4-Hydroxybenzaldehyde Restricts the Intracellular Growth of Toxoplasma gondii by Inducing SIRT1-Mediated Autophagy in Macrophages

Toxoplasma gondii is an intracellular protozoan parasite that infects approximately one third of the human popu- lation worldwide. Considering the toxicity and side effects of anti-toxoplasma medications, it is important to develop effec- tive drug alternatives with fewer and less severe off-target effects. In this study, we found that 4-hydroxybenzaldehyde (4- HBA) induced autophagy and the expression of NAD-dependent protein deacetylase sirtuin-1 (SIRT1) in primary murine bone marrow-derived macrophages (BMDMs). Interestingly, treatment of BMDMs with 4-HBA significantly reduced the number of macrophages infected with T. gondii and the proliferation of T. gondii in infected cells. This effect was impaired by pretreating the macrophages with 3-methyladenine or wortmannin (selective autophagy inhibitors) or with sirtinol or EX527 (SIRT1 inhibitors). Moreover, we found that pharmacological inhibition of SIRT1 prevented 4-HBA-mediated expres- sion of LC3-phosphatidylethanolamine conjugate (LC3-II) and the colocalization of T. gondii parasitophorous vacuoles with autophagosomes in BMDMs. These data suggest that 4-HBA promotes antiparasitic host responses by activating SIRT1- mediated autophagy, and 4-HBA might be a promising therapeutic alternative for the treatment of toxoplasmosis.

A Novel Rapid Fungal Promoter Analysis System Using the Phosphopantetheinyl Transferase Gene, npgA, in Aspergillus nidulans

To develop a convenient promoter analysis system for fungi, a null-pigment mutant (NPG) of Aspergillus nidulans was used with the 4′-phosphopantetheinyl transferase (PPTase) gene, npgA, which restores the normal pigmentation in A. nidulans, as a new reporter gene. The functional organization of serially deleted promoter regions of the A. nidulans trpC gene and the Cryphonectria parasitica crp gene in filamentous fungi was representatively investigated to establish a novel fungal promoter assay system that depends on color complementation of the NPG mutant with the PPTase npgA gene. Several promoter regions of the trpC and crp genes were fused to the npgA gene containing the 1,034-bp open reading frame and the 966-bp 3’ downstream region from the TAA, and the constructed fusions were introduced into the NPG mutant in A. nidulans to evaluate color recovery due to the transcriptional activity of the sequence elements. Serial deletion of the trpC and crp promoter regions in this PPTase reporter assay system reaffirmed results in previous reports by using the fungal transformation step without a laborious verification process. This approach suggests a more rapid and convenient system than conventional analyses for fungal gene expression studies.

Viral Effects of a dsRNA Mycovirus (PoV-ASI2792) on the Vegetative Growth of the Edible Mushroom Pleurotus ostreatus

A double-stranded RNA (dsRNA) mycovirus was detected in malformed fruiting bodies of Pleurotus ostreatus strain ASI2792, one of bottle cultivated commercial strains of the edible oyster mushroom. The partial RNA-dependent RNA polymerase (RdRp) gene of the P. ostreatus ASI2792 mycovirus (PoV-ASI2792) was cloned, and a cDNA sequences alignment revealed that the sequence was identical to the RdRp gene of a known PoSV found in the P. ostreatus strain. To investigate the symptoms of PoV-ASI2792 infection by comparing the isogenic virus-free P. ostreatus strains with a virus-infected strain, isogenic virus-cured P. ostreatus strains were obtained by the mycelial fragmentation method for virus curing. The absence of virus was verified with gel electrophoresis after dsRNA-specific virus purification and Northern blot analysis using a partial RdRp cDNA of PoV-ASI2792. The growth rate and mycelial dry weight of virus-infected P. ostreatus strain with PoV-ASI2792 mycovirus were compared to those of three virus-free isogenic strains on 10 different media. The virus-cured strains showed distinctly higher mycelial growth rates and dry weights on all kinds of experimental culture media, with at least a 2.2-fold higher mycelial growth rate on mushroom complete media (MCM) and Hamada media, and a 2.7-fold higher mycelial dry weight on MCM and yeastmalt-glucose agar media than those of the virus-infected strain. These results suggest that the infection of PoV mycovirus has a deleterious effect on the vegetative growth of P. ostreatus.

What Is an Optimal Dose of Sufentanil for Attenuating the Hemodynamic Response to Tracheal Intubation?

BACKGROUND: Laryngoscopy and tracheal intubation often cause hemodynamic changes such as hypertension and tachycardia. This study was carried out to determine the optimal dose of sufentanil for attenuating the hemodynamic changes that occur during the induction of anesthesia with propofol. METHODS: The authors examined 100 ASA class 1-2 patients, who were scheduled for elective surgery anddivided randomly into 4 groups. Anesthesia was induced with propofol (5.0microg/kg target controlled infusion). Three minutes later, rocuronium 1.2 mg/kg was administered. Group 1 (CON group) received no sufentanil, and groups 2, 3 and 4 (SO3, SO5, SO7 groups) received 0.3, 0.5, 0.7 microg/kg, sufentanil, respectively. The hemodynamic changes and BIS were measured at preinduction, 1 and 3 minutes after propofol infusion, and 1 and 3 minutes after sufentanil infusion, intubation, and post-intubation period for 10 minutes. RESULTS: In the SO3, SO5, SO7 groups, the systolic and diastolic and mean arterial pressure did notincrease compared with that at preinduction. However, in the SO7 group, the systolic and diastolic and mean arterial pressure decreased significantly 1 minute after intubation. In the SO3 group, the heart rate increased significantly after intubation compared with preinduction. On the other hand, the heart rate did not increase after intubation in the SO5 and SO7 groups. CONCLUSIONS: When anesthesia is induced with propofol TCI (5.0 microg/ml, the authors suggest that the recommended dosage of sufentanil for attenuating the hemodynamic changes accompanying a laryngoscopy and tracheal intubation be approximately 0.5microg/kg.