Short Time Course Research Articles

Similarities and distinctions in the activation of the Candida glabrata Pdr1 regulatory pathway by azole and non-azole drugs.

Incidences of fluconazole (FLC) resistance among Candida glabrata clinical isolates is a growing issue in clinics. The pleiotropic drug response (PDR) network in C. glabrata confers azole resistance and is defined primarily by the Zn2Cys6 zinc cluster-containing transcription factor Pdr1 and target genes such as CDR1, that encodes an ATP-binding cassette transporter protein thought to act as a FLC efflux pump. Mutations in the PDR1 gene that render the transcription factor hyperactive are the most common cause of fluconazole resistance among clinical isolates. The phenothiazine class drug fluphenazine and a molecular derivative, CWHM-974, which both exhibit antifungal properties, have been shown to induce the expression of Cdr1 in Candida spp. We have used a firefly luciferase reporter gene driven by the CDR1 promoter to demonstrate two distinct patterns of CDR1 promoter activation kinetics: gradual promoter activation kinetics that occur in response to ergosterol limitations imposed by exposure to azole and polyene class antifungals and a robust and rapid CDR1 induction occurring in response to the stress imposed by fluphenazines. We can attribute these different patterns of CDR1 induction as proceeding through the promoter region of this gene since this is the only segment of the gene included in the luciferase reporter construct. Genetic analysis indicates that the signaling pathways responsible for phenothiazine and azole induction of CDR1 overlap but are not identical. The short time course of phenothiazine induction suggests that these compounds may act more directly on the Pdr1 protein to stimulate its activity.

O-263 The risk of hypogonadism after microdissection testicular sperm extraction (micro TESE) in non-obstructive azoospermia (NOA) patients with various etiologies

Abstract Study question What is the risk factor of hypogonadism after micro TESE in NOA patients? Summary answer Klinefelter syndrome (KS), past history of cryptorchidism, preoperative testicular volume <8 ml or testosterone <287 ng/dL are significant risk factors of hypogonadism after micro TESE. What is known already Micro TESE combined with intracytoplasmic sperm injection (ICSI) is currently standard procedure to treat infertility in cases of NOA. Although many studies have reported the effectiveness of the treatment concerning about sperm retrieval rates (SRR) and the live birth rates, data on safety, especially on the risk of hypogonadism, are limited. Most previous studies, including our past study, regarding the risk of hypogonadism after TESE have been limited to relatively few in number of patients and the investigations only of the changes of mean serum testosterone levels. Study design, size, duration This study retrospectively investigated 535 NOA patients including 336 cases of unexplained NOA, 91 KS, 16 other chromosomal abnormalities, 31 azoospermia factor (AZF)c deletions, 31 past history of cryptorchidism, and 30 post anticancer chemotherapy, who had undergone micro TESE at our institution from September 2013 to August 2018. All of them had been examined serum follicle-stimulating hormone (FSH), luteinizing hormone (LH), and testosterone levels before and at least once in 1-3 months after micro TESE. Participants/materials, setting, methods In this study, hypogonadism was defined as total testosterone <250 ng/dL, which is established by the Japanese Society of Men’s Health as diagnostic criteria for late onset hypogonadism syndrome, and/or starting testosterone replacement therapy (TRT). We assessed risk factors of hypogonadism after micro TESE using indicators such as age, preoperative testicular volume and testosterone levels, past medical history, unilateral or bilateral, first time or not micro TESE. Main results and the role of chance In total of 535 men, FSH and LH after micro TESE significantly increased vs baseline (26.2 vs 29.9 IU/L, 9.6 vs 12.8 IU/L, respectively). On the other hand, testosterone levels significantly decreased after surgery (351.2 vs 297.5 ng/dL). Although postoperative hypogonadism was found in 42% (227/535), many of whom were cases of KS or preoperative testosterone levels <250 ng/dL. Sixteen men with KS, 3 men with unexplained NOA, and 2 men with past history of cryptorchidism started TRT because they complained clinical symptoms after micro TESE. Of the 21 men who started TRT, only 5 had new-onset low testosterone levels after micro TESE. According to the assessment of any association of various indicators and low postoperative total testosterone levels using multiple logistic regression analysis, significant risk factors of hypogonadism early after micro TESE were cases of KS (OR 11.4, 95% CI 4.4-29.9), preoperative testosterone levels <287 ng/dL (OR 7.5, 95% CI 4.7-12.0), preoperative testicular volume <8 ml (OR 3.9, 95% CI 2.3-6.4), past history of cryptorchidism (OR 2.9, 95% CI 1.1-7.8). Limitations, reasons for caution Most examinations of postoperative hormone levels were performed in short-term period after micro TESE, so we could only assess the risk of postoperative hypogonadism in a short time course in this cohort study. Wider implications of the findings The strength of the current study is that the risk factors of hypogonadism after micro TESE was assessed in a large population of NOA men with various etiologies. The results of this study provide useful clinical information about the risk of postoperative hypogonadism for NOA patients considering micro TESE. Trial registration number not applicable

Poster 151: Femoral Cartilage T2* Relaxation Times Correlate With Inflammatory Biomarker MCP-1 Levels In Serum, But Not Synovial Fluid, After ACL Injury In A Skeletally Immature Porcine Model

Poster 151: Femoral Cartilage T2* Relaxation Times Correlate With Inflammatory Biomarker MCP-1 Levels In Serum, But Not Synovial Fluid, After ACL Injury In A Skeletally Immature Porcine Model

Pedagogical grammar as the framework of TEFL research. Part 14. Age and foreign language acquisition: experimental data

The paper presents a comparative analysis of various groups of data (language acquisition speed, quality of language acquisition, comparative acquisition efficiency depending on the period of exposure, adult-child, and younger-children versus older-childrenʼs differences in acquisition) obtained in different experiments over a substantial time span. As it follows from the experimental studies analysis, there is conflicting data regarding the existence of a “critical age” for the foreign language acquisition and its specific limits. Much of the data can be interpreted in various ways. Differences between adults and children, older children and younger children can be explained not only by the existence of the “critical age”, but also by other reasons. The advantage of older students may be explained by their better memory, which allows them to learn a large number of clichés with which they successfully communicate, even having a very limited supply of language material. In general, basing on the available data, it is hardly possible to state that there is any age boundary in human life beyond which the foreign language acquisition is impossible or difficult to any serious degree. Experimental data, while contradictory, do not give rise to a straightforward conclusion about the advantage of any age group in any aspect of language. In short-time courses, adults progress faster but on longer stretches, children close the gap and even outpace adults. There are quite a few differences in language acquisition between adults and children, as well as older and younger children, but these differences are probably not due to the existence of a “critical age”, but to other factors discussed in the paper. It is likely that adults can learn language just as effectively as children provided the factors that impede their learning (strong instrumental and integrative motivation, absence of unfavorable affective factors, etc.) are eliminated. However, this assumption requires further research.

Bioconversion of a Peanut Oil Processing By-Product into a Novel α-Glucosidase Inhibitor: Hemi-Pyocyanin

Hemi-pyocyanin (HPC) is a heterocyclic nitrogenous compound with some reported potential medical effects. The current report aimed to investigate the potential use of organic industrial waste for the production of HPC via microbial fermentation. The novel antidiabetic activity of HPC was also accessed and reported in this work. A peanut oil processing by-product (groundnut cake) was screened as the best substrate for Pseudomonas aeruginosa TUN03 conversion to obtain high-yield HPC. This compound was further produced in a 14 L bioreactor system on a large scale (6 L per pilot) and reached higher productivity (35.1 μg/mL) in a shorter time course of cultivation (8 h) compared to fermentation on a small scale in flasks (19.5 μg/mL; 3 days of fermentation). On assessing its activity, HPC demonstrated potent inhibition against α-glucosidase, an antidiabetic enzyme, with a low IC50 value (0.572 mg/mL) and a maximum inhibition rate of 100%. In an in silico study, HPC was found to inhibit α-glucosidase with a good binding energy score (−9.0 kcal/mol) via interaction with amino acids Lys156, Leu313, and Arg315 at the active site, and three bonds (1 H-acceptor and 2 pi-H) were generated. The data from five Lipkin’s rules and ADMET-based pharmacokinetics and pharmacology revealed that HPC possesses drug-like properties and good ADMET properties within the required allotted limitations. The data obtained in the current work highlighted the potential application of groundnut cakes for the eco-friendly and scaled-up production of HPC, a new anti-α-glucosidase agent that should be further investigated for type 2 diabetes management.

Vitamin D Increases Irisin Serum Levels and the Expression of Its Precursor in Skeletal Muscle

Irisin is a myokine synthesized by skeletal muscle, which performs key actions on whole-body metabolism. Previous studies have hypothesized a relationship between irisin and vitamin D, but the pathway has not been thoroughly investigated. The purpose of the study was to evaluate whether vitamin D supplementation affected irisin serum levels in a cohort of 19 postmenopausal women with primary hyperparathyroidism (PHPT) treated with cholecalciferol for six months. In parallel, to understand the possible link between vitamin D and irisin, we analyzed the expression of the irisin precursor, Fndc5, in the C2C12 myoblast cell line treated with a biologically active form of vitamin D, 1α,25-dihydroxyvitamin D3 (1,25(OH)2D3). Our results demonstrate that vitamin D supplementation resulted in a significant increase in irisin serum levels (p = 0.031) in PHPT patients. In vitro, we show that vitamin D treatment on myoblasts enhanced Fndc5 mRNA after 48 h (p = 0.013), while it increased mRNAs of sirtuin 1 (Sirt1) (p = 0.041) and peroxisome proliferator-activated receptor γ coactivator 1α (Pgc1α) (p = 0.017) over a shorter time course. Overall, our data suggest that vitamin-D-induced modulation of Fndc5/irisin occurs through up-regulation of Sirt1, which together with Pgc1α, is an important regulator of numerous metabolic processes in skeletal muscle.

Review of air quality monitoring : Case study of Peenya Industrial area

Increased levels of industrial activity, high growth rates in automobiles and emissions from various sources have caused deterioration in the ambient air around us leaving a deep, harmful impact on our health. In this study we take Peenya industrial area , Bangalore to study meteorological monitoring with different techniques and methodologies, by using advanced sophisticated instruments equipped with digital display and data loggers to produce very accurate data which enables us to provide reports within a short course of time. We are targeting environmental monitoring with respect meteorological monitoring , ambient air and indoor air quality for road projects, hospitals & school/ college premises, pharma industries, MNC’s and software companies to take necessary precautionary measurements on what could be basis to reduce the levels of pollutants which cause various health hazards to human beings.

PAX3-FOXO1 coordinates enhancer architecture, eRNA transcription, and RNA polymerase pause release at select gene targets

Transcriptional control is a highly dynamic process that changes rapidly in response to various cellular and extracellular cues, making it difficult to define the mechanism of transcription factor function using slow genetic methods. We used a chemical-genetic approach to rapidly degrade a canonical transcriptional activator, PAX3-FOXO1, to define the mechanism by which it regulates gene expression programs. By coupling rapid protein degradation with the analysis of nascent transcription over short time courses and integrating CUT&RUN, ATAC-seq, and eRNA analysis with deep proteomic analysis, we defined PAX3-FOXO1 function at a small network of direct transcriptional targets. PAX3-FOXO1 degradation impaired RNA polymerase pause release and transcription elongation at most regulated gene targets. Moreover, the activity of PAX3-FOXO1 at enhancers controlling this core network was surprisingly selective, affecting single elements in super-enhancers. This combinatorial analysis indicated that PAX3-FOXO1 was continuously required to maintain chromatin accessibility and enhancer architecture at regulated enhancers.

Combinatorial PI3K/AKT Pathway Inhibition As a Therapeutic Approach in Diffuse Large B-Cell Lymphoma

Background. Diffuse large B-cell lymphoma (DLBCL) is the most frequent type of non-Hodgkin lymphoma (NHL, ~ 25-30% of NHL cases). Currently, DLBCL treatment is based on non-specific chemotherapy combination (cyclophosphamide, doxorubicin, vincristine, and prednisone) with anti-CD20 monoclonal antibody. Although the DLBCL survival dramatically improved during last decades, approximately 40 % of DLBCL patients could not be cured with the currently used therapeutic approaches. Novel approaches are, therefore, needed to improve DLBCL survival and to decrease treatment related toxicity. Since many DLBCL tumors depend on phosphatidylinositol 3-kinase /protein kinase B (PI3K/AKT) signaling pathway, it represents one of the possible novel therapeutic targets. Despite promising results in various cancers, PI3K/AKT pathway inhibitors show only partial efficacy, low specificity, and resistance development in lymphomas. Since the PI3K/AKT activity is highly regulated, we hypothesize that the PI3K/AKT pathway needs to be inhibited at multiple levels simultaneously to achieve its full and lasting inhibition. Methods. To provide a proof of principle of multilevel vertical PI3K/AKT inhibition, we tested the effect of four inhibitors targeting the PI3K/AKT pathway at four different levels: PI3Kδ inhibitor idelalisib, PDPK1 inhibitor GSK2334470, AKT inhibitor ipatasertib, and mTOR inhibitor rapamycin on a spectrum of commonly used DLBCL model cell lines. Precise measurement of AKT activity in living cells was done using our recently published genetically encoded Förster Resonance Energy Transferred (FRET) based AKT activity reporter including novel methodology for flow cytometry-based FRET measurement (PMID: 34128311) allowing medium throughput and measurement in kinetics setting. Results. First, we measured how single inhibitors affect the AKT activity. The AKT activity IC50 concentrations were in 100 nM range except for rapamycin that had no AKT inhibitory effect. These concentrations were approximately 10 times lower in comparison to IC50 values calculated from net cell growth inhibition assessing the overall inhibitor toxicity. This suggests compensatory mechanisms and/or off target effects of PI3K/AKT inhibitors. Precise measurement of AKT activity allowed us to identify the minimal concentration of each inhibitor at which AKT activity was fully inhibited. Kinetics and short time course experiments showed that maximal AKT inhibition is achieved within hours of inhibitors treatment initiation. Importantly, pharmacological inhibition of one or two members of PI3K/AKT signaling cascade did not lead to lasting AKT activity inhibition. Cellular contra-regulatory mechanisms compensated for the decrease of AKT activity within 24 hours even in the presence of the inhibitors. Moreover, we detected a massive increase in AKT activity following release of the cells from media with PI3K/AKT pathway inhibitors. This further illustrates the powerful cellular contra-regulatory mechanisms and highlights the need for combinatorial approach. Combination of at least three inhibitors was necessary to achieve lasting AKT activity decrease (long time course experiment, Fig. 1A). Cross titration experiments further confirmed synergistic effect between tested PI3K/AKT pathway inhibitors on cell growth inhibition and allowed to determine the concentration ratios of their maximal synergism. Importantly, testing of single inhibitors and two, three, or all four inhibitors in fixed concentration combinations identified a substantial decrease in cell growth/toxicity IC50 concentrations of individual inhibitors (Fig. 1B). Conclusions. Concept of inhibiting more members of PI3K/AKT pathway was suggested before using combinations and/or dual inhibitors of AKT and mTOR, however, our data suggest that more than two inhibitors are needed for lasting PI3K/AKT inhibition. Moreover, we provided a proof of principle for further multilevel PI3K/AKT signaling inhibition testing, including in vivo cell line and/or patient derived xenograft models. Our data suggest that proposed combinations should allow significant decrease of inhibitors concentrations limiting off-target effect. This approach might overcome above mentioned issues of PI3K/AKT pathway inhibition and allow full therapeutical utilization of this pathway inhibition possibly applicable to other lymphoma types. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Need for cognition is associated with the interaction of reward and task-load on effort: A verification and extension study

Here, we work to provide nuance around the assumption that people will work for rewards. We examine whether individuals' inherent tendency to mobilize cognitive effort (need for cognition, NFC) moderates this effect. We re-analyzed our existing data to verify an effect reported by Sandra and Otto (2018) regarding the association between NFC and reward-induced cognitive effort expenditure, using a more ecological cognitive task design and adding a psychophysiological measure of effort. Specifically, distinct from their short time course visual task-switching paradigm, we used a relatively long course auditory comprehension task paradigm. We found that, consistent with the original study, increased cognitive effort in response to incentive reward depends on individual differences in cognitive motivation (need for cognition). We also found that, to observe consistent phenomena, different indices of effort (behavioral and psychophysiological) need to be considered when evaluating the relationship between the effort expenditure and cognitive motivation. Pupil dilation showed an advantage over reaction time in revealing mental effort mobilized over a prolonged cognitive task. Our results suggest that assessing cognitive motivation when planning a behavior-change program involving reward feedback for positive performance could help to optimize individuals' effort investment.

Evaluation of high-intensity interval training on MMP-9 and Galectin-3 indices in male judokas of Shushtar City

Interval exercises are used as an effective approach to improving fitness in a short course of time. The effect of such exercises on functional and physiological adaptations as well as inflammatory responses in athletes is still unclear. The effect of such exercises on functional and physiological adaptations as well as inflammatory responses in athletes is still unclear. Therefore, the present study aimed to investigate high-intensity interval training on MMP-9 and Galectin-3 indices in male judokas in Shushtar city. Materials and methods: For this purpose, among male judokas in Shushtar, 12 people were randomly selected (with an age range of 18-25 years, mean weight of 64.5 ± 6.52, and body mass index of 22.03 ± 2.15) to conduct the research. Subjects underwent a training session that included a combination of judo and bodybuilding exercises under the supervision of an instructor and by controlling the intensity of the exercises by measuring the heart rate during the training. Blood samples were taken from the subjects in 3 shifts (before the camp, after one training course, and after seven training sessions) to measure the levels of metalloproteinase matrix-9 and glactin-3.

Abstract 2362: Pax3-foxo1 coordinates enhancer architecture, eRNA transcription, and RNA polymerase pause release at select gene targets

Abstract Background: Gene expression networks are perturbed in every human malignancy. One common way that this occurs is through chromosomal translocations such as the the t(2;13)(q35;q14), which creates the oncogenic transcription factor, PAX3-FOXO1, in alveolar rhabdomyosarcoma (aRMS). Identification of the direct transcriptional targets and mechanisms of control by translocated transcription factors is critical to our understanding of disease etiology and the development of targeted therapeutic approaches. However, defining the genes directly controlled by DNA binding proteins and the mechanisms of regulation at those direct target genes is challenging, because traditional genetic approaches to inactivate transcription factors cannot distinguish direct from indirect changes in gene expression. Methods: A CRISPR-based approach was used to engineer the endogenous PAX3-FOXO1 locus to incorporate the FKBP12F36V degron tag. This allowed degradation of PAX3-FOXO1 within 2 hr of treatment with the proteolysis targeting chimera, dTAG-47. We performed short time courses to analyze nascent transcription (precision nuclear run-on sequencing; PRO-seq), chromatin accessibility (ATAC-seq), and genome-wide mapping of transcriptional complexes (CUT&RUN). This identified a small cohort of high-confidence PAX3-FOXO1 gene targets and associated PAX3-FOXO1-regulated enhancer elements. In addition, we used CRISPR editing to add APEX2 and 3XFLAG tags to PAX3-FOXO1 to enable deep proteomics to identify PAX3-FOXO1-associated transcriptional complexes in aRMS cell lines. Results: Degradation of PAX3-FOXO1-FKBP12F36V led to growth inhibition, hallmarks of myogenic differentiation, and reduced growth in soft agar. PAX3-FOXO1 binding was detected at over 44,000 sites throughout the genome using CUT&RUN, and loss of chromatin bound PAX3-FOXO1 was observed within 2-4hr of PAX3-FOXO1 degradation. PRO-seq analysis defined a core transcriptional network that rapidly collapsed upon PAX3-FOXO1 degradation. Unexpectedly, loss of PAX3-FOXO1 impaired RNA polymerase pause release and transcription elongation at regulated gene targets. Deep proteomic analysis showed that PAX3-FOXO1 was in close proximity to many transcriptional regulatory complexes, but appeared to coordinate the activity of these complexes at only a few hundred sites. Moreover, the continued presence of PAX3-FOXO1 at these elements was required to maintain chromatin accessibility. Conclusion: This work demonstrates the utility of using rapid degradation of endogenous transcription factors to define their direct transcription targets and the mechanisms by which these factors control gene expression. Overall, this work provides a detailed mechanism by which PAX3-FOXO1 maintains an oncogenic transcriptional regulatory network, and emphasizes the utility of PAX3-FOXO1 as a therapeutic target in aRMS. Citation Format: Susu Zhang, Jing Wang, Qi Liu, Hayes McDonald, Monica Bomber, Hillary Layden, Jacob Ellis, Scott Borinstein, Scott Hiebert, Kristy Stengel. Pax3-foxo1 coordinates enhancer architecture, eRNA transcription, and RNA polymerase pause release at select gene targets [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2362.

The Delay of Raphanus raphanistrum subsp. sativus (L.) Domin Seed Germination Induced by Coumarin Is Mediated by a Lower Ability to Sustain the Energetic Metabolism.

In the present study, the mode of action of coumarin using the germination process as a target was investigated. A dose–response curve, built using a range of concentrations from 0 to 800 µM, allowed us to identify a key concentration (400 µM) inhibiting the germination process, reducing its speed without compromising seed development. Successively, short time-course (0–48 h) experiments were carried out to evaluate the biochemical and metabolic processes involved in coumarin-induced germination delay. The results pointed out that coumarin delayed K+, Ca2+, and Mg2+ reabsorption, suggesting a late membrane reorganisation. Similarly, seed respiration was inhibited during the first 24 h but recovered after 48 h. Those results agreed with ATP levels, which followed the same trend. In addition, the untargeted metabolomic analysis allowed to identify, among the pathways significantly impacted by the treatment, amino acids metabolism, the TCA cycle, and the glyoxylate pathway. The results highlighted that coumarin was able to interact with membranes reorganisation, delaying them and reducing the production of ATP, as also supported by pathway analysis and cell respiration. The in vivo 31P-NMR analysis supported the hypothesis that the concentration chosen was able to affect plant metabolism, maintaining, on the other hand, its viability, which is extremely important for studying natural compounds’ mode of action.

Modelling biophoton emission kinetics based on the initial intensity value in Helianthus annuus plants exposed to different types of stress

Biophoton radiation also referred to as ultra-weak photon emission (UPE) is used to denote a spontaneous and permanent photon emission associated with oxidative processes in cells and seems to universally occur in all living systems as a result of the generation of reactive oxygen species (ROS) that are produced under stress conditions. The measurement of this biophoton emission allows for a non-invasive approach in monitoring phenological stages throughout plant development which has direct relevance in agriculture research. In this study, the emission of photons emanating from sunflower (Helianthus annuus, L.) plants exposed to biotic and abiotic stress has been investigated. In healthy plants raised under controlled growth conditions UPE was low whereas in stressed individuals it considerably increased; particularly upon water stress. The kinetics of the signal is shown to reveal an exponential decay with characteristic dynamics, which appears to reflect different physiological states concomitantly setting in upon stress. The dynamics of the signal decay is shown to vary according to the type of stress applied (biotic vs. abiotic) hence suggesting a putative relationship between the kinetic traits of change in the signal intensity-decay and stress. Intriguingly, the determination of the change in the intensity of biophoton emission that ensued in a short time course was possible by using the initial biophoton emission intensity. The predictability level of the equations demonstrated the applicability of the model in a corroborative manner when employing it in independent UPE-measurements, thus permitting to forecast the intensity change in a very accurate way over a short time course. Our findings allow the notion that albeit stress confers complex and complicated changes on oxidative metabolism in biological systems, the employment of biophoton imaging offers a feasible method making it possible to monitor oxidative processes triggered by stress in a non-invasive and label-free way which has versatile applications especially in precision agriculture.

Meal-to-meal and day-to-day macronutrient variation in an ad libitum vending food paradigm

BackgroundTheory posits that macronutrient intake is regulated by protein consumption and adequate intake of protein results in consumption of less carbohydrates and fat. The current study investigates the effect of protein intake on calorie and macronutrient content using an ad libitum vending machine paradigm. MethodsHealthy volunteers (n = 287; 177 m; Age = 36 ± 11; BMI = 32 ± 8) were admitted to our clinical research unit. Macronutrient meal content (grams) and energy intake (Kcal) were quantified by specialized food processing software and collected on an hourly basis over a three-day period using a validated ad libitum vending machine paradigm. Body composition was assessed by DXA. Lagged multi-level models accounting for age, sex, race/ethnicity, fat and fat free mass indices were fitted to examine the impact of prior macronutrient content on subsequent meals. ResultsProtein intake was associated with decreased energy intake (Kcal; B = −1.67 kcal, p = 0.0048), lower protein and carbohydrate intake (B = −0.08 g, p = 0.0006; B = −0.21 g, p = 0.0003, respectively) at subsequent meals. Daily Macronutrient intake and subsequent intake were positively associated. ConclusionsDietary protein exhibits a negative regulatory effect on a short-term meal-to-meal rather than day-to-day basis. In the setting of readily available food, protein intake impacts energy intake only over very short time courses.

Anti-Mycobacterial Drug Resistance in Japan: How to Approach This Problem?

Mycobacteriosis is mainly caused by two groups of species: Mycobacterium tuberculosis and non-tuberculosis mycobacteria (NTM). The pathogens cause not only respiratory infections, but also general diseases. The common problem in these pathogens as of today is drug resistance. Tuberculosis (TB) is a major public health concern. A major challenge in the treatment of TB is anti-mycobacterial drug resistance (AMR), including multidrug-resistant tuberculosis and extensively drug-resistant tuberculosis. Recently, the success rate of the treatment of drug-resistant tuberculosis (DR-TB) has improved significantly with the introduction of new and repurposed drugs, especially in industrialized countries such as Japan. However, long-term treatment and the adverse events associated with the treatment of DR-TB are still problematic. To solve these problems, optimal treatment regimens designed/tailor-made for each patient are necessary, regardless of the location in the world. In contrast to TB, NTM infections are environmentally oriented. Mycobacterium avium-intracellulare complex (MAC) and Mycobacterium abscessus species (MABS) are the major causes of NTM infections in Japan. These bacteria are naturally resistant to a wide variation of antimicrobial agents. Macrolides, represented by clarithromycin (CLR) and amikacin (AMK), show relatively good correlation with treatment success. However, the efficacies of potential drugs for the treatment of macrolide-resistant MAC and MABS are currently under evaluation. Thus, it is particularly difficult to construct an effective treatment regimen for macrolide-resistant MAC and MABS. AMR in NTM infections are rather serious in Japan, even when compared with challenges associated with DR-TB. Given the AMR problems in TB and NTM, the appropriate use of drugs based on accurate drug susceptibility testing and the development of new compounds/regimens that are strongly bactericidal in a short-time course will be highly expected.

Development and evaluation of IL-6 overexpressing mesenchymal stem cells (MSCs).

Mesenchymal stem/stromal cell (MSC) therapy has been investigated in multiple diseases and conditions. Although the mechanisms of MSC-based therapies are not fully understood, we and others have shown interleukin 6 (IL-6) to be an important factor in MSC function. IL-6 contributes to many biological events, such as immune response, neurogenesis, and bone remodeling. In our study, we tested the feasibility of engineering MSCs by IL-6 mRNA transfection (eMSCs-IL6) and evaluated the optimal time to harvest them after transfection. We then assessed the functional characteristics of eMSCs-IL6. Quantitative real-time PCR and ELISA results have shown that mature IL-6 mRNA was efficiently transfected into MSCs using a lipofectamine based method. The IL-6 mRNA and protein overexpression peaked after 1day of transfection and the secreted IL-6 protein was sustained for at least 6days. A short time course experiment demonstrated that 4h after transfection was the best time point to harvest and freeze eMSCs-IL6 for future studies. In addition, eMSCs-IL6 maintained their characteristics as defined by International Society for Cell& Gene Therapy. The immunosuppressive capacity of conditioned culture medium (CCM) from eMSCs-IL6 (CCM-IL6) was significantly enhanced compared to naïve MSCs conditioned culture medium (CCM-control). Our studies established for the first time the feasibility of efficiently generating IL-6 overexpressing MSCs which have enhanced immunosuppressive capacity. This is providing a novel approach to improve the efficacy of MSCs for potential application in regenerative medicine.

The effect of participation of interventional radiology team in a primary trauma survey on patient outcome

PurposeThe purpose of this study was to examine the survival benefits of a workflow in which an interventional radiology (IR) team participates in a primary trauma survey on patients with hemodynamically unstable trauma. Materials and methodsA retrospective observational study was conducted between 2012 and 2019 at a single institution. Patients who underwent an IR procedure as the initial hemostasis were assigned to the hemodynamically stable group (HSG) or hemodynamically unstable group (HUG). The primary and secondary outcomes were survival at hospital discharge compared with the probability of survival (Ps) and the time course. ResultsA total of 160 patients (100 men, 60 women; median age, 57.5 years [interquartile range (IQR): 31.5–72 years]) with an injury severity score of 24 (IQR: 13.75–34) were included. A total of 125 patients were included in the HSG group and 35 patients in the HUG group. The observational survival rate was significantly greater than the Ps rate by 4.9% (95% confidence interval [CI]: 1.6–8.4%; P = 0.005) in HSG and by 24.6% in HUG (95% CI: 16.9–32.3%; P < 0.001). The observational survival rate was significantly greater than Ps in HUG than in HSG (P < 0.001). The median time to initiate IR procedures and the median procedure time in HUG were 54 min [IQR: 45–66 min] and 48 min [IQR: 30–85 min], respectively; both were significantly shorter than those in the HSG. ConclusionA trauma workflow utilizing an IR team in a primary survey is associated with improved survival of patients with hemodynamically unstable trauma when compared with Ps with a shorter time course.

690 Acute post-implantation enlargement of transcatheter self-expandable valve: insights from a single-centre prospective registry

Abstract Aims Transcatheter implantation of a balloon-expandable aortic valve is associated with an acute stent recoil . Conversely, the acute effects of nitinol-based stent frame- self expandable valve on post-deployment dimensions have not been reported. Accordingly, the aim of this study was to assess the occurrence and degree of acute prosthetic dimension changes after Evolute R valve implantation. Methods A total of 58 consecutive patients undergoing transcatheter aortic valve intervention (TAVI) with a widely used nitinol based self-expandable device (Evolute R, Medtronic, Minneapolis, USA) were included in this study. We measured valve diameters at three different sections: a) distal (aortic) level, b) central (annulus) level, and c) proximal (ventricular) level. Valve expansion was calculated as the difference between the diameters calculated immediately after valve deployment (A) and at the end of the procedure (B). The absolute and relative acute stent recoil were defined as B-A and (B-A)/B*100, respectively. A linear regression model was performed to test the association between the degree of valve extension, with baseline, as well as procedural characteristics. Results Final stent diameters were significantly higher compared to those achieved immediately after valve implantation in proximal (20.87±3.20 mm versus 20.37±3.27 mm, P &amp;lt;0.001), central (19.58±1.63 mm versus 19.12±1.75 mm, P &amp;lt;0.001) as well as distal (26.99±1.53 mm versus 26.41±1.57 mm, P &amp;lt;0.001) segments. The mean absolute and relative enlargement of the valve was respectively 0.50±0.51 mm and 2.48±2.57% in the proximal, 0.46±0.57 mm and 2.39±2.96% in the central and 0.58±0.59 mm and 2.14±2.14% in the distal segment. Among baseline and procedural characteristics assessed in the logistic regression model no association was observed between pre-dilation and valve expansion across all the segments (proximal: r2=0.004, P=0.885, central: r2=0.004, P=0.637, and distal r2=0.05, P=0.10). Of interest, post dilation affected only the expansion of the central segment (r2=0.241, P&amp;lt;0.001), while no association was observed for the proximal (r2=0.059, P=0.065) and distal (r2=0.0002, P=0.916) parts. The expansion of the proximal segment was associated with higher maximum/minimum diameter ratio measured by CT (r2=0.08, P=0.045), while no association was observed for the central (r2= 0.020; P=0.992) and distal (r2=1.111, P=0.683) segments. Conclusions This is the first study that documented a significant degree of post-deployment recoil of the Evolute R self-expandable valve, that was consistent across all the segments. The stent-valve expansion of the proximal segment was associated with a greater annulus dissymmetry reported by the maximum and minimum diameter ratio assessed by CT scan. However, further studies are required to assess the short and long-term time-course of self-expandable valve enlargement and the clinical relevance of this finding.

Physical Activity and Nutritional Influence on Immune Function: An Important Strategy to Improve Immunity and Health Status.

Physical activity (PA) and nutrition are the essential components of a healthy lifestyle, as they can influence energy balance, promote functional ability of various systems and improve immunity. Infections and their associated symptoms are the common and frequent challenges to human health that are causing severe economic and social consequences around the world. During aging, human immune system undergoes dramatic aging-related changes/dysfunctions known as immunosenescence. Clinically, immunosenescence refers to the gradual deterioration of immune system that increases exposure to infections, and reduces vaccine efficacy. Such phenomenon is linked to impaired immune responses that lead to dysfunction of multiple organs, while lack of physical activity, progressive loss of muscle mass, and concomitant decline in muscle strength facilitate immunosenescence and inflammation. In the present review, we have discussed the role of nutrition and PA, which can boost the immune system alone and synergistically. Evidence suggests that long-term PA is beneficial in improving immune system and preventing various infections. We have further discussed several nutritional strategies for improving the immune system. Unfortunately, the available evidence shows conflicting results. In terms of interaction with food intake, PA does not tend to increase energy intake during a short time course. However, overcoming nutritional deficiencies appears to be the most practical recommendation. Through the balanced nutritious diet intake one can fulfill the bodily requirement of optimal nutrition that significantly impacts the immune system. Supplementation of a single nutrient as food is generally not advisable. Rather incorporating various fruits and vegetables, whole grains, proteins and probiotics may ensure adequate nutrient intake. Therefore, multi-nutrient supplements may benefit people having deficiency in spite of sufficient diet. Along with PA, supplementation of probiotics, bovine colostrum, plant-derived products and functional foods may provide additional benefits in improving the immune system.