Demonstration of antibodies to Bacillus piliformis in SPF colonies and experimental transplacental infection by Bacillus piliformis in mice

1978 ◽  
Vol 12 (1) ◽  
pp. 23-26 ◽  
Author(s):  
A. Schaich Fries

Antibodies to Bacillus piliformis were demonstrated by the immunofluorescence antibody technique in sera from mice and rabbits from SPF breeding colonies. Mice in various stages of pregnancy were experimentally infected with Bacillus piliformis and killed 2 to 3 days later. The organism was demonstrated in the uterus, foetal membranes and in the liver of the foetuses. Infection was not limited to any particular stage of pregnancy.

2020 ◽  
Vol 0 ◽  
pp. 1-5
Author(s):  
Parvathy Santhosh ◽  
Kidangazhiathmana Ajithkumar

Anti-nuclear antibodies (ANAs) are a group of antibodies that are characteristically associated with connective tissue diseases (CTDs). Indirect immunofluorescence antibody technique, having a high sensitivity, is the most common technique used for detection, results of which are expressed in terms of the pattern of fluorescence, substrate used, and the titer of a positive test. Other methods include solid-phase assays. ANA test must be performed only when there is a clinical suspicion of an autoimmune CTD. ANA should not be used as a screening tool for asymptomatic individuals. It is essential in clinical practice to be aware of when to order ANA testing, and how to correctly interpret the test results.


Development ◽  
1970 ◽  
Vol 24 (2) ◽  
pp. 313-330
Author(s):  
A. E. Wild

Fluorescent protein tracing, involving F.I.T.C.-conjugated proteins and the fluorescent antibody technique, was employed to study the sites and mechanism of transport of a variety of proteins across the rabbit foetal membranes. No evidence was found for an intercellular transmission of proteins across the yolk sac splanchnopleure to the exocoel, but all proteins were shown to become localized in absorptive vesicles in the yolk-sac endoderm. Different proteins became similarly localized in absorptive vesicles having different sizes and profiles. Characteristic broken vesicles were present, and more than one protein was demonstrated in each absorptive vesicle. The yolk-sac endoderm was confirmed as the selective site for transmission of proteins to the foetal circulation, since only proteins readily detected in the foetal serum were present in, and below, the basement membrane. The paraplacental chorion was shown to be the site for transmission of proteins to the exocoel and the process to be one of diffusion. Unlike normal proteins, F.l.T.C. conjugates readily became localized within macrophages present in the paraplacental chorion and yolk-sac vascular mesenchyme. These findings are discussed in the light of differences previously shown to occur between transmission of proteins to the foetal fluids and to the foetal blood, and in the light of a current hypothesis to account for selection of proteins by the yolk-sac endoderm.


Author(s):  
Ichiro Yamamoto ◽  
Toshiaki Tachibana ◽  
Hiroko Maruyama ◽  
Noriyuki Komatsu ◽  
Hiroyuki Kuramoto ◽  
...  

We have paid attention to the alteration of glycosyltransferase in carcinoma cells, because it might be related to the malignancy of the cells. In this connection, localization of β1-4 galactosyl transferase (β1-4 Gal T) in human endometrial carcinoma cells was examined immunocytochemically using two kinds of cell lines, each of which showed different degree of differentiation.An antibody was purified from the rabbit antiserum against the synthetic peptide, IFNRLVFRGMSC (W89) of human β1-4 Gal T coupled with KLH (keyhole limpet hemocyanine) by protein A column and peptide-affinity column chromatography. The anti-W89 serum reacts to the C-terminus of human β 1-4 Gal T and to both membrane-bound and soluble forms of the enzyme. Cell line of well differentiated endometrial adenocarcinoma (I) and that of poorly differentiated endometrial adenocarcinoma (50B) were cultivated respectively in MEM medium containing 15% FCS and 2 mM glutamine for 4 d at 37°C under 5% CO2. The cells were fixed in a mixture of 4% paraformaldehyde and 0.1% glutaraldehyde in 0.1 M Soerensen’s phosphate buffer (pH 7.4) at 4°C for 30 min, washed with PBS, then freezed and thawed. The indirect method of the peroxidase- labeled antibody technique was used for immunocytochemistry of both LM and TEM on the cell lines. The cells were dehydrated in ethanol and embedded in TAAB 812. Ultrathin sections were observed under a TEM, JEM-100S.


Author(s):  
Ray A. Weigand ◽  
Gregory C. Varjabedian

We previously described the intracellular localization of murine mammary tumor virus (MuMTV) p28 protein in thin sections (1). In that study, MuMTV containing cells fixed in 3% paraformaldehyde plus 0.05% glutaraldehyde were labelled after thin sectioning using ferritin-antiferritin in an unlabelled antibody technique. We now describe the labelling of murine leukemia virus (MuLV) particles using the unlabelled antibody technique coupled to ferritin-Fab antiferritin. Cultures of R-MuLV in NIH/3T3 cells were grown to 90% confluence (2), fixed with 2% paraformaldehyde plus 0.5% glutaraldehyde in 0.1 M cacodylate at pH 7.2, postfixed with buffered 17 OsO4, dehydrated with a series of etha-nols, and embedded in Epon. Thin sections were collected on nickel grids, incubated in 107 H2O2, rinsed in HEPES buffered saline, and subjected to the immunoferritin labelling procedure. The procedure included preincubation in 27 egg albumin, a four hour incubation in goat antisera against purified gp69/71 of MuLV (3) (primary antibody), incubation in F(ab’)2 fragments of rabbit antisera to goat IgG (secondary antibody), incubation in apoferritin, incubation in ferritin-Fab ferritin, and a brief fixation with 2% glutaraldehyde. The sections were stained with uranyl acetate and examined in a Siemens IA electron microscope at an accelerating voltage of 60 KV.


1995 ◽  
Vol 31 (5-6) ◽  
pp. 439-442 ◽  
Author(s):  
D. C. Johnson ◽  
K. A. Reynolds ◽  
C. P. Gerba ◽  
I. L. Pepper ◽  
J. B. Rose

Raw sewage disposal in marine waters is a common practice in many countries. This practice raises health risk concerns of possible transmission of Giardia and Cryptosporidium. Both of these protozoa have been shown to be transmitted by recreational swimming. To date no studies have determined the efficiency of their detection and concentration in marine waters. This study evaluated the efficiency of their detection in tap water and from marine waters in Hawaii with two different filter types. This study compared a polypropylene fiber cartridge filter, DPPPY (1.0 μm nominal porosity) (Cuno, Meriden, CT) which is typically used for parasite detection and the Filterite negatively charged filter (0.45μm) (Filtemp Sales, Inc., Phoenix, AZ). The latter would allow for both viruses and parasites to be concentrated simultaneously. The organisms were removed from the filter by passing the eluent through the filters in the opposite direction of collection and detected by indirect immunofluorescence antibody staining specific for Giardia and Cryptosporidium. Processing was simpler and faster with the Filterite filter and the overall efficiency for both Giardia and Cryptosporidium detection was greater. These methods are currently being used for the detection of the oocysts and cysts at bathing beaches in Hawaii impacted by marine sewage discharge.


2000 ◽  
Vol 41 (4-5) ◽  
pp. 301-308 ◽  
Author(s):  
N. Noda ◽  
H. Ikuta ◽  
Y. Ebie ◽  
A. Hirata ◽  
S. Tsuneda ◽  
...  

Fluorescent antibody technique by the monoclonal antibody method is very useful and helpful for the rapid quantification and in situ detection of the specific bacteria like nitrifiers in a mixed baxterial habitat such as a biofilm. In this study, twelve monoclonal antibodies against Nitrosomonas europaea (IFO14298) and sixteen against Nitrobacter winogradskyi (IFO14297) were raised from splenocytes of mice (BALB/c). It was found that these antibodies exhibited little cross reactivity against various kinds of heterotrophic bacteria. The direct cell count method using monoclonal antibodies could exactly detect and rapidly quantify N. europaea and N. winogradskyi. Moreover, the distribution of N. europaea and N. winogradskyi in a biofilm could be examined by in situ fluorescent antibody technique. It was shown that most of N. winogradskyi existed near the surface part and most of N. europaea existed at the inner part of the polyethylene glycol (PEG) gel pellet, which had entrapped activated sludge and used in a landfill leachate treatment reactor. It was suggested that this monoclonal antibody method was utilized for estimating and controlling the population of nitrifying bacteria as a quick and favorable tool.


Science ◽  
1964 ◽  
Vol 145 (3635) ◽  
pp. 943-945 ◽  
Author(s):  
G. C. Brown ◽  
H. F. Maassab ◽  
J. A. Veronelli ◽  
T. J. Francis

2000 ◽  
Vol 279 (6) ◽  
pp. R2015-R2021 ◽  
Author(s):  
Philippe Bauer ◽  
Tomas Welbourne ◽  
Takeharu Shigematsu ◽  
Janice Russell ◽  
D. Neil Granger

Although bacterial endotoxins [lipopolysaccharide (LPS)] can confer tissue resistance to subsequent inflammatory insults, the mechanisms that underlie this LPS-preconditioning (LPS-PC) response remain poorly defined. The dual-radiolabeled monoclonal antibody technique was used to examine whether LPS-PC alters the upregulation (protein) of E- and P-selectins after subsequent LPS challenge. In the gut of wild-type (C57BL/6J) mice, LPS-PC was associated with a reduction in E- (66%) and P-selectin (33%) expression. A similar reduction in E-selectin expression was observed in mutant mice that were genetically deficient in either the endothelial or inducible isoform of nitric oxide synthase or that overexpressed the human gene for Cu/Zn superoxide dismutase. Severe combined immunodeficient mice, genetically devoid of lymphocytes, did exhibit partial inhibition of the LPS-PC response. We conclude that 1) LPS-PC can be demonstrated for E- and P-selectins in some vascular beds (e.g., gut), 2) the mechanism(s) underlying this blunted selectin response does not include a major role for either nitric oxide and superoxide, and 3) circulating lymphocytes may contribute to the LPS-PC response.


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