scholarly journals LR White is preferable to Unicryl for immunogold detection of fixationsensitive nuclear antigens

10.4081/1748 ◽  
2010 ◽  
Vol 46 (4) ◽  
pp. 359 ◽  
Author(s):  
V. V. Philimonenko ◽  
J. Janácek ◽  
P. Hozák

Author(s):  
Julie A. Martini ◽  
Robert H. Doremus

Tracy and Doremus have demonstrated chemical bonding between bone and hydroxylapatite with transmission electron microscopy. Now researchers ponder how to improve upon this bond in turn improving the life expectancy and biocompatibility of implantable orthopedic devices.This report focuses on a study of the- chemical influences on the interfacial integrity and strength. Pure hydroxylapatite (HAP), magnesium doped HAP, strontium doped HAP, bioglass and medical grade titanium cylinders were implanted into the tibial cortices of New Zealand white rabbits. After 12 weeks, the implants were retrieved for a scanning electron microscopy study coupled with energy dispersive spectroscopy.Following sacrifice and careful retrieval, the samples were dehydrated through a graduated series starting with 50% ethanol and continuing through 60, 70, 80, 90, 95, and 100% ethanol over a period of two days. The samples were embedded in LR White. Again a graduated series was used with solutions of 50, 75 and 100% LR White diluted in ethanol.



Author(s):  
David L. Spector ◽  
Robert J. Derby

Studies in our laboratory are involved in evaluating the structural and functional organization of the mammalian cell nucleus. Since several major classes (U1, U2, U4/U6, U5) of small nuclear ribonucleoprotein particles (snRNPs) play a crucial role in the processing of pre-mRNA molecules, we have been interested in the localization of these particles within the cell nucleus. Using pre-embedding immunoperoxidase labeling combined with 3-dimensional reconstruction, we have recently shown that nuclear regions enriched in snRNPs form a reticular network within the nucleoplasm which extends between the nucleolar surface and the nuclear envelope. In the present study we were inte rested in extending these nuclear localizations using cell preparation techniques which avoid slow penetration of fixatives, chemical crosslinking of potential antigens and solvent extraction. CHOC 400 cells were cryofixed using a CF 100 ultra rapid cooling device (LifeCell Corp.). After cryofixation cells were molecular distillation dried, vapor osmicated, in filtra ted in 100% Spurr resin in vacuo and polymerized in molds a t 60°C. Using this procedure we were able to evaluate the distribution of snRNPs in resin embedded cells which had not been chemically fixed, incubated in cryoprotectants or extracted with solvents.





2021 ◽  
Vol 19 (1) ◽  
pp. 237-244
Author(s):  
Youssef EL Hassouni ◽  
Mohammed Bourhia ◽  
Ahmed Bari ◽  
Riaz Ullah ◽  
Hafiz Majid Mahmood ◽  
...  

Abstract Autoimmune diseases are pathological conditions in which the immune system mistakenly attacks its own tissues. This study evaluates the performance of two techniques, which are identifiers of autoantibody specifics: immunoblot and immunodot. This study was conducted in 300 patients of whom 62 were tested positive for antinuclear antibodies. The patients were initially screened for antinuclear antibodies using indirect immunofluorescence. Then, the identification of specific autoantibodies such as anti-extractable nuclear antigens (ENAs) was carried out using the immunoblot and immunodot techniques. The results showed that immunoblot and immunodot did not present a significant difference in their sensitivity against anti-SSA/52, SSB, CENP-B, PCNA, U1-snRNP, Jo-1, Pm-scl, and Mi-2 (p > 0.05). However, the two techniques showed a significant difference in their sensitivity toward autoantibodies anti-DNAn, anti-histone, anti-SmD1, and anti-ds-DNA (p < 0.05). The immunoblot data were in complete accordance with the immunodot data (100%) regarding the detection of autoantibodies such as anti SSA/52, SSB, CENP-B, PCNA, U1-snRP, Jo-1, Pm-scl, and Mi-2, 80% regarding SmD1, and 75% concerning ds-DNA. We should certainly pay closer attention to the efficiency of the techniques used in the diagnosis of autoimmune diseases.



Viruses ◽  
2021 ◽  
Vol 13 (2) ◽  
pp. 175
Author(s):  
Antony Idam Mamimandjiami ◽  
Augustin Mouinga-Ondémé ◽  
Jill-Léa Ramassamy ◽  
Délia Doreen Djuicy ◽  
Philippe V. Afonso ◽  
...  

Human herpesvirus 8 (HHV-8) is the etiological agent of all forms of Kaposi’s sarcoma (KS). K1 gene studies have identified five major molecular genotypes with geographical clustering. This study described the epidemiology of HHV-8 and its molecular diversity in Gabon among Bantu and Pygmy adult rural populations and KS patients. Plasma antibodies against latency-associated nuclear antigens (LANA) were searched by indirect immunofluorescence. Buffy coat DNA samples were subjected to polymerase chain reaction (PCR) to obtain a K1 gene fragment. We studied 1020 persons; 91% were Bantus and 9% Pygmies. HHV-8 seroprevalence was 48.3% and 36.5% at the 1:40 and 1:160 dilution thresholds, respectively, although the seroprevalence of HHV-8 is probably higher in Gabon. These seroprevalences did not differ by sex, age, ethnicity or province. The detection rate of HHV-8 K1 sequence was 2.6% by PCR. Most of the 31 HHV-8 strains belonged to the B genotype (24), while the remaining clustered within the A5 subgroup (6) and one belonged to the F genotype. Additionally, we reviewed the K1 molecular diversity of published HHV-8 strains in Africa. This study demonstrated a high seroprevalence of HHV-8 in rural adult populations in Gabon and the presence of genetically diverse strains with B, A and also F genotypes.





1985 ◽  
Vol 6 (11) ◽  
pp. 314-315 ◽  
Author(s):  
D.I Stott
Keyword(s):  


2021 ◽  
Vol 17 (5) ◽  
pp. 250-257
Author(s):  
Yasser Emad ◽  
Yasser Ragab ◽  
Nevin Hammam ◽  
Nashwa El-Shaarawy ◽  
Ossama Ibrahim ◽  
...  


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