Human Milk Exosomes Induce ZO-1 Expression via Inhibition of REDD1 Expression in Human Intestinal Epithelial Cells.

Human milk exosomes (HMEs) enhance intestinal barrier function and contribute to an improvement in inflammation and mucosal injury, such as necrotizing enteritis (NEC), in infants. Here, we aimed to elucidate the intracellular factors involved in HME-induced expression of zonula occludens-1 (ZO-1), a tight junction protein, in Caco-2 human intestinal epithelial cells. HME treatment for 72 h significantly increased transepithelial electrical resistance in these cells. The mean ZO-1 protein levels in cells treated with HME for 72 h were significantly higher than those in the control cells. The mRNA and protein levels of regulated in development and DNA damage response 1 (REDD1) in HME-treated cells were significantly lower than those in the control cells. Although HME treatment did not increase the mechanistic target of rapamycin (mTOR) level in Caco-2 cells, it significantly increased the phosphorylated mTOR (p-mTOR) level and p-mTOR/mTOR ratio. The ZO-1 protein levels in cells treated with an inducer of REDD1, cobalt chloride (CoCl2) alone were significantly lower than those in the control cells. However, ZO-1 protein levels in cells co-treated with HME and CoCl2 were significantly higher than those in cells treated with CoCl2 alone. Additionally, REDD1 protein levels in cells treated with CoCl2 alone were significantly higher than those in the control cells. However, REDD1 protein levels in cells co-treated with HME and CoCl2 were significantly lower than those in cells treated with CoCl2 alone. This HME-mediated effect may contribute to the development of barrier function in the infant intestine and protect infants from diseases.

Early Changes in Postural Balance Following Inverted V-Shaped High Tibial Osteotomy in Patients With Knee Osteoarthritis.

Patients with knee osteoarthritis and varus knee deformity have impaired postural balance, resulting in decreased walking performance and an increased risk of falls. This study aimed to investigate the early changes in the postural balance following inverted V-shaped high tibial osteotomy (HTO). Fifteen patients with medial knee osteoarthritis were recruited. Postural balance was assessed using the center-of-pressure (COP) data during single-leg standing before and 6 weeks after inverted V-shaped HTO. The maximum range, mean velocity, and area of COP movements in the anteroposterior and mediolateral directions were analyzed. Preoperative and postoperative visual analog scale for knee pain was assessed. The maximum range of COP in the mediolateral direction decreased (P = .017), whereas the mean velocity of COP in the anteroposterior direction increased 6 weeks postoperatively (P = .011). The visual analog scale score for knee pain significantly improved at 6 weeks postoperatively (P = .006). Valgus correction with inverted V-shaped HTO resulted in improved postural balance in the mediolateral direction and good short-term clinical outcomes early following surgery. Early rehabilitation after inverted V-shaped HTO should focus on postural balance in the anteroposterior direction.

Norepinephrine transporter expressed on mammary epithelial cells incorporates norepinephrine in milk into the cells.

Mammary epithelial cells synthesize and secrete norepinephrine (NE) into breast milk to regulate ß-casein expression through the adrenergic ß2 receptor. We investigated the expression, localization, and roles of NE transporter (NET) in the mammary epithelium during lactation. mRNA and protein levels of NET were determined in primary normal human mammary epithelial cells (pHMECs) and non-malignant human mammary epithelial MCF-12A cells. In nursing CD1 mice, NET localized to the apical membranes of the mammary epithelium. The intracellular NE content of pHMECs incubated with NE increased. Although the ß-casein concentration in milk was slightly higher at day 10 than at day 2 of lactation, the NE concentration and lactation-related proteins were only slightly changed on days 2-10. Restraint stress increased the NE concentration in milk from nursing mice and NET protein levels were significantly higher than in non-stressed nursing mice. NET is expressed on the apical membrane of mammary epithelial cells and incorporates NE in milk into cells, potentially regulating the NE concentration in milk.

Analysis of risk factors for skin disorders caused by anti-epidermal growth factor receptor antibody drugs and examination of methods for their avoidance.

WHAT IS KNOWN AND OBJECTIVE: Cancer drug treatment is often discontinued because of skin disorder aggravation. However, information on risk factors for skin disorders caused by anti-epidermal growth factor receptor (EGFR) antibody drugs is limited. The aim of this study was to analyse the factors associated with skin disorders caused by anti-EGFR antibody drugs and establish a method to minimize such aggravations. METHODS: We retrospectively examined 67 colorectal cancer patients treated with anti-EGFR antibody drugs for the first time. RESULTS AND DISCUSSION: A higher proportion of males than females experienced drug withdrawal, dose reduction or treatment discontinuation. The multiple logistic regression analysis revealed body weight as a risk factor affecting drug withdrawal, dose reduction or treatment discontinuation because of an acneiform rash. An examination of methods to avoid the aggravation of skin disorders revealed the acneiform rash grade in patients who received prophylactic minocycline was significantly lower than that in patients who did not receive prophylactic minocycline. Furthermore, among patients with grade 1 acneiform rash at the initiation of minocycline, the proportion of those who withdrew, required dose reduction or discontinued treatment was lower than that among patients with grade 2 acneiform rash. WHAT IS NEW AND CONCLUSION: High body weight was identified as a novel factor for skin disorder aggravation caused by anti-EGFR antibody drugs. The aggravation of skin disorders during cancer treatment with anti-EGFR antibody drugs can potentially be avoided by carefully observing the onset of acneiform rash in affected patients with high body weight and using minocycline prophylactically or as an early-stage intervention.

Dynamic postural control correlates with activities of daily living and quality of life in patients with knee osteoarthritis.

BACKGROUND: Knee osteoarthritis (OA) negatively affects dynamic postural control, which is a basic function that individuals use to perform activities of daily living (ADL). The purpose of this study was to investigate the associations of center of pressure (COP) control during the transition from double-leg to single-leg standing with subjective assessments of ADL and quality of life (QOL) in patients with knee OA. METHODS: Thirty-six patients (29 females) with moderate-to-severe knee OA participated. Dynamic postural control was evaluated during the transition from double-leg to single-leg standing. Each patient stood on a force plate, lifted the less affected limb as fast as possible, and maintained single-leg standing with the more affected limb. The COP movements corresponding to anticipatory postural adjustment (APA) and transitional phases were assessed. The maximum displacement and peak velocity of the COP movements in the medial-lateral direction were calculated. The Knee Injury and Osteoarthritis Outcome Score (KOOS) was used for the subjective assessment of ADL and QOL. Pearson's product correlation analysis was performed to investigate the associations of COP movements in the APA and transitional phases with KOOS-ADL and KOOS-QOL. RESULTS: In the APA phase, the maximum COP displacement was significantly correlated with KOOS-ADL (r = -0.353, P = 0.035) and KOOS-QOL (r = -0.379, P = 0.023). In the transitional phase, the maximum COP displacement and peak COP velocity were significantly correlated with KOOS-ADL (maximum displacement: r = 0.352, P = 0.035; peak velocity: r = 0.438, P = 0.008) and with KOOS-QOL (maximum displacement: r = 0.357, P = 0.032; peak velocity: r = 0.343, P = 0.040). CONCLUSIONS: The present study showed that smaller COP movements in the APA phase and smaller and slower COP movements in the transitional phase correlated with poorer ADL and QOL conditions in patients with knee OA. These findings suggest that poor dynamic postural control is associated with poor ADL and QOL conditions in patients with moderate-to-severe medial knee OA. Conservative treatment for patients with knee OA may need to focus on dynamic postural control during the transition from double-leg to single-leg standing.

Cancer Cachexia May Hinder Pain Control When Using Fentanyl Patch.

The objective of this study was to evaluate the influence of cancer cachexia on pain control in cancer patients receiving a transdermal fentanyl patch (FP) and to investigate whether dry skin was a factor related to cancer cachexia and uncontrolled pain. We retrospectively reviewed the medical records of 77 patients receiving FP treatment for the first time, who were classified into cancer cachexia and non-cancer-cachexia groups, according to European Palliative Care Research Collaborative criteria. On day 7 after FP administration, the mean FP dose and morphine equivalent dose (MED) in the cancer cachexia group were significantly higher than in the non-cancer-cachexia group. Additionally, in the cancer cachexia group, there was a significantly larger degree of variation in pain intensity over 7 d than in the non-cachexia group. In patients who were switched from FP to morphine injection, the mean pain intensity and MED on day 3 after morphine injection were significantly lower than those immediately before morphine injection. Subsequently, to investigate whether dry skin was involved in poor pain control in the cancer cachexia group, transepidermal water loss (TEWL) was compared between 15 additional patients classified into cancer cachexia and non-cancer cachexia groups; the mean TEWL in the cancer cachexia group was found to be significantly lower. Our data suggest that cancer cachexia may be a risk factor for poor pain control in patients receiving FP treatment, and that uncontrolled pain in FP treatment may be caused by the inhibition of fentanyl transdermal absorption due to dry skin.

Abdominal draw-in maneuver changes neuromuscular responses to sudden release from trunk loading in patients with non-specific chronic low back pain.

BACKGROUND: Abdominal draw-in maneuver (ADIM) has been recommended to achieve appropriate trunk muscle response for patients with non-specific chronic low back pain (CLBP). However, it has remained unclear whether the intervention with ADIM could change the trunk muscle response to sudden release from loading, which is considered to contribute mechanical circumstances to low back pain. The purpose of the present study was to investigate the effects of the intervention with ADIM on electromyography (EMG) activities of trunk muscles following sudden release from loading. METHODS: Seventeen subjects with non-specific CLBP participated. Subjects resisted trunk flexion or extension loading in semi-seated position, and then the loading was suddenly released. EMG recordings of 6 trunk muscles were acquired using a wireless surface EMG system. Onset and offset times were calculated from the EMG data. The intervention with ADIM was provided for 4 weeks. The onset and offset times were compared between pre- and post-intervention with ADIM. RESULTS: At the post-intervention, the onset of trunk flexors following release from trunk flexion loading became significantly earlier than pre-intervention (P = 0.028). The offset of flexors following release from trunk extension loading of post-intervention was significantly earlier than that of pre-intervention (P = 0.001). CONCLUSIONS: We showed that the intervention with ADIM changed the EMG activity of trunk flexors in response to sudden release from loading. These results suggest a possibility that ADIM might be effective to improve the neuromuscular control of trunk flexors for the treatment of young patients with non-specific CLBP.

Deterioration of Glycemic Control Contributes to the Prevalence of Proteinuria among Bevacizumab-Treated Cancer Patients with Type 2 Diabetes Mellitus.

The objective of this study was to investigate whether improving glycemic control reduces the prevalence and progression of proteinuria among bevacizumab (BEV)-treated cancer patients with type 2 diabetes mellitus (DM). We retrospectively reviewed the medical records of 55 patients with type 2 DM who were treated with BEV between July 1 2011 and May 31 2018 at Iwate Medical University Hospital. The patients were classified based on changes in glycated hemoglobin (HbA1c) level during the 3 months following BEV administration into the "HbA1c elevated" group (+0.5% or above, n=24) and the "HbA1c non-elevated" group (indicating no change or decrease; n=31). At 3 months following BEV administration, the means of HbA1c and its change rate in the 'HbA1c elevated' group was significantly higher than that in the 'HbA1c non-elevated' group, and the prevalence of proteinuria in the 'HbA1c elevated' group was significantly higher than that in the 'HbA1c non-elevated' group. Additionally, our subjects were classified into the proteinuria group and non-proteinuria group. The mean HbA1c level in the proteinuria group was significantly higher than that in the non-proteinuria group at 3 months following BEV administration. Furthermore, the mean rates of change of HbA1c level in patients experiencing grades 1 and 2 proteinuria were +9.97±2.26 and +14.0±3.82%, respectively. These values were significantly higher than those of patients with no proteinuria (-2.15±1.96%). Our results suggest that deterioration of glycemic control contributes to the prevalence of proteinuria among BEV-treated cancer patients with type 2 DM.

Analysis of serotonin concentrations in human milk by high-performance liquid chromatography with fluorescence detection.

Serotonin (5-hydroxytryptamine, 5-HT) plays an important role in milk volume homeostasis in the mammary gland during lactation; 5-HT in milk may also affect infant development. However, there are few reports on 5-HT concentrations in human breast milk. To address this issue, we developed a simple method based on high-performance liquid chromatography with fluorescence detection (HPLC-FD) for measuring 5-HT concentrations in human breast milk. Breast milk samples were provided by four healthy Japanese women. Calibration curves for 5-HT in each sample were prepared with the standard addition method between 5 and 1000 ng/ml, and all had correlation coefficients >0.999. The recovery of 5-HT was 96.1%-101.0%, with a coefficient of variation of 3.39%-8.62%. The range of 5-HT concentrations estimated from the calibration curves was 11.1-51.1 ng/ml. Thus, the HPLC-FD method described here can effectively extract 5-HT from human breast milk with high reproducibility.

Serotonin suppresses ß-casein expression via PTP1B activation in human mammary epithelial cells.

Serotonin (5-hydroxytriptamine, 5-HT) has an important role in milk volume homeostasis within the mammary gland during lactation. We have previously shown that the expression of ß-casein, a differentiation marker in mammary epithelial cells, is suppressed via 5-HT-mediated inhibition of signal transduction and activator of transcription 5 (STAT5) phosphorylation in the human mammary epithelial MCF-12A cell line. In addition, the reduction of ß-casein in turn was associated with 5-HT7 receptor expression in the cells. The objective of this study was to determine the mechanisms underlying the 5-HT-mediated suppression of ß-casein and STAT5 phosphorylation. The ß-casein level and phosphorylated STAT5 (pSTAT5)/STAT5 ratio in the cells co-treated with 5-HT and a protein kinase A (PKA) inhibitor (KT5720) were significantly higher than those of cells treated with 5-HT alone. Exposure to 100 µM db-cAMP for 6 h significantly decreased the protein levels of ß-casein and pSTAT5 and the pSTAT5/STAT5 ratio, and significantly increased PTP1B protein levels. In the cells co-treated with 5-HT and an extracellular signal-regulated kinase1/2 (ERK) inhibitor (FR180294) or Akt inhibitor (124005), the ß-casein level and pSTAT5/STAT5 ratio were equal to those of cells treated with 5-HT alone. Treatment with 5-HT significantly induced PTP1B protein levels, whereas its increase was inhibited by KT5720. In addition, the PTP1B inhibitor sc-222227 increased the expression levels of ß-casein and the pSTAT5/STAT5 ratio. Our observations indicate that PTP1B directly regulates STAT5 phosphorylation and that its activation via the cAMP/PKA pathway downstream of the 5-HT7 receptor is involved in the suppression of ß-casein expression in MCF-12A cells.

The relationship between the load on the knee joint during walking and the biomechanical characteristics of single-leg standing.

[Purpose] The purpose of the present study was to investigate the relationship between the external knee adduction moment (KAM) during walking and the biomechanical characteristics of single-leg standing in healthy subjects. [Subjects and Methods] Twenty-eight healthy subjects were recruited for this study. Data were collected while the subjects performed walking and single-leg standing using a motion analysis system with six digital video cameras and two force plates. Pearson's correlation coefficient was used to quantify the relationship between peak KAM during walking and single-leg standing. To determine whether the kinematic behavior of the pelvis and trunk during single-leg standing are associated with peak KAM during walking, Pearson's correlation coefficients were calculated and stepwise linear regression was performed. [Results] The peak KAM during single-leg standing was significantly correlated with that during walking. The peak KAM during walking was significantly correlated with the peak lateral lean of the trunk and the peak lateral tilt of the pelvis during single-leg standing. The results of stepwise linear regression analysis show the peak KAM during walking was partially explained by the peak lateral lean of the trunk during single-leg standing. [Conclusion] Our findings suggest that single-leg standing might be a useful method for predicting the peak KAM during walking.

Serotonin regulates ß-casein expression via 5-HT7 receptors in human mammary epithelial MCF-12A cells.

We previously reported that serotonin (5-hydroxytryptamine; 5-HT) suppresses ß-casein expression, a differentiation marker in mammary epithelial cells, via inhibition of the signal transducer and activator of transcription 5 (STAT5) phosphorylation in the human mammary epithelial cell line, MCF-12A. In this study, we investigated the expression pattern of the different 5-HT receptor subtypes in MCF-12A cells, and identified the receptors involved in 5-HT-mediated suppression of ß-casein protein expression. ß-Casein mRNA expression was inhibited by 30 µM 5-HT in a time-dependent manner. Treatment with 30 µM 5-HT for 72 h decreased ß-casein protein levels and STAT5 phosphorylation (pSTAT5). The cells expressed four 5-HT receptors subtypes (5-HTR1D, 2B, 3A, and 7) at the mRNA and protein level, and their expression was elevated by prolactin (PRL) treatment. Additionally, the mRNA levels of 5-HTR1D and 5-HTR7 were significantly higher than the other 5-HT receptors in the cells. Tryptophan hydroxylase 1 mRNA was detectable in the cells in the absence of PRL, and PRL treatment significantly increased its expression. ß-Casein and pSTAT5/STAT5 levels in the cells co-treated with 5-HT and a selective 5-HTR1D inhibitor, BRL15572, were equal to those observed in cells treated with 5-HT alone. However, in the cells co-treated with 5-HT and a selective 5-HTR7 inhibitor, SB269970, ß-casein and pSTAT5/STAT5 levels increased in a SB269970 concentration-dependent manner. In conclusion, we showed that 5-HT regulates ß-casein expression via 5-HTR7 in MCF-12A human mammary epithelial cells.

[DEVELOPMENT OF MEDICAL EDUCATIONAL PROGRAM FOR PEDIATRICIANS TO IMPROVE THEIR BEHAVIOR IN THE MANAGEMENT OF FOOD ALLERGY].

BACKGROUND: Many children are suffering from food allergy, however, not all of them are treated properly by pediatricians based on the latest practice guideline. The guideline does not recommend excessive elimination diet, but suggests initial appropriate skincare treatment before examining candidate allergens for food-allergy-patients with atopic dermatitis. We developed and evaluated a curriculum for continuing medical education for pediatrician about food allergy. METHODS: We provided a medical educational program for pediatricians at the Division of Allergy in National Center for Child Health and Development, from January 2013 to January 2014. The program was aimed to improve their behavior in the management of food allergy, if they excessively eliminated diet from the patients and did not treat atopic dermatitis appropriately. The program was constituted of SBOs and educational strategy. Participant's self evaluation of learning and that of behavior were carried out before and after the training, and their scores were analyzed for the program evaluation. RESULTS: Thirty-five participants completed the program. All the scores rating 22 items for evaluation of learning were increased. Five items out of 7 about evaluation of behavior were significantly improved. CONCLUSION: We developed a medical educational program about food allergy for pediatricians. After training based on this program, the participants' knowledge and skills were improved and their behavior was desirably modified.

Serotonin suppresses ß-casein expression via inhibition of the signal transducer and activator of transcription 5 (STAT5) protein phosphorylation in human mammary epithelial cells MCF-12A.

Serotonin (5-hydroxytryptamine; 5-HT) has an important physiological role in controlling lactation, namely, milk volume homeostasis, within mammary glands. The objectives of this study were to evaluate whether exogenous 5-HT can suppress ß-casein expression, a differentiation marker, produced in human mammary epithelial cells, and to determine whether 5-HT can attenuate ß-casein signaling via the prolactin (PRL) receptor (PRLr) and Janus kinase 2/signal transducer and activator of transcription 5 (STAT5) pathway. PRL treatment increased the mRNA level of ß-casein in the MCF-12A human mammary epithelial cell line, and the highest level occurred at days 7 and 14 of culture. In contrast, PRLr expression was not affected significantly by PRL treatment. PRL treatment in MCF-12A cells increased levels of ß-casein and phosphorylated STAT5 (pSTAT5) proteins in a concentration-dependent manner, with a slight increase of STAT5 protein. ß-Casein expression was inhibited by 0.1 mM 5-HT in a time-dependent manner. Additionally, treatment with 0.1 mM 5-HT for 72 h decreased protein levels of ß-casein and pSTAT5, with a slight decrease in STAT5 levels. These results suggest that exogenous 5-HT can inhibit STAT5 phosphorylation, resulting in a decrease in ß-Casein expression. In conclusion, we showed that exogenous 5-HT decreased ß-casein expression in MCF-12A human mammary epithelial cells, and that 5-HT was responsible for inhibiting phosphorylation of STAT5, resulting in a decline in lactational function.

A retrospective study on the influence of nutritional status on pain management in cancer patients using the transdermal fentanyl patch.

It is unknown whether nutritional status influences pain intensity in cancer patients receiving a transdermal fentanyl patch (FP). This study aimed to determine whether nutritional status is associated with pain intensity and to evaluate the influence of changes in nutritional status on pain intensity in cancer patients receiving transdermal FP treatment. We included 92 patients receiving transdermal FP treatment for the first time with switching from oxycodone. The patients were classified into low- and normal-nutrition groups based on their nutritional status, which was assessed according to the Nutrition Risk Screening 2002 (NRS 2002) parameters. The pain intensity of each patient was evaluated by a numeric rating scale (11-point scale from 0 to 10). NRS 2002 score and pain intensity were obtained on day 3 after the FP was applied to the skin. Pain intensities were significantly higher among patients in the low-nutrition group than among patients in the normal-nutrition group. NRS 2002 scores showed a significant positive correlation with the pain intensities. In 52 of 92 patients, who were evaluated using the NRS 2002 score and pain intensity on day 30 after FP application, the changes in NRS 2002 scores were significantly related to changes in pain intensities (odds ratio, 30.0; 95% confidence interval, 4.48-200.97; p=0.0005). These results suggest that an increase in the NRS 2002 score is a risk factor for an increase in pain intensity in cancer patients receiving FP treatment. Malnutrition may lead to poor pain management in cancer patients receiving FP treatment.

Identification of a novel benzimidazole derivative as a highly potent NPY Y5 receptor antagonist with an anti-obesity profile.

Optimization of HTS hit 1 for NPY Y5 receptor binding affinity, CYP450 inhibition, solubility and metabolic stability led to the identification of some orally available oxygen-linker derivatives for in vivo study. Among them, derivative 4i inhibited food intake induced by the NPY Y5 selective agonist, and chronic oral administration of 4i in DIO mice caused a dose-dependent reduction of body weight gain.

Effect of toe-out gait modification on patellofemoral joint loading.

BACKGROUND: Toe-out gait has been proposed as a conservative treatment to reduce medial tibiofemoral joint loading. However, patellofemoral joint loading during toe-out gait is not yet understood. RESEARCH QUESTION: Does the toe-out gait modification affect patellofemoral joint loading? METHODS: Sixteen healthy adults were enrolled in this study. The natural gait and toe-out gait were measured using a three-dimensional motion analysis and a force plate. The knee flexion angle and external knee flexion moment during the stance phase were calculated. Thus, dynamic knee joint stiffness, a proxy of patellofemoral joint loading, was defined as a linear regression of the knee flexion moment and knee flexion angle during the early stance. Additionally, the peak patellofemoral compressive force during the early stance was calculated using a musculoskeletal simulation. A paired t-test was used to compare these biomechanical parameters during the natural gait and toe-out gait. RESULTS: The toe-out gait significantly increased the peak patellofemoral compressive force (mean difference = 0.37 BW, P = 0.017) and dynamic knee joint stiffness (mean difference = 0.07%BW*Ht/°, P = 0.001). The 1st peak of the knee flexion moment also significantly increased in the toe-out gait (mean difference = 1.01%BW*Ht, P = 0.003); however, the knee flexion angle did not change significantly (initial contact: mean difference = 1.7°, P = 0.078; peak: mean difference = 1.3°, P = 0.224). SIGNIFICANCE: Toe-out gait increased the patellofemoral compressive force and dynamic knee joint stiffness because of increasing knee flexion moment, but not the knee flexion angle. When the toe-out gait is adapted, clinicians should pay attention to an increase in the patellofemoral joint loading.

Identification of a novel and orally available benzimidazole derivative as an NPY Y5 receptor antagonist with in vivo efficacy.

Optimization of lead compound 2 is described, mainly focusing on modification at the C-2 position of the benzimidazole core. Replacement of the phenyl linker of 2 with saturated rings resulted in identification of compound 8b which combines high Y5 receptor binding affinity with a good ADME profile leading to in vivo efficacy.

Design, synthesis and identification of novel benzimidazole derivatives as highly potent NPY Y5 receptor antagonists with attractive in vitro ADME profiles.

Optimization of our HTS hit 1, mainly focused on modification at the C-2 position of the benzimidazole core, is described. Elimination of the flexible and metabolically labile -S-CH(2)- part and utilization of less lipophilic pyridone substructure led to identification of novel NPY Y5 receptor antagonists 6, which have low to sub-nanomolar Y5 receptor binding affinity with improved CYP450 inhibition profiles, good solubilities and high metabolic stabilities.