Valproic Acid Inhibits Chronic Toxoplasma Infection and Associated Brain Inflammation in Mice.

Individuals infected with Toxoplasma gondii are prone to psychobehavioral disorders, most notably schizophrenia and bipolar disorder. Valproic acid reportedly inhibits the proliferation of T. gondii tachyzoites in vitro. However, animals treated with the drug neither lived longer during acute infection nor had fewer brain cysts upon chronic infection. In this study, a quantitative real-time PCR (qPCR) method was applied to quantify copy numbers of BAG1 (a bradyzoite-specific protein), REP529 DNA (a repetitive DNA fragment of the parasite), and SAG1 (a highly expressed tachyzoite-specific surface protein) in the brains of chronically infected mice treated with valproic acid. The treatment inhibited the infection and decreased BAG1, SAG1, and REP529 copy numbers in mice brains (P < 0.0001), comparable to the effects of trimethoprim-sulfamethoxazole (TMP-SMZ), the common medication for toxoplasmosis treatment. Moreover, valproic acid decreased brain tumor necrosis factor alpha (TNF-α) expression (P < 0.0001) comparably to TMP-SMZ. Histological examination of mouse brains showed marked reductions in cyst establishment, perivascular infiltration of lymphocytes, and glial nodules to the same levels as those in the TMP-SMZ group. Our results provide direct evidence for the efficacy of valproic acid, a mood-stabilizing and antipsychotic drug, against chronic Toxoplasma infection. These results might help modulate therapeutic regimens for neuropsychiatric patients and aid in the design of more effective anti-Toxoplasma drugs.

Development of a Quantitative Real-Time PCR Assay for Detection of Toxoplasma gondii in Brain Samples.

BACKGROUND: Toxoplasmosis is a worldwide-distributed infection that can cause serious diseases, mainly in congenitally infected and immunodeficient individuals. PCR assays play an indispensable role in the detection of Toxoplasma gondii in different biological samples. METHODS: This study was conducted in the Parasitology Department at Pasteur Institute of Iran (Tehran) during 2016-2018. We designed a highly sensitive quantitative real-time PCR (RT-qPCR) targeted REP-529, a noncoding repetitive DNA. We cloned the amplicon in a plasmid (pTZREP-529) and used it to generate the standard curve. The Toxoplasma RT-qPCR characteristics, i.e., detection limit, specificity, linear dynamic range, linearity, intra-, and inter-assay precisions, were determined. The detection limit of the assay was one plasmid copy number (PCN) per reaction (about 0.004 T. gondii genome), and the linear dynamic range was equal to 6 logs (1× 101 to 1× 107 PCN per reaction). RESULTS: The assay showed no signal when genomic DNA of Plasmodium falciparum, Leishmania major, and Trichomonas vaginallis were used. The standard curve was drawn using dilutions of pTZREP-529 plasmid spiked with genomic DNA from a mouse brain, and test characteristics were shown unaffected. Applying the Toxoplasma RT-qPCR, we showed brain cysts were significantly decreased in mice vaccinated with GRA2 antigen of Toxoplasma formulated in Monophosphoryl Lipid A (MPL) adjuvant. CONCLUSION: We have developed a quantitative, specific, and highly sensitive PCR for detecting T. gondii in biological samples.

Two-step optimization process for grass hydrolysate application as biodiesel feedstock with novel quality characteristics.

A major obstacle to biodiesel commercialization is supplying feedback which increases production costs. The potential of some oleaginous yeast for conversion of waste materials to biodiesel feedstock can overcome this problem. In this study, a potential oleaginous yeast strain was used for single-cell oil (SCO) production. Two sets of experiments were designed for the optimization process. According to the results obtained from the first experiment, lipid production and lipid content of this strain increased from 1.96 g/L and 22.6% to 3.85 g/L and 35.18% by optimization of grass hydrolysis, respectively. The results of the second experiment indicate an increase in SCO production and lipid content to 7.28 g/L and 56.39%, respectively. These results were obtained when HNO3 was used for substrate pre-treatment. Lipid analysis by gas chromatography-mass spectrometry showed a suitable and high potential of fatty acid profile for biodiesel production, which was then confirmed by evaluating the physicochemical properties of the biodiesel obtained in compliance with the US and EU standards. Consumption of microbial oil and low-cost substrate can compensate the high costs of feedstock in biodiesel production.