Prevalence of antibodies to specific infectious agents in bovine fetuses from a slaughterhouse in Minnesota

Sera from 486 bovine fetuses, approximately 60 to 270 days of gestation, were collected at slaughter and tested for the presence of immunoglobulins (Ig). One hundred ten (27%) of the sera were positive for IgG and/or IgM. The earliest age at which fetuses tested positive for IgM and IgG was estimated to be 100 and 120 days, respectively. Ig concentration increased with increased age of the fetus. Sera that were positive for Ig were tested for the presence of specific antibodies to five different infectious agents. Bovine parvovirus antibodies were found in 99 of 110 sera (90%) by hemagglutination inhibition (HI) test. However, only 35 (31.8%) of these sera were positive by serum neutralization (SN) test. Antibodies to parainfluenza-3 virus were detected in 30 sera (27%) by HI test and in 20 sera (18%) by SN test. Five (4%) sera contained SN antibodies to bovine viral diarrhea virus. Only one (0.9%) serum sample contained SN antibodies to infectious bovine rhinotracheitis virus. None of the sera had antibodies against five Leptospira spp. Results of this study suggest that bovine parvovirus may be a potential cause of reproductive problems in cattle.     

The effect on kaolin adsorption of serum on the virus neutralization and enzyme-linked immunosorbent assays of antibody to bovine herpesvirus 1

Two procedures have been used for measuring antibody titres to bovine herpes virus 1 (BHV1): the serum neutralization (SN) test and enzyme-linked immunosorbent assay (ELISA). One hundred and thirty-two sera selected for their low SN titres were tested both unadsorbed and after adsorption with kaolin to determine the effect of kaolin on the titres. With ELISA, the titres of unadsorbed and kaolin adsorbed were not significantly different but with the SN test many treated sera, originally with weak positive titres, became negative after kaolin adsorption. Thus, if the ELISA results are specific for BHV1 antibody then the SN test findings suggest that treatment of sera with kaolin, rather than removing a viral inhibitor, removes a substance from the serum which potentiates SN antibody. This in turn indicates that low SN titres (reciprocal of titre less than or equal to 4, for instance) are probably specific for BHV1 SN antibody whether or not they are abolished by kaolin treatment of the serum.     

Microtiter Tests for Detecting Antibody in Bovine Serum to Parainfluenza 3 Virus, Infectious Bovine Rhinotracheitis Virus, and Bovine Virus Diarrhea Virus

Microneutralization tests for detection of antibody in bovine serum to three bovine viruses are described. The Madin-Darby bovine kidney cell line was used with parainfluenza 3 virus (PI 3), whereas serially cultivated bovine embryonic kidney cells were used for infectious bovine rhinotracheitis virus and bovine virus diarrhea virus. Comparison of micro-hemagglutination-inhibition (HI) with micro-serum-neutralization (SN) tests for PI 3 showed the SN test to be more sensitive, more specific, and therefore more useful than the HI test for detecting antibody. Although the effect of trypsin-periodate treatment of serum was to reduce the HI titer of numerous sera by a twofold dilution, sufficient evidence could not be found to indicate that nonspecific HI inhibitors to PI 3 are present in bovine sera.     

The dependence of the mutagenicity of methanesulphonic acid esters in S. typhimurium TA100 on the alkylation mechanism

Four different model nucleophiles, 4-(p-nitrobenzyl)pyridine (NBP), N-methylmercaptoimidazole (MMI), trifluoroacetic acid (TFA) and H2O were tested with 22 methanesulphonates of widely varying structures for their suitability to predict mutagenic activities in S. typhimurium TA100. The soft nucleophiles NBP (N-alkylation) and MMI (S-alkylation) revealed as highly sensitive for SN2 reactivities whereas TFA (solvolysis at the O-atom) and H2O (hydrolysis) were very sensitive for SN1 reactivities. No correlation between the NBP or the MMI test and the Ames test was found. Quite good correlations could be demonstrated for the TFA test and the hydrolysis rates: with rising activities in the TFA solvolysis the mutagenic potencies were increasing up to a maximum at i-propyl methanesulphonate. After that due to the fast hydrolysis the mutagenicities were decreasing again despite increasing TFA solvolysis rates. In general the secondary methanesulphonates exerted high SN1 reactivities and distinct mutagenic activities, whereas the primary compounds showed no or very low SN1 reactivities and low mutagenic potentials. The "activated" compounds cyclopropylmethyl methanesulphonate, benzyl methanesulphonate and allyl methanesulphonate exerted high SN1 and SN2 reactivities. Methyl methanesulphonate displayed a high mutagenicity in spite of its lack in SN1 reactivity. This is probably due to the induction of the error prone repair (pkM 101 plasmid in TA100). The relation between the alkylating reactivities (SN1 and SN2) and the molecular mechanisms leading to back mutation is discussed.     

Effects of environmental stress and nutlet morph on proportion and within-flower number-combination of morphs produced by the fruit-dimorphic species Lappula duplicicarpa (Boraginaceae)

Phenotypic plasticity in species with heteromorphic diaspores is considered to be a plastic bet-hedging strategy in environmentally fluctuating habitats. The primary aim of this study was to determine the influence of level of abiotic (soil moisture and nutrient availability) and biotic (pure and mixed density) factors on the long-glochid (LN; high dispersal, low dormancy)/short-glochid (SN; low dispersal, high dormancy) nutlet ratio and on nutlet combination of the fruit-dimorphic cold desert winter annual/spring ephemeral Lappula duplicicarpa derived from different nutlet morphs. We hypothesized that stress would decrease the LN: SN ratio. All levels of the four treatments resulted in significant variation in the LN: SN ratio and in the proportion of each nutlet combination. Reproductive biomass varied less than plant mass in plants from both morphs. In favorable environments, the LN: SN ratio and number of SN–3LN and 4LN combinations were relatively high, while in unfavorable environments they were relatively low, thus supporting our hypothesis. Relative allocation to reproductive biomass was significantly negatively correlated with allocation to vegetative biomass. The downward shift that occurred in the LN: SN diaspore ratio in response to stressful growth conditions in Lappula duplicicarpa is assumed to be a bet-hedging strategy.     

Protective effects of sodium nitrite preconditioning against alcohol-induced acute liver injury in mice

The purpose of the present study was to investigate the effect of sodium nitrite (SN) on alcohol-induced acute liver injury in mice. Forty male C57bL/6 mice were randomly divided into 4 groups. Acute alcohol-induced liver injury group were injected intraperitoneal (ip) with alcohol (4.5 g/kg); SN preconditioning group were pretreated with SN (16 mg/kg, ip) for 12 h, and received alcohol (4.5 g/kg, ip) injection; Control and SN groups were treated with saline and SN, respectively. After the treatments, liver index (liver/body weight ratio) was determined. Colorimetric technique was performed to measure the serum alanine transaminase (ALT), aspartate transaminase (AST), liver superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT) activities, as well as malondialdehyde (MDA) content. The pathological index of liver tissue was assayed by HE and TUNEL fluorometric staining. Using Western blot and immunohistochemistry staining, the expression of hypoxia-inducible factor-1α (HIF-1α) protein was detected. The results showed that, compared with acute alcohol-induced liver injury group, pretreatment with low doses of SN decreased liver index and serum levels of ALT and AST, weakened acute alcohol-induced hepatocyte necrosis, improved pathological changes in liver tissue, increased live tissue SOD, GSH-Px and CAT activities, reduced MDA content and apoptosis index of hepatocytes, and up-regulated HIF-1α protein level in liver tissue. These results suggest that the pretreatment of SN can protect hepatocytes against alcohol-induced acute injury, and the protective mechanism involves inhibition of oxidative stress and up-regulation of HIF-1α protein level.     

Prevalence of feline viral antibodies in random-source laboratory cats

Over a period of 1973 to 1979, a serologic survey of virus infections was conducted on feline sera collected in four universities which located in different prefectures; Obihiro, Saitama, Kanagawa and Tokyo. A significant hemagglutination-inhibition (HI) antibody titer of 1 : 8 or higher to feline panleukopenia virus (FPLV) was detected in 130 (58%) of the 226 sera used. No remarkable difference in the HI antibody prevalence in cats to FPLV was recognized by years or localities. Of a total of 188 cats tested, 99 (53%) presented positive serum neutralizing (SN) antibody titers to the No. 1 strain of feline calicivirus (FCV). Especially in Kanagawa, 17 (77%) of the 22 cats had positive SN titers. However, only 42 (22%) of the 188 sera showed positive SN titers to the Kyoritsu strain of FCV. Such lower positivity in the cats was observed with 13% in the SN test to human reovirus type 3 (Reo-3). The incidence of positive SN antibodies to feline rhinotracheitis virus (FRV) also remained in low values of 20 to 27% with the exception of high percentage of 86 in Tokyo. The dissemination of FPLV, FRV, FCV and Reo-3 was briefly discussed in relation with the age distribution of viral antibodies in cats.     

The peculiar nature of the guanidine hydrochloride-induced two-state denaturation of staphylococcal nuclease: a calorimetric study.

This work determines the ratio of DeltaH(vH) /DeltaH(cal) for staphylococcal nuclease (SN) denaturation in guanidine hydrochloride (GdnHCl) to test whether GdnHCl-induced denaturation is two-state. Heats of mixing of SN as a function of [GdnHCl] were determined at pH 7.0 and 25 degrees C. The resulting plot of DeltaH(mix) vs [GdnHCl] exhibits a sigmoid shaped curve with linear pre- and post-denaturational base lines. Extending the pre- and post-denaturational lines to zero [GdnHCl] gives a calorimetric DeltaH (DeltaH(cal)) of 24.1 +/- 1.0 kcal/mol, for SN denaturation in the limit of zero GdnHCl concentration. Guanidine hydrochloride-induced denaturation Gibbs energy changes in the limit of zero denaturant concentration (DeltaG degrees (N)(-)(D)) at pH 7. 0 were determined for SN from fluorescence measurements at fixed temperatures over the range from 15 to 35 degrees C. Analysis of the resulting temperature-dependent DeltaG degrees (N)(-)(D) data defines a van't Hoff denaturation enthalpy change (DeltaH(vH)) of 26. 4 +/- 2.8 kcal/mol. The model-dependent van't Hoff DeltaH(vH) divided by the model-independent DeltaH(cal) gives a ratio of 1.1 +/- 0.1 for DeltaH(vH)/DeltaH(cal), a result that rules out the presence of thermodynamically important intermediate states in the GdnHCl-induced denaturation of SN. The likelihood that GdnHCl-induced SN denaturation involves a special type of two-state denaturation, known as a variable two-state process, is discussed in terms of the thermodynamic implications of the process.     

The effect of sodium nucleinate on allergic and immunological reactions

Experiments on immunized rabbits and guinea pigs indicated that sodium nucleinate (SN) was capable of weakening or entirely eliminating anaphylactic and skin reactions of delayed type hypersensitivity to repeated administration of staphyloanatoxin, APDT vaccine. The findings on patients with the infectious form of bronchial asthma and chronic rheumatism showed that sodium nucleinate attenuated reactions to the subcutaneous administration of staphylococcal and streptococcal allergens. The treatment of patients suffering from infectious-allergic bronchial asthma and rheumatism with SN resulted in the recovery of deficient T cells, T-suppressors, normalization of immunoglobulin concentrations. In children with acute glomerulonephritis sodium nucleinate normalized decreased T-suppressor cells and increased IgG and circulating immune complexes (CIC), resulting in a pronounced remission of disease. The mechanism of desensitization and elimination of CIC by SN has not been explored, however, the parameters of SN-induced immunomodulation are known rather completely. It is suggested that SN brings about accumulation in the cell of cyclic AMP which diminished membrane permeability, activates monoaminooxidase resulting in the degradation of histamine and other biogenic amines, enhances the synthesis of endogenous corticosteroids with their desensitizing properties. All these effects contribute to the elimination of delayed type hypersensitivity. The role of SN in the inhibition of delayed type hypersensitivity remains obscure.     

Serologic detection of adenoviral hemorrhagic disease in black-tailed deer in California

An enzyme-linked immunosorbent assay (ELISA) and a serum neutralization (SN) test were developed to measure serum antibodies against the adenovirus causing hemorrhagic disease in free-ranging and captive experimentally-infected black-tailed deer (Odocoilenus hemionus columbianus) in California (USA). There was a strong (rho = 0.874) and significant (P < 0.0001) correlation between ELISA and SN titers, although the SN assay was more sensitive than the ELISA.     

Tetrahydrophthalazine derivative "sodium nucleinate" exert its anti-inflammatory effects through inhibition of oxidative burst in human monocytes

We described the use of a new chemical substance Sodium nucleinate (SN) as an immunomodulatory substance exhibiting antiinflammatory properties. Sodium nucleinate (SN) registrated in Russian Federation as Tamerit, is 2-amino-1,2,3,4-tetrahydrophthalazine-1,4-dione sodium salt dihydrate, derivative of well known chemical substance luminol. To comprehend the mechanisms of SN immunomodulatory activity, we examined the SN modulation of the oxidative burst responses of whole blood human monocytes and polimorphonuclear cells (PMC) stimulated with phorbol 12-myristate 13-acetate (PMA) or E. coli suspension in vitro. SN did not inhibit the proportion of neutrophils and monocytes phagocytosing E. coli. Oxidative burst responses of monocytes stimulated with PMA were strongly inhibited at SN concentration ranging from 10-500 mg/ml, less efficient inhibitor was SN in E. coli stimulated monocytes (inhibition range was from 50-500 mg/ml SN). SN inhibited PMC oxidative burst only in range 100-500 mg/ml SN. In conclusion, we found SN as an efficient inhibitor of oxidative burst in monocytes. Since ROS generation in monocytes/macrophages has been found to be important for LPS-driven production of several proinflammatory cytokines, SN may exsert its antiinflammatory effects through monocyte/macrophage oxidative burst inhibition.     

Phase behaviour and morphology of binary mixtures of DPPC with stearonitrile, stearic acid, and octadecanol at the air-water interface

The behaviour of dipalmitoylphosphatidylcholine (DPPC), mixed with stearonitrile (SN), was investigated at the air-water interface by surface pressure-area (pi-A) measurements and by direct visualisation of monolayers by Brewster angle microscopy (BAM). The pi-A-X diagram of system DPPC/SN was compared with the corresponding diagrams of systems DPPC/stearic acid (SA) and DPPC/octadecanol (OD) at 20 degrees C. Monolayers of the three systems reach the closest packing of alkyl chains in the 0.4-0.6 range of XDPPC. Thermodynamic analysis indicates miscibility in the three binary systems with negative deviations from the ideal behaviour. Morphological features of system DPPC/SN change significantly with XDPPC and temperature in the range 10-30 degrees C. At 10 and 20 degrees C mixed monolayers form condensed states from low pi all over the composition range. At 30 degrees C, the liquid-expanded (LE)--liquid-condensed (LC) phase transition occurs at increasing pi with XDPPC. The shape and size of condensed domains change with XDPPC and pi. Contrarily to the behaviour of pure components, mixed monolayers of DPPC/SN exhibit orientational order in the 0.2-0.6 mol fraction range of DPPC. BAM observation confirmed the partial miscibility indicated by GE data in a limited range of compositions at 30 degrees C.     

外源NH_4~+对穗花狐尾藻根系形态和养分吸收的影响

在温室内进行静态实验,以原沉积物(CK)和分别添加0.24%和0.48%氯化铵(SN1和SN2)的沉积物作为底质培养沉水植物,探讨了高NH4+环境中穗花狐尾藻的根系形态特征及营养物质积累与干物质分配策略。结果表明,穗花狐尾藻主根直径(0.432-0.518mm)与主根/侧根比率(1.50-4.39)均随着沉积物NH4+含量升高呈现增大趋势,但单株主根总长(31.64-171.67cm)则在高NH4+环境中显著变短;其中,SN2处理中穗花狐尾藻主根数量(8.17条/株)显著低于SN1(14.67条/株)和CK(14.33条/株)处理,而SN1与CK处理之间差异不显著。SN1和SN2处理中穗花狐尾藻植株全氮含量(55.98和55.19mg/g)均显著高于CK(42.89mg/g)处理,而SN1和SN2处理中穗花狐尾藻植株全磷含量(1.63和1.53mg/g)则比CK处理中(3.71mg/g)显著降低。穗花狐尾藻植株干物质积累量(168.17-405.81mg/株)和全磷积累量(0.25-1.51mg/株)均随沉积物NH4+含量升高而显著下降,但植株全氮积累量(9.12-21.08mg/株)则表现为:SN1CKSN2。全氮和全磷在叶片中的分配率都随着沉积物NH+含量升高而显著降低,而在根系和茎秆中分配率则显著增加。     

Substance P and secretoneurin in vitreous aspirates of patients with various vitreoretinal diseases

By means of highly sensitive radioimmunoassays, the levels of substance P (SP) and secretoneurin (SN) were detected in vitreous aspirates of patients with macular holes which served as controls, in patients with nonproliferative diabetic retinopathy (DR), active proliferative diabetic retinopathy (active PDR), inactive PDR, rhegmatogenous retinal detachment and proliferative vitreoretinopathy (PVR). Furthermore, SN-like immunoreactivities were characterized by reversed phase-HPLC. The concentration of SN was more than 20-fold higher in macular holes when compared with SP and reversed phase HPLC revealed evidence that the vitreous levels of SN represent authentic SN. SN was significantly decreased in patients with nonproliferative DR, active PDR and inactive PDR by more than 70% which seems to result from a reduced expression and/or secretion from the cilary epithelium and a reduced release from the retina both due to diabetes mellitus. By contrast SP was increased in rhegmatogenous retinal detachment most obviously due to an enhanced outflow of the peptide through retinal breaks. Despite their proangiogenic activities, SP and SN are unlikely to be involved in the pathogenesis of neovascularizations in DR because of their unchanged and reduced levels, respectively, but the low levels of both peptides may facilitate the regression of vasoproliferations following laser photocoagulation.     

Pair bonding prevents reinforcing effects of testosterone in male California mice in an unfamiliar environment

Testosterone (T) can be released by stimuli such as social interactions, and thereby influence future social behaviours. Because the reinforcing effects of T can induce preferences for specific environmental locations, T has the potential to alter behaviour through space use. In a monogamous species, this T pulse may contribute differently to space use in sexually naive (SN) and pair-bonded (PB) males: SN males may be more likely to explore new areas to set up a territory than PB males, which are more likely to defend an existing, established territory. In this study, we test for variation in T-driven space use by examining variation in the formation of conditioned place preferences (CPPs) in SN and PB male California mice. In the three-chambered CPP apparatus, subcutaneous administrations of physiological levels of T were used to repeatedly condition SN and PB males to a side chamber, which is an unfamiliar/neutral environment. The final tests revealed that T-induced CPPs in the side chamber are developed in SN, but not PB males. This study fills a gap in our knowledge about plasticity in the rewarding nature of T pulses, based on past social experience.     

Genotoxicity of monofunctional methanesulphonates in the SOS chromotest as a function of alkylation mechanisms. A comparison with the mutagenicity in S. typhimurium TA100

17 monofunctional methanesulphonates of widely varying structures were investigated in the SOS chromotest using the E. coli strain PQ37. All compounds tested were positive in this assay. The monofunctional methanesulphonates in general possess low SOSiP values. Five of the compounds tested i.e. iBMS, NpMS, 2 PhPMS, PkMS and 1,3-DC12PMS (for abbreviations see Table 1) did not show increasing beta-galactosidase activity and both the positive induction factors and the positive SOSiP values resulted from the toxicity correction as performed according to Quillardet and Hofnung (1985). In general methanesulphonates with a higher SN1 reactivity, in particular the secondary compounds, showed clear genotoxic activities whereas those possessing low SN1 reactivities (primary compounds) induced a low SOS repair indicating that the alkylation of O-atoms in the DNA bases contributes more to the induction of SOS repair in strain PQ37 than N-alkylations. The only exception was methyl methanesulphonate (MMS) which possessed a very high SN2 reactivity but a rather low SN1 reactivity. It had the highest SOSiP value of all tested methanesulphonates. No dependence of the genotoxicity on the SN2 reactivity could be found in this series. In general the phenyl-substituted methanesulphonates showed higher SOSiP values, which is presumably due to their relatively high SN1 reactivities and their relatively long life times in aqueous systems. There is a clear relationship between SN1 reactivities and the SOSiP values: the SOSiP values increase with rising SN1 reactivities reaching a maximum at iPMS after which the genotoxicities decrease due to the decreasing life times. The compounds with very high SN1 reactivities also possess very high hydrolysis rates. A good correlation could be established between the mutagenicities in S. typhimurium TA100 and the SOS chromotest (strain PQ37). Only 4 small deviations from this correlation could be found. The reasons for these deviations are discussed.     

Therapeutic effect of Gag-nuclease fusion protein on retrovirus-infected cell cultures.

Capsid-targeted viral inactivation is a novel protein-based strategy for the treatment of viral infections. Virus particles are inactivated by targeting toxic fusion proteins to virions, where they destroy viral components from within. We have fused Staphylococcus nuclease (SN) to the C-terminal end of Moloney murine leukemia virus Gag and demonstrated that expression of this fusion protein in chronically infected chicken embryo fibroblasts resulted in its incorporation into virions and subsequent inactivation of the virus particles by degradation of viral RNA. Release of particles incorporating Gag-SN fusion proteins into the extracellular milieu activates the nuclease and results in destruction of the virion from within. By comparing the effects of incorporated SN and SN*, an enzymatically inactive missense mutant form of SN, on the infectivity of virus particles, we have clearly demonstrated that nucleolytic activity is the antiviral mechanism. Expression of Gag-SN fusion proteins as a therapeutic agent causes a stable reduction of infectious titers by 20- to 60-fold. The antiviral effect of capsid-targeted viral inactivation in our model system, using both prophylactic and therapeutic approaches, suggests that a similar anti-human immunodeficiency virus strategy might be successful.     

衣壳蛋白靶向灭活策略应用于抗登革病毒感染的研究

衣壳蛋白靶向病毒灭活是近年来新兴的抗病毒策略。为探索该策略在抗登革病毒感染中的应用 ,首先建立了稳定表达登革 2型病毒衣壳蛋白 (D2C)与葡萄球菌核酸酶 (SN)融合蛋白D2C_SN的哺乳动物细胞系 ,然后以登革病毒攻击上述细胞系 ,研究表达的融合蛋白D2C_SN对产生的子代病毒颗粒感染性的影响。结果表明融合蛋白D2C_SN能够在病毒装配过程中与野生型衣壳蛋白共组装入子代病毒颗粒内部 ,并导致病毒基因组的降解。与正常BHK细胞相比较 ,融合蛋白D2C_SN可导致产生的子代病毒感染性滴度降低 10 3～ 10 4 ,显示出很强的抗病毒效果