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Query: UNIPROT:P30536 (PBS)
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The shortening of the time interval between the onset of oestrus and ovulation in sows by the transcervical administration of seminal plasma was investigated in 23 German Landrace gilts, using the technique of single horn infusions (Mariensee model) in combination with the transcutaneous sonographic monitoring of ovaries. Preparative surgery comprised the detachment of the left uterine horn from the corpus, leaving the caudal end open to the peritoneal cavity but sealing the corpus wound. The left ovary was loosely tied to the ventral abdominal wall for better sonographic distinction. The animals were used in two to four consecutive cycles. After detection of oestrus by the teaser boar, the patent (right) horns were filled by transcervical infusion of 100 ml of a variety of test solutions. Ovulation was probed by transcutaneous sonography at intervals of 4 h thereafter. Native seminal plasma provoked ovulation in the ipsilateral ovary of the treated horn 10.7 h earlier than in the contralateral ovary. This effect was reduced to 7.3 h after charcoal treatment of seminal plasma; addition of 10 micrograms oestradiol restored the effect in full, while 10 micrograms of oestradiol in PBS shortened the time interval to only 3.3 h versus the control ovary. Little effect was seen with oestrone sulfate, none with prostaglandins in PBS or with PBS alone. The preliminary characterization of the nonsteroidal component of seminal plasma advancing ipsilateral ovulation after transcervical infusion suggests a proteinaceous nature. The activity resides in the 1-10 kDa fraction separated by ultrafiltration and is lost after treatment with pronase.
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PMID:Advanced ovulation in gilts by the intrauterine application of a low molecular mass pronase-sensitive fraction of boar seminal plasma. 856 67

Mycoplasma gallisepticum- or M. synoviae-challenged chickens were monitored with serological assays (serum plate agglutination, hemagglutination inhibition, and enzyme-linked immunosorbent assay) and polymerase chain reaction (PCR). The tracheal swabs from M. gallisepticum-challenged chickens received three different treatments (phosphate-buffered saline [PBS], Frey's broth, or 10 mM Tris-HCl/250 mM ethylenediaminetetraacetic acid/ 2.5% sodium dodecyl sulfate [STE]) prior to DNA purification. A nonphenolic method for DNA extraction was utilized. The best PCR results were obtained with PBS swab treatment. The nonphenolic method for DNA extraction was compared with a phenolic method in an experiment with tracheal swabs from M. synoviae-challenged chickens and commercial flocks. Both methods gave comparable results.
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PMID:Polymerase chain reaction optimization for Mycoplasma gallisepticum and M. synoviae diagnosis. 871 37

It is generally accepted that luminal surfaces of adult microvascular endothelia present an anionic barrier that limits passage of anionic macromolecules. To assess the ontogeny of the barrier, temporal and spatial expression of endothelial anionic sites was evaluated in the chorioallantoic membrane of chicken embryos from days 4.5 to 18 of incubation. After an initial flush, the vessels were perfused with cationic ferritin (CF, 1.0 mg/ml in PBS) for 2 min. Following a second flush to remove unbound CF, the chick chorioallontoic membranes (CAMs) were fixed and processed for electron microscopy. Continuous CF binding was revealed on the luminal endothelium, the junctional clefts and the plasmalemmal vesicles from days 4.5 to 14. However, by day 18, anionic sites had become discontinuous. Prior perfusion with protamine sulfate abolished CF binding and facilitated native ferritin binding. Further ultrastructural evaluation, using peroxidase labeled LFA lectin, revealed sialic acid moieties in patches on the CAM endothelium. Thus, in early chick embryogenesis, the CAM endothelium displays a continuous pattern of luminal anionic sites comprised in part of sialic acid. As the CAM ages, endothelial anionic sites become reduced. That the expression of endothelial anionic domains remained constant despite changes in CAM microvascular permeability in early development (Rizzo et al., 1995a) serves to suggest a minimal role for anionic domains in the development of microvascular permselectivity during normal angiogenesis.
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PMID:Distribution of anionic sites on microvascular endothelium of the chick chorioallantoic membrane. 876 Aug 58

Monoclonal anti-nucleosome antibodies (mAbs) complexed to nucleosomal antigens can bind to DNA and to heparan sulfate (HS) in ELISA and to the GBM in vivo in a rat renal perfusion system, whereas non-complexed mAbs do not bind [1]. In this study, we analyzed whether heparin (HEP) or N-desulfated/acetylated heparins (DSA-HEP), structurally and functionally strongly related to HS, are able to prevent the binding of these complexed mAbs to DNA and to HS in vitro and to rat GBM in vivo. In ELISA the binding of nucleosome complexed antinucleosome antibodies to DNA and HS was inhibited dose-dependently by HEP, DSA-HEP and low molecular weight (LMW) DSA-HEP. Intravenous injection of nucleosome/anti-nucleosome immune complexes without heparin/heparinoids in BALB/c mice led to GBM binding, while simultaneous injection of heparin/heparinoids with complexed antibodies or pretreatment with heparin subcutaneously prior to injection of complexes prevented this binding. Subsequently, we tested the preventive effect of HEP, DSA-HEP and LMW-DSA-HEP on progression of renal disease in MRL/lpr mice. Treatment was started at an age of eight weeks in a dose of 50 micrograms daily. With all three drugs albuminuria was significantly delayed compared to PBS treated controls (cumulative incidence of proteinuria at 20 weeks in controls 60% vs. 13%, 14% and 6% respectively for HEP, DSA-HEP and LMW-DSA-HEP; P < 0.05). At week 21 the glomerulonephritis was histologically less severe in heparin/heparinoid treated animals (P = 0.02). In immunofluorescence the amount of immunoglobulin and C3 deposits in the glomerular capillary wall tended to be less in heparin/heparinoid treated mice compared to PBS treated controls (P = 0.07). Furthermore, at 20 weeks anti-HS levels in plasma of heparin/heparinoid treated mice were significantly lower (P < 0.05). We conclude that interaction of heparin or heparin analogs with HS reactive immune complexes containing nucleosomal antigens prevents the binding of these immune complexes to the GBM and delays nephritis in MRL/lpr mice.
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PMID:Heparin and heparinoids prevent the binding of immune complexes containing nucleosomal antigens to the GBM and delay nephritis in MRL/lpr mice. 891 22

Interporous hydroxyapatite ceramic (Ca10(PO4)6(OH)2) has excellent bio-compatibility and interlinked pore structure, antibiotics could be loaded into pores in vacuum system. To confirm penetration of the agent to the HAb (2 cm3 cubic block), the aminoglycoside antibiotic (Isepamicin Sulfate; ISP) dissolved in eosin dye at various vacuum pressures. In ISP slow release study, the blocks were placed in 5 ml of PBS at a temperature of 37 degrees C. The PBS was replaced every 48 h and samples containing released ISP were stored until assay. All were found to release the drug maintaining a mean concentration of 0.41 microg ml(-1) even after 18 days of nine exchanges. This concentration of antibiotic exceeded the minimum inhibitory concentration against the common causative organisms of osteomyelitis. The results suggest that HAb impregnated with antibiotics using a simple vacuum system may serve as a valuable new method of administering local chemotherapy, primarily when used as a strut graft for bone defects.
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PMID:Synthesis of antibiotic-loaded interporous hydroxyapatite blocks by vacuum method and in vitro drug release testing. 966 58

Experiments were conducted to improve survival of mouse spermatozoa through the cryopreservation process. In the first experiment, percentages of motile spermatozoa and fertilizing capacities of spermatozoa were evaluated when mouse spermatozoa were cryopreserved using three previously reported cryopreservation media: (1) 18% raffinose in 3% skim milk; (2) Tes/Tris medium containing 25% egg yolk and 1.25% glycerol; and (3) PBS containing 18% raffinose and 1.75% glycerol, each at three different cooling rates (-3, -10, and -50 degrees C/min). Spermatozoa frozen in the skim milk/raffinose medium exhibited the highest percentage of motile spermatozoa (39%) when cells were frozen at -10 degrees C/min (P<0.05). The second experiment evaluated the effects of modifying the Tes/Tris/egg yolk medium, comparing different concentrations of egg yolk, BSA, and sodium dodecyl sulfate. Reducing egg yolk from 25% of the medium volume to 5%, increased percentages of motile spermatozoa after cryopreservation from 29 to 36% (P<0.05). Addition of 1% BSA and sodium dodecyl sulfate to medium containing 5% egg yolk further improved percentages of motile spermatozoa after freezing. In the final experiment, 20% whole egg was substituted for 5% egg yolk and 1% BSA used in previous experiments and resulted in percentages of motile spermatozoa (51%) equal to that of the skim milk-raffinose medium. However, fertility rates were higher (68%) than for spermatozoa frozen in the skim milk-raffinose medium (P < 0.05) and were comparable to the fertility rates of fresh spermatozoa (77%; P>0.05). In conclusion, freezing mouse spermatozoa in a medium containing 20% whole egg, 0.035% sodium dodecyl sulfate, and 1.25% glycerol using a cooling rate of -10 degrees C/min preserves the motility and fertilization capacity of mouse spermatozoa.
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PMID:Fertilizing potential of mouse spermatozoa cryopreserved in a medium containing whole eggs. 1067 48

The ability of adult Haemonchus placei intestinal homogenate to confer protection against homologous challenge infection was evaluated. Calves were immunized twice with 100 microg H. placei intestinal protein in 5% dextran-sulfate/PBS (vaccinates) or PBS alone (controls) and were challenged with approximately 3300 infective H. placei larvae. There was no significant difference between groups in the total number of nematodes recovered but significantly fewer (p < 0.001) adult females were recovered from vaccinates. The proportion of fourth-stage larvae in vaccinates was significantly greater (p < or = 0.05) than in controls. Lengths of adult male and female nematodes were significantly shorter (p < 0.001) in vaccinated calves, and the numbers of eggs present in the uteri of female nematodes from vaccinates were significantly decreased (p < 0.001). Counts of nematode eggs per gram of feces (EPG) of vaccinates were significantly less than that for controls on Days 29-49 post-challenge (p < or = 0.05). Vaccinates had significant increases in serum IgG1 and IgG2 log(10) titers (p < or = 0.05) but not in serum IgM. EPG, numbers of females, and size of males and females were negatively correlated with increased mean post-challenge IgG1 and IgG2 titers. Reduction in binding of periodate-treated gut homogenate by immune serum indicated a carbohydrate specific component in the immune response.
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PMID:Vaccination of calves with Haemonchus placei intestinal homogenate. 1071 62

This study was performed to isolate a vaccine strain of S. aureus from clinical or subclinical mastitis and to choose the most optimal adjuvant for immune response of alpha toxin and capsular polysaccharide (CPS) of field strain. Of thirty strains of S. aureus isolated from milk of clinical or subclinical mastitis, V112 strain isolated from milk of gangrenous mastitis was used in this vaccine. Twenty one of rabbits were allocated into 5 groups based on adjuvants and immunized twice every 2 weeks for 8 weeks. This vaccine was composed of alpha toxin (10 hemolytic units) and formalinized whole cells (1 x 10(11) cells/ml. Five rabbits received PBS solution as a control group. The highest antibody titers against alpha toxin and CPS were observed in dextran sulfate- and aluminium hydroxide-adjuvant group at 8 weeks after immunization, respectively. These results of the study showed that one adjuvant could not induce strong and long-term immune response of alpha toxin and CPS antigens. Therefore, the use of combined adjuvants in subunit vaccine may be useful and feasible.
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PMID:Effects of adjuvants on the immune response of staphylococcal alpha toxin and capsular polysaccharide (CPS) in rabbit. 1077 May 93

The protective capacity of vaccination with Haemonchus placei whole gut homogenate against challenge with the non-blood-feeding nematode Ostertagia ostertagi was evaluated in calves. Ten helminth-free calves were randomly assigned to two groups. Group 1 received 100microg H. placei intestinal homogenate in the adjuvant 5% dextran sulfate/PBS, while Group 2 received the adjuvant alone. Injections were administered subcutaneously on Days 0 and 28. All calves were challenged with approximately 26,100 O. ostertagi larvae on Day 42. Serum antibody response and counts of nematode eggs per gram of feces (EPG) were determined throughout the study. Calves were necropsied at 5.5 weeks post-challenge for recovery of nematodes. Although significant increases were detected in both serum IgG(1) and IgG(2) of Group 1 calves (p<0.05), there was no significant difference in the total number of O. ostertagi recovered from the two groups (p>0.05). Lengths of adult nematodes were not significantly different between groups nor were the numbers of eggs present in adult females recovered from each group significantly different (p>0.05). There were also no significant differences between groups regarding fecal egg counts (p>0.05). Results suggest either: (1) the antigens targeted by the induced antibodies were not present in O. ostertagi; (2) the antigens targeted by the induced antibodies were present, but not essential to O. ostertagi survival; or (3) the antigen was present and essential, but amount of antibody ingested was insufficient to cause damage to the nematode.
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PMID:Ostertagia ostertagi challenge of calves vaccinated with Haemonchus placei intestinal homogenate. 1082 16

The staining method developed by Christian Gram was introduced as a simple and highly selective tool for demonstrating myxosporean and coccidian sporogonic stages. When using standard blood staining procedures for those enigmatic parasites it is sometimes difficult to distinguish them from fish host tissue. They clearly exhibit a partial gram-positive reaction in histological sections, but staining is variable in air dried fish organ imprints. To visualize the gram-negative background of different host tissue components in histological sections, the conventional safranin counterstain of the gram protocol may be modified as follows: after application of 2% crystal violet (basic violet 3) and Lugol's solution, sections are stained with 0.1% nuclear fast red-5% aluminum sulfate and 0.35% aniline blue (acid blue 22) dissolved in saturated aqueous picric acid. Replacement of the gram-specific dye crystal violet with 2% malachite green gave similar results in organ imprints containing myxospores or coccidia, but only in sections containing myxosporea. Staining for 1 min with an aqueous solution of 0.5% malachite green and followed 1 min washing was sufficient for rapidly demonstrating the parasite spores in organ imprints of both myxosores and oocysts. With regard to the role of acid mucopolysaccharides and other carbohydrates in the gram reaction of spores, alcian blue 8GX staining was compared to the binding of FITC-labeled WGA, GS I and GS II. Each lectin was applied at 20 microl/ml PBS, HEPES for 1 hr. Whereas WGA yielded a nonspecific pattern like the alcian blue staining, GS II resulted in a pattern similar to the gram staining results. This binding was weak in untreated specimens, but was significantly enhanced when digested first within trypsin overnight in a humid chamber at 37 degrees C. The binding of GS II to both myxosporidian and coccidian spores suggests that they are both composed of polymers containing N-acetyl-D-glucosamine residues. Furthermore, the results suggest that this hexosamine plays a key role in the gram reaction.
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PMID:Gram staining and lectin binding properties of Myxosporea and Sporozoea. 1144 Feb 98


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