The use of immunostimulants in fish larval aquaculture

The production of fish larvae is often hampered by high mortality rates, and it is believed that most of this economic loss due to infectious diseases is ca. 10% in Western European aquaculture sector. The development of strategies to control the pathogen load and immuno-prophylactic measures must be addressed further to realise the economic "potential" production of marine fish larvae and thus improve the overall production of adult fish. The innate defence includes both humoral and cellular defence mechanisms such as the complement system and the processes played by granulocytes and macrophages. A set of different substances such as beta-glucans, bacterial products, and plant constituents may directly initiate activation of the innate defence mechanisms acting on receptors and triggering intracellular gene activation that may result in production of anti-microbial molecules. These immunostimulants are often obtained from bacterial sources, brown or red algae and terrestrial fungi are also exploited as source of novel potentiating substances. The use of immunostimulants, as dietary supplements, can improve the innate defence of animals providing resistance to pathogens during periods of high stress, such as grading, reproduction, sea transfer and vaccination. The immunomodulation of larval fish has been proposed as a potential method for improving larval survival by increasing the innate responses of the developing animals until its adaptive immune response is sufficiently developed to mount an effective response to the pathogen. To this end it has been proposed that the delivery of immunostimulants as a dietary supplement to larval fish could be of considerable benefit in boosting the animals innate defences with little detriment to the developing animal. Conversely, there is a school of thought that raises the concern of immunomodulating a neotanous animal before its immune system is fully formed as this may adversely affect the development of a normal immune response.     

Influence of copper on the early post-implantation mouse embryo: An in vivo and in vitro study

Summary Female mice were injected intravenously with copper sulphate on either the 7th day (early egg cylinder stage of development), the 8th day (late egg cylinder stage), or the 9th day (early somite stage of development), and examined on the 10th day of gestation. Injection on the 7th day was found to be embryo-lethal; when females were injected on the 8th day, the majority of the surviving embryos exhibited anomalies of the neural tube and/or the heart, while injection on the 9th day resulted in a very low incidence of anomalies. The most common malformations seen on the 10th day involved failure of closure of the neural tube in the head region of the embryo, and various types of anomalies of cardiac rotation and shape. When additional females injected on the 8th day were examined on the 12th day, a high proportion of the fetuses examined had developed exencephaly.A further group of embryos from untreated females were explanted on the 9th day and cultured in vitro in various concentrations of copper sulphate. The lowest levels tested had little obvious effect on neural tube closure. Intermediate doses resulted in, retarded and anomalous embryonic development, while the highest levels employed resulted in neural tube and cardiac anomalies similar to those produced in vivo.The results demonstrate both the direct toxic effect of copper on embryonic development and that the stage of embryonic development at the time of exposure determines both the nature and the extent of the effect.     

中国婴儿中脊灰母传抗体水平对两种疫苗免疫效果的影响

为了了解2月龄婴儿中针对脊髓灰质炎病毒的中和抗体水平,并探讨母传抗体对脊髓灰质炎减毒活疫苗(OPV)和灭活疫苗(IPV)免疫效果的影响。对416名2月龄婴儿分别接种OPV和IPV,采集免疫前后血清,用微量中和法检测血清中Ⅰ、Ⅱ、Ⅲ型脊髓灰质炎病毒中和抗体滴度,评价抗体GMT水平及4倍增长情况。检测结果显示,2月龄婴儿母传抗体Ⅰ、Ⅱ、Ⅲ型阳性率分别为45%、38.2%和17.5%,抗体GMT水平为9.0、8.1和5.2。经接种两组疫苗后,母传抗体阳性者与阴性者免后抗体GMT水平相比,OPV组无明显差异,IPV组阳性者略低于阴性者。在免前抗体滴度<1∶32人群中,OPV组免后抗体滴度4倍增长率及几何滴度增长倍数分别为:Ⅰ型93.6%、71.2;Ⅱ型98.2%、43.7;Ⅲ型91.7%、47.9;IPV组免后抗体滴度4倍增长率及几何滴度增长倍数分别为:Ⅰ型82%、9.4;Ⅱ型62.8%、5.1;Ⅲ型95.6%、11.7;在免前抗体滴度1∶32～1∶128人群中,OPV组Ⅰ型92.3%、23;Ⅱ型86.4%、13.9;Ⅲ型55.6%、4.1;IPV组Ⅰ型48%、2.5;Ⅱ型15%、0.9;Ⅲ型55.6%、2.7。目前中国2月龄婴儿免前脊灰抗体阳性率较高,尤其是Ⅰ、Ⅱ型。脊灰母传抗体对两种疫苗免疫效果有一定干扰,对IPV疫苗的影响较为明显。     

Influence of melatonin on mammary gland growth: in vivo and in vitro studies

Our objective was to determine whether melatonin influenced mammary growth in response to mammogenic hormones. Prepubertal female BALB/c mice were injected for 9 days with 1 microgram of 17 beta-estradiol and 1 mg of progesterone or 17 beta-estradiol/progesterone plus 50, 100, or 200 micrograms of melatonin. Area of the parenchyma and total DNA content of the second thoracic gland were similar between controls and melatonin-injected mice. However, micrograms of DNA/100 mg of mammary tissue were lower in animals treated with 17 beta-estradiol/progesterone plus 200 micrograms of melatonin than in controls. Triglyceride content of mammary glands from animals treated with 100 or 200 micrograms of melatonin/day increased relative to controls. In an in vitro experiment, thoracic mammary glands of 21-day-old mice were cultured for 6 days in a mammogenic milieu of hormones (17 beta-estradiol/progesterone, aldosterone, bovine prolactin, growth hormone, and insulin) with 0 (control), 10(-6), 10(-9), or 10(-12) M melatonin. Relative to controls, 10(-12) M melatonin increased and 10(-6) M melatonin decreased mammary DNA and uptake of [methyl-3H]thymidine. We conclude that high doses of melatonin reduce mammary development in normal mice and that some of this effect may be mediated directly at the mammary tissue.     

Influence of captopril on the in vitro and in vivo effects of leu- and met-enkephalins

Captopril (10(-6) g/ml) enhanced and prolonged the inhibitory effect of leu- and met-enkephalins on the contractions of the isolated guinea-pig ileum section induced by electrical stimulation. In mice, the drug (25 mg/kg, subcutaneously) increased the degree and duration of the analgetic effect of enkephalins. It is concluded that the analgetic effect of captopril is related to the influence on the activity of the endogenous antinociceptive system.     

In vitro and in vivo release of cytostatic factors from Lactobacillus casei-elicited peritoneal macrophages after stimulation with tumor cells and immunostimulants

Summary The effect of tumor cells and immunostimulants on the release of cytostatic factors (CF) from Lactobacillus casei YIT 9018 (LC)-, Corynebacterium parvum (CP)- or peptone-elicited peritoneal macrophages (PM) was investigated in vitro and in vivo. Significant release of CF into the culture medium from PM elicited with LC was induced by seven of eight mitomycin C-pretreated tumor cell lines and not by normal spleen cells, while no CF was released extracellularly from peptone-elicited PM given the same stimulus. CF were released from LC-elicited PM (LCEPM) after stimulation with LC, bacille Calmette-Guérin, streptococcal preparation OK-432, fucoidan or lipopolysaccharide, and LC but not CP induced CF production in the peritoneal cavities of LC- or CP-primed mice. The release of CF from LCEPM after stimulation with mitomycin C-pretreated 3T12-3 cells was inhibited by D-mannose and not by L-fucose. L-Rhamnose and mannose 6-phosphate, but not D-mannose or L-fucose, caused the release of CF from the PM.It was suggested that the release of CF from activated PM is caused by stimulation by some tumor cells, sugars, or bacterial immunostimulants, D-Mannose and L-rhamnose on the surface of tumor cells or bacteria, respectively, may play an important role in the release of CF from activated macrophages.     

槲皮素体内外抗氧化作用和比较研究

目的：测定槲皮素的体外总抗氧化力,进一步观察槲皮素灌胃后大鼠外周血总抗氧化力的变化,并与芦丁、维生素C、维生素E相比较,方法：总抗氧化力采用Fe^3 还原法,槲皮素,芦丁分析采用紫外分光光度法及高效液相色谱法,结果：相同浓度条件下槲皮素的体外总抗氧化力显著强于芦丁,与传统的抗氧化剂维生素C、维生素E相当。槲皮素40mg/kg灌胃1h后大鼠外周血总抗氧化及槲皮素含量（紫外分光光度法）较灌胃前升高最为明显,维生素C也有显著提高外周血总抗氧化力的作用,芦丁与维生素E未表现出显著作用。血浆高效液相分析表明槲皮灌胃后未出现明显的槲皮素收峰,而与其峰相邻的两个未知峰的面积增大。结论：槲皮素的体外抗氧化作用强于芦丁,与传统的抗氧化剂维生素C、维生素E相当,槲皮素吸收后经代谢形成衍生物,提高血浆总抗氧化力的程度与维生素C相近。     

Syringomyelia hydrodynamics: an in vitro study based on in vivo measurements

A simplified in vitro model of the spinal canal, based on in vivo magnetic resonance imaging, was used to examine the hydrodynamics of the human spinal cord and subarachnoid space with syringomyelia. In vivo magnetic resonance imaging (MRI) measurements of subarachnoid (SAS) geometry and cerebrospinal fluid velocity were acquired in a patient with syringomyelia and used to aid in the in vitro model design and experiment. The in vitro model contained a fluid-filled coaxial elastic tube to represent a syrinx. A computer controlled pulsatile pump was used to subject the in vitro model to a CSF flow waveform representative of that measured in vivo. Fluid velocity was measured at three axial locations within the in vitro model using the same MRI scanner as the patient study. Pressure and syrinx wall motion measurements were conducted external to the MR scanner using the same model and flow input. Transducers measured unsteady pressure both in the SAS and intra-syrinx at four axial locations in the model A laser Doppler vibrometer recorded the syrinx wall motion at 18 axial locations and three polar positions. Results indicated that the peak-to-peak amplitude of the SAS flow waveform in vivo was approximately tenfold that of the syrinx and in phase (SAS approximately 5.2 +/- 0.6 ml/s, syrinx approximately 0.5 +/- 0.3 ml/s). The in vitro flow waveform approximated the in vivo peak-to-peak magnitude (SAS approximately 4.6 +/- 0.2 ml/s, syrinx approximately 0.4 +/- 0.3 ml/s). Peak-to-peak in vitro pressure variation in both the SAS and syrinx was approximately 6 mm Hg. Syrinx pressure waveform lead the SAS pressure waveform by approximately 40 ms. Syrinx pressure was found to be less than the SAS for approximately 200 ms during the 860-ms flow cycle. Unsteady pulse wave velocity in the syrinx was computed to be a maximum of approximately 25 m/s. LDV measurements indicated that spinal cord wall motion was nonaxisymmetric with a maximum displacement of approximately 140 microm, which is below the resolution limit of MRI. Agreement between in vivo and in vitro MR measurements demonstrates that the hydrodynamics in the fluid filled coaxial elastic tube system are similar to those present in a single patient with syringomyelia. The presented in vitro study of spinal cord wall motion, and complex unsteady pressure and flow environment within the syrinx and SAS, provides insight into the complex biomechanical forces present in syringomyelia.     

Some safety aspects of Salmonella vaccines for poultry: in vivo study of the genetic stability of three Salmonella typhimurium live vaccines

Live vaccine strains of Salmonella should be avirulent, immunogenic and genetically stable. Some isolates of three commercially available live vaccine strains of Salmonella typhimurium, sampled during a study on their persistence in a vaccinated flock of chickens, were analyzed for genetic stability using macrorestriction analysis of their genome. Two out of the three vaccine strains showed genetic instabilities. Two of the 51 isolates of Zoosaloral vaccine strain and nine of the 32 analyzed isolates of chi(3985), a genetically modified organism, were variants and showed different macrorestriction profiles.     

Influence of RGD-containing oligopeptide-coated surface on bone formation in vitro and in vivo

New bone formation on an RGD-containing oligopeptide-coated surface in vitro and in vivo was investigated. The surface showed two-fold higher osteoblastic cell adhesion and differentiation in vitro, and revealed statistically significant in vivo bone formation compared with the control (P < 0.05).     

Hyperoside ameliorates the progression of osteoarthritis: An in vitro and in vivo study

BackgroundOsteoarthritis (OA) is a common degenerative joint disease. The pathogenesis of OA is closely related to inflammatory responses and apoptosis of chondrocytes. Hyperoside (Hyp), a natural flavonoid compound, exerts multiple bioactivities in various diseases.PurposeOur study aims to investigate the anti-arthritic effects of Hyp and delineate the potential mechanism at the cellular level.MethodsMurine chondrocytes were stimulated with interleukin-1β (IL-1β) with or without Hyp treatment. CCK-8 assay was used to evaluate the cytotoxic effect of Hyp. DCFH-DA was used to detect intracellular ROS. Annexin V-FITC/PI method was applied to examine apoptosis of chondrocytes. The anti-arthritic effects of Hyp and related mechanisms were investigated by examining and analyzing relative markers through quantitative PCR, western blot analysis and immunofluorescent staining. C57BL/6 mice were performed the destabilized medial meniscus (DMM) surgery to establish OA model and then injected intraperitoneally with Hyp (20 mg/kg)) for 4 or 8 weeks. Finally, mice were sacrificed and knee joints were collected for histological observation and analysis.ResultsHyp inhibited IL-1β-induced expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). Additionally, Hyp attenuated IL-1β-induced destruction of the extracellular matrix (ECM) by downregulating the expression of MMPs and ADAMTS5, and meanwhile upregulating the expression of collagen II, aggrecan, and SOX9. Also, Hyp pretreatment reduced IL-1β-induced overproduction of ROS and apoptosis of chondrocytes. Mechanistically, Hypexerted anti-inflammatory effects by partly suppressing the PI3K/AKT/NF-κB and the MAPK signaling pathways, enhancing the Nrf2/HO-1 to limit the activation of NF-κB. Moreover, Hyp played an anti-apoptotic effect via the Nrf2/ROS/BAX/Bcl-xlaxis. In vivo, cartilage degradation was attenuated with a lower OARSI score in Hyp-treated group compared to the DMM group.ConclusionOur study demonstrated that anti-arthritic effects of Hyp in vitro and in vivo, indicating Hyp might serve as a potential agent for the treatment of OA.     

Influence of nonsteroidal anti-inflammatory preparations on the in vitro and in vivo effects of sodium arachidonate

It has been demonstrated on isolated guinea-pig ileum and rats that nonsteroid antiinflammatory drugs (acetylsalicylic acid, ibuprofen, diclofenac sodium, butadione, and indomethacin) antagonized spasmogenic and inflammatory effects of sodium arachidonate, but not of other mediators of inflammation such as histamine, serotonin, bradykinin and PGE2. "Antiarachidonic" potency of nonsteroid antiinflammatory drugs correlated well with their antiinflammatory activity and their ability to inhibit endogenous PG biosynthesis. This method determining the antagonism to arachidonic acid effects in simple in vitro and in vivo models can be useful for screening nonsteroid antiinflammatory drug potential.     

Thymic accessory cell complexes in vitro and in vivo: morphological study

Summary Murine thymic macrophages and interdigitating cells, also called thymic accessory cells, were characterized by means of light- and electron microscopy. The cells were studied in suspension, during isolation by enzymatic digestion and in vivo. They were observed as isolated cells or as components of multicellular complexes, some of which were rosettes and were composed of lymphoid cells centered on each type of accessory cell. We also noted other cell complexes including macrophages that resembled classical epithelial nurse cells. We consider that multicellular complexes represent lymphostromal associations already existing in vivo, because we observed them at the periphery of thymic pieces undergoing enzymatic treatment. The heterogeneity of macrophages that we observed in vitro was also noted in vivo. In vivo macrophages were of three types: classical phagocytic cells distributed throughout the gland, cortical elongated cells in close contact with lymphoid blast cells, and atypical nurse cells containing mitotic cells and located in the inner cortex. The morphological aspects of the latter two cell types suggest that cortical macrophages in vivo have other roles: they can be interpreted as images of positive or negative cell selection. We also believe that rosettes are formed by elongated cortical macrophages when they are enzymatically isolated from the thymus.Part of this work was presented at the Second Thymus Workshop, Rolduc, The Netherlands, April 1989     

Influence of specific mutations on the thermal stability of the td group I intron in vitro and on its splicing efficiency in vivo: a comparative study.

Group I introns constitute excellent systems for analyzing the relationship between RNA tertiary folding and catalysis. Within a hierarchical framework interpretation of RNA folding, secondary structure motifs subtend RNA three-dimensional (3D) architecture. Thus, mutations in two-dimensional motifs are expected to have effects different from those disrupting 3D contacts. Using UV spectroscopy, we have studied the influence of nucleotide substitutions, in both secondary and tertiary structure elements, on the thermal stability of the tertiary folding of the bacteriophage T4 td group I intron. Further, we present a quantitative analysis of the relationship between the splicing efficiency in vivo and the stability of the intron structure as monitored by UV melting curves. We conclude that the stability of the tertiary structure of a group I intron as measured by UV melting is generally a good indication of its ability to splice in vivo.