High-throughput screening of small-molecules libraries identified antibacterials against clinically relevant multidrug-resistant A. baumannii and K. pneumoniae.

BACKGROUND: The current pipeline for new antibiotics fails to fully address the significant threat posed by drug-resistant Gram-negative bacteria that have been identified by the World Health Organization (WHO) as a global health priority. New antibacterials acting through novel mechanisms of action are urgently needed. We aimed to identify new chemical entities (NCEs) with activity against Klebsiella pneumoniae and Acinetobacter baumannii that could be developed into a new treatment for drug-resistant infections. METHODS: We developed a high-throughput phenotypic screen and selection cascade for generation of hit compounds active against multidrug-resistant (MDR) strains of K. pneumoniae and A. baumannii. We screened compound libraries selected from the proprietary collections of three pharmaceutical companies that had exited antibacterial drug discovery but continued to accumulate new compounds to their collection. Compounds from two out of three libraries were selected using "eNTRy rules" criteria associated with increased likelihood of intracellular accumulation in Escherichia coli. FINDINGS: We identified 72 compounds with confirmed activity against K. pneumoniae and/or drug-resistant A. baumannii. Two new chemical series with activity against XDR A. baumannii were identified meeting our criteria of potency (EC50 ≤50 µM) and absence of cytotoxicity (HepG2 CC50 ≥100 µM and red blood cell lysis HC50 ≥100 µM). The activity of close analogues of the two chemical series was also determined against A. baumannii clinical isolates. INTERPRETATION: This work provides proof of principle for the screening strategy developed to identify NCEs with antibacterial activity against multidrug-resistant critical priority pathogens such as K. pneumoniae and A. baumannii. The screening and hit selection cascade established here provide an excellent foundation for further screening of new compound libraries to identify high quality starting points for new antibacterial lead generation projects. FUNDING: BMBF and GARDP.

An experimental verification of fungal overgrowth in temporary houses at the site of the Great East Japan Earthquake.

Fungal contamination in the indoor air of prefabricated temporary houses at the site of the Great East Japan Earthquake revealed extremely high levels compared to those found in conventional residences. We experimentally investigated fungal growth levels on different interior materials to support fungal overgrowth in prefabricated temporary houses. Three species each of allergenic fungi and invasive fungi observed in temporary housing were selected for inoculation tests with various interior materials. The experiments with fungal inoculation were conducted in conformance with standards for industrial products described in the Japanese" JIS Z 2911:2018 Methods of test for fungus resistance" with small modifications. After incubation, visual and stereomicroscopic assessments were performed to determine fungal growth levels. The viability of the fungi varied according to the interior material type. Our findings demonstrate the importance of antifungal measures in indoor environments and the need for additional research on the growth levels of fungal species on various interior materials.

Development of progressive multifocal leukoencephalopathy after cord blood transplantation in a patient with refractory angioimmunoblastic T-cell lymphoma.

A patient undergoing cord blood transplantation for refractory angioimmunoblastic T-cell lymphoma was subsequently managed with long-term immunosuppressants for chronic graft-versus-host disease (GVHD). On day 591 post-transplant, she exhibited disorientation and cognitive dysfunction. Magnetic resonance imaging (MRI) of the brain revealed two hyperintense foci in the white matter, suggestive of progressive multifocal leukoencephalopathy (PML). However, we did not include PML in the differential diagnosis at that time. Unfortunately, she developed progressive cognitive impairment, and repeated brain MRIs showed a progression in lesion size. She was still taking immunosuppressants to control her GVHD, therefore we suspected PML. The diagnosis of PML was confirmed through the detection of a John Cunningham (JC) virus in the cerebrospinal fluid on day 640 post-transplant. This report highlights the critical need to consider PML in differential diagnoses for post-allogeneic transplant patients, especially those who exhibit progressive neurological symptoms while on prolonged immunosuppressant therapy.

Imeglimin improves systemic metabolism by targeting brown adipose tissue and gut microbiota in obese model mice.

Imeglimin is a recently developed anti-diabetic drug that could concurrently promote insulin secretion and insulin sensitivity, while its mechanisms of action are not fully understood. Here we show that imeglimin administration could protect mice from high fat diet-induced weight gain with enhanced energy expenditure and attenuated whitening of brown adipose tissue. Imeglimin administration led to significant alteration of gut microbiota, which included an increase of Akkermansia genus, with attenuation of obesity-associated gut pathologies. Ablation of microbiota by antibiotic treatment partially abrogated the insulin sensitizing effects of imeglimin, while not affecting its actions on body weight gain or brown adipose tissue. Collectively, our results characterize imeglimin as a potential agent promoting energy expenditure and gut integrity, providing new insights into its mechanisms of action.

Pentafluorosulfanyl-substituted biaryl derivatives as MATE-type transporter inhibitors targeting drug-resistant bacteria.

Multidrug and toxin extrusion (MATE) inhibitors improve the antimicrobial susceptibility of drug-resistant bacteria by preventing the efflux of administered antibiotics. In this study, we optimized the chemical structure of a previously identified bacterial-selective MATE inhibitor 1 (EC50 > 30 µM) to improve its activity further. Compound 1 was divided into three fragments (aromatic part, linker part, and guanidine part), and each part was individually optimized. Compound 31 (EC50 = 1.8 µM), a novel pentafluorosulfanyl-containing molecule synthesized following optimized parts, showed antimicrobial activity against MATE-expressing strains at concentrations lower than conventional inhibitor 1 when co-administrated with norfloxacin. Furthermore, 31 was not cytotoxic at effective concentrations. This suggests that compound 31 can be a promising candidate for combating bacterial infections, particularly those resistant to conventional antibiotics by MATE expression.

The Feasibility of Using a Deep Learning-Based Model to Determine Cardiac Computed Tomographic Contrast Dose.

PURPOSE: This study aimed to predict contrast effects in cardiac computed tomography (CT) from CT localizer radiographs using a deep learning (DL) model and to compare the prediction performance of the DL model with that of conventional models based on patients' physical size. METHODS: This retrospective study included 473 (256 men and 217 women) cardiac CT scans between May 2014 and August 2017. We developed and evaluated DL models that predict milligrams of iodine per enhancement of the aorta from CT localizer radiographs. To assess the model performance, we calculated and compared Pearson correlation coefficient ( r ) between the actual iodine dose that was necessary to obtain a contrast effect of 1 HU (iodine dose per contrast effect [IDCE]) and IDCE predicted by DL, body weight, lean body weight, and body surface area of patients. RESULTS: The model was tested on 52 cases for the male group (mean [SD] age, 63.7 ± 11.4) and 44 cases for the female group (mean [SD] age, 69.8 ± 11.6). Correlation coefficients between the actual and predicted IDCE were 0.607 for the male group and 0.412 for the female group, which were higher than the correlation coefficients between the actual IDCE and body weight (0.539 for male, 0.290 for female), lean body weight (0.563 for male, 0.352 for female), and body surface area (0.587 for male, 0.349 for female). CONCLUSIONS: The performance for predicting contrast effects by analyzing CT localizer radiographs with the DL model was at least comparable with conventional methods using the patient's body size, notwithstanding that no additional measurements other than CT localizer radiographs were required.

Preliminary assessment of automated radiology report generation with generative pre-trained transformers: comparing results to radiologist-generated reports.

PURPOSE: In this preliminary study, we aimed to evaluate the potential of the generative pre-trained transformer (GPT) series for generating radiology reports from concise imaging findings and compare its performance with radiologist-generated reports. METHODS: This retrospective study involved 28 patients who underwent computed tomography (CT) scans and had a diagnosed disease with typical imaging findings. Radiology reports were generated using GPT-2, GPT-3.5, and GPT-4 based on the patient's age, gender, disease site, and imaging findings. We calculated the top-1, top-5 accuracy, and mean average precision (MAP) of differential diagnoses for GPT-2, GPT-3.5, GPT-4, and radiologists. Two board-certified radiologists evaluated the grammar and readability, image findings, impression, differential diagnosis, and overall quality of all reports using a 4-point scale. RESULTS: Top-1 and Top-5 accuracies for the different diagnoses were highest for radiologists, followed by GPT-4, GPT-3.5, and GPT-2, in that order (Top-1: 1.00, 0.54, 0.54, and 0.21, respectively; Top-5: 1.00, 0.96, 0.89, and 0.54, respectively). There were no significant differences in qualitative scores about grammar and readability, image findings, and overall quality between radiologists and GPT-3.5 or GPT-4 (p > 0.05). However, qualitative scores of the GPT series in impression and differential diagnosis scores were significantly lower than those of radiologists (p < 0.05). CONCLUSIONS: Our preliminary study suggests that GPT-3.5 and GPT-4 have the possibility to generate radiology reports with high readability and reasonable image findings from very short keywords; however, concerns persist regarding the accuracy of impressions and differential diagnoses, thereby requiring verification by radiologists.

Donor Lymphocyte Infusion for Relapsed Acute Leukemia or Myelodysplastic Syndrome after Hematopoietic Stem Cell Transplantation: A Single-Institute Retrospective Analysis.

Objective The prognosis of the patients who relapsed after allogeneic hematopoietic stem cell transplantation (allo-HSCT) is poor, and therapeutic options are limited. In the present study, we investigated the efficacy and factors associated with the survival in patients with acute leukemia or myelodysplastic syndrome (MDS) who relapsed following allo-HSCT and were treated with donor lymphocyte infusion (DLI) in real-world practice. Patients Twenty-nine patients with acute myeloid leukemia21, acute lymphoid leukemia4 or MDS4 were enrolled. Eleven patients were diagnosed with hematological relapse, and 18 were diagnosed with molecular or cytogenetic relapse. Results The median injection number and median total number of infused CD3+ T cells were 2 and 5.0×107/kg, respectively. The cumulative incidence of acute graft-versus-host disease (aGVHD) of grade ≥II at 4 months after the initiation of DLI was 31.0%. Extensive chronic graft-versus-host disease (cGVHD) occurred in 3 (10.3%) patients. The overall response rate was 51.7%, including 3 cases of hematological complete remission (CR) and 12 cases of molecular/cytogenetic CR. Cumulative relapse rates at 24 and 60 months following DLI in patients who achieved CR were 21.4% and 30.0%, respectively. The overall survival rates at 1, 2 and 3 years after DLI were 41.4%, 37.9% and 30.3%, respectively. Molecular/cytogenetic relapse, a longer interval from HSCT to relapse, and concomitant chemotherapy with 5-azacytidine (Aza) were significantly associated with a relatively long survival following DLI. Conclusion These results indicated that DLI was beneficial for patients with acute leukemia or MDS who relapsed after allo-HSCT and suggested that DLI in combination with Aza for molecular or cytogenetic relapse might result in favorable outcomes.

Impact of Visually Induced Motion Sickness from VR Depending on Viewing Patterns, View Movement, and Background Motion.

The impact of visually induced motion sickness from virtual reality (VR) because of viewing patterns, view movements, and background global motion was investigated experimentally through classification into four categories.Each of the ten subjects underwent watching four patterns with bio-signal measurements, such as electrocardiogram and respiration, answering a subjective questionnaire.The results of the subjective evaluation of biological effects indicate that the VR viewing pattern has a significantly lower influence on visually induced motion sickness than other patterns, such as viewing with global background motion or sight motion by operating the controller.Clinical Relevance- This study aimed to reduce the effects of visually induced motion sickness in VR viewing, particularly to reduce the burden on the eyes and body.

Gut insulin action protects from hepatocarcinogenesis in diabetic mice comorbid with nonalcoholic steatohepatitis.

Diabetes is known to increase the risk of nonalcoholic steatohepatitis (NASH) and hepatocellular carcinoma (HCC). Here we treat male STAM (STelic Animal Model) mice, which develop diabetes, NASH and HCC associated with dysbiosis upon low-dose streptozotocin and high-fat diet (HFD), with insulin or phlorizin. Although both treatments ameliorate hyperglycemia and NASH, insulin treatment alone lead to suppression of HCC accompanied by improvement of dysbiosis and restoration of antimicrobial peptide production. There are some similarities in changes of microflora from insulin-treated patients comorbid with diabetes and NASH. Insulin treatment, however, fails to suppress HCC in the male STAM mice lacking insulin receptor specifically in intestinal epithelial cells (ieIRKO), which show dysbiosis and impaired gut barrier function. Furthermore, male ieIRKO mice are prone to develop HCC merely on HFD. These data suggest that impaired gut insulin signaling increases the risk of HCC, which can be countered by restoration of insulin action in diabetes.

[Efficacy of Bright Greenish-Yellow Fluorescence Sorting on Mycotoxin-Contaminated Nutmeg Selection and Characterization of Fungal Flora Related to Mycotoxin Production].

Spices have been known to be highly contaminated commodities with mycotoxins. The Codex Alimentarius reports that nutmeg is particularly contaminated with aflatoxins (AFs) and ochratoxin A (OTA). To eliminate contaminated commodities, visual sorting and bright greenish-yellow fluorescence (BGYF) sorting are used as low-cost technologies in production engineering. In Indonesia, nutmeg is mainly sorted by visual sorting and classified into three grades according to the Indonesian national standards, with importers further defining their own brand as imported products. In this study, we evaluate the efficacy of BGYF sorting as a further selection method to reduce AFs and OTA using the importer's own brand. Further, the level of these mycotoxins and the relationship between fungal flora and mycotoxin contamination were examined. These results showed that BGYF sorting effectively reduces AFs as well as OTA. In addition, BGYF-positive groups were infected by Aspergillus sections Flavi, Nigri, and Circumdati.

The utility of automatic segmentation of kidney MRI in chronic kidney disease using a 3D convolutional neural network.

We developed a 3D convolutional neural network (CNN)-based automatic kidney segmentation method for patients with chronic kidney disease (CKD) using MRI Dixon-based T1-weighted in-phase (IP)/opposed-phase (OP)/water-only (WO) images. The dataset comprised 100 participants with renal dysfunction (RD; eGFR < 45 mL/min/1.73 m2) and 70 without (non-RD; eGFR ≥ 45 mL/min/1.73 m2). The model was applied to the right, left, and both kidneys; it was first evaluated on the non-RD group data and subsequently on the combined data of the RD and non-RD groups. For bilateral kidney segmentation of the non-RD group, the best performance was obtained when using IP image, with a Dice score of 0.902 ± 0.034, average surface distance of 1.46 ± 0.75 mm, and a difference of - 27 ± 21 mL between ground-truth and automatically computed volume. Slightly worse results were obtained for the combined data of the RD and non-RD groups and for unilateral kidney segmentation, particularly when segmenting the right kidney from the OP images. Our 3D CNN-assisted automatic segmentation tools can be utilized in future studies on total kidney volume measurements and various image analyses of a large number of patients with CKD.

A case of Hodgkin lymphoma-type Richter syndrome presenting as small-intestinal perforation.

Hodgkin lymphoma type of Richter syndrome (HL-type RS) is a rare disease that arises in patients with chronic lymphocytic leukemia (CLL). HL-type RS lesions can manifest in various sites and are often accompanied by related symptoms. This is the first case report to describe diagnosis of HL-type RS after emergency surgery for gastrointestinal perforation caused by the development of a HL-type RS lesion. A 47-year-old man diagnosed with CLL three years prior began treatment with ibrutinib due to worsening anemia and splenomegaly two months prior to the emergency department presentation. Although splenomegaly improved, lymphocytopenia, anemia, and a newly arising mesenteric lymphadenopathy continued to worsen. He presented to the emergency department with abdominal pain, and subsequent surgery revealed small intestinal perforation and mesenteric lymphadenopathy with HL-type RS confirmed by histopathological examination of the resected small intestine. He subsequently received brentuximab vedotin, doxorubicin, vinblastine, and dacarbazine (A + AVD), which effectively managed the HL-type RS. If CLL clinical presentation deviates from the typical course, an early tissue biopsy should be considered to evaluate for HL-type RS. Given the adoption of the A + AVD regimen as the standard treatment for Hodgkin lymphoma, further research is needed to evaluate its efficacy in HL-type RS.

Generative adversarial network based digital stain conversion for generating RGB EVG stained image from hyperspectral H&E stained image.

Significance: Quantification of elastic fiber in the tissue specimen is an important aspect of diagnosing different diseases. Though hematoxylin and eosin (H&E) staining is a routinely used and less expensive tissue staining technique, elastic and collagen fibers cannot be differentiated using it. So, in conventional pathology, special staining technique, such as Verhoeff's van Gieson (EVG), is applied physically for this purpose. However, the procedure of EVG staining is very expensive and time-consuming. Aim: The goal of our study is to propose a deep-learning-based computerized method for the generation of RGB EVG stained tissue from hyperspectral H&E stained one to save the time and cost of conventional EVG staining procedure. Approach: H&E stained hyperspectral image and EVG stained RGB whole slide image of human pancreatic tissue have been leveraged for this experiment. CycleGAN-based deep learning model has been proposed for digital stain conversion while images from source and target domains are of different modalities (hyperspectral and RGB) with different channel dimensions. A set of three basis functions have been introduced for calculating one of the losses of the proposed method, which retains the relevant features of EVG stained image within the reduced channel dimension of the H&E stained one. Results: The experimental results showed that a set of three basis functions including linear discriminant function and transmittance spectrum of eosin and hematoxylin better retained the essential properties of the elastic fiber to be discriminated from collagen fiber within the reduced dimension of the hyperspectral H&E stained image. Also, only a smaller number of paired training data with our proposed training method contributed significantly to the generation of more realistic EVG stained image with more precise identification of elastic fiber. Conclusions: RGB EVG stained image is generated from hyperspectral H&E stained image for which our model has performed two types of image conversion simultaneously: hyperspectral to RGB and H&E to EVG. The experimental results show that the intentionally designed set of three basis functions contains more relevant information and prove the effectiveness of our proposed method in generating realistic RGB EVG stained image from hyperspectral H&E stained one.

Determination of the biological origin of enzyme preparations using SDS-PAGE and peptide mass fingerprinting.

Enzymes are mainly extracted from the culture broth of microorganisms. Various commercially available enzyme preparations (EPs) are derived from different microorganisms, and the source of the EP should be the same as that mentioned in the manufacture's information. The development of analytical methods that can determine the origin of the final products is important for ensuring that the EPs are nontoxic, especially when used as food additives. In this study, various EPs were subjected to SDS-PAGE, and the main protein bands were excised. After in-gel digestion, the generated peptides were analysed using MALDI-TOF MS, and protein identification was performed by searching the set of peptide masses against protein databases. In total, 36 EPs including amylase, ß-galactosidase, cellulase, hemicellulase and protease were analysed, and the information about the enzyme sources was obtained for 30 EPs. Among these, the biological sources determined for 25 EPs were consistent with the manufacturer's information; for the remaining five, enzymes produced by closely-related species were shown as matching proteins due to high sequence similarity. Six enzymes derived from four microorganisms could not be identified because their protein sequences were not registered in the database. As these databases are expanded, this approach of using SDS-PAGE and peptide mass fingerprinting (PMF) can determine the biological origin of enzymes rapidly and contribute to ensuring the safety of EPs.

Long-term follow-up after R-High CHOP/CHASER/LEED with Auto-PBSCT in untreated mantle cell lymphoma-Final analysis of JCOG0406.

Progression-free survival after R-High CHOP/CHASER/LEED with auto-PBSCT in untreated mantle cell lymphoma in JCOG0406 study. A continuous pattern of relapse was observed.

Deep learning-based reconstruction and 3D hybrid profile order technique for MRCP at 3T: evaluation of image quality and acquisition time.

OBJECTIVES: To evaluate the image quality of the 3D hybrid profile order technique and deep-learning-based reconstruction (DLR) for 3D magnetic resonance cholangiopancreatography (MRCP) within a single breath-hold (BH) at 3 T magnetic resonance imaging (MRI). METHODS: This retrospective study included 32 patients with biliary and pancreatic disorders. BH images were reconstructed with and without DLR. The signal-to-noise ratio (SNR), contrast, contrast-to-noise ratio (CNR) between the common bile duct (CBD) and periductal tissues, and full width at half maximum (FWHM) of CBD on 3D-MRCP were evaluated quantitatively. Two radiologists scored image noise, contrast, artifacts, blur, and overall image quality of the three image types using a 4-point scale. Quantitative and qualitative scores were compared using the Friedman test and post hoc Nemenyi test. RESULTS: The SNR and CNR were not significantly different when under respiratory gating- and BH-MRCP without DLR. However, they were significantly higher under BH with DLR than under respiratory gating (SNR, p = 0.013; CNR, p = 0.027). The contrast and FWHM of MRCP under BH with and without DLR were lower than those under respiratory gating (contrast, p < 0.001; FWHM, p = 0.015). Qualitative scores for noise, blur, and overall image quality were higher under BH with DLR than those under respiratory gating (blur, p = 0.003; overall, p = 0.008). CONCLUSIONS: The combination of the 3D hybrid profile order technique and DLR is useful for MRCP within a single BH and does not lead to the deterioration of image quality and space resolution at 3 T MRI. CLINICAL RELEVANCE STATEMENT: Considering its advantages, this sequence might become the standard protocol for MRCP in clinical practice, at least at 3.0 T. KEY POINTS: • The 3D hybrid profile order can achieve MRCP within a single breath-hold without a decrease in spatial resolution. • The DLR significantly improved the CNR and SNR of BH-MRCP. • The 3D hybrid profile order technique with DLR reduces the deterioration of image quality in MRCP within a single breath-hold.

Activation of AMP-activated protein kinase (AMPK) through inhibiting interaction with prohibitins.

5'-Adenosine monophosphate-activated protein kinase (AMPK) is a potential therapeutic target for various medical conditions. We here identify a small-molecule compound (RX-375) that activates AMPK and inhibits fatty acid synthesis in cultured human hepatocytes. RX-375 does not bind to AMPK but interacts with prohibitins (PHB1 and PHB2), which were found to form a complex with AMPK. RX-375 induced dissociation of this complex, and PHBs knockdown resulted in AMPK activation, in the cultured cells. Administration of RX-375 to obese mice activated AMPK and ameliorated steatosis in the liver. High-throughput screening based on disruption of the AMPK-PHB interaction identified a second small-molecule compound that activates AMPK, confirming the importance of this interaction in the regulation of AMPK. Our results thus indicate that PHBs are previously unrecognized negative regulators of AMPK, and that compounds that prevent the AMPK-PHB interaction constitute a class of AMPK activator.