[Evaluation of Hygiene in Dry Powder Steroid Drug Inhalers Used by Elderly Asthma Patients].

Dry powder inhalers (DPI) are frequently used by asthmatic patients, and the usage rate increases every year. The pharmacists at our hospital provided initial inhalation instructions on how the inhaler must be used but did not elaborate on the cleaning of the device. Therefore, the cleaning status of the inhaler is unknown, and there is a possibility of bacterial growth. This study investigated the cleaning status and hygiene of steroid drug inhalers used by elderly asthma patients. We administered a questionnaire to investigate the inhaler cleaning status after inhalation, and conducted a cross-sectional survey on hygiene using ATP measurement and bacterial culture examination. Considering the responses by 53 patients, it became clear that the ATP values of patients who answered "never cleaned" after inhalation were significantly higher than those who answered "cleaned every time". Moreover, some bacteria were detected in 62% of inhalers; 4 patients' inhalers contained bacteria other than normal oral microbial flora. In conclusion, because the inhalers used by elderly patients are in poor hygienic conditions, we must give cleaning instructions accordingly. We believe that it is necessary to give proper medical instructions along with instructions on the cleaning method with dry cloth.

Robust circadian rhythms in organoid cultures from PERIOD2::LUCIFERASE mouse small intestine.

Disruption of circadian rhythms is a risk factor for several human gastrointestinal (GI) diseases, ranging from diarrhea to ulcers to cancer. Four-dimensional tissue culture models that faithfully mimic the circadian clock of the GI epithelium would provide an invaluable tool to understand circadian regulation of GI health and disease. We hypothesized that rhythmicity of a key circadian component, PERIOD2 (PER2), would diminish along a continuum from ex vivo intestinal organoids (epithelial 'miniguts'), nontransformed mouse small intestinal epithelial (MSIE) cells and transformed human colorectal adenocarcinoma (Caco-2) cells. Here, we show that bioluminescent jejunal explants from PERIOD2::LUCIFERASE (PER2::LUC) mice displayed robust circadian rhythms for >72 hours post-excision. Circadian rhythms in primary or passaged PER2::LUC jejunal organoids were similarly robust; they also synchronized upon serum shock and persisted beyond 2 weeks in culture. Remarkably, unshocked organoids autonomously synchronized rhythms within 12 hours of recording. The onset of this autonomous synchronization was slowed by >2 hours in the presence of the glucocorticoid receptor antagonist RU486 (20 µM). Doubling standard concentrations of the organoid growth factors EGF, Noggin and R-spondin enhanced PER2 oscillations, whereas subtraction of these factors individually at 24 hours following serum shock produced no detectable effects on PER2 oscillations. Growth factor pulses induced modest phase delays in unshocked, but not serum-shocked, organoids. Circadian oscillations of PER2::LUC bioluminescence aligned with Per2 mRNA expression upon analysis using quantitative PCR. Concordant findings of robust circadian rhythms in bioluminescent jejunal explants and organoids provide further evidence for a peripheral clock that is intrinsic to the intestinal epithelium. The rhythmic and organotypic features of organoids should offer unprecedented advantages as a resource for elucidating the role of circadian rhythms in GI stem cell dynamics, epithelial homeostasis and disease.

Influence of visual information and test paradigm on clinical olfactory test results.

OBJECTIVE: The goal of this study was to determine if visual information and test paradigms affect clinical olfactory test results. METHODS: Three hundred and ninety-seven Japanese patients with complaints of olfactory dysfunction were administered both a new clinical olfactory test, the Odor Stick Identification Test for Japanese (OSIT-J), and the Japanese benchmark olfactory test, T&T olfactometry. Four different methods were used to administer the OSIT-J combining paradigms using word or picture-word alternatives with the four-plus alternative method based on a top-down strategy or the two-step identification method based on a bottom-up strategy. OSIT-J scores were compared for the different methods, referring to benchmark scores obtained with T&T olfactometry. RESULTS: OSIT-J scores using picture-word alternatives and the four-plus alternative method showed a stronger correlation with T&T olfactometry test scores than those using word alternatives and the two-step identification method, respectively. The average OSIT-J scores of the four-plus alternative method using picture-word alternatives were significantly higher than those using word alternatives in anosmic and severely hyposmic patients. The time required to administer the OSIT-J using both picture-word alternatives and the four-plus alternative method was the shortest of the four OSIT-J methods. CONCLUSIONS: Visual information and test paradigms may affect clinical olfactory test results. The OSIT-J method using picture-word alternatives and the four-plus alternative method may be the most suitable for clinical practice.

Thermodynamic instability of siRNA duplex is a prerequisite for dependable prediction of siRNA activities.

We developed a simple algorithm, i-Score (inhibitory-Score), to predict active siRNAs by applying a linear regression model to 2431 siRNAs. Our algorithm is exclusively comprised of nucleotide (nt) preferences at each position, and no other parameters are taken into account. Using a validation dataset comprised of 419 siRNAs, we found that the prediction accuracy of i-Score is as good as those of s-Biopredsi, ThermoComposition21 and DSIR, which employ a neural network model or more parameters in a linear regression model. Reynolds and Katoh also predict active siRNAs efficiently, but the numbers of siRNAs predicted to be active are less than one-eighth of that of i-Score. We additionally found that exclusion of thermostable siRNAs, whose whole stacking energy (DeltaG) is less than -34.6 kcal/mol, improves the prediction accuracy in i-Score, s-Biopredsi, ThermoComposition21 and DSIR. We also developed a universal target vector, pSELL, with which we can assay an siRNA activity of any sequence in either the sense or antisense direction. We assayed 86 siRNAs in HEK293 cells using pSELL, and validated applicability of i-Score and the whole DeltaG value in designing siRNAs.

Cerebellar activation of cortical motor regions: comparisons across mammals.

This chapter discusses the nature of mammalian cerebello-thalamo-cortical projections. These play an important role in motor control, but with species differences evident in the innervation patterns of thalamo-cortical (T-C) fibers relaying cerebellar inputs. A phylogenetic comparison of the mode of cerebellar activation of cortical motor regions reveals that cerebellar inputs are relayed by the deep and superficial T-C projections in the cat, but predominately by the latter in rat and monkey. Another difference across mammals is the nature of cerebellar activation of cortical neurons. Fast-conducting pyramidal tract neurons in the cat routinely receive fast-rising EPSPs from the deep cerebellar nuclei (CN). This tendency is not observed in rat and monkey, however. These findings suggest that the responsiveness of cortical output neurons also shows species differences, these having bearing on the development of species-specific motor skills. Common to all three species, however, are fast-rising and large CN-EPSP responses of layer III pyramidal neurons to CN input. It is argued that layer III pyramidal neurons modulate cerebellar input to layer V pyramidal neurons, which latter cells provide command signals from the motor cortex to the lower centers.