Site-Selective Incorporation of a Functional Group into Lys175 in the Vicinity of the Active Site of Chymotrypsin by Using Peptidyl α-Aminoalkylphosphonate Diphenyl Ester-Derivatives.

We previously reported that Lys175 in the region of the active site of chymotrypsin (Csin) could be site-selectively modified by using an N-hydroxy succinimide (NHS) ester of the peptidyl derivative containing 1-amino-2-ethylphenylphosphonate diphenyl ester [NHS-Suc-Ala-Ala-PheP(OPh)2]. In this study, the Lys175-selective modification method was expanded to incorporate functional groups into Lys 175 in Csin. Two types of peptidyl phosphonate derivatives with the dansyl group (Dan) as a functional molecule, Dan-ß-Ala-[Asp(NHS) or Glu(NHS)]-Ala-Ala-(R)-PheP(OPh)2 (DanD and DanE, respectively), were synthesized, and their action was evaluated when modifying Lys175 in Csin. Ion-exchange chromatography (IEC), fluorescence spectroscopy, and LC-MS/MS were used to analyze the products from the reaction of Csin with DanD or DanE. By IEC and LC-MS/MS, the results showed that DanE reacted with Csin more effectively than DanD to produce the modified Csin (DanMCsin) bearing Dan at Lys175. DanMCsin exhibited an enzymatic activity corresponding to 1/120 of Csin against Suc-Ala-Ala-Phe-pNA. In addition, an effect of Lys175 modification on the access of the proteinaceous Bowman-Birk inhibitor to the active site of DanMCsin was investigated. In conclusion, by using a peptidyl derivative containing 1-amino-2-ethylphenylphosphonate diphenyl ester, we demonstrated that a functional group could be incorporated into Lys175 in Csin.

Exploration of an Inflection Point of Ventilation Parameters with Anaerobic Threshold Using Strucchange.

(1) Background: When measuring anaerobic work threshold (AT), the conventional V-slope method includes the subjectivity of the examiner, which cannot be eliminated completely. Therefore, we implemented an engineering method using strucchange to objectively search for the inflection point of AT. (2) Methods: Seventeen subjects (15 men and 2 women) were included in the study. The subjects rode an ergometer and performed a ramp load test for 18 min and 30 s. (3) Results: In VE (Ventilation), 11 out of 12 subjects had the same results with 95% confidence intervals for the AT by the strucchange and respiratory metabolic apparatus. In VCO2 (Carbon dioxide emissions), 9 out of 12 subjects had the same results with 95% confidence intervals for the AT with the strucchange and respiratory metabolic apparatus. In VE, 3 out of 12 subjects showed the same results for respiratory metabolic analysis and the AT by the V-slope method. In VCO2, 3 out of 12 subjects showed the same results for the respiratory metabolic analysis and AT by the V-slope method in VCO2. (4) Conclusions: Strucchange was more objective and significant in identifying the AT than the V-slope method.

JUNB governs a feed-forward network of TGFß signaling that aggravates breast cancer invasion.

It is well established that transforming growth factor-ß (TGFß) switches its function from being a tumor suppressor to a tumor promoter during the course of tumorigenesis, which involves both cell-intrinsic and environment-mediated mechanisms. We are interested in breast cancer cells, in which SMAD mutations are rare and interactions between SMAD and other transcription factors define pro-oncogenic events. Here, we have performed chromatin immunoprecipitation (ChIP)-sequencing analyses which indicate that the genome-wide landscape of SMAD2/3 binding is altered after prolonged TGFß stimulation. De novo motif analyses of the SMAD2/3 binding regions predict enrichment of binding motifs for activator protein (AP)1 in addition to SMAD motifs. TGFß-induced expression of the AP1 component JUNB was required for expression of many late invasion-mediating genes, creating a feed-forward regulatory network. Moreover, we found that several components in the WNT pathway were enriched among the late TGFß-target genes, including the invasion-inducing WNT7 proteins. Consistently, overexpression of WNT7A or WNT7B enhanced and potentiated TGFß-induced breast cancer cell invasion, while inhibition of the WNT pathway reduced this process. Our study thereby helps to explain how accumulation of pro-oncogenic stimuli switches and stabilizes TGFß-induced cellular phenotypes of epithelial cells.

Palbociclib enhances activin-SMAD-induced cytostasis in estrogen receptor-positive breast cancer.

Cyclin-dependent kinase (CDK) 4 and CDK6 inhibitors are effective therapeutic options for hormone receptor (HR)-positive, human epidermal growth factor receptor 2 (HER2)-negative advanced breast cancer. Although CDK4/6 inhibitors mainly target the cyclin D-CDK4/6-retinoblastoma tumor suppressor protein (RB) axis, little is known about the clinical impact of inhibiting phosphorylation of other CDK4/6 target proteins. Here, we focused on other CDK4/6 targets, SMAD proteins. We showed that a CDK4/6 inhibitor palbociclib and activin-SMAD2 signaling cooperatively inhibited cell cycle progression of a luminal-type breast cancer cell line T47D. Palbociclib enhanced SMAD2 binding to the genome by inhibiting CDK4/6-mediated linker phosphorylation of the SMAD2 protein. We also showed that cyclin G2 plays essential roles in SMAD2-dependent cytostatic response. Moreover, comparison of the SMAD2 ChIP-seq data of T47D cells with those of Hs578T (triple-negative breast cancer cells) indicated that palbociclib augmented different SMAD2-mediated functions based on cell type, and enhanced SMAD2 binding to the target regions on the genome without affecting its binding pattern. In summary, palbociclib enhances the cytostatic effects of the activin-SMAD2 signaling pathway, whereas it possibly strengthens the tumor-promoting aspect in aggressive breast cancer.

[Reliability and validity of the Japanese translation of Brief Self-Control Scale (BSCS-J)].

Self-control refers to the ability to execute goal-oriented behavior despite the presence of temptation(s) to do otherwise. Since self-control has a wide-range impact on our daily lives, it is of critical importance to assess individual differences of self-control with a highly reliable and valid, yet simple, measure. Toward this end, three studies were conducted to test reliability and validity of the Japanese-translated version of Brief Self-Control Scale (Tangney, Baumeister, & Boone, 2004). The scale showed good internal consistency (Study 1) and retest reliability (Study 2). The. total score of the scale was correlated with the self-reported indices of self-control (e.g., daily experience of ego-depletion, study hours) and performance in the Stop Signal Task (Study 3), indicating its high converging validity.

[An attempt to construct a Japanese version of the Implicit Positive and Negative Affect Test (IPANAT)].

The Implicit Positive and Negative Affect Test (IPANAT) is an instrument for the indirect assessment of positive and negative affect. A Japanese version of the IPANAT was developed and its reliability and validity were examined. In Study 1, factor analysis identified two independent factors that could be interpreted as implicit positive and negative affect, which corresponded to the original version. The Japanese IPANAT also had sufficient internal consistency and acceptable test-retest reliability. In Study 2, we demonstrated that the Japanese IPANAT was associated with explicit state affect (e.g., PANAS), extraversion, and neuroticism, which indicated its adequate construct validity. In Study 3, we examined the extent to which the Japanese IPANAT was sensitive to changes in affect by assessing a set of IPANAT items after the presentation of positive, negative, or neutral photographs. The results indicated that the Japanese IPANAT was sufficiently sensitive to changes in affect resulting from affective stimuli. Taken together, these studies suggest that the Japanese version of the IPANAT is a useful instrument for the indirect assessment of positive and negative affect.

Survey of the association between tooth extraction and development of medication-related osteonecrosis of the jaw in patients undergoing cancer treatment with bone-modifying agents.

OBJECTIVE: This study aimed to verify whether tooth extraction before the administration of bone-modifying agents (BMA) was effective in preventing the onset of medication-related osteonecrosis of the jaw (MRONJ). MATERIALS AND METHODS: This retrospective study included patients with a history of receiving BMA for cancer treatment. The patients were classified into three groups based on the timing of tooth extraction: no tooth extraction before the onset of MRONJ, tooth extraction before the administration of BMA, and tooth extraction after the administration of BMA. The incidence of MRONJ was compared between the groups. Fisher's exact test and Bonferroni correction were used to test for differences in proportions between the three groups. RESULTS: The total number of subjects was 123. Twenty-four patients (19.5%) developed MRONJ. The incidence rates were 12.3% (10/81), 17.9% (5/28), and 64.3% (9/14) in the non-extraction group, the extraction before BMA administration group, and the extraction after BMA administration group, respectively, showing statistically significant differences between the extraction after BMA administration group and the non-extraction groups and between the extraction after BMA administration group and the extraction before BMA administration group (p < 0.001, p = 0.0049). On the other hand, there was no statistically significant difference in incidence between the non-extraction and the extraction before BMA administration group (p = 0.5274). CONCLUSIONS: Tooth extraction before the administration of BMA is effective in preventing the onset of MRONJ in patients receiving BMA for cancer treatment. Prevention of MRONJ development in patients receiving BMA for cancer treatment contributes to the maintenance of patients' quality of life.

[The effect of priming of a diet goal on the implicit evaluations of goal and temptation relevant targets].

This research investigated how to overcome temptations and protect high-order goals while pursuing a goal. We hypothesized that in order to promote self-regulation, individuals non-cousciously engage in asymmetric evaluative responses to goal-relevant and temptation-relevant stimuli. In an experiment, we manipulated either diet goal or academic goal. Then, we measured evaluations of either sugary drinks (e.g., Coke, Fanta) or healthy drinks (e.g., Healthya Green Tea, Black Oolong Tea). The results showed that participants who activated a diet goal had significantly more positive evaluations of healthy drinks than sugary drinks. In addition, this tendency was moderated by the means of dieting (i.e., only participants who cut down on sweets when going on a diet). The role of non-consious asymmetric evaluations for self-regulation is discussed.

Utility of cyclic variation of heart rate score as a screening tool for sleep-disordered breathing in patients with heart failure.

STUDY OBJECTIVES: Patients with sleep-disordered breathing have cyclic variation of heart rate (CVHR) in response to respiratory events. However, limited data are available regarding the utility of CVHR as a screening tool for sleep-disordered breathing in patients with mixed heart failure (HF) and those without HF. METHODS: We enrolled consecutive patients with and without HF who underwent full polysomnographies with simultaneous Holter electrocardiogram monitoring. We determined the temporal position of the individual dips comprising the CVHR score using time-domain methods. RESULTS: The data of 101 patients, including 70 with and 31 without HF, were analyzed. The CVHR score was significantly correlated with the apnea-hypopnea index (r = .667, P < .001) and limits of agreement between the apnea-hypopnea index and CVHR score were -21.8 to 35.2. The receiver operating characteristic analysis demonstrated that the CVHR score (best cut-off of 23.5 events/h) identified severe sleep-disordered breathing with a sensitivity of 83.3%, specificity of 79.5%, and the area under the curve of 0.856. In addition, there was no interaction between the presence or absence of HF and the apnea-hypopnea index-CVHR score relationship (P = .323). CONCLUSIONS: The CVHR score, determined by Holter electrocardiogram monitoring, is a useful tool for evaluating sleep-disordered breathing even in patients with mixed HF and patients without HF. CITATION: Yatsu S, Kasai T, Naito R, et al. Utility of cyclic variation of heart rate score as a screening tool for sleep-disordered breathing in patients with heart failure. J Clin Sleep Med. 2021;17(11):2187-2196.

Is there an undiscovered neurocircuit for regulating GH secretion? -Pitfalls of GHRP-2 and ITT as GH provocative tests-.

GH secretion is mainly regulated at the hypothalamus by a dual interplay between growth hormone releasing hormone (GHRH) and somatostatin, which are modulated by various factors. We examined the regulatory mechanism of GH secretion in an apparently healthy young man without decreased IGF-1 concentration and nocturnal GH secretion, but who showed low responses to insulin tolerance (ITT) and to GHRP-2 tests. The patient also had no GH response to acute aerobic exercise. However, he had normal secretion of pituitary hormone based on hypothalamic releasing hormone tests combined with CRH, GRH as GHRH, LH-RH and TRH. In addition, he had a GH response without paradoxical secretion to TRH stimulation as well as an ACTH response to subcutaneous glucagon stimulation, and AVP secretion responded to 5% hypertonic saline infusion, though it was not adequately stimulated by ITT. MRI showed no structural abnormalities in the hypothalamus-pituitary gland. These findings indicate that this subject may have an undiscovered neurocircuit for regulating GH secretion, as well as other neurohormones, to maintain homeostasis, even though there were low responses of the hormones to ITT and GHRP-2 stimuli, probably via altered secretion of hypothalamic hormones.

The ALK-1/SMAD/ATOH8 axis attenuates hypoxic responses and protects against the development of pulmonary arterial hypertension.

Dysregulated bone morphogenetic protein (BMP) signaling in endothelial cells (ECs) is implicated in vascular diseases such as pulmonary arterial hypertension (PAH). Here, we showed that the transcription factor ATOH8 was a direct target of SMAD1/5 and was induced in a manner dependent on BMP but independent of Notch, another critical signaling pathway in ECs. In zebrafish and mice, inactivation of Atoh8 did not cause an arteriovenous malformation-like phenotype, which may arise because of dysregulated Notch signaling. In contrast, Atoh8-deficient mice exhibited a phenotype mimicking PAH, which included increased pulmonary arterial pressure and right ventricular hypertrophy. Moreover, ATOH8 expression was decreased in PAH patient lungs. We showed that in cells, ATOH8 interacted with hypoxia-inducible factor 2α (HIF-2α) and decreased its abundance, leading to reduced induction of HIF-2α target genes in response to hypoxia. Together, these findings suggest that the BMP receptor type II/ALK-1/SMAD/ATOH8 axis may attenuate hypoxic responses in ECs in the pulmonary circulation and may help prevent the development of PAH.

Development and Clinical Application of a Novel Autonomic Transient Response-Based Screening System for Major Depressive Disorder Using a Fingertip Photoplethysmographic Sensor.

Over 350 million people across the world suffer from major depressive disorder (MDD). More than 10% of MDD patients have suicide intent, while it has been reported that more than 40% patients did not consult their doctors for MDD. In order to increase consultation rate of potential MDD patients, we developed a novel MDD screening system which can be used at home without help of health-care professionals. Using a fingertip photoplethysmograph (PPG) sensor as a substitute of electrocardiograph (ECG), the system discriminates MDD patients from healthy subjects using autonomic nerve transient responses induced by a mental task (random number generation) via logistic regression analysis. The nine logistic regression variables are averages of heart rate (HR), high frequency (HF) component of heart rate variability (HRV), and the low frequency (LF)/HF ratio of HRV before, during, and after the mental task. We conducted a clinical test of the proposed system. Participants were 6 MDD patients (4 females and 2 males, aged 23-60 years) from Shizuoka Saiseikai General Hospital psychiatry outpatient unit and 14 healthy volunteers from University of Electro-Communications (6 females and 8 males, aged 21-63 years). The average PPG- and ECG (as a reference)-derived HR, HF and LF/HF were significantly correlated with each other (HR; r = 1.00, p < 0.0001, HF; r = 0.98, p < 0.0001, LF/HF; r = 0.98, p < 0.0001). Leave-one-out cross validation (LOOCV) revealed 83% sensitivity and 93% specificity. The proposed system appears promising for future MDD self-screening at home and are expected to encourage psychiatric visits for potential MDD patients.

Simple and objective screening of major depressive disorder by heart rate variability analysis during paced respiration and mental task conditions.

Since objective biomarkers for major depressive disorder (MDD) are not readily available, clinical psychiatrists diagnose patients with MDD subjectively based on clinical interviews and diagnostic criteria. It often raises various concerns, including false responses by patients, subjective factors, and inexperience of the attendants leading to incorrect diagnosis. Here, we developed a self-monitoring system for simple and objective screening of MDD using a photoplethysmography (PPG) sensor and a 24-GHz microwave radar, which was based on the analysis of heart rate variability (HRV) during paced respiration and mental task conditions. In our previous study, we assessed the reactivity of HRV measurements during a mental task (random number generation) condition in patients with MDD and healthy control subjects. The HRV indices are less reactive in patients with MDD compared to healthy subjects during the mental task, which enabled us to identify the patients at risk for depression. In this study, the reactivity of HRV was measured not only in the mental task but also during paced respiration (i.e., 5-s inhalation and 5-s exhalation) conditions, thereby assessing more detailed autonomic nervous system (ANS) activity via HRV indices. To investigate the effect of paced respiration on MDD screening, we compared the ANS activity via HRV indices in with/without paced respiration conditions in 28 drug-naïve patients with MDD and 27 healthy control subjects. The result showed that ANS significantly activated during the paced respiration condition (p<;0.05). The sensitivity in detecting patients with MDD was 86% under paced respiration and mental task conditions, which was higher than the sensitivity (68%) under mental task condition alone.