Clarithromycin overcomes stromal cell-mediated drug resistance against proteasome inhibitors in myeloma cells via autophagy flux blockage leading to high NOXA expression

by Shota Moriya, Hiromi Kazama, Hirotsugu Hino, Naoharu Takano, Masaki Hiramoto, Shin Aizawa, Keisuke Miyazawa

We previously reported that macrolide antibiotics, such as clarithromycin (CAM), blocked autophagy flux, and simultaneous proteasome and autophagy inhibition by bortezomib (BTZ) plus CAM resulted in enhanced apoptosis induction in multiple myeloma (MM) cells via increased endoplasmic reticulum (ER) stress loading. However, in actual therapeutic settings, cell adhesion-mediated drug resistance between bone marrow stromal cells (BMSC) and MM cells has been known to be a barrier to treatment. To investigate whether CAM could enhance BTZ-induced cytotoxicity in MM cells under direct cell adhesion with BMSC, we established a co-culture system of EGFP-labeled MM cells with BMSC. The cytotoxic effect of BTZ on MM cells was diminished by its interaction with BMSC; however, the attenuated cytotoxicity was recovered by the co-administration of CAM, which upregulates ER stress loading and NOXA expression. Knockout of NOXA in MM cells canceled the enhanced cell death by CAM, indicating that NOXA is a key molecule for cell death induction by the co-administration of CAM. Since NOXA is degraded by autophagy as well as proteasomes, blocking autophagy with CAM resulted in the sustained upregulation of NOXA in MM cells co-cultured with BMSC in the presence of BTZ. Our data suggest that BMSC-associated BTZ resistance is mediated by the attenuation of ER stress loading. However, the addition of CAM overcomes BMSC-associated resistance via upregulation of NOXA by concomitantly blocking autophagy-mediated NOXA degradation and transcriptional activation of NOXA by ER stress loading.

Uncertainty in the association between socio-demographic characteristics and mental health

by Nataliya Rybnikova, Dani Broitman, Murielle Mary-Krause, Maria Melchior, Yakov Ben-Haim

Questionnaires are among the most basic and widespread tools to assess the mental health of a population in epidemiological and public health studies. Their most obvious advantage (firsthand self-report) is also the source of their main problems: the raw data requires interpretation, and are a snapshot of the specific sample’s status at a given time. Efforts to deal with both issues created a bi-dimensional space defined by two orthogonal axes, in which most of the quantitative mental health research can be located. Methods aimed to assure that mental health diagnoses are solidly grounded on existing raw data are part of the individual validity axis. Tools allowing the generalization of the results across the entire population compose the collective validity axis. This paper raises a different question. Since one goal of mental health assessments is to obtain results that can be generalized to some extent, an important question is how robust is a questionnaire result when applied to a different population or to the same population at a different time. In this case, there is deep uncertainty, without any a priori probabilistic information. The main claim of this paper is that this task requires the development of a new robustness to deep uncertainty axis, defining a three-dimensional research space. We demonstrate the analysis of deep uncertainty using the concept of robustness in info-gap decision theory. Based on data from questionnaires collected before and during the Covid-19 pandemic, we first locate a mental health assessment in the space defined by the individual validity axis and the collective validity axis. Then we develop a model of info-gap robustness to uncertainty in mental health assessment, showing how the robustness to deep uncertainty axis interacts with the other two axes, highlighting the contributions and the limitations of this approach. The ability to measure robustness to deep uncertainty in the mental health realm is important particularly in troubled and changing times. In this paper, we provide the basic methodological building blocks of the suggested approach using the outbreak of Covid-19 as a recent example.

Do residential patterns affect women’s labor market performance? An empirical study based on CHFS data

by Siyan Zhou, Qing Wang

Based on China Household Finance Survey (CHFS) data from 2019, this paper explores the impact of the residential pattern of coresidence with parents on the labor market performance of women in married families with minor children. The study finds that coresidence with parents significantly increases the possibility of female labor market participation and positively impacts women’s employment income. To overcome the potential endogeneity problem of residential patterns, this paper uses the Heckman two-step method and the conditional mixed process estimation method (CMP method) for regression, and the conclusions remain robust. The mechanism analysis shows that coresidence with parents has both grandchild care and elderly care factors, which have a spillover effect and a crowding-out effect on female labor market performance, respectively. Since the spillover effect is more significant than the crowding-out effect, coresidence with parents positively impacts women’s labor market performance. The heterogeneity analysis shows that in terms of labor force participation rate, coresidence with parents has a more significant impact on women in families with children aged 0–6, women in families without boys, and women in families with employed husbands. In terms of income, coresidence with parents has a more significant impact on women in families with employed husbands. This study provides a new perspective for promoting female labor market performance and can serve as a reference for future policy formulation.

In silico exploration of <i>Serratia</i> sp. BRL41 genome for detecting prodigiosin Biosynthetic Gene Cluster (BGC) and in vitro antimicrobial activity assessment of secreted prodigiosin

by Farhana Boby, Md. Nurul Huda Bhuiyan, Barun Kanti Saha, Subarna Sandhani Dey, Anik Kumar Saha, Md Jahidul Islam, Mahci Al Bashera, Shyama Prosad Moulick, Farhana Jahan, Md. Asad Uz Zaman, Sanjana Fatema Chowdhury, Showti Raheel Naser, Md. Salim Khan, Md. Murshed Hasan Sarkar

The raising concern of drug resistance, having substantial impacts on public health, has instigated the search of new natural compounds with substantial medicinal activity. In order to find out a natural solution, the current study has utilized prodigiosin, a linear tripyrrole red pigment, as an active ingredient to control bacterial proliferation and prevent cellular oxidation caused by ROS (Reactive Oxygen Species). A prodigiosin-producing bacterium BRL41 was isolated from the ancient Barhind soil of BCSIR Rajshahi Laboratories, Bangladesh, and its morphological and biochemical characteristics were investigated. Whole genome sequencing data of the isolate revealed its identity as Serratia sp. and conferred the presence of prodigiosin gene cluster in the bacterial genome. “Prodigiosin NRPS”, among the 10 analyzed gene clusters, showed 100% similarity with query sequences where pigC, pigH, pigI, and pigJ were identified as fundamental genes for prodigiosin biosynthesis. Some other prominent clusters for synthesis of ririwpeptides, yersinopine, trichrysobactin were also found in the chromosome of BRL41, whilst the rest displayed less similarity with query sequences. Except some first-generation beta-lactam resistance genes, no virulence and resistance genes were found in the genome of BRL41. Structural illumination of the extracted red pigment by spectrophotometric scanning, Thin-Layer Chromatography (TLC), Fourier Transform Infrared Spectroscopy (FTIR), and change of color at different pH solutions verified the identity of the isolated compound as prodigiosin. Serratia sp. BRL41 attained its maximum productivity 564.74 units/cell at temperature 30˚C and pH 7.5 in two-fold diluted nutrient broth medium. The compound exhibited promising antibacterial activity against Gram-positive and Gram-negative bacteria with MIC (Minimum Inhibitory Concentration) and MBC (Minimum Bactericidal Concentration) values ranged from 3.9 to15.62 μg/mL and 7.81 to 31.25 μg/mL respectively. At concentration 500 μg/mL, except in Salmonella enterica ATCC-10708, prodigiosin significantly diminished biofilm formed by Listeria monocytogens ATCC-3193, Pseudomonas aeruginosa ATCC-9027, Escherichia coli (environmental isolate), Staphylococcus aureus (environmental isolate). Cellular glutathione level (GSH) was elevated upon application of 250 and 500 μg/mL pigment where 125 μg/mL failed to show any free radical scavenging activity. Additionally, release of cellular components in growth media of both Gram-positive and Gram-negative bacteria were facilitated by the extract that might be associated with cell membrane destabilization. Therefore, the overall findings of antimicrobial, antibiofilm and antioxidant activities suggest that in time to come prodigiosin might be a potential natural source to treat various diseases and infections.

Online search interest in long-term symptoms of coronavirus disease 2019 during the COVID-19 pandemic in Japan: Infodemiology study using the most visited search engine in Japan

by Kosuke Ishizuka, Taiju Miyagami, Tomoya Tsuchida, Mizue Saita, Yoshiyuki Ohira, Toshio Naito

Prolonged symptoms that occur after COVID-19 (long-COVID) vary from mild, which do not interfere with daily life, to severe, which require long-term social support. This study assessed the secular trend in online searches on long-COVID in Japan. We conducted an observational study using data provided by Yahoo! JAPAN on the monthly search volume of query terms related to long-COVID from January 2020 to December 2022, including the search volume of the query “コロナ後遺症” (long-COVID in Japanese). The number of new cases of COVID-19 by month was used as a control for search trends, and the symptoms retrieved in conjunction with long-COVID were compared. Trends in online searches for each symptom of long-COVID were analyzed. The symptoms of long-COVID were classified according to “Component 1—Symptoms and Complaints” of the International Classification of Primary Care, 2nd edition (ICPC-2). Interest in long-COVID increased in response to peaks in the number of new cases of COVID-19 in Japan. The most frequent symptom searches with long-COVID were hair loss/baldness (3,530, 21,400, and 33,600 searches in 2020, 2021, and 2022, respectively), cough (340, 7,900 and 138,910 searches in 2020, 2021, and 2022, respectively), disturbance of smell/taste (230, 13,340, and 44,160 searches in 2020, 2021, and 2022, respectively), and headache (580, 6,180, and 42,870 searches in 2020, 2021, and 2022, respectively). In addition, the ranking of interest in “weakness/tiredness, general” in long-COVID increased each year (not in the top 10 in 2020, seventh in 2021, and second in 2022), and the absolute number of searches also increased. To our knowledge, this is the first study to investigate secular trends in online interest in long-COVID in the world. Continued monitoring of online interest in long-COVID is necessary to prepare for a possible increase in the number of patients with long-COVID.