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Fatty acid binding protein 4 (FABP4) is a secreted adipokine linked to obesity and progression of a variety of cancers. Obesity increases extracellular FABP4 (eFABP4) levels in animal models and in obese breast cancer patients compared with lean healthy controls. Using MCF-7 and T47D breast cancer epithelial cells, we show herein that eFABP4 stimulates cellular proliferation in a time and concentration dependent manner while the non-fatty acid-binding mutant, R126Q, failed to potentiate growth. When E0771 murine breast cancer cells were injected into mice, FABP4 null animals exhibited delayed tumor growth and enhanced survival compared with injections into control C57Bl/6J animals. eFABP4 treatment of MCF-7 cells resulted in a significant increase in phosphorylation of extracellular signal-regulated kinase 1/2 (pERK), transcriptional activation of nuclear factor E2-related factor 2 (NRF2) and corresponding gene targets ALDH1A1, CYP1A1, HMOX1, SOD1 and decreased oxidative stress, while R126Q treatment did not show any effects. Proximity-labeling employing an APEX2–FABP4 fusion protein revealed several proteins functioning in desmosomes as eFABP4 receptor candidates including desmoglein (DSG), desmocollin, junction plankoglobin, desomoplankin, and cytokeratins. AlphaFold modeling predicted an interaction between eFABP4, and the extracellular cadherin repeats of DSG2 and pull-down and immunoprecipitation assays confirmed complex formation that was potentiated by oleic acid. Silencing of DSG2 in MCF-7 cells attenuated eFABP4 effects on cellular proliferation, pERK levels, and ALDH1A1 expression compared with controls. Implications: These results suggest desmosomal proteins, and in particular desmoglein 2, may function as receptors of eFABP4 and provide new insight into the development and progression of obesity-associated cancers.

S1. DSC and DSG isoforms expressed in MCF-7 cells.

Fatty acid binding protein 4 (FABP4) is a secreted adipokine linked to obesity and progression of a variety of cancers. Obesity increases extracellular FABP4 (eFABP4) levels in animal models and in obese breast cancer patients compared with lean healthy controls. Using MCF-7 and T47D breast cancer epithelial cells, we show herein that eFABP4 stimulates cellular proliferation in a time and concentration dependent manner while the non-fatty acid-binding mutant, R126Q, failed to potentiate growth. When E0771 murine breast cancer cells were injected into mice, FABP4 null animals exhibited delayed tumor growth and enhanced survival compared with injections into control C57Bl/6J animals. eFABP4 treatment of MCF-7 cells resulted in a significant increase in phosphorylation of extracellular signal-regulated kinase 1/2 (pERK), transcriptional activation of nuclear factor E2-related factor 2 (NRF2) and corresponding gene targets ALDH1A1, CYP1A1, HMOX1, SOD1 and decreased oxidative stress, while R126Q treatment did not show any effects. Proximity-labeling employing an APEX2–FABP4 fusion protein revealed several proteins functioning in desmosomes as eFABP4 receptor candidates including desmoglein (DSG), desmocollin, junction plankoglobin, desomoplankin, and cytokeratins. AlphaFold modeling predicted an interaction between eFABP4, and the extracellular cadherin repeats of DSG2 and pull-down and immunoprecipitation assays confirmed complex formation that was potentiated by oleic acid. Silencing of DSG2 in MCF-7 cells attenuated eFABP4 effects on cellular proliferation, pERK levels, and ALDH1A1 expression compared with controls.Implications:These results suggest desmosomal proteins, and in particular desmoglein 2, may function as receptors of eFABP4 and provide new insight into the development and progression of obesity-associated cancers.

S2. Predicted orientation of the F57 side chain in the apoFABP4 form.

Acetaminophen (APAP)-induced liver injury (AILI) is the most common cause of acute liver failure. Although the mechanisms that trigger AILI are well known, it is less understood how to halt AILI progression and facilitate liver recovery. Therefore, it is necessary to understand the pathophysiology of APAP hepatotoxicity in patients and to examine predictive/preventive markers. In a clinical study, we had a case in which aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels increased in a patient with a low ratio of APAP glucuronide concentration (AP-G)/APAP plasma concentration. Then a reverse translational study was conducted for clarifying this clinical question. The relationship between plasma AP-G/APAP concentration ratio and the levels of AST and ALT was examined by in vivo and in vitro experiments. In in vivo experiments, 10-week-old rats showed lower UGT activity, lower AP-G/APAP concentration ratios, and higher AST and ALT levels than 5-week-old rats. This suggests an inverse correlation between the AP-G/APAP concentration ratio and the AST, ALT levels in APAP-treated rats. Furthermore, as a result of the in vitro experiment, it was confirmed that the cell viability decreased when the AP-G/APAP concentration ratio in the culture medium decreased. Since the decrease in the plasma AP-G/APAP concentration ratio appears earlier than the increase of AST and ALT levels, the ratio might be a presymptomatic marker of AILI. When APAP is used for a long time, it is recommended to perform therapeutic drug monitoring of the AP-G/APAP concentration ratio, which is a predictive/preventive marker of AILI.

We have investigated the morphology of dipalmitoyl phosphatidyl choline (DPPC)-dimyristoyl PC (DMPC) mixed monolayer formed on the water surface by the dropping method using surface tension measurement (STm), Brewster angle microscopy (BAM), and infrared external reflection spectroscopy (IERS). The limiting molecular area (A0) obtained by STm showed that the negative deviation at < mole fraction (xDMPC)= 0.4 whereas the positive deviation at > xDMPC =0.6 when DMPC was added to the DPPC monolayer. BAM images also showed that the condensed rigid monolayer at xDMPC < 0.4 changed to the expandable monolayer at xDMPC > 0.5. Moreover, IER spectra showed that the alkyl chains of both DPPC and DMPC molecules are in trans conformation in mixed monolayer at xDMPC =0.2 whereas gauche conformation existed at xDMPC = 0.8. Above findings have justified the transition of flexible gauche conformation alkyl chains of DMPC molecule into rigid trans conformation by the addition of DPPC molecule at xDMPC < 0.4 whereas alkyl chains retained flexible gauche conformation in both DPPC and DMPC molecules at xDMPC > 0.6.

Supplementary Methods and Materials, Figure Legends 1-4 from Combined Transductional Untargeting/Retargeting and Transcriptional Restriction Enhances Adenovirus Gene Targeting and Therapy for Hepatic Colorectal Cancer Tumors

Supplementary Figure 1 from Combined Transductional Untargeting/Retargeting and Transcriptional Restriction Enhances Adenovirus Gene Targeting and Therapy for Hepatic Colorectal Cancer Tumors

Supplementary Figure 3 from Combined Transductional Untargeting/Retargeting and Transcriptional Restriction Enhances Adenovirus Gene Targeting and Therapy for Hepatic Colorectal Cancer Tumors

Supplementary Figure 2 from Combined Transductional Untargeting/Retargeting and Transcriptional Restriction Enhances Adenovirus Gene Targeting and Therapy for Hepatic Colorectal Cancer Tumors

Supplementary Figure 4 from Combined Transductional Untargeting/Retargeting and Transcriptional Restriction Enhances Adenovirus Gene Targeting and Therapy for Hepatic Colorectal Cancer Tumors

In this study, eight types of bacteria were cultivated, including Staphylococcus aureus. The infrared absorption spectra of the gas surrounding cultured bacteria were recorded at a resolution of 0.5 cm−1 over the wavenumber range of 7500–500 cm−1. From these spectra, we searched for the infrared wavenumbers at which characteristic absorptions of the gas surrounding Staphylococcus aureus could be measured. This paper reports two wavenumber regions, 6516–6506 cm−1 and 2166–2158 cm−1. A decision tree–based machine learning algorithm was used to search for these wavenumber regions. The peak intensity or the absorbance difference was calculated for each region, and the ratio between them was obtained. When these ratios were used as training data, decision trees were created to classify the gas surrounding Staphylococcus aureus and the gas surrounding other bacteria into different groups. These decision trees show the potential effectiveness of using absorbance measurement at two wavenumber regions in finding Staphylococcus aureus. Graphical abstract

The electron spin resonance (ESR) spectroscopy technique was used to study various organic radicals, such as 2,2,6,6-tetramethyl-1-piperidinyloxyl (TEMPO), 4-hydroxy-TEMPO (TEMPOL), 2-X-nitronylnitroxide (2-X-NN, X = Ph, NO2Ph, or cyclohexyl), 4-Y-benzonitronylnitroxide (4-Y-PhBzNN, Y = Ph or NO2Ph), and 2-Z-iminonitroxide (2-Z-IN, Z = Ph or NO2Ph) dispersed in a polymethylmethacrylate (PMMA) matrix. The experiments were conducted at room temperature. The complex nature of the recorded ESR spectra could be attributed to the superposition of the rotational diffusion component of TEMPO (or TEMPOL) in the nanospace of the PMMA matrix with the rigid-limit component. A single component of the rigid-limit was observed for 2-X-NN and 4-Y-PhBzNN radicals dispersed in the PMMA matrix. The isotropic components of g and hyperfine ( A ) tensor, estimated by analyzing the solution spectra, were used to determine the g and A components of 4-Y-PhBzNN. Only the rotational diffusion component was observed for the 2-Z-IN radical. These results demonstrated that the PMMA matrix contains cylindrical nanospaces. Various radicals other than TEMPO derivatives could be used in the ESR spin probe technique as probe molecules for determining the structures, sizes, and shapes of the nanospaces.

Esophageal adenocarcinoma (EAC) has increased in incidence in Western countries, and its poor prognosis necessitates the development of novel therapeutics. We previously reported the potential of conditionally replicative adenoviruses (CRAd) as a novel therapeutic treatment for this disease. To further augment the therapeutic effectiveness of our cyclooxygenase-2 (Cox2) controlled CRAd in EAC, we inserted an interferon alpha (IFN) transgene into the viral genome that is expressed upon viral replication. In this manuscript, we analyze the cytotoxic and oncolytic effects of an IFN-expressing oncolytic adenovirus in EAC and the role of the Cox2 promoter in providing for selective replication in human tissues. An infectivity-enhanced IFN-expressing CRAd (5/3 Cox2 CRAd ΔE3 ADP IFN) and other control viruses were first tested in vitro with cell lines. For the in vivo study, EAC xenografts in nude mice were treated with a single intratumoral dose of virus. An ex vivo analysis with live tissue slices was conducted using surgically resected EAC patient specimens. Expression of IFN significantly enhanced the cytotoxic and oncolytic effect of a Cox2-promoter controlled CRAd. This virus showed significant tumor growth suppression in a xenograft model. Furthermore, in human EAC samples, the promoter-controlled virus demonstrated selective replication in cancerous tissues, leaving normal esophageal tissue unaffected. An IFN-expressing CRAd driven by the Cox2 promoter has strong oncolytic effects as well as cancer-specific replication. Our novel vector possesses critical characteristics that make it a potential candidate for clinical translation to treat EAC.

Context Mechanisms underlying pituitary corticotroph adenoma adrenocorticotropin (ACTH) production are poorly understood, yet circulating ACTH levels closely correlate with adenoma phenotype and clinical outcomes. Objective We characterized the 5′ ends of proopiomelanocortin (POMC) gene transcripts, which encode the precursor polypeptide for ACTH, in order to investigate additional regulatory mechanisms of POMC gene transcription and ACTH production. Methods We examined 11 normal human pituitary tissues, 32 ACTH-secreting tumors, as well as 6 silent corticotroph adenomas (SCAs) that immunostain for but do not secrete ACTH. Results We identified a novel regulatory region located near the intron 2/exon 3 junction in the human POMC gene, which functions as a second promoter and an enhancer. In vitro experiments demonstrated that CREB binds the second promoter and regulates its transcriptional activity. The second promoter is highly methylated in SCAs, partially demethylated in normal pituitary tissue, and highly demethylated in pituitary and ectopic ACTH-secreting tumors. In contrast, the first promoter is demethylated in all POMC-expressing cells and is highly demethylated only in pituitary ACTH-secreting tumors harboring the ubiquitin-specific protease 8 (USP8) mutation. Demethylation patterns of the second promoter correlate with clinical phenotypes of Cushing disease. Conclusion We identified a second POMC promoter regulated by methylation status in ACTH-secreting pituitary tumors. Our findings open new avenues for elucidating subcellular regulation of the hypothalamic-pituitary-adrenal axis and suggest the second POMC promoter may be a target for therapeutic intervention to suppress excess ACTH production.

The use of 4 mm thickness Mini-C(T) specimens for Master Curve evaluation is drawing attention as a possible alternative / supplemental surveillance test method to current Charpy-based approaches in on-going efforts to update JEAC4201[1]. Obtaining a “valid” To per ASTM E1921 [2] and JEAC4216 [3] is a challenge for the Mini-C(T) because its tests must be performed at a low temperature relative to To to stay below the plasticity limit (KJclimit) of the testing standards while remaining above another limit on minimum test temperature (To – 50 °C) imposed by the testing standards. Minimizing the number of censored data caused by the violation of the KJclimit requirement is important in practical surveillance testing applications, since only a limited number (e.g., 12) of specimens may be available. Therefore, finding the appropriate test temperature within the first few specimens tested is essential. In this paper, the authors develop a KJc test temperature selection algorithm to maximize the chance of obtaining an E1921 and JEAC4216-valid To value with 12 or fewer tests. By the combined consideration of rejecting fewer specimens for being conducted below the low-temperature limit and having fewer specimens censored due to excess plasticity, we assumed that the test temperature of the next specimen in a series could be identified as the intersection of the KJclimit curve and the 98% tolerance bound of the KJc distribution, which is determined from an estimate of To made using already tested specimens. The appropriateness of this assumption is confirmed by a series of Monte-Carlo calculations, which demonstrates that this approach produces a valid To by testing 12 Mini-C(T)s > 98 % of the time. The study validated this testing reliability for a realistic range of yield stress and To values for reactor pressure vessel steels.

The Master Curve and its index temperature To have, over the last three decades, gained more and more acceptance in Codes, Standards, and the regulatory frameworks of different countries. For example, the Japanese Electric Association Code (JEAC) 4206 permits use of To-based index temperatures. In the United States RTTo is directly part of Section XI of the ASME Code while use of To along with extended Master Curve techniques is permitted via Code Case N-830-1. Similarly, To is permitted within both the German and the Swiss regulatory frameworks. One barrier to wider use of Master Curve/ To-based methods is the need to perform fracture toughness (KJc) tests using often limited supplies of irradiated or archive materials to determine To. While recent advances in testing technology permit the use of “mini” compact tension specimens, four of which can be machined from one-half of a broken Charpy specimen, the testing and associated engineering and regulatory analysis still represents an additional cost. Fortunately, a variety of methods exist to estimate To from prior information before any additional KJc testing is conducted. This paper will review these methods, which rely on existing information about the material type, limited but not yet valid KJc data sets, and/or Charpy data that is known through reactor pressure vessel surveillance programs. While not a replacement for direct To testing of plant materials these methods can estimate the likely range of To, which can help determine the materials for which additional testing is likely to produce the most benefit.

Background: Health consciousness in daily life is very important because lifestyle habits, such as dietary and exercise habits, sleep, and smoking, are involved in the development and progression of lifestyle-related diseases, such as cancer, heart diseases, stroke, liver diseases, kidney diseases, and diabetes. Purpose: In this study, we surveyed to see if the health consciousness increased and what behaviors in daily life changed in students of a nursing school (Seiwa Nursing College, Tokyo, Japan) after they learned medical care knowledge required to become a nurse. Results: After learning medical care knowledge for 3 years, the students were found to have a higher level of health consciousness in the third year than they did in the first year. They applied the medical care knowledge they learned in daily life. All data were collected in 2019. Conclusions: The medical care knowledge learned in a nursing school was demonstrated to serve as information to increase the health consciousness.

Compression of cross-section data used for high-resolution core analysis is performed using a dimensionality reduction technique based on the singular value decomposition (SVD) and low-rank approxi...

In this study, basic research on the formation of a film for improving the corrosion resistance of a Mg alloy (AZ31) surface was carried out by the addition of dilute phosphoric acid during water jet peening (WJP) and multifunction cavitation (MFC). These mechanochemical (MC) processes are referred to as MC-WJP and MC-MFC, respectively. MC-WJP and MC-MFC processing formed a film on the AZ31 alloy surface that contained phosphate compounds, including oxides, hydroxides, and carbides. Because MFC or WJP processing forms fine cracks on the alloy surface, we concluded that a thick film was formed on the “MC” surfaces because phosphoric acid adhered to these cracks. The film formed on the surface processed by MC-MFC was thicker than that formed on the MC-WJP processed surface because the former process produces higher-temperature cavitation bubbles. Keywords: Multifunction cavitation, Water jet peening, Mg alloy, Corrosion resistance, Phosphate film, GD-OES

INTRODUCTION. Esophageal adenocarcinoma (EAC) has increased in incidence in Western countries, and its poor prognosis necessitates the development of novel therapeutics. We previously reported the potential of conditionally replicative adenoviruses (CRAd) as a novel therapeutic treatment for this disease. To further augment the therapeutic effectiveness of our cyclooxygenase-2 (Cox2) controlled CRAd in EAC, we inserted an interferon alpha (IFN) transgene into the viral genome that is expressed upon viral replication. In this manuscript, we analyze the cytotoxic and oncolytic effects of an IFN-expressing oncolytic adenovirus in EAC and the role of the Cox2 promoter in providing for selective replication in human tissues. METHODS. An infectivity-enhanced IFN-expressing CRAd (5/3 Cox2 CRAd ΔE3 ADP IFN) and other control viruses were first tested in vitro with cell lines. For the in vivo study, EAC xenografts in nude mice were treated with a single intratumoral dose of virus. An ex vivo analysis with live tissue slices was conducted using surgically resected EAC patient specimens. RESULTS. Expression of IFN significantly enhanced the cytotoxic and oncolytic effect of a Cox2-promoter controlled CRAd. This virus showed significant tumor growth suppression in a xenograft model. Furthermore, in human EAC samples, the promoter-controlled virus demonstrated selective replication in cancerous tissues, leaving normal esophageal tissue unaffected. CONCLUSION. An IFN-expressing CRAd driven by the Cox2 promoter has strong oncolytic effects as well as cancer-specific replication. Our novel vector possesses critical characteristics that make it a potential candidate for clinical translation to treat EAC.

Obesity and diabetes are associated with an increased incidence of pancreatic cancer. Fatty acid binding protein 4 (FABP4), noted to be higher in patients with severe obesity, is linked to the development and progression of several cancers, and its level in the circulation decreases after bariatric surgery.In this paper, we evaluate the role of FABP4 in pancreatic cancer progression.University Hospital and Laboratories, United States.When Panc-1 (human) and Pan02 (mouse) pancreatic cancer cells were treated with FABP4 or the-single-point mutant FABP4 (R126Q, fatty acid binding site mutant), only FABP4 stimulated cellular proliferation. The transcriptional activity of nuclear factor E2-related factor 2 (NRF2) was increased in response to FABP4 but not the R126Q. FABP4 treatment also led to downregulation of reactive oxygen species (ROS) activity. Consistent with induced cell propagation by FABP4, the growth of Pan02 tumor was decreased in FABP4-null animals compared with C57BL/6J controls.These results suggest that FABP4 increases pancreatic cancer proliferation via activation of NRF2 and downregulation of ROS activity.

Gas samples were collected from the air surrounding single and mixed laboratory cultures, and preliminary data on human breath samples were also obtained. The infrared spectra of a variety of gasses were measured at high resolution (0.5 cm