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Athraa Harjan Mohsen Alduhaidhawi,1 Sundus Nsaif AlHuchaimi,2 Thikra Abdullah Al- Mayah,2 Mushtak TS Al-Ouqaili,3 Samar Sami Alkafaas,4 Saravanan Muthupandian,5,6 Morteza Saki7 1Department of Biology, Faculty of Science, University of Kufa, Najaf, Iraq; 2Department of Parasitology, Faculty of Medicine, University of Kufa, Najaf, Iraq; 3Department of Microbiology, College of Medicine, University of Anbar, Ramadi, Al-Anbar Governorate, Iraq; 4Department of Chemistry, Division of Biochemistry, Faculty of Science, Tanta University, Tanta, Egypt; 5Department of Microbiology and Immunology, Division of Biomedical Sciences, School of Medicine, College of Health Sciences, Mekelle University, Mekelle, 1871, Ethiopia; 6Department of Pharmacology, AMR and Nanomedicine Laboratory, Center for Transdisciplinary Research, Saveetha Dental College, Saveetha Institute of Medical and Technical Sciences (SIMATS), Chennai, 60007, India; 7Department of Microbiology, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, IranCorrespondence: Saravanan Muthupandian, Department of Microbiology and Immunology, Division of Biomedical Sciences, School of Medicine, College of Health Sciences, Mekelle University, Mekelle, 1871, Ethiopia, Tel +919443077097, Email bioinfosaran@gmail.com Morteza Saki, Department of Microbiology, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran, Tel +989364221187, Email mortezasaki1981@gmail.comPurpose: This study aimed to evaluate the presence of CRISPR-Cas system genes and their possible association with antibiotic resistance patterns of Enterococcus faecalis and Enterococcus faecium species isolated from hospital wastewater (HWW) samples of several hospitals.Methods: HWW samples (200 mL) were collected from wastewater discharged from different hospitals from October 2020 to March 2021. The isolation and identification of enterococci species were performed by standard bacteriology tests and polymerase chain reaction (PCR). Antibiotic resistance was determined using the disc diffusion. The presence of various CRISPR-Cas systems was investigated by PCR. The association of the occurrence of CRISPR-Cas systems with antibiotic resistance was analyzed with appropriate statistical tests.Results: In total, 85 different enterococci species were isolated and identified using phenotypic methods. The results of PCR confirmed the prevalence of 50 (58.8%) E. faecalis and 35 (41.2%) E. faecium, respectively. In total, 54 (63.5%) of 85 isolates showed the presence of CRISPR-Cas loci. The incidence of CRISPR-Cas was more common in E. faecalis. CRISPR1, CRISPR2, and CRISPR3 were present in 35 (41.2%), 47 (55.3%), and 30 (35.3%) enterococci isolates, respectively. The CRISPR-Cas positive isolates showed significant lower resistance rates against vancomycin, ampicillin, chloramphenicol, erythromycin, rifampin, teicoplanin, tetracycline, imipenem, tigecycline, and trimethoprim-sulfamethoxazole in comparison with CRISPR-Cas negative isolates. The results showed that the presence of CRISPR-Cas genes was lower in multidrug-resistant (MDR) isolates (53.1%, n = 26/49) compared to the non-MDR enterococci isolates (77.8%, n = 28/36) (P = 0.023).Conclusion: This study revealed the higher prevalence of E. faecalis than E. faecium in HWWs. Also, the lack of CRISPR-Cas genes was associated with more antibiotic resistance rates and multidrug resistance in E. faecalis and E. faecium isolates with HWW origin.Keywords: antibiotic resistance, CRISPR-Cas, Enterococcus faecalis, Enterococcus faecium, hospital wastewater, HWW

Effat Abbasi Montazeri,1,2,* Azar Dokht Khosravi,1,2,* Morteza Saki,1– 3 Mehrandokht Sirous,2,4 Bijan Keikhaei,5 Sakineh Seyed-Mohammadi1– 3 1Infectious and Tropical Diseases Research Center, Health Research Institute, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran; 2Department of Microbiology, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran; 3Student Research Committee, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran; 4Department of Microbiology and Parasitology, Faculty of Medicine, Bushehr University of Medical Sciences, Bushehr, Iran; 5Thalassemia and Hemoglobinopathy Research Center, Health Institute, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran*These authors contributed equally to this workCorrespondence: Morteza Saki; Sakineh Seyed-Mohammadi Tel +98 9364221187; +98 9352655807Fax +98 6133332036Email mortezasaki1981@gmail.com; sakinehseyedmohammadi@gmail.comIntroduction: This study aimed to evaluate the frequency rate of extended-spectrum beta-lactamase-producing Enterobacteriaceae (ESBL-PE) causing bloodstream infections (BSIs) in cancer patients referred to one of the major referral hospitals in Ahvaz city, southwest Iran.Materials and Methods: In this study, 1700 blood cultures were collected from 610 cancer patients suspected to have BSI from October 2016 to August 2017 referred to the Shafa cancer hospital, Ahvaz, southwest of Iran. The blood culture bottles were incubated aerobically at 35– 37ºC for 24 hours and then sub-cultured on routine microbiology culture media. The bacterial colonies were identified using standard tests. The antibiotic susceptibility testing was achieved by the disc-diffusion method. The phenotypic detection of ESBLs was carried out by the combination disc-diffusion test (CDDT). Finally, the polymerase chain reaction (PCR) was performed to investigate the presence of blaTEM, blaCTX, blaSHV, and blaPER genes.Results: The prevalence of BSI in cancer patients was 16.4% (100/610). Gram-negative rods with rate of 74% (74/100) were the most prevalent bacteria. The frequency of Enterobacteriaceae family was 21% including Escherichia coli (n: 8), Klebsiella pneumoniae (n: 6), Enterobacter spp. (n: 5), Citrobacter freundii (n: 1), and Serratia marcescens (n: 1). All isolates were multidrug-resistant (resistance to three or more antibiotics). The results of CDDT showed that 42.8% (9/21) of Enterobacteriaceae isolates had a positive ESBL test of which 100% (9/9) indicated positive band for at least one of the ESBL genes by PCR method. The blaCTX-M and blaTEM genes were detected in 38% (8/21) and 23.8% (5/21) of isolates, respectively, while the blaSHV and blaPER were not detected in any isolates.Conclusion: Based on the results, surveillance, and antibiotic stewardship programs should be implemented for cancer patients to prevent the spread of more ESBL-PE that have limited therapeutically choices.Keywords: extended-spectrum beta-lactamase, ESBL, cancer patients, Enterobacteriaceae, hematological malignancies, Iran

Mojtaba Moosavian,1,2 Sakineh Seyed-Mohammadi,2,3 Morteza Saki,2,3 Fatemeh Shahi,2,3 Mahtab Khoshkholgh Sima,2 Davoud Afshar,4 Sara Barati5 1Infectious and Tropical Diseases Research Center, Health Research Institute, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran; 2Department of Microbiology, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran; 3Student Research Committee, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran; 4Department of Microbiology and Virology, School of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran; 5Department of Pathobiology, School of Veterinary, University of Shahid Chamran, Ahvaz, Iran Background: Legionnaires’ disease is an important public health problem that can cause substantial mortality and morbidity. Legionnaires’ disease-risk estimation may be compromised by uncertainties in Legionella-detection methods. The aim of this study was the detection of L. pneumophila in respiratory specimens of hospitalized patients with respiratory symptoms by culture, PCR, and loop-mediated isothermal amplification (LAMP) methods.Methods: Sputum and bronchoalveolar lavage samples were obtained from patients with pneumonia admitted to teaching hospitals in Ahvaz, Iran from June 2016 to March 2017. Isolation of Legionella spp. was done by culturing the samples directly onto buffered charcoal–yeast extract and modified Wadowsky–Yee agar medium. Then, PCR and LAMP assays were performed for detection of L. pneumophila via its mip gene in respiratory specimens.Results: A total of 100 respiratory specimens were collected. Our results showed that 1% of the samples were culture positive for Legionella spp., and 3% and 7% of samples were positive for L. pneumophila using the mip gene on PCR and LAMP assays, respectively.Conclusion: Legionnaires’ disease should be considered in the diagnosis of pulmonary infectious diseases. Also, the LAMP assay is a faster method with higher sensitivity and specificity than conventional methods, such as PCR and culture, for laboratory diagnosis of Legionnaires’ disease. Keywords: Legionella pneumophila, LAMP, PCR, mip gene

Ahmad Farajzadeh Sheikh,1,2 Mojtaba Moosavian,1,2 Mahtab Abdi,2 Mohsen Heidary,3,4 Fatemeh Shahi,2 Nabi Jomehzadeh,5 Sakineh Seyed-Mohammadi,1,2,6 Morteza Saki,1,2,6 Saeed Khoshnood1,2,6 1Infectious and Tropical Diseases Research Center, Health Research Institute, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran; 2Department of Microbiology, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran; 3Department of Microbiology, Faculty of Medicine, Iran University of Medical Sciences,Tehran, Iran; 4Student Research Committee, School of Allied Medical Sciences, Iran University of Medical Sciences, Tehran, Iran; 5Abadan School of Medical Sciences, Abadan, Iran; 6Student Research Committee, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran Introduction: Shigellosis is a significant global human health problem, and Shigella is in charge of almost 165 million cases of this disease annually, of whom 163 million cases are in developing countries and 1.5 million cases are in developed countries. The main aims of the current survey were to identify Shigella spp. isolated from diarrheal patients by conventional biochemical tests, determine the antimicrobial susceptibility profiles by disk diffusion method, and detect the ipaH gene using the PCR assay.Methods: The bacterial isolates were identified as Shigella spp. by microbiological tests and were serogrouped by the slide agglutination test. Antimicrobial susceptibility testing was performed using the disk diffusion method. PCR was performed to detect the ipaH gene.Results: The Shigella strains were isolated from 522 patients with various diarrhea, including bloody diarrhea (3%), mucoid plus bloody diarrhea (1.9%), mucoid diarrhea (3.2%), and watery diarrhea (3.2%). Overall, 69 (13.2%) isolates were positive for Shigella spp., of which 34 (49.3%) serotypes were identified as Shigella flexneri, 22 (31.9%) serotypes were identified as Shigella sonnei, 9 (13%) serotypes were identified as Shigella boydii, and 4 (5.8%) serotypes were identified as Shigella dysenteriae. Antibiotic susceptibility tests revealed that the highest resistance percentage was related to ampicillin (82%) and trimethoprim-sulfamethoxazole (77%), and ciprofloxacin and ceftriaxone were the best antibiotics against Shigella isolates.Conclusion: We concluded that Shigella spp. can be considered as an etiological agent of diarrhea in southwest Iran. Since the drug resistance pattern of Shigella differs geographically and over time within a country, continuous and regular surveillance program is necessary. Keywords: Shigella, diarrhea, resistance, Iran

Mansour Amin,1,2,* Mehrandokht Sirous,2,* Hazhir Javaherizadeh,3–5 Mohammad Motamedifar,6,7 Morteza Saki,2,8 Hojat Veisi,2 Saeedeh Ebrahimi,1,9 Sakineh Seyed-Mohammadi,2,8,* Mohammad Hashemzadeh1,2 1Infectious and Tropical Diseases Research Center, Health Research Institute, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran; 2Department of Microbiology, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran; 3Abuzar Children’s Hospital, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran; 4Alimentary Tract Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran; 5Nursing Care Research Center in Chronic Diseases, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran; 6Department of Bacteriology and Virology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran; 7Shiraz HIV/AIDS Research Center, Institute of Health, Shiraz University of Medical Sciences, Shiraz, Iran; 8Student Research Committee, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran; 9Department of Virology, Faculty of Mdicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran *These authors contributed equally to this work Introduction: Enteroaggregative Escherichia coli (EAEC) has been implicated as an emerging cause of traveler’s diarrhea, persistent diarrhea among children, and immunocompromised patients. The present study aimed to investigate the prevalence of antibiotic resistance, extended-spectrum β-lactamase (ESBL) production, and virulence factors of EAEC isolates obtained from Iranian children suffered from diarrhea. Materials and methods: In this cross-sectional study, from March 2015 to February 2016, 32 EAEC isolates were collected from fecal samples of children aged

Statement of Problem: Fixed orthodontic appliances in the oral cavity make tooth cleaning procedures more complicated. Objectives: This study aimed to compare the efficacy of computerized oral hygiene instruction with verbal technique among fixed orthodontic patients referred to the evening clinic of Orthodontics of Shiraz Dental School. Materials and Methods: A single-blind study was performed in Orthodontic Department of Shiraz, Islamic Republic of Iran, from January to May 2015 following the demonstrated exclusion and inclusion criteria. The sample size was considered 60 patients with 30 subjects in each group. Bleeding on probing and plaque indices and dental knowledge were assessed in the subjects to determine pre-intervention status. A questionnaire was designed for dental knowledge evaluation. The patients were randomly assigned into the computerized and verbal groups. Three weeks after the oral hygiene instruction, indices of bleeding on probing and plaque index and the dental knowledge were evaluated to investigate post-intervention outcome. The two groups were compared by chi-square and student t tests. The pre- and post- intervention scores in each group were compared using paired t-test. Results: In the computerized group, the mean score for plaque index and bleeding on probing index was significantly decreased while dental health knowledge was significantly increased after oral hygiene instruction, in contrast to the verbal group. Conclusions: Within the limitations of the current study, computerized oral hygiene instruction is proposed to be more effective in providing optimal oral health status compared to the conventional method in fixed orthodontic patients.

Long-period comets are planetesimal remnants constraining the environment and volatiles of the protoplanetary disc. We report the discovery of hyperbolic long-period comet C/2022 E3 Zwicky Transient Facility (ZTF), which has a perihelion ∼1.11 au, an eccentricity ≳1 and an inclination ∼109°, from images taken with the Palomar 48-inch telescope during morning twilight on 2022 March 2. Additionally, we report the characterization of C/2022 E3 (ZTF) from observations taken with the Palomar 200-inch, the Palomar 60-inch, and the NASA Infrared Telescope Facility in early 2023 February to 2023 March when the comet passed within ∼0.28 au of the Earth and reached a visible magnitude of ∼5. We measure g – r = 0.70 ± 0.01, r – i = 0.20 ± 0.01, i – z = 0.06 ± 0.01, z – J = 0.90 ± 0.01, J – H = 0.38 ± 0.01, and H – K = 0.15 ± 0.01 colours for the comet from observations. We measure the A(0°)fρ (0.8 μm) in a 6500 km radius from the nucleus of 1483 ± 40 cm, and CN, C3, and C2 production of 5.43 ± 0.11 × 1025, 2.01 ± 0.04 × 1024, and 3.08 ± 0.5 × 1025 mol s−1, similar to other long-period comets. We additionally observe the appearance of jet-like structures at a scale of ∼4000 km in wide-field g -band images, which may be caused by the presence of CN gas in the near-nucleus coma. [ABSTRACT FROM AUTHOR]

Polycystic ovary syndrome (PCOS) is associated with an increased risk of psychological distress as well as enhanced responses to psychosocial stress. Recently, it was hypothesized that PCOS patients may be at high risk of novel COVID-19 infections and worse clinical presentations during such infections. Here, we evaluated the effects of PCOS on stress responses to bacterial and viral mimetics using dihydrotestosterone-induced PCOS model rats. Lipopolysaccharide (LPS; a bacterial mimetic) or polyinosinic-polycytidylic acid (Poly-IC; a viral mimetic) was injected into PCOS model rats (PCOS) and non-PCOS rats (control), and the rats' stress responses were evaluated. In the PCOS group, the rats’ anorectic and febrile responses to LPS injection were enhanced, whereas their anorectic and febrile responses to Poly-IC injection were unaltered. The PCOS group also exhibited greater changes in peripheral cytokine levels in response to LPS, but not Poly-IC. On the contrary, after the injection of Poly-IC depressed locomotor activity was more evident in the PCOS group, whereas no such changes were observed after LPS injection. These findings indicate that although the stress responses of PCOS model rats to infection may be enhanced, the patterns of change in stress responses and their underlying mechanisms may differ between bacterial and viral infections.

Abstract Aims Sodium‐glucose co‐transporter 2 (SGLT2) inhibitors have shown potential therapeutic benefits in heart failure (HF). However, data on their real‐world usage and benefits in acute decompensated heart failure (ADHF) are limited. Methods and results We conducted a post hoc analysis of real‐world data from 1108 patients with ADHF admitted to Nihon University Itabashi Hospital (Tokyo, Japan) between 2018 and 2022. Patients were divided into two groups based on the prescription of SGLT2 inhibitors during hospitalization: an SGLT2 inhibitor group (SGLT2i group) (n = 289) and a non‐SGLT2i group (n = 819). The primary endpoints were death and rehospitalization for HF after discharge. The median age was 76 [interquartile range (IQR): 66, 83] years, and 732 patients (66%) were male. Data showed an increasing trend in the prescription of SGLT2 inhibitors since 2021. During a median follow‐up period of 366 days (IQR: 116, 614), 458 (41.3%) patients reached the primary endpoint. The Kaplan–Meier analysis showed that the SGLT2i group had a significantly lower rate of composite events than the non‐SGLT2i group, both overall (log‐rank test, P

TCRαβ+ CD4− CD8− double-negative T (DNT) cells are minor populations in peripheral blood, and their roles have mostly been discussed in inflammation and autoimmunity. However, the functions of DNT cells in tumor microenvironment remain to be elucidated. We investigated their characteristics, possible origins and functions in colorectal cancer tissues as well as their corresponding tumor-draining lymph nodes. We found a significant enrichment of DNT cells in tumor tissues compared with their corresponding lymph nodes, especially in tumors with lower T cell infiltration. T cell receptor (TCR) sequence analysis of CD4+ T, CD8+ T and DNT cells indicated that TCR sequences detected in DNT cells were found in CD8+ T cells, but rarely in CD4+ T cells, suggesting that a part of DNT cells was likely to be originated from CD8+ T cells. Through a single-cell transcriptomic analysis of DNT cells, we found that a DNT cell cluster, which showed similar phenotypes to central memory CD8+ T cells with low expression of effector and exhaustion markers, revealed some specific gene expression patterns, including higher GZMK expression. Moreover, in flow cytometry analysis, we found that DNT cells lost production of cytotoxic mediators. These findings imply that DNT cells might function as negative regulators of anti-tumor immune responses in tumor microenvironment.

How the microbiome regulates responses of systemic innate immune cells is unclear. In the present study, our purpose was to document a novel mechanism by which the microbiome mediates crosstalk with the systemic innate immune system. We have identified a family of microbiome Bacteroidota-derived lipopeptides—the serine-glycine (S/G) lipids, which are TLR2 ligands, access the systemic circulation, and regulate proinflammatory responses of splenic monocytes. To document the role of these lipids in regulating systemic immunity, we used oral gavage with an antibiotic to decrease the production of these lipids and administered exogenously purified lipids to increase the systemic level of these lipids. We found that decreasing systemic S/G lipids by decreasing microbiome Bacteroidota significantly enhanced splenic monocyte proinflammatory responses. Replenishing systemic levels of S/G lipids via exogenous administration returned splenic monocyte responses to control levels. Transcriptomic analysis demonstrated that S/G lipids regulate monocyte proinflammatory responses at the level of gene expression of a small set of upstream inhibitors of TLR and NF-κB pathways that include Trem2 and Irf4. Consistent with enhancement in proinflammatory cytokine responses, decreasing S/G lipids lowered gene expression of specific pathway inhibitors. Replenishing S/G lipids normalized gene expression of these inhibitors. In conclusion, our results suggest that microbiome-derived S/G lipids normally establish a level of buffered signaling activation necessary for well-regulated innate immune responses in systemic monocytes. By regulating gene expression of inflammatory pathway inhibitors such as Trem2, S/G lipids merit broader investigation into the potential dysfunction of other innate immune cells, such as microglia, in diseases such as Alzheimer’s disease.

Background A preoperative intervention called “prehabilitation” has become increasingly important to prevent postoperative complications. Prescribing tailor-made programs according to patient characteristics is essential to enhance the benefits of this intervention. However, data on the physical characteristics of preoperative gastrointestinal cancer patients with myopenia or myosteatosis are lacking, especially for Asian individuals. This study aimed to determine the characteristics of physical function in preoperative gastrointestinal cancer patients with skeletal muscle abnormalities. Materials and Methods This single-center cross-sectional study included patients who underwent open or laparoscopic surgery for primary colorectal or gastric cancer between October 2016 and August 2020. Skeletal muscle mass and density were measured using preoperative computed tomography images at the level of the third lumbar vertebra. Skeletal muscle abnormalities (myopenia / myosteatosis) were defined using sex- and age-specific thresholds based on previous studies. The assessment of preoperative physical function included handgrip strength, isometric knee-extensor muscle force, and 6-minute walking distance. Patients were divided into the following four groups; 1) normal group; 2) myopenia / non-myosteatosis group; 3) non-myopenia / myosteatosis group; 4) myopenia / myosteatosis group. Preoperative physical functions were compared between the normal and other groups. Results Overall, 126 patients with a median age of 69 years were included in the analysis. The prevalence of myopenia and myosteatosis in the patients was 63% and 52%, respectively. Patients with both skeletal muscle abnormalities (myopenia and myosteatosis) had poorer physical function than normal patients. Conclusions More than half of the Japanese preoperative patients with gastrointestinal cancer had either myopenia or myosteatosis, and the patients with both myopenia and myosteatosis were considered as a “high risk for severe sarcopenia” population. This information can be used to prescribe an optimal, customized treatment to preoperative patients with gastrointestinal cancer.

Previous studies have suggested that the effects of androgens on body weight (BW) and appetite are affected by the estrogen milieu in females; however, the mechanism underlying these effects remains unclear. We hypothesized that androgens may affect endogenous oxytocin (OT), which is a hypothalamic anorectic factor, and that these effects of androgens may be altered by the estrogen milieu in females. To investigate this hypothesis, in the present study, we examined the effects of testosterone on peripheral and central OT levels in ovariectomized female rats that did or did not receive estradiol supplementation. Ovariectomized female rats were randomly divided into non-estradiol-supplemented or estradiol-supplemented groups, and half of the rats in each group were concurrently supplemented with testosterone (i.e., rats were divided into four groups, n = 7 per each group). We also measured peripheral and central OT receptor (OTR) gene expression levels. As a result, we found that testosterone increased serum and hypothalamic OT levels and OT receptor mRNA levels in non-estradiol-supplemented rats, whereas it had no effects on these factors in estradiol-supplemented rats. In addition, testosterone reduced food intake, BW gain, and fat weight in non-estradiol-supplemented rats, whereas it did not have any effects on BW, appetite, or fat weight in estradiol-supplemented rats. These findings indicate that the effects of androgens on OT may be affected by the estrogen milieu, and elevated OT levels may be related to the blunting of appetite and prevention of obesity under estrogen-deficient conditions.

Feline infectious peritonitis (FIP) is a lethal disease caused by a pathogenic coronavirus, feline infectious peritonitis virus (FIPV), in cats. Effective vaccines have been unsuccessful due to the frequent mutation of FIPV and antibody-dependent enhancement (ADE) caused by vaccine-induced IgG antibodies (Abs). This study examined the induction of pan-coronavirus IgM Ab in mice and its ameliorating effects in feline FIP using CoV-mMAP8, an octavalent dendrimer composed of multiple antigenic peptides. The 11-amino acid peptide (SAIEDLLFNKV) was designed as the highly conserved region of the fusion peptide at the N-terminus of S2’ subunit of the spike protein found in human and animal coronaviruses and was then conjugated to an octavalent dendrimer to form CoV-mMAP8. After a total of three injections of CoV-mMAP8 into Balb/c mice with α-galactosylceramide (α-GC) co-administered in the second injection, serum titers of IgM Abs increased against the peptide, recombinant spike proteins of SARS-CoV-2 and MERS-CoV, and crude viral antigens of canine coronavirus, porcine endemic diarrhea virus, and FIPV. In contrast, serum titers of IgG Abs did not significantly increase against any antigens. When CoV-mMAP8 was injected into three cats experimentally infected with FIPV, hyperthermia was improved within seven days after the injection with ameliorating inflammatory markers such as the platelet-to-lymphocyte ratio and the systemic immune-inflammatory index. One cat that showed recurrent hyperthermia received an additional injection of CoV-mMAP8, and clinical improvement was observed again. Postmortem examinations confirmed chronic lesions of FIP in all the cats, providing evidence that FIPV had been successfully infected and treated with CoV-mMAP8 in all the cats. Based on the induction of pan-coronavirus IgM Abs in mice and ameliorating effects in FIP of cats, it is assumed that CoV-mMAP8 has the potential to overcome the challenges posed by variants and ADE in FIPV. The mutational compatibility of CoV-mMAP8 can make it a viable universal vaccine for various coronaviruses beyond FIPV.

Background Nonobstructive general angioscopy (NOGA) can identify vulnerable plaques in the aortic lumen that serve as potential risk factors for cardiovascular events such as embolism. However, the association between computed tomography (CT) images and vulnerable plaques detected on NOGA remains unknown. Methods and Results We investigated 101 patients (67±11 years; women, 13.8%) who underwent NOGA and contrast‐enhanced CT before or after 90 days in our hospital. On CT images, the aortic wall thickness, aortic wall area (AWA), and AWA in the vascular area were measured at the thickest point from the 6th to the 12th thoracic vertebral levels. Furthermore, the association between these measurements and the presence or absence of NOGA‐derived aortic plaque ruptures (PRs) at the same vertebral level was assessed. NOGA detected aortic PRs in the aortic lumens at 145 (22.1%) of the 656 vertebral levels. The presence of PRs was significantly associated with greater aortic wall thickness (3.3±1.7 mm versus 2.1±1.2 mm), AWA (1.33±0.68 cm2 versus 0.89±0.49 cm2), and AWA in the vascular area (23.2%±9.3% versus 17.2%±7.6%) (P

The immunogenicity of cancer cells is influenced by several factors, including the expression of the major histocompatibility complex class I (MHC-I), antigen expression, and the repertoire of proteasome-produced epitope peptides. The malignant pleural mesothelioma cell line ACC-MEOS-4 (MESO-4) expresses high levels of MHC-I and Wilms tumor 1 (WT1) tumor antigens. Using a functional T cell reporter assay specific for the HLA-A*24:02 restricted WT1 epitope (WT1235, CMTWNQMNL), we searched for factors that augmented the immunogenicity of MESO-4, focusing on proteasomes, which have a central role in the antigen processing machinery. ONX-0914, a selective inhibitor of the immunoproteasome subunit β5i, enhanced immunogenicity dose-dependently at low concentrations without cytotoxicity. In addition, CD8+ T lymphocytes recognizing WT1 showed greater cytotoxicity against MESO-4 pre-treated with ONX-0914. MESO-4 expresses a standard proteasome (SP) and immunoproteasome (IP). Notably, IP has distinct catalytic activity from SP, favoring the generation of antigenic peptides with high affinity for MHC-I in antigen-presenting cells and cancer cells. In vitro, immunoproteasome digestion assay and mass spectrometry analysis showed that IP cleaved WT1235 internally after the hydrophobic residues. Importantly, this internal cleavage of the WT1235 epitope was mitigated by ONX-0914. These results suggest that ONX-0914 prevents the internal destructive cleavage of WT1235 by IP, thereby promoting the specific presentation of the WT1 epitope by MESO-4. In conclusion, selective IP inhibitors might offer a means to modulate cancer cell immunogenicity by directing the presentation of particular tumor epitopes.

Treating ocular involvement in eosinophilic granulomatosis with polyangiitis (EGPA) can be challenging. We present the case of a 37-year-old woman with EGPA who had severe bilateral visual field defects. Laboratory results showed leukocytosis (17,500 WBC/μL, 25.8 % eosinophils), negative MPO-ANCA titer, and elevated PR3-ANCA level (33.2 IU/mL). Diffusion-weighted MRI revealed bilateral hyperintense occipital lesions, which were more prominent on the left. Her therapy initially included a steroid pulse, followed by daily prednisolone, but her visual field defects remained refractory. The addition of intravenous cyclophosphamide (5 courses) and intravenous immunoglobulin decreased her optic neuropathy and resolved her visual field defects.

Abstract Background This work aims to develop a deep learning model, assessing atlantoaxial subluxation (AAS) in rheumatoid arthritis (RA), which can often be ambiguous in clinical practice. Methods We collected 4691 X-ray images of the cervical spine of the 906 patients with RA. Among these images, 3480 were used for training the deep learning model, 803 were used for validating the model during the training process, and the remaining 408 were used for testing the performance of the trained model. The two-dimensional key points’ detection model of Deep High-Resolution Representation Learning for Human Pose Estimation was adopted as the base convolutional neural network model. The model inferred four coordinates to calculate the atlantodental interval (ADI) and space available for the spinal cord (SAC). Finally, these values were compared with those by clinicians to evaluate the performance of the model. Results Among the 408 cervical images for testing the performance, the trained model correctly identified the four coordinates in 99.5% of the dataset. The values of ADI and SAC were positively correlated among the model and two clinicians. The sensitivity of AAS diagnosis with ADI or SAC by the model was 0.86 and 0.97 respectively. The specificity of that was 0.57 and 0.5 respectively. Conclusions We present the development of a deep learning model for the evaluation of cervical lesions of patients with RA. The model was demonstrably shown to be useful for quantitative evaluation.

Abstract Estrogens play important roles in uterine growth and homeostasis through estrogen receptors (ESR1 and ESR2). To address the role of ESR1-mediated tissue events in the murine uterus, we analyzed mice with a mesenchymal tissue-specific knockout of Esr1. Isl1-driven Cre expression generated Esr1 deletion in the uterine stroma and endometrium (Isl-Esr1KO). We showed that overall structure of the Isl1-Esr1KO mouse uterus developed normally, but estrogen responsiveness and subsequent growth were defective, suggesting that mesenchymal ESR1 is necessary for both epithelial and mesenchymal cell proliferation. Furthermore, RNA-seq analysis revealed that the majority of estrogen-induced genes were regulated by stromal ESR1. In control mice, E2 administration induced 9476 up-regulated differentially expressed genes (DEGs), whereas only 1801 up-regulated DEGs were induced by E2 in Isl1-Esr1KO mice. We further showed that stromal ESR1-regulated genes in the mouse uterus included several growth factors and cytokines, which are potential factors that regulate epithelial and stromal tissue interaction, and also genes involved in lipid homeostasis. Therefore, we infer that stromal ESR1 expression is indispensable for most estrogen actions in the mouse uterus and the current results provide new insights into estrogen-mediated homeostasis in female reproductive organs.

Abstract Background New-onset anisocoria is an important clinical clue to life-threatening intracranial injury. Anisocoria alone without impairment of extraocular muscles is a rare presentation of moderate traumatic brain injury (TBI). Case presentation A 79-year-old woman was transported to hospital soon after falling off a bicycle. Glasgow Coma Scale score on arrival was 11 (E3V3M5). On examination at admission, she was found to be drowsy. Bruising was seen around the right eye and pupil diameters differed (right, 4.5 mm; left, 3.0 mm; both reactive to light). Computed tomography of the head revealed hemorrhagic contusion in the left temporal lobe and left pretectal area of the midbrain, right clavicular fracture, and pulmonary contusion with fractures of the 3rd and 4th ribs. Magnetic resonance imaging confirmed hemorrhagic contusion of the midbrain. The patient achieved full recovery of motor and mental functions with conservative treatment and was discharged on hospital day 17. Conclusion We encountered a case of anisocoria without major extraocular muscle impairment due to moderate TBI with midbrain contusion.

A method for line interference reduction to be used in signal-averaged electrocardiography (SAECG) systems is proposed and its performance is analyzed. This new method is an adaptation of a previously reported technique for removal of line interference from conventional electrocardiograms. It involves the recording of a line reference signal simultaneous with the lead signals, so that a shifted and scaled version of it can be used to subtract line interference from the leads. It is shown that this fine interference subtraction method can reduce line interference effectively and without introducing any additional noise into the ECG signal. It is also shown that Late Potential diagnostic decisions are not altered when this filter is applied. It is recommended that this technique be used in SAECG when line interference is unavoidable.

Abstract Background The mainstay of treatment for eosinophilic granulomatosis with polyangiitis (EGPA) is systemic corticosteroid therapy; some patients also receive intravenous immunoglobulins, other immunosuppressive agents, and biologics. Mepolizumab, an anti-interleukin-5 monoclonal antibody, induces remission and decreases the daily corticosteroid dose; however, the clinical efficacy of mepolizumab in EGPA and the prognosis with long-term treatment with this drug are unknown. Methods Seventy-one EGPA patients were treated at Hiratsuka City Hospital, Japan, between April 2018 and March 2022. We administered mepolizumab for a mean of 2.8 ± 1.7 years to 43 patients in whom remission could not be induced by conventional treatment. After excluding 18 patients who had received mepolizumab for less than 3 years, we classified 15 patients into a “super-responder group” (the daily dose of corticosteroids or other immunosuppressant could be decreased, or the interval between IVIG treatments could be prolonged) and 10 patients into a “responder group” (neither of these changes was achievable). Eosinophil numbers, serum IgG levels, daily doses of corticosteroids and other immunosuppressants, Birmingham Vasculitis Activity Score (BVAS), and relapse frequency before and after mepolizumab initiation were determined. Results Blood eosinophil count at diagnosis and the lowest serum IgG level before mepolizumab treatment were significantly higher in super-responders than in responders (p

Abstract Aims Acute decompensated heart failure (ADHF) presents with pulmonary congestion, which is caused by an increased pulmonary arterial wedge pressure (PAWP). PAWP is strongly associated with prognosis, but its quantitative evaluation is often difficult. Our prior work demonstrated that a deep learning approach based on chest radiographs can calculate estimated PAWP (ePAWP) in patients with cardiovascular disease. Therefore, the present study aimed to assess the prognostic value of ePAWP and compare it with other indices of haemodynamic congestion. Methods and results We conducted a post hoc analysis of a single‐centre, prospective, observational heart failure registry and analysed data from 534 patients admitted for ADHF between January 2018 and December 2019. The deep learning approach was used to calculate ePAWP from chest radiographs at admission and discharge. Patients were divided into three groups based on the ePAWP tertiles at discharge, as follows: first tertile group (ePAWP ≤ 11.2 mm Hg, n = 178), second tertile group (11.2

This qualitative case study examines the learning that occurred when a small group of middle grade youths embarked upon a personal excursion during a game-based math walk. Math walks are an informal learning activity where learners and facilitators explore mathematical concepts embedded in everyday spaces. The MathExplorer app is a location-based mobile game designed to enhance and gamify math walks. In our broader research, we investigated a group of 18 middle grade learners who used MathExplorer to engage in math walks at a local nature preserve. While most youths in this study used the game as planned by the researchers, one group deviated from the plan and devised new ways of playing the game and participating in the math walks. We see this deviation, or personal excursion, as a source of insight for research on game-based math walks. To understand the learning that took place during this personal excursion, we draw upon sociocultural and self-directed theories of learning. Using methods of interaction analysis and embodied action conversation framework, we analyzed the small groups’ discussion, movement, and game-use to understand: (1) the point at which the students departed from the planned use of MathExplorer; and (2) the learning that took place after this departure. The findings include how the youth explicitly incorporate mathematics into game play through an activity-as-planned, and how the youth embark on a personal excursion relating to game mechanics and gamification, with an implicit focus on mathematics. We discuss the importance of personal excursions for designing informal mathematics learning experiences.

BackgroundThis study aimed to examine the clinical role of non-gated computed tomography (CT) in ruling out fatal chest pain in patients with non-ST-elevation acute coronary syndrome (NSTE-ACS), with a focus on the time of arrival at the hospital to coronary angiography (CAG) and peak creatine kinase (CK) levels.MethodsWe retrospectively examined 196 NSTE-ACS patients who were admitted with urgently diagnosed NSTE-ACS and underwent percutaneous coronary intervention between March 2019 and October 2022. The patients were divided into three groups, namely, non-CT group, CT and defect− group, and CT and defect+ group, based on whether they underwent a CT scan and the presence or absence of perfusion defects on the CT image.ResultsAfter the initial admission for NSTE-ACS, 40 patients (20.4%) underwent non-gated CT prior to CAG. Among these 40 patients, 27 had a perfusion defect on the CT scan. The incidence of contrast-induced nephropathy was not different among the three groups. The CT and defect+ group had a shorter arrival-to-CAG time than that of the non-CT group. In NSTE-ACS patients with elevated CK levels, the CT and defect+ group had lower peak CK levels than those in the non-CT group.ConclusionNSTE-ACS patients with perfusion defects on non-gated CT had a shorter time from arrival to CAG, which might be associated with a lower peak CK. Non-gated CT might be useful for early diagnosis and early revascularization in the clinical setting of NSTE-ACS.

Abstract Herein, we describe a highly potent gravitropic bending inhibitor with no concomitant growth inhibition. Previously, we reported that (2Z,4E)-5-phenylpenta-2,4-dienoic acid (ku-76) selectively inhibits root gravitropic bending of lettuce radicles at 5 μM. Based on the structure–activity relationship study of ku-76 as a lead compound, we designed and synthesized various C4-substituted analogs of ku-76. Among the analogs, 4-phenylethynyl analog exhibited the highest potency for gravitropic bending inhibition, which was effective at only 0.01 μM. Remarkably, 4-phenylethynyl analog is much more potent than the known inhibitor, NPA. Substitution in the para position on the aromatic ring of 4-phenylethynyl group was tolerated without diminished activity. In addition, evaluation using Arabidopsis indicated that 4-phenylethynyl analog inhibits gravitropism by affecting auxin distribution in the root tips. Based on the effects on Arabidopsis phenotypes, 4-phenylethynyl analog may be a novel inhibitor that differs in action from the previously reported auxin transport inhibitors.

It was reported that the consumption of Saccharina japonica (SJ) lowers blood pressure (BP) in hypertensive rats. Hypertension is related to gut microbiota, and hypertensive patients develop dysbiosis. It was reported that the intake of dietary fiber and polysaccharides contained in SJ changes gut microbiota and increases short-chain fatty acids (SCFAs). The present study examined the effect of BP lowering by SJ in spontaneously hypertensive rats (SHRs) and observed changes in gut microbiota composition and SCFAs concentration. Male SHRs and Wistar Kyoto rats (WKYs) were fed a diet containing 5% SJ or a control diet for six weeks. We measured systolic BP (SBP) weekly, as well as mean arterial BP (MAP), the 16S rRNA gene, and SCFAs in the cecal contents at the end of the period. As a result, the intake of SJ significantly decreased SBP and MAP in SHRs. As well, it significantly changed the microbial diversity by altering the gut microbiota composition. Particularly, it increased the abundance of Bacteroides acidifaciens, which may be associated with the antihypertensive effect of SJ. Thus, SJ intake suppressed the increase in BP and altered the gut microbiota composition, although it did not significantly change the SCFAs concentration in the cecal contents.

Although IgG-free immunosensors are in high demand owing to ethical concerns, the development of convenient immunosensors that alternatively integrate recombinantly produced antibody fragments, such as single-chain variable fragments (scFvs), remains challenging. The low affinity of antibody fragments, unlike IgG, caused by monovalent binding to targets often leads to decreased sensitivity. We improved the affinity owing to the bivalent effect by fabricating a bivalent antibody–enzyme complex (AEC) composed of two scFvs and a single glucose dehydrogenase, and developed a rapid and convenient scFv-employed electrochemical detection system for the C-reactive protein (CRP), which is a homopentameric protein biomarker of systemic inflammation. The development of a point-of-care testing (POCT) system is highly desirable; however, no scFv-based CRP-POCT immunosensors have been developed. As expected, the bivalent AEC showed higher affinity than the single scFv and contributed to the high sensitivity of CRP detection. The electrochemical CRP detection using scFv-immobilized magnetic beads and the bivalent AEC as capture and detection antibodies, respectively, was achieved in 20 min without washing steps in human serum and the linear range was 1–10 nM with the limit of detection of 2.9 nM, which has potential to meet the criteria required for POCT application in rapidity, convenience, and hand-held detection devices without employing IgGs.

Introduction: Variants in the galactosidase alpha (GLA) gene cause Fabry disease (FD), an X-linked lysosomal storage disorder caused by α-galactosidase A (α-GAL) deficiency. Recently, disease-modifying therapies have been developed, and simple diagnostic biomarkers for FD are required to initiate these therapies in the early stages of the disease. Detection of urinary mulberry bodies and cells (MBs/MCs) is beneficial for diagnosing FD. However, few studies have evaluated the diagnostic accuracy of urinary MBs/MCs in FD. Herein, we retrospectively evaluated the diagnostic ability of urinary MBs/MCs for FD. Methods: We analyzed the medical records of 189 consecutive patients (125 males and 64 females) who underwent MBs/MCs testing. Out of these, two female patients had already been diagnosed with FD at the time of testing, and the remaining 187 patients were suspected of having FD and underwent both GLA gene sequencing and/or α-GalA enzymatic testing. Results: Genetic testing did not confirm the diagnosis in 50 females (26.5%); hence, they were excluded from the evaluation. Two patients were previously diagnosed with FD, and sixteen were newly diagnosed. Among these 18 patients, 15, including two who had already developed HCM at diagnosis, remained undiagnosed until targeted genetic screening of at-risk family members of patients with FD was performed. The accuracy of urinary MBs/MCs testing exhibited a sensitivity of 0.944, specificity of 1, positive predictive value of 1, and negative predictive value of 0.992. Conclusions: MBs/MCs testing is highly accurate in diagnosing FD and should be considered during the initial evaluation prior to genetic testing, particularly in female patients.

Abstract Aims The Model for End‐stage Liver Disease eXcluding International normalized ratio (MELD‐XI) is an established scoring system that reflects hepatorenal function. However, little is known about the prognostic value of changes in MELD‐XI score during hospitalization in acute decompensated heart failure (ADHF). Methods and results We prospectively analysed 536 patients admitted for ADHF between January 2018 and December 2019. In the MELD‐XI, 9.44 is the lowest possible score and considered to be normal, and values above 9.44 are classified as high. We calculated MELD‐XI scores at admission and discharge and used them to divide patients into four groups depending on whether the score was high (>9.44) or normal (9.44) at each time point as follows: normal score at both measurements (persistently normal group, n = 99), high score at admission and normal score at discharge (high‐to‐normal group, n = 108), normal score at admission and high score at discharge (normal‐to‐high group, n = 24), and high score at both measurements (persistently high group, n = 305). The persistently high group had higher blood urea nitrogen, creatinine, and N‐terminal pro‐brain natriuretic peptide levels at both admission and discharge and significantly higher left ventricular end‐diastolic, left atrial, right ventricular end‐diastolic, and maximal inferior vena cava diameters at discharge. During the median follow‐up period of 369 days (Q1, Q3: 97, 576), 231 (43.1%) patients reached the primary endpoint (a composite of all‐cause death or re‐hospitalization for heart failure). The Kaplan–Meier analysis revealed a significantly higher composite event rate in the persistently high group than in the persistently normal and high‐to‐normal groups (log‐rank test, P

Abstract Background Pegfilgrastim (PEG) is a sustained-duration pegylated form of filgrastim, a granulocyte-colony stimulating factor agent that is widely used as prophylaxis against febrile neutropenia during chemotherapy. We report the case of a breast cancer patient who developed PEG-induced vasculitis complicated by subarachnoid hemorrhage (SAH) and review the relevant literature. Case presentation A 48-year-old woman had undergone surgery for breast cancer and was receiving docetaxel and cyclophosphamide as adjuvant chemotherapy (docetaxel 75 mg/m2, cyclophosphamide 600 mg/m2); on day 4 of treatment, PEG had been administered. On day 14, she was admitted to hospital with fever, general malaise, and neck pain, and her C-reactive protein level was found to be high (12.65 mg/dL). Although infection was initially suspected, antimicrobial treatment was ineffective and other laboratory test results were negative for this. Contrast-enhanced computed tomography on day 22 showed thickened vessel walls in the left subclavian artery, the origin of the common carotid artery, and the thoracoabdominal aorta. On day 26, magnetic resonance imaging of the head to investigate possible causes of headache showed signs consistent with SAH, and magnetic resonance angiography images showed irregularity in the basilar artery wall; the findings of both studies were considered to be due to PEG-induced vasculitis. Once treatment with prednisolone 40 mg/day had started, the wall thickening and irregularity improved. Conclusion Although an uncommon adverse effect, vasculitis affecting vessels of various sizes may be caused by PEG. To the best of our knowledge, this report is the first to describe a case of G-CSF-induced vasculitis complicated by SAH. In cases of persistent high fever and elevated inflammatory response after PEG administration and in the absence of infection, clinicians should consider the possibility of drug-induced vasculitis.

Informal science education researchers have become increasingly interested in how out-of-school spaces that offer STEM (science, technology, engineering, and math) programs inform learners’ STEM achievement, interests, and affective outcomes. Studies have found that these spaces can offer critical learning and developmental opportunities for underrepresented racially minoritized (URM) students (Black, Latinx, low socioeconomic status) in STEM subjects. Shifting away from the leaky STEM pipeline analogy, researchers have posited contemporary understandings to explain why the minoritization of URM girls persists. Informal learning environments such as STEM summer camps are being studied to assess how URM girls experience and interact with STEM in novel ways. These environments can inform the research field about how URM girls’ perceptions of their STEM identities, abilities, efficacy, and belonging in STEM develop as they engage in those spaces. This mixed-method study used a multiple-case-study approach to examine how aspects of URM middle school girls’ STEM identities positively changed after participating in a one-week, sleep-away, single-gender STEM summer camp held at a university in the Southwestern U.S. Drawing on intersectionality and STEM identity, we used ecological systems theory to design our research study, examining how URM middle school girls narrate their STEM identities in this informal learning environment. Using quantitative analyses and deductive coding methods, we explored how elements of girls’ STEM identities were shaped during and after their participation in the STEM summer camp. Findings from our study highlight (1) quantitative changes in girl participants’ STEM identities, sense of belonging in STEM, and perceived STEM ability belief, (2) qualitative results supporting our quantitative findings, and (3) how the intersectionality of participants’ race and gender played a role in their STEM identities. This study points to the potential of STEM informal learning camps as a way of developing and fostering URM girls’ STEM identities.

In recent years, the effects of androgens on metabolic and body weight regulation systems and their underlying mechanisms have been gradually revealed in females. In women and experimental animals of reproductive age, androgen excess can adversely affect metabolic functioning, appetite, and body weight regulation. In addition, excess androgens can increase the risk of metabolic disorders, such as obesity, insulin resistance, and diabetes. These unfavorable effects of androgens are induced by alterations in the actions of hypothalamic appetite-regulatory factors, reductions in energy expenditure, insulin resistance in skeletal muscle, and β-cell dysfunction. Interestingly, these unfavorable effects of androgens on metabolic and body-weight regulation systems are neither observed nor evident in ovariectomized animals and post-menopausal women, indicating that the adverse effects of androgens might be dependent on the estrogen milieu. Recent findings may provide novel sex- and age-specific strategies for treating metabolic diseases.

Abstract The influence of the so‐called “Airbnb effect” on local housing markets has grown into a global concern for affordability crisis, gentrification, and displacement. We analyzed the impacts of short‐term rental (STR) on the rental housing market and explored the relative factors for the rent gaps in Tokyo and Osaka: two major metropolitan areas in Japan. This research builds upon the rent gap theory by Neil Smith and clarifies the rent gaps by following previous work. Our results demonstrate that landlords in Osaka, more depopulated with unoccupied housings, have already removed a significant number of residential rental units from the housing market, and those of Tokyo, less depopulated with less unoccupied housings, is likely to evict their tenants to shift more profitable touristic short‐term rentals when their long‐term rentals’ rent revenue decreases. We concluded that the rent gaps affected by the depopulated societal context require tougher limits on touristic short‐term rentals to mitigate “transnational displacement.” The future of community livability depends upon municipalities’ ordinance as well as their strategies: how to leverage unoccupied housing to survive an era of shrinking economy.