胰岛功能衰竭,3 d后血糖由(8.95±1.80) mmol·L^-1升高至(22.84±4.90) mmol·L^-1,血浆胰岛素由(81.76±2.12) mU·L^-1降至(58.92±18.20)mU·L^-1。正常大鼠外周血白细胞未见iNOS-mRNA表达,而STZ诱导的糖尿病大鼠外周血白细胞iNOS-mRNA表达高度增强。结论：STZ诱导的胰岛β细胞损伤与白细胞的iNOS-mRNA表达存在正相关关系。本实验为检测外周血白细胞某些信号通路提供了简便易行的原位杂交试验方法。     

雌激素对大鼠心内神经节神经生长因子表达的影响

目的:观察神经生长因子(NGF)在卵巢切除大鼠心房后壁心内神经节的表达及雌激素的调节作用.方法:实验动物分正常组、卵巢切除组、卵巢切除后雌二醇替代组,各组动物审温下饲养3周后,进行 NGF 免疫组织化学显色.结果:各组大鼠心内神经节均存在 NGF 阳性神经元,该类神经细胞胞体多数呈圆形或椭圆形,大、中、小型细胞均有发现.正常组和卵巢切除后雌二醇替代组 NGF 免疫阳性神经无约占全部神经节细胞总数的比例分别为 61%和64%,但卵巢切除组大鼠心内神经节 NGF 阳性神经元的数量明显减少,表达明显降低,NGF 免疫阳性细胞约占全部神经节细胞总数的比例为28%.结论:心内神经节细胞内含 NGF;雌激素可能影响心内神经节细胞的 NGF 表达.     

Isolation and characterization of cells of a primaryculture of rat renal glomeruli

I. M. Sechenov First Moscow Medical Institute. Institute of Medical Genetics, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR N. P. Bochkov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 109, No. 1, pp. 57–60, January, 1990.     

Altered EphA5 mRNA expression in rat brain with a single methamphetamine treatment

Methamphetamine is a potent and indirect dopaminergic agonist which can cause chronic brain dysfunctions including drug abuse, drug dependence and drug-induced psychosis. Methamphetamine is known to trigger molecular mechanisms involved in associative learning and memory, and thereby alter patterns of synaptic connectivity. The persistent risk of relapse in methamphetamine abuse, dependence and psychosis may be caused by such alterations in synaptic connectivity. EphA5 receptors constitute large families of tyrosine kinase receptor and are expressed almost exclusively in the nervous system, especially in the limbic structures. Recent studies suggest EphA5 to be important in the topographic projection, development, and plasticity of limbic structures, and to be involved in dopaminergic neurotransmission. We used in situ hybridization to examine whether methamphetamine alters EphA5 mRNA expression in the brains of adult male Wister rats. EphA5 mRNA was widely distributed in the medial frontal cortex, cingulate cortex, piriform cortex, hippocampus, habenular nucleus and amygdala. Compared to baseline expression at 0 h, EphA5 mRNA was significantly decreased (by 20%) in the medial frontal cortex at 24 h, significantly increased (by 30%) in the amygdala at 9 and 24 h, significantly but transiently decreased (by 30%) in the habenular nucleus at 1 h after a single injection of methamphetamine. Methamphetamine did not change EphA5 mRNA expression in the cingulate cortex, piriform cortex or hippocampus. Our results that methamphetamine altered EphA5 mRNA expression in rat brain suggest methamphetamine could affect patterns of synaptic connectivity, which might be responsible for methamphetamine-induced chronic brain dysfunctions.     

Isolation and primary characterization of a new bacteriophage of obligate methylotrophic bacteria

Ø KISR 1, a new bacteriophage of obligate methylotrophic bacteria, was isolated from a local pilot plant. The phage belongs to the Podoviridae family of viruses characterized by a short non-contractile tail and double-stranded DNA. The size of phage genome was determined on the basis of electron micrographs and restriction enzyme analysis to be about 46 kb. The host range of Ø KISR 1 is restricted to three very closely related methanol-utilizing KISR strains belonging to the genus Methylophilus. The phage does not originate from the methylotrophic KISR strains which were tested for lysogeny.     

Isolation and characterization of Sedlec virus, a new bunyavirus from birds

A pathogenic agent designated AV 172 was isolated from the blood of a Reed Warbler (Acrocephalus scirpaceus) out of 767 samples from birds belonging to 35 species and 14 families. The birds (largely wetland passerines) were captured in the reed-belt littoral of Nesyt fishpond in southern Moravia, Czechoslovakia, during the years 1984 to 1987. Virus AV 172 has been found to represent probably a new species (designated virus "Sedlec") of family Bunyaviridae. Sedlec virus is pathogenic to suckling and adult mice when inoculated intracerebrally (i.c.) but not intraperitoneally (i.p.) and its ether-sensitive spherical particles measures 90-100 nm.     

Isolation and characterization of a human cytomegalovirus glycoprotein containing a high content of O-linked oligosaccharides

Summary Several disulfide linked glycoprotein complexes were extracted from human cytomegalovirus with a non-ionic detergent and separated by anion exchange high performance liquid chromatography (HPLC). One complex had a molecular weight of 93,000 and was classified as gCII-93. Another complex had a molecular weight greater than 200,000 and was classified as gCII-200. Both complexes immunoprecipitated with a monoclonal antibody (9E10). A third set of complexes (classified as gC-I) immunoprecipitated with another monoclonal antibody (41C2). Isolated complexes were reduced, alkylated, and individual glycoproteins separated by gel-filtration HPLC. Glycoproteins with molecular weights of 50–52,000 from gCII-93 and gCII-200 appeared to be the same glycoprotein since they could be immunoprecipitated by 9E10 and had identical peptide maps. The amino sugar content of these glycoproteins was compared to that of higher molecular weight glycoproteins obtained from gCII-200 and to a glycoprotein with a molecular weight of 93,000 (gp93 (I)) from gCI. Glycoproteins with molecular weights of 50–52,000 from gCII-93 and gCII-200 contained similar amounts of galactosamine (GalN), glucosamine (GlcN) and sialic acid. However, they contained 2–3 times more GalN than any other glycoprotein from gCII-200 and 10 times more GalN than was detected in gp93 (I). All glycoproteins from gCII-93 or gCII-200 also contained more sialic acid when compared to gp93 (I). GalN in these glycoproteins was present in O-linked oligosaccharides. This was demonstrated by release of low molecular weight oligosaccharides from high molecular weight glycopeptides by mild base hydrolysis and the conversion of GalN to galactosaminitol. Thus, gp 52(II) appears to have a unique phenotype marked by a high amount of O-linked oligosaccharides.     

Isolation, expression and immunological characterization of a calcium-binding protein from Parietaria pollen

The diagnosis and therapy of allergic disorders are usually performed with crude extracts which are a heterogeneous mixture of proteins with different allergenic potency. The knowledge of the allergenic composition is a key step for diagnostic and therapeutic options. Parietaria judaica pollen represents one of the main sources of allergens in the Mediterranean area and its major allergens have already been identified (Par j 1 and Par j 2). In addition, inhibition studies performed using a calcium-binding protein (CBP) from grass pollen (Phl p 7) showed the presence of a homologue of this cross-reactive allergen in the Parietaria extract. Screening of a cDNA library allowed us to isolate a 480bp cDNA containing the information for an 87 AA long protein with high level of homology to calcium-binding proteins from other allergenic sources. It was expressed as a recombinant allergen in Escherichia coli and purified by affinity chromatography. Its expression allowed us to study the prevalence of this allergen in a population of allergic patients in southern Europe. Immunoblotting and inhibition studies showed that this allergen shares a pattern of IgE epitopes in common with other 2-EF-hand calcium-binding proteins from botanically non-related species. The immunological properties of the Pj CBP were investigated by CD63 activation assay and CFDA-SE staining. In conclusion, DNA recombinant technology allowed the isolation, expression and immunological characterization of a cross-reactive calcium-binding protein allergen from Parietaria judaica pollen.     

Isolation and characterization of a new virus infecting the blue-green alga Plectonema boryanum

Isolation of the first long, contractile-tailed virus infecting Plectonema is reported. The virus has the shortest latent period of the known blue-green algae viruses.     

Isolation and characterization of a new strain of Ehrlichia chaffeensis from a patient with nearly fatal monocytic ehrlichiosis.

Ehrlichia chaffeensis is the causative agent of human monocytic ehrlichiosis, a disease that ranges in severity from asymptomatic infection to death. Only one isolate of E. chaffeensis has been made, the Arkansas strain, upon which all characterizations of the agent of human monocytic ehrlichiosis have been based. We report the isolation and characterization of a new strain of E. chaffeensis, the 91HE17 strain, which was cultivated from a patient with a nearly fatal illness. The new isolate grows best in culture with careful control of pH. The two isolates are nearly identical as determined by light and electron microscopy and have significant antigenic identity in fluorescent-antibody and immunoblot assays using polyclonal antisera and the E. chaffeensis-specific monoclonal antibody 1A9. Isolate 91HE17 had 99.9% nucleotide sequence identity with the Arkansas strain in the 16S rRNA gene. Parts of the Escherichia coli GroE operon homologs had identical restriction enzyme digestion patterns, and a 425-bp region of the GroEL gene had at least 99.8% sequence identity between the E. chaffeensis Arkansas and 91HE17 strains. Isolate 91HE17 lacked an epitope identified in E. chaffeensis Arkansas by the monoclonal antibody 6A1. This new E. chaffeensis isolate is very similar to the Arkansas strain and provides the opportunity to substantiate the existence of diversity among ehrlichiae which infect humans. Specific factors which differ among strains may then be compared to assess their potential contributions toward cellular pathogenicity and ultimately toward the development of disease in humans.     

Isolation and immunological characterization of a group of new B lymphomas from CBA mice

We have isolated and characterized a new series of B lymphoma which occurred spontaneously in a group of CBA/N mice that were transferred with spleen or lymph node cells from 24-month-old CBA/Ca mice. Tumor cell lines from six CBA/N mice that received spleen cells were rescued and designated as BKS-2, BKS-3, BKS-4, BKS-5, BKS-6, and BKS-7. Also, tumor cells from a recipient of lymph node cells were rescued and the resulting cell line was designated BKL. These tumor cells expressed membrane immunoglobulin (mu, kappa), major histocompatibility complex Class I and Class II molecules, B220, Lyb8, Fc receptors, J11d, interleukin 2 receptors, and Ly1. All of the tumors did not express the T cell specific markers Thy 1.2, L3T4, and Lyt2.1. They appeared to be clonal in origin, since they exhibited common rearrangements at both heavy and light chain immunoglobulin loci. Phenotypically, these lymphomas appeared to be analogous to immature B cells. Also, these lymphomas displayed different functional reactivities when treated with various B cell mitogens and growth factors in vitro. Anti-mu antibodies which normally induce B cell growth inhibited the proliferation of these lymphoma cells in vitro, whereas they responded to lipopolysaccharide, T cell-derived growth factors, and interleukin 5 by enhanced proliferation. These tumor cells expressed constitutively high levels of c-myc mRNA.