Chlamydial Infection-Dependent Synthesis of Sphingomyelin as a Novel Anti-Chlamydial Target of Ceramide Mimetic Compounds

The obligate intracellular bacterium Chlamydia trachomatis is the major causative agent of bacterial sexually transmitted diseases worldwide. In infected cells, the ceramide transport protein (CERT) is recruited to inclusions, where C. trachomatis replicates using host-synthesized ceramide. The ceramide is converted to sphingomyelin (SM) by a chlamydial infection-dependent SM synthesis (cidSM-synthesis) pathway, which occurs even in the absence of the SM synthases (SMS)-1 and -2 of host cells. The ceramide mimetic compound (1R,3S)-HPA-12 and the nonmimetic compound E16A, both of which are potent inhibitors of CERT, repressed the proliferation of C. trachomatis in HeLa cells. Unexpectedly, (1R,3R)-HPA-12, a ceramide mimetic compound that lacks CERT inhibitory activity, also exhibited potent anti-chlamydial activity. Using endogenous SMS-knockout mutant HeLa cells, we revealed that (1R,3R)-HPA-12 mildly inhibited cidSM-synthesis. In addition, LC-MS analysis revealed that (1R,3R)-HPA-12 is converted to a phosphocholine-conjugated metabolite in an infection-dependent manner. Imaging analysis with a fluorescent analog of ceramide suggested that cidSM-synthesis occurs in the bacterial bodies and/or inclusions. Collectively, these results suggested that (1R,3R)-HPA-12 exerts its anti-chlamydia activity not only as an inhibitor of cidSM-synthesis, but also via putative toxic effects of its phosphocholine adduct, which is most likely produced by the cidSM-synthesis route

Juvenile hormone acid O-methyltransferase in Drosophila melanogaster

Juvenile hormone (JH) acid O-methyltransferase (JHAMT) is the enzyme that transfers a methyl groupfrom S-adenosyl-L-methionine (SAM) to the carboxyl group of JH acids to produce active JHs in thecorpora allata. While the JHAMT gene was originally identified and characterized in the silkwormBombyx mori, no orthologs from other insects have been studied until now. Here we report on thefunctional characterization of the CG17330/DmJHAMT gene in the fruit fly Drosophila melanogaster.Recombinant DmJHAMT protein expressed in Escherichia coli catalyzes the conversion of farnesoic acidand JH III acid to their cognate methyl esters in the presence of SAM. DmJHAMT is predominantlyexpressed in corpora allata, and its developmental expression profile correlates with changes in the JHtiter. While a transgenic RNA interference against DmJHAMT has no visible effect, overexpression ofDmJHAMT results in a pharate adult lethal phenotype, similar to that obtained with application of JHanalogs, suggesting that the temporal regulation of DmJHAMT is critical for Drosophila development

Longitudinal Study on Quality of Life and Psychosocial Conditions in Light of Responses to Illness-Related Information in Postoperative Cancer Patients

Objective: Illness-related information can be significant for cancer patients after gastrointestinal (GI) surgery in terms of their performing adaptive tasks. This study longitudinally investigated the health outcomes of Japanese patients who read a booklet about cancer patients' problems and adaption tasks and evaluated the association between the responses to the booklet and the patients' health outcomes. Methods: A questionnaire survey about quality of life (QOL), fatigue, anxiety, cognitive plight, and resilience was administered to postoperative patients with GI cancer 1 week after their discharge from hospital and 6 months after surgery. The questionnaires were returned by email. Results: The mean age of the 32 patients at 1 week was 60.9 years; nearly 68.8% of them were men. As a whole, only two variables, QOL and anxiety, were significantly improved at 6 months over those at 1 week. Three statements were taken to gauge the responses to the booklet. In the two-way ANOVA that took QOL and responses to the booklet as independent variables, the post hoc test found that QOL was significantly improved in patients who agreed with the statement “I vaguely understood the content” or “I will deal with my tasks as described in the scenarios” but not in patients who agreed with the statement “The scenarios reflect my situation.” The anxiety in patients who agreed with the statement “The scenarios reflect my situation” was high at both survey points. Conclusions: This study suggests that associations between the responses to the informational booklet and patients' health outcomes partially indicate the directional property of how to support their information usage

Infection of Epstein–Barr Virus in Type III Latency Modulates Biogenesis of Exosomes and the Expression Profile of Exosomal miRNAs in the Burkitt Lymphoma Mutu Cell Lines

Infection of Epstein-Barr virus (EBV), a ubiquitous human gamma herpesvirus, is associated with various malignancies in B lymphocytes and epithelial cells. EBV encodes 49 microRNAs in two separated regions, termed the BART and BHRF1 loci. Although accumulating evidence demonstrates that EBV infection regulates the profile of microRNAs in the cells, little is known about the microRNAs in exosomes released from infected cells. Here, we characterized the expression profile of intracellular and exosomal microRNAs in EBV-negative, and two related EBV-infected Burkitt lymphoma cell lines having type I and type III latency by next-generation sequencing. We found that the biogenesis of exosomes is upregulated in type III latently infected cells compared with EBV-negative and type I latently infected cells. We also observed that viral and several specific host microRNAs were predominantly incorporated in the exosomes released from the cells in type III latency. We confirmed that multiple viral microRNAs were transferred to the epithelial cells cocultured with EBV-infected B cells. Our findings indicate that EBV infection, in particular in type III latency, modulates the biogenesis of exosomes and the profile of exosomal microRNAs, potentially contributing to phenotypic changes in cells receiving these exosomes

Longitudinal Study on Quality of Life and Psychosocial Conditions in Light of Responses to Illness-Related Information in Postoperative Cancer Patients

Objective: Illness-related information can be significant for cancer patients after gastrointestinal (GI) surgery in terms of their performing adaptive tasks. This study longitudinally investigated the health outcomes of Japanese patients who read a booklet about cancer patients\u27 problems and adaption tasks and evaluated the association between the responses to the booklet and the patients\u27 health outcomes. Methods: A questionnaire survey about quality of life (QOL), fatigue, anxiety, cognitive plight, and resilience was administered to postoperative patients with GI cancer 1 week after their discharge from hospital and 6 months after surgery. The questionnaires were returned by email. Results: The mean age of the 32 patients at 1 week was 60.9 years; nearly 68.8% of them were men. As a whole, only two variables, QOL and anxiety, were significantly improved at 6 months over those at 1 week. Three statements were taken to gauge the responses to the booklet. In the two-way ANOVA that took QOL and responses to the booklet as independent variables, the post hoc test found that QOL was significantly improved in patients who agreed with the statement "I vaguely understood the content" or "I will deal with my tasks as described in the scenarios" but not in patients who agreed with the statement "The scenarios reflect my situation." The anxiety in patients who agreed with the statement "The scenarios reflect my situation" was high at both survey points. Conclusions: This study suggests that associations between the responses to the informational booklet and patients\u27 health outcomes partially indicate the directional property of how to support their information usage

Relationship between the physical and psychosocial conditions of postoperative gastrointestinal cancer patients and their responses to an informational material

Objective: Postoperative patients with gastrointestinal (GI) cancer have multiple adaptation tasks and care needs to improve their quality of life (QOL). Whether their supportive care needs differ according to their physical and psychosocial conditions is unclear. This study investigated patients\u27 (1) physical and psychosocial conditions (QOL, fatigue, anxiety, cognitive plight, and resilience) and (2) responses to an informational booklet describing cancer patients\u27 problems and adaptation tasks, and examined the association between the two factors. Methods: A questionnaire survey was conducted to postoperative patients with GI cancer. Results: The mean age of the 69 respondents was 63 years; 59.4% of the respondents were men. Nine patients who did not read the booklet showed high fatigue and cognitive plight and low QOL. The patients (36.2%) who chose “I vaguely understood the content” showed low scores for resilience and cognitive plight while those (8.5%) who chose “I will deal with my tasks as described in the scenarios” showed high scores for both of these variables. Conclusions: The condition of some patients continued to be highly affected by their cancer. In terms of understanding the contents of the booklet, resilience was significant, and cognitive plight did not necessarily have a negative impact. The provision of information by means of a booklet might not be suitable for patients who are highly affected by their cancer. Patients may need additional support to be able to make good use of the information provided in such a booklet