Evaluation of GENECUBE Mycoplasma for the detection of macrolide-resistant Mycoplasma pneumoniae.

Introduction. Resistance against macrolide antibiotics in Mycoplasma pneumoniae is becoming non-negligible in terms of both appropriate therapy and diagnostic stewardship. Molecular methods have attractive features for the identification of Mycoplasma pneumoniae as well as its resistance-associated mutations of 23S ribosomal RNA (rRNA).Hypothesis/Gap Statement. The automated molecular diagnostic sytem can identify macrolide-resistant M. pneumoniae.Aim. To assess the performance of an automated molecular diagnostic system, GENECUBE Mycoplasma, in the detection of macrolide resistance-associated mutations.Methodology. To evaluate whether the system can distinguish mutant from wild-type 23S rRNA, synthetic oligonucleotides mimicking known mutations (high-level macrolide resistance, mutation in positions 2063 and 2064; low-level macrolide resistance, mutation in position 2067) were assayed. To evaluate clinical oropharyngeal samples, purified nucleic acids were obtained from M. pneumoniae-positive samples by using the GENECUBE system from nine hospitals. After confirmation by re-evaluation of M. pneumoniae positivity, Sanger-based sequencing of 23S rRNA and mutant typing using GENECUBE Mycoplasma were performed.Results. The system reproducibly identified all synthetic oligonucleotides associated with high-level macrolide resistance. Detection errors were only observed for A2067G (in 2 of the 10 measurements). The point mutation in 23S rRNA was detected in 67 (26.9 %) of 249 confirmed M. pneumoniae-positive clinical samples. The mutations at positions 2063, 2064 and 2617 were observed in 65 (97.0 %), 2 (3.0 %) and 0 (0.0 %) of the 67 samples, respectively. The mutations at positions 2063 and 2064 were A2063G and A2064G, respectively. The results from mutant typing using GENECUBE Mycoplasma were in full agreement with the results from sequence-based typing.Conclusion. GENECUBE Mycoplasma is a reliable test for the identification of clinically significant macrolide-resistant M. pneumoniae.

[Development and effectiveness of care preventive supporter training programs based on a literature review].

Objective　Many preventive care supporter (e.g. kaigo-yobo supporter) training programs, conducted to train community residents, are developed by municipalities. However, it is not necessary that only municipalities can train people effectively or efficiently. In this paper, we initially reviewed the relevant literature and clarified the definitions of concepts like "program contents" and "evaluation indicators," while also planning our own training programs. Later, we developed a program based on the review and examined the results.Methods　The literature of the training program was examined, and the training program was developed based on the result. Four researchers and three public health nurses from a community general support center, in the Otsuchi Town of Iwate Prefecture, developed a training program from June to September 2017. The training program developed was then conducted from October to November 2017. To evaluate the participants' satisfaction with the program, a self-report survey was conducted. To evaluate the outcomes of the program, we measured their degree of comprehension of their community's challenges, before and after the program.Results　The training program was divided into two parts following the literature review. In the first part, the content of the supporters' activities following the program was determined (Type A), and, in the second, the same content was evaluated by the participants within the program (Type B). Type A consisted of various aspects including both concrete knowledge and skills needed to conduct care preventiveactivities after the program. In Type B, there were many aspects-including both lectures and exercises-that aimed to increase the participants' awareness of community challenges, as well as inspection to learn about pioneering activities which helped them consider concrete care preventive activities following the program. In Otsuchi Town, we found it to be imperative for participants to consider how to respond to various situations and accordingly plan the training program for use in Type B. To evaluate the results, 12 participants were analyzed. Participants included two men and ten women, with an average age of 71.4±10.0 years [range: 53-88]. Comprehension levels of community challenges (3.1→4.1, P=0.046), as well as the confidence to actively involve themselves in their own preventive care strategies (3.4→4.0, P=0.035), significantly increased after involvement in the program. However, their confidence to work for community preventive care support groups (3.1→3.5, P=0.227) did not increase significantly.Conclusion　We clarified certain viewpoints, such as the purpose, content, and evaluation indices of community care training programs, by reviewing the relevant literature. Based on the discovered viewpoints, we were then able to obtain certain results through implementing our own training programs, thereby significantly increasing participant comprehension and confidence levels.

Some pharmaceutical and inclusion properties of 2-hydroxybutyl-ß-cyclodextrin derivative.

2-Hydroxybutyl-ß-cyclodextrins (HB-ß-CyDs) with different degrees of substitution (D.S.) were prepared and their physicochemical and biological properties and solubilizing abilities were studied and compared with those of 2-hydroxypropyl-ß-cyclodextrin (HP-ß-CyD). The surface activity of HB-ß-CyD was higher than that of HP-ß-CyD (D.S. 5.6) and increased with its concentration and D.S. The moisture sorption of HB-ß-CyD (D.S. 5.5) was less than that of HP-ß-CyD (D.S. 5.6), because of the introduction of hydrophobic hydroxybutyl groups in a molecule. The hemolytic activity (rabbit erythrocytes) decreased in the order of 2,6-di-O-methyl-ß-cyclodextrin (DM-ß-CyD)>methyl-ß-cyclodextrin (M-ß-CyD)>HB-ß-CyD (D.S. 5.5)>ß-CyD>HP-ß-CyD (D.S. 5.6). The hemolytic activity of HB-ß-CyD increased with D.S. and HB-ß-CyD induced echinocyte (or crenation), as well as DM-ß-CyD does. It was suggested from the solubility study of membrane components that HB-ß-CyD interacted predominantly with cholesterol in erythrocytes, resulting in the hemolysis. The inclusion ability of HB-ß-CyD was higher than that of HP-ß-CyD (D.S. 5.6), especially for poorly water-soluble drugs with long linear structures such as biphenylylacetic acid and flurbiprofen (FP). For example, HB-ß-CyD formed the inclusion complex with FP in a molar ratio of 1:1, by including the biphenyl moiety in the host cavity. The dissolution rate of FP/HB-ß-CyD (D.S. 5.5) complex was faster than that of HP-ß-CyD (D.S. 5.6) complex. The results suggested that HB-ß-CyDs have considerable pharmaceutical potential and can work as a fast-dissolving carrier for poorly water-soluble drugs.

Serotonergic modulation of absence-like seizures in groggy rats: a novel rat model of absence epilepsy.

To explore the role of the serotonergic system in modulating absence seizures, we examined the effects of 5-HT(1A) and 5-HT(2) agonists on the incidence of spike-and-wave discharges (SWD) in Groggy (GRY) rats, a novel rat model of absence-like epilepsy. GRY rats exhibited spontaneous absence-like seizures characterized by the incidence of sudden immobile posture and synchronously-associated SWD. The total duration of SWD in GRY rats was about 300 - 400 s/15-min observation period under the control conditions. However, the incidence of SWD was markedly reduced either by the 5-HT(1A) agonist (±)-8-hydroxy-2-(di-n-propylamino)-tetralin [(±)8-OH-DPAT] or the 5-HT(2) agonist (±)-1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane [(±)DOI]. The 5-HT reuptake inhibitors, fluoxetine and clomipramine, also inhibited the SWD generation. In addition, the inhibitory effects of (±)8-OH-DPAT and (±)DOI were reversed by WAY-100135 (5-HT(1A) antagonist) and ritanserin (5-HT(2) antagonist), respectively. The present results suggest that the serotonergic system negatively regulates the incidence of absence seizures by stimulation of 5-HT(1A) and 5-HT(2) receptors.