scholarly journals Multilocus Genotyping of Pneumocystis jirovecii from Deceased Cuban AIDS Patients Using Formalin-Fixed and Paraffin-Embedded Tissues

2021 ◽  
Vol 7 (12) ◽  
pp. 1042
Author(s):  
Vicente Friaza ◽  
Yaxsier de Armas ◽  
Virginia Capó ◽  
Rubén Morilla ◽  
Arturo Plascencia-Hernández ◽  
...  
Keyword(s):  
Mitochondrial Gene ◽  
Pcr Amplification ◽  
Pneumocystis Jirovecii ◽  
Nucleotide Sequencing ◽  
Genotype 3 ◽  
Aids Patients ◽  
Tissue Samples ◽  
Epidemiology Studies ◽  
Formalin Fixed

The results of the genotypic characterization of Pneumocystis jirovecii are described in lung tissue samples from 41 Cubans who died of AIDS with pneumocystosis between 1995 and 2008. Histological sections of the lung preserved as formalin-fixed and paraffin-embedded tissue were examined. PCR amplification and nucleotide sequencing of the two mitochondrial genes (large and small) of the pathogen allowed verification of a predominance of genotype 3 (85T/248C) of the large mitochondrial gene and genotype 3 (160A/196T) of the small mitochondrial gene over a period of 14 years (1995–2008). These results suggest that the 85T/248C//160A/196T genotype circulates with the highest frequency (81.3%) among AIDS patients in Cuba. Multilocus analysis indicates a limited circulation of pathogen genotypes on the island with the existence of a clonal genotype with an epidemic structure. Furthermore, it appears that circulating strains of P. jirovecii have not developed mutations related to sulfonamide resistance. Taken together, the data in this study revealed important elements about pneumocystosis in Cuban patients dying of AIDS and the usefulness of formalin-fixed and paraffin-embedded samples to carry out molecular epidemiology studies of P. jirovecii.

scholarly journals Characterization of Serologically NontypeableActinobacillus actinomycetemcomitans Isolates

1998 ◽  
Vol 36 (7) ◽  
pp. 2019-2022 ◽  
Author(s):  
Susanna Paju ◽  
Maria Saarela ◽  
Satu Alaluusua ◽  
Paula Fives-Taylor ◽  
Sirkka Asikainen
Keyword(s):  
Restriction Analysis ◽  
Pcr Amplification ◽  
Actinobacillus Actinomycetemcomitans ◽  
Genotype 3 ◽  
Banding Patterns ◽  
Arbitrarily Primed Pcr

Our previous studies have shown that Actinobacillus actinomycetemcomitans isolates of a given arbitrarily primed PCR (AP-PCR) genotype belong to the same serotype (of serotypes a through e). In the present study we investigated whether the AP-PCR genotypes of nonserotypeable A. actinomycetemcomitans isolates match those of the serotypeable isolates. The isolates were additionally characterized by restriction analysis of the apaH PCR amplification products. The material included 75 nonserotypeable and 18 serotypeable A. actinomycetemcomitans isolates from 34 epidemiologically unrelated subjects. The serotypeable isolates were obtained from subjects who also harbored nonserotypeable isolates. Eight AP-PCR genotypes were distinguished among the isolates; six genotypes matched those detected in our previous studies, whereas two genotypes were new. Intraindividually, the A. actinomycetemcomitans isolates produced identical AP-PCR banding patterns, regardless of whether they were serotypeable or nonserotypeable, in 22 of 23 subjects participating with multiple isolates. AP-PCR genotype 3, corresponding to serotype c, was by far the most common among the nonserotypeable isolates (62% of subjects). Results obtained with the apaH restriction analysis confirmed the results obtained with AP-PCR for 31 of the 34 subjects. The results suggest that nonserotypeable A. actinomycetemcomitans isolates originate from serotypeable isolates, especially from serotype c isolates, and the likelihood of the existence of additional serotypes is small.

scholarly journals Clinical, pathological and molecular evaluations and CT scan screening of coenurosis (Coenurus cerebralis) in sheep and calves

2017 ◽  
Vol 26 (1) ◽  
pp. 3-9 ◽  
Author(s):  
Abdullah Gazioglu ◽  
Sami Simsek ◽  
Omer Kizil ◽  
Ali Osman Ceribasi ◽  
Harun Kaya Kesik ◽  
...  
Keyword(s):  
Ct Scan ◽  
Mitochondrial Gene ◽  
Pcr Amplification ◽  
Cerebellar Hemisphere ◽  
Left Frontal Lobe ◽  
Coenurus Cerebralis ◽  
Hematoxylin And Eosin ◽  
The Right ◽  
Brain Cysts

Abstract The aims of this study were to diagnose coenurosis by means of computerized tomography (CT) scan imaging and molecular characterization of the CO1 gene using the polymerase chain reaction (PCR). Sheep and calves were necropsied, and CT scans on the cephalic region were performed on the animals. Sections of brain tissue infected with parasites were then stained with hematoxylin and eosin for microscopic examination. Material collected from brain cysts was fixed in 70% ethanol. PCR amplification was carried out using the CO1 mitochondrial gene. A total of 60 calves and 80 sheep were examined clinically and, of these, 15 calves and 38 sheep showed signs of depression, with counterclockwise circling movements and altered head carriage. Four sheep and one calf were necropsied, and C. cerebralis cysts were detected in all of them. A hypodense cyst was monitored in the right cerebellar hemisphere on a CT scan on one sheep. A cyst was found in the left frontal lobe on a CT scan on one calf. Microscopically, C. cerebralis cysts were surrounded by a fibrous or epithelial wall that presented necrosis on cerebral sections of both the sheep and the cattle. The CO1-PCR assay yielded a 446 bp band, which was sequenced and phylogenetically analyzed: the results confirmed the presence of T. multiceps. This study reports the first use of CT imaging on naturally infected calves and sheep for diagnosing coenurosis.

scholarly journals Detection of a putative novel adenovirus by PCR amplification, sequencing and phylogenetic characterisation of two gene fragments from formalin-fixed paraffin-embedded tissues of a cat diagnosed with disseminated adenovirus disease

2017 ◽  
Vol 65 (4) ◽  
pp. 574-584 ◽  
Author(s):  
Béla Lakatos ◽  
Ákos Hornyák ◽  
Zoltán Demeter ◽  
Petra Forgách ◽  
Frances Kennedy ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Polymerase ◽  
Pcr Amplification ◽  
Adenovirus Infection ◽  
Chain Reaction ◽  
Tissue Samples ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Polymerase Chain ◽  
Formalin Fixed

Adenoviral nucleic acid was detected by polymerase chain reaction (PCR) in formalin-fixed paraffin-embedded tissue samples of a cat that had suffered from disseminated adenovirus infection. The identity of the amplified products from the hexon and DNA-dependent DNA polymerase genes was confirmed by DNA sequencing. The sequences were clearly distinguishable from corresponding hexon and polymerase sequences of other mastadenoviruses, including human adenoviruses. These results suggest the possible existence of a distinct feline adenovirus.

Characterization of a cDNA for Rhesus Monkey Protein C Inhibitor – Evidence for N-Terminal Involvement in Heparin Stimulation

1995 ◽  
Vol 74 (04) ◽  
pp. 1079-1087 ◽  
Author(s):  
Klaus-P Radtke ◽  
José A Fernández ◽  
Bruno O Villoutreix ◽  
Judith S Greengard ◽  
John H Griffin
Keyword(s):  
Rhesus Monkey ◽  
Protein C ◽  
Activated Protein C ◽  
Pcr Amplification ◽  
Amino Acid Sequences ◽  
Heparin Binding ◽  
Reactive Center ◽  
Protein C Inhibitor ◽  
Polymerase Chain

SummarycDNAs for protein C inhibitor (PCI) were cloned from human and rhesus monkey 1 liver RNAs by reverse transcription and polymerase chain reaction (PCR) amplification. Sequencing showed that rhesus monkey and human PCI cDNAs were 93% identical. Predicted amino acid sequences differed at 26 of 387 residues. Pour of these differences (T352M, N359S, R362K, L3631) were in the reactive center loop that is important for inhibitory specificity, and two were in the N-terminal helix (M8T, E13K) that is implicated in glycosaminoglycan binding. PCI in human or rhesus monkey plasma showed comparable inhibitory activity towards human activated protein C in the presence of 10 U/ml heparin. However, maximal acceleration of the inhibition of activated protein C required 5-fold lower heparin concentration for rhesus monkey than for human plasma, consistent with the interpretation that the additional positive charge (E13K) in a putative-heparin binding region increased the affinity for heparin.

Streptococcus pluranimalium meningoencephalitis in a horse

2021 ◽  
pp. 104063872110234
Author(s):  
Dah-Jiun Fu ◽  
Akhilesh Ramachandran ◽  
Craig Miller
Keyword(s):  
Ribosomal Rna ◽  
Cross Sections ◽  
Nucleotide Sequencing ◽  
Formalin Fixed Paraffin ◽  
Quarter Horse ◽  
Formalin Fixed Paraffin Embedded ◽  
History Of ◽  
The Brain ◽  
Gram Positive Cocci ◽  
Formalin Fixed

A 3-y-old, female Quarter Horse with a history of acute neurologic signs was found dead and was submitted for postmortem examination. Areas of petechial and ecchymotic hemorrhage were present on cross-sections of the cerebrum, cerebellum, and brainstem. Histologic examination of the brain revealed severe, purulent meningoencephalitis and vasculitis with a myriad of intralesional gram-positive cocci. Streptococcus pluranimalium was identified from formalin-fixed, paraffin-embedded tissue obtained from sites with active lesions by PCR and nucleotide sequencing of bacterial 16S ribosomal RNA. S. pluranimalium should be considered as a cause of meningoencephalitis in a horse.

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