Detection of human sarcocystosis using dried blood on filter papers: An immunofluorescent antibody test

Sarcocystosis, a parasitic infection caused by a protozoa belonging to the genus Sarcocystis, is found worldwide in both and animals. Sarcocystis spp., require two animal hosts to complete their life cycle. The infection has gathered more global attention after recent outbreaks, especially amongst wester travellers to Malaysia. Other than sporadic cases and the current outbreaks, little information is available regarding human Sarcocystis infection in Malaysia. The present study aims to determine the prevalence of sarcocystosis among humans using an immunofluorescent antibody (IFA) test applied to dried blood on filter papers. A total of 200 blood samples were collected on filter papers from autopsy cases at two Malaysian hospitals: Sungai Buloh Hospital (peninsular Malaysia) and Queen Elizabeth Hospital (Malaysian Borneo). Antigens were prepared from bradyzoites harvested from positive goats’ muscle samples. Of the 200 samples, 32 (16%) had Sarcocystis antibodies that showed positive fluorescence reactions on filter papers. There was no significant difference (t-test, p value > 0.05) in prevalence rates between samples collected from autopsies at peninsular Malaysia and Malaysian Borneo. The results demonstrated that the filter paper technique can be used as one of the alternative serological tests in the diagnostic of human sarcocystosis. © 2019, Malaysian Society for Parasitology. All rights reserved

Genome-Wide Analysis of Copy Number Variation Identifies Candidate Gene Loci Associated with the Progression of Non-Alcoholic Fatty Liver Disease

<div><p>Between 10 and 25% of individuals with non-alcoholic fatty liver disease (NAFLD) develop hepatic fibrosis leading to cirrhosis and hepatocellular carcinoma (HCC). To investigate the molecular basis of disease progression, we performed a genome-wide analysis of copy number variation (CNV) in a total of 49 patients with NAFLD [10 simple steatosis and 39 non-alcoholic steatohepatitis (NASH)] and 49 matched controls using high-density comparative genomic hybridization (CGH) microarrays. A total of 11 CNVs were found to be unique to individuals with simple steatosis, whilst 22 were common between simple steatosis and NASH, and 224 were unique to NASH. We postulated that these CNVs could be involved in the pathogenesis of NAFLD progression. After stringent filtering, we identified four rare and/or novel CNVs that may influence the pathogenesis of NASH. Two of these CNVs, located at 13q12.11 and 12q13.2 respectively, harbour the exportin 4 (<i>XPO4</i>) and phosphodiesterase 1B (<i>PDE1B</i>) genes which are already known to be involved in the etiology of liver cirrhosis and HCC. Cross-comparison of the genes located at these four CNV loci with genes already known to be associated with NAFLD yielded a set of genes associated with shared biological processes including cell death, the key process involved in ‘second hit’ hepatic injury. To our knowledge, this pilot study is the first to provide CNV information of potential relevance to the NAFLD spectrum. These data could prove invaluable in predicting patients at risk of developing NAFLD and more importantly, those who will subsequently progress to NASH.</p></div

Rare and novel CNVs in NASH.

<p>Start = first base-pair location in the copy number region, End = last base-pair location in the copy number region.</p

Enriched GO terms associated with NASH.

<p>GO_BP, Gene Ontology biological process; GO_CC, Gene Ontology cellular component.</p><p>*Modified Fisher’s Exact test, <i>P</i>-Value ≤0.05.</p

Size range distribution of the CNVs.

<p>The CNVs ranged in size from 5.77</p

qPCR validation performed on selected genomic regions.

<p>(<b>A</b>) Results for the region 11q11. (<b>B</b>) Results for the region 13q12.11. A copy number of around 2 was deemed to be indicative of wild-type status (i.e. no CNV), a copy number of 1 was indicative of one copy lost, whereas a copy number of 3 or above was held to indicate copy number gain(s). The error bars represent the standard error among four replicates.</p

Top regions of copy number gains and losses in NASH.

<p>Start = first base-pair location in the copy number region, End = last base-pair location in the copy number region.</p