Honokiol Blocks Store Operated Calcium Entry in CHO Cells Expressing the M3 Muscarinic Receptor: Honokiol and Muscarinic Signaling

Background: Honokiol, a cell-permeable phenolic compound derived from the bark of magnolia trees and present in Asian herbal teas, has a unique array of pharmacological actions, including the inhibition of multiple autonomic responses. We determined the effects of honokiol on calcium signaling underlying transmission mediated by human M3 muscarinic receptors expressed in Chinese hamster ovary (CHO) cells. Receptor binding was determined in radiolabelled ligand binding assays; changes in intracellular calcium concentrations were determined using a fura-2 ratiometric imaging protocol; cytotoxicity was determined using a dye reduction assay. Results: Honokiol had a potent (EC50 ≈ 5 μmol/l) inhibitory effect on store operated calcium entry (SOCE) that was induced by activation of the M3 receptors. This effect was specific, rapid and partially reversible, and was seen at concentrations not associated with cytotoxicity, inhibition of IP3 receptor-mediated calcium release, depletion of ER calcium stores, or disruption of M3 receptor binding. Conclusions: It is likely that an inhibition of SOCE contributes to honokiol disruption of parasympathetic motor functions, as well as many of its beneficial pharmacological properties

The relationship between preoperative American Society of Anesthesiologists Physical Status Classification scores and functional recovery following hip-fracture surgery

Abstract Background Little is known about the relationship of the American Society of Anesthesiologists Physical Status Classification scores (ASA scores) on patient outcomes following hip fracture surgery in Asian countries. Therefore, this study explored the association of patients’ preoperative ASA scores on trajectories of recovery in physical functioning and health outcomes during the first year following postoperative discharge for older adults with hip-fracture surgery in Taiwan. Methods The data for this study was generated from three prior studies. Participants (N = 226) were older hip-fracture patients from an observational study (n = 86) and two clinical trials (n = 61 and n = 79). Participants were recruited from the trauma wards of one medical center in northern Taiwan and data was collected prior to discharge and at 1, 3, 6, and 12 months after hospital discharge. Participants were grouped as ASA class 1–2 (50.5%; ASA Class 1, n = 7; ASA Class 2, n = 107) and ASA class 3 (49.5%, n = 112). Measures for mortality, service utilization, activities of daily living (ADL), measured by the Chinese Barthel Index, and health related quality of life, measured by Medical Outcomes Study Short Form-36, were assessed for the two groups. Generalized estimating equations (GEE) were used to analyze the changes over time for the two groups. Results During the first year following hip-fracture surgery, ASA class 1–2 participants had significantly fewer rehospitalizations (6%, p = .02) and better scores for mental health (mean = 70.29, standard deviation = 19.03) at 6- and 12-months following discharge than those classified as ASA 3. In addition, recovery of walking ability (70%, p = .001) and general health (adjusted mean = 58.31, p = .003) was also significantly better than ASA 3 participants. Conclusions There was a significant association of hip-fracture patients classified as ASA 1–2 with better recovery and service utilization during the first year following surgery. Interventions for hip fractured patients with high ASA scores should be developed to improve recovery and quality of life.https://deepblue.lib.umich.edu/bitstream/2027.42/138818/1/12891_2017_Article_1768.pd

Robust estimation of bacterial cell count from optical density

Optical density (OD) is widely used to estimate the density of cells in liquid culture, but cannot be compared between instruments without a standardized calibration protocol and is challenging to relate to actual cell count. We address this with an interlaboratory study comparing three simple, low-cost, and highly accessible OD calibration protocols across 244 laboratories, applied to eight strains of constitutive GFP-expressing E. coli. Based on our results, we recommend calibrating OD to estimated cell count using serial dilution of silica microspheres, which produces highly precise calibration (95.5% of residuals <1.2-fold), is easily assessed for quality control, also assesses instrument effective linear range, and can be combined with fluorescence calibration to obtain units of Molecules of Equivalent Fluorescein (MEFL) per cell, allowing direct comparison and data fusion with flow cytometry measurements: in our study, fluorescence per cell measurements showed only a 1.07-fold mean difference between plate reader and flow cytometry data

Relationship between physical activity and perceptions of ageing from the perspective of healthy ageing among older people with frailty with chronic disease: a cross-sectional study

Abstract Background In Taiwan, the 2019 Elderly Frailty Assessment found that 11.2% of older people have frailty problems. Some researchers have found that older persons’ negative perspectives on ageing aggravate the progression of frailty, thereby increasing their risk of disability. This study aimed to investigate associations of physical activity and perceptions of ageing on perspectives of healthy ageing in older people with frailty and chronic diseases and to compare the differences in their frailty status. Methods This study used a descriptive cross-sectional design. Participants were recruited from community long-term care stations. The inclusion criteria were (1) no severe cognitive impairment and ability to communicate in Mandarin and Taiwanese; (2) over 65 years old; (3) at least one chronic disease; and (4) at least one debilitating item in the Study of Osteoporotic Fracture index. A total of 312 participants were recruited. The Brief Ageing Perceptions Questionnaire Chinese version, Healthy Ageing Perspectives Questionnaire, and Physical Activity Scale for the Elderly Chinese Version were used for measurement. Results The study results found that demographic variables, perceptions of ageing, and physical activity were significantly correlated with perspectives on healthy ageing, including age, Activities of Daily Living, education, all domains of perceptions of ageing, and household- and work-related physical activity. With regard to the frailty status level, prefrailty was better than frailty from the perspective of healthy ageing in older people with chronic disease (t = 5.35, p < 0.05). Hierarchical regression analysis was used to predict the healthy ageing perspectives of older persons with chronic disease involving a chronic time-line, positive control, health-related changes, and work-related activities. Those domains could predict 21% of the variance in healthy ageing perspectives. Conclusion It is suggested that in community long-term care stations, health care providers can arrange activities to improve the perception of ageing that are acceptable for older people with frailty and chronic diseases and encourage older people to participate in service activities to achieve a sense of social participation

The effect of transcranial direct current stimulation on bilateral asymmetry and joint angles of the lower limb for females when crossing obstacles

Abstract Background Gait asymmetry is often accompanied by the bilateral asymmetry of the lower limbs. The transcranial direct current stimulation (tDCS) technique is widely used in different populations and scenarios as a potential tool to improve lower limb postural control. However, whether cerebral cortex bilateral tDCS has an interventional effect on postural control as well as bilateral symmetry when crossing obstacles in healthy female remains unknown. Methods Twenty healthy females were recruited in this prospective study. Each participant walked and crossed a height-adjustable obstacle. Two-way repeated ANOVA was used to evaluate the effect of group (tDCS and sham-tDCS) and height (30%, 20%, and 10% leg length) on the spatiotemporal and maximum joint angle parameters for lower limb crossing obstacles. The Bonferroni post-hoc test and paired t-test were used to determine the significance of the interaction effect or main effect. The statistically significant differences were set at p < 0.05. Results The Swing time (SW) gait asymmetry (GA), Stance time (ST) GA, leading limb hip-knee-ankle maximum joint angles and trailing limb hip-knee maximum joint angles decreased in the tDCS condition compared to the sham-tDCS condition at 30%, 20% leg’s length crossing height except for 10% leg’s length, whereas there was a significant decrease in SW/ST GA between the tDCS condition and the sham-tDCS condition at 30%, 20%, 10% leg’s length crossing height (P < 0.05). Conclusion We conclude that tDCS intervention is effective to reduce bilateral asymmetry in spatio-temporal parameters and enhance dynamic balance in female participants during obstacle crossing when the heights of the obstacles were above 10% of the leg’s length. Trial registration No ChiCTR2100053942 (date of registration on December 04, 2021). Prospectively registered in the Chinese Clinical Trial Registry

Biphenols Block Calcium Entry in Response to Activation of the M3 Muscarinic Receptor

Cell-permeable biphenols, such as honokiol, present in herbal teas, inhibit autonomic responses. The effects of biphenols on calcium signaling activated by human M3 muscarinic receptors was studied in Chinese hamster ovary (CHO) cells. Receptor binding was determined by radiolabelled ligand binding; intracellular calcium concentrations were determined using a fura-2 ratiometric imaging protocol; cytotoxicity was determined using a dye reduction assay. Activation of M3 muscarinic receptors with carbachol induced a biphasic increase in [Ca2+]i: an initial, IP3- mediated release of Ca2+ from endoplasmic reticulum stores followed by a sustained phase of steady Ca2+ entry (i.e., store operated calcium entry, SOCE). Biphneols had potent (EC50 ≈ μM) inhibitory effects on SOCE that was induced by the activation of the M3 receptor. SOCE was also examined by acute exposure to thapsigarin (2 μM) in a calcium-free medium, followed by reintroduction of calcium. Again, SOCE was severely depressed by biphenols. The effect of honokiol on SOCE was specific, rapid and partially reversible, and was seen at concentrations not associated with cytotoxicity, inhibition of IP3 receptor-mediated calcium release, or depletion of ER calcium stores. Moreover, biphenols did not disrupt the ligand binding properties of M3 receptors. An inhibition of SOCE probably contributes to biphenol disruption of parasympathetic functions