Detection of Pancreatitis and Protein-Losing Gastroenteropathy Associated With Systemic Lupus Erythematosus by Protein Leakage Scintigraphy.

Systemic lupus erythematosus (SLE) rarely causes pancreatitis and/or protein-leakage gastroenteropathy as a gastrointestinal (GI) manifestation. Here, we describe the incidental detection of SLE-associated pancreatitis by protein leakage scintigraphy.

Protein nanoarrays using the annexin A5 two-dimensional crystal on supported lipid bilayers.

Protein nanoarrays are regularly ordered patterns of proteins fixed on a solid surface with a periodicity on the order of nanometers. They have significant potential applications as highly sensitive bioassays and biosensors. While several researchers have demonstrated the fabrication of protein nanoarrays with lithographic techniques and programmed DNA nanostructures, it has been difficult to fabricate a protein nanoarray containing a massive number of proteins on the surface. We now report the fabrication of nanoarrays of streptavidin molecules using a two-dimensional (2D) crystal of annexin A5 as a template on supported lipid bilayers that are widely used as cell membranes. The 2D crystal of annexin A5 has a six-fold symmetry with a period of about 18 nm. There is a hollow of a diameter of about 10 nm in the unit cell, surrounded by six trimers of annexin A5. We found that a hollow accommodates up to three streptavidin molecules with their orientation controlled, and confirmed that the molecules in the hollow maintain their specific binding capability to biotinylated molecules, which demonstrates that the fabricated nanoarray serves as an effective biosensing platform. This methodology can be directly applied to the fabrication of nanoarrays containing a massive number of any other protein molecules.

Fundamental and higher eigenmodes of qPlus sensors with a long probe for vertical-lateral bimodal atomic force microscopy.

The detection of vertical and lateral forces at the nanoscale by atomic force microscopy (AFM) reveals various mechanical properties on surfaces. The qPlus sensor is a widely used force sensor, which is built from a quartz tuning fork (QTF) and a sharpened metal probe, capable of high-resolution imaging in viscous liquids such as lubricant oils. Although a simultaneous detection technique of vertical and lateral forces by using a qPlus sensor is required in the field of nanotribology, it has still been difficult because the torsional oscillations of QTFs cannot be detected. In this paper, we propose a method to simultaneously detect vertical and lateral force components by using a qPlus sensor with a long probe. The first three eigenmodes of the qPlus sensor with a long probe are theoretically studied by solving a set of equations of motion for the QTF prong and probe. The calculation results were in good agreement with the experimental results. It was found that the tip oscillates laterally in the second and third modes. Finally, we performed friction anisotropy measurements on a polymer film by using a bimodal AFM utilizing the qPlus sensor with a long probe to confirm the lateral force detection.

Molecular-Resolution Imaging of Interfacial Solvation of Electrolytes for Lithium-Ion Batteries by Frequency Modulation Atomic Force Microscopy.

Solvation structures formed by ions and solvent molecules at solid/electrolyte interfaces affect the energy storage performance of electrochemical devices, such as lithium-ion batteries. In this study, the molecular-scale solvation structures of an electrolyte, a solution of lithium bis(trifluoromethanesulfonyl)imide (LiTFSI) in propylene carbonate (PC) at the electrolyte-mica interface, were measured using frequency-modulation atomic force microscopy (FM-AFM). The spacing of the characteristic force oscillation in the force versus distance curves increased with increasing ion concentration, suggesting an increase in the effective size of molecules at the interface. Molecular dynamics simulations showed that the effective size of molecular assemblies, namely, solvated ions formed at the interface, increased with increasing ion concentrations, which was consistent with the experimental results. Knowledge of molecular-scale structures of solid/electrolyte interfaces obtained by a combination of FM-AFM and molecular dynamics simulations is important in the design of electrolytes for future energy devices and in improving their properties.

Nanomechanics of self-assembled surfactants revealed by frequency-modulation atomic force microscopy.

Surfactants play a critical role in bottom-up nanotechnologies due to their peculiar nature of controlling the interfacial energy. Since their operational mechanism originates from the molecular-scale formation and disruption processes of molecular assemblies (i.e., micelles), conventional static-mode atomic force microscopy has made a significant contribution to unravel the detailed molecular pictures. Recently, we have successfully developed a local solvation measurement technique based on three-dimensional frequency-modulation atomic force microscopy, whose spatial resolution is not limited by jump-to-contact. Here, using this novel technique, we investigate molecular nanomechanics in the formation and disruption processes of micelles formed on a hydrophobic surface. Furthermore, an experiment employing a hetero-nanostructure reveals that the nanomechanics depends on the form of the molecular assembly. Namely, the hemifusion and disruption processes are peculiar to the micellar surface and cause a higher energy dissipation than the monolayer surface. The technique established in this study will be used as a generic technology for further development of bottom-up nanotechnologies.

Structured Water Molecules on Membrane Proteins Resolved by Atomic Force Microscopy.

Water structuring on the outer surface of protein molecules called the hydration shell is essential as well as the internal water structures for higher-order structuring of protein molecules and their biological activities in vivo. We now show the molecular-scale hydration structure measurements of native purple membrane patches composed of proton pump proteins by a noninvasive three-dimensional force mapping technique based on frequency modulation atomic force microscopy. We successfully resolved the ordered water molecules localized near the proton uptake channels on the cytoplasmic side of the individual bacteriorhodopsin proteins in the purple membrane. We demonstrate that the three-dimensional force mapping can be widely applicable for molecular-scale investigations of the solid-liquid interfaces of various soft nanomaterials.

Low-Background Tip-Enhanced Raman Spectroscopy Enabled by a Plasmon Thin-Film Waveguide Probe.

Tip-enhanced Raman spectroscopy (TERS) is a nano-optical approach to extract spatially resolved chemical information with nanometer precision. However, in the case of direct-illumination TERS, which is often employed in commercial TERS instruments, strong fluorescence or far-field Raman signals from the illuminated areas may be excited as a background. They may overwhelm the near-field TERS signal and dramatically decrease the near-field to far-field signal contrast of TERS spectra. It is still challenging for TERS to study the surface of fluorescent materials or a bulk sample that cannot be placed on an Au/Ag substrate. In this study, we developed an indirect-illumination TERS probe that allows a laser to be focused on a flat interface of a thin-film waveguide located far away from the region generating the TERS signal. Surface plasmon polaritons are generated stably on the waveguide and eventually accumulated at the tip apex, thereby producing a spatially and energetically confined hotspot to ensure stable and high-resolution TERS measurements with a low background. With this thin-film waveguide probe, TERS spectra with obvious contrast from a diamond plate can be acquired. Furthermore, the TERS technique based on this probe exhibits excellent TERS signal stability, a long lifetime, and good spatial resolution. This technique is expected to have commercial potential and enable further popularization and development of TERS technology as a powerful analytical method.

Surface charge density measurement of a single protein molecule with a controlled orientation by AFM.

The spatial distribution of functional groups causes a charge distribution that often has a close relationship with its biofunctions. To understand them of the protein molecules, measurements of the charge distribution under physiological conditions are desired. Atomic force microscopy (AFM) has been utilized to measure the surface charge density by measuring the electric double layer (EDL) force caused by the overlap of the EDLs on the surfaces of the AFM tip and the biomolecule. Here, we demonstrated the surface charge density measurement of a single streptavidin (SA) protein molecule by the three-dimensional force mapping method based on frequency modulation AFM (FM-AFM). The SA has a strong affinity to biotin because of the electrostatic interactions between the molecules. Therefore, the surface charge density measurements of the biotin-binding sites and other surface areas of the molecule have been anticipated. However, the surface charge density of the surfaces other than the biotin-binding side has never been measured. We demonstrate the surface charge density measurement of the top surface of the single SA molecule, which is perpendicular to the biotin-binding sides, with a controlled orientation using DNA origami as a template by FM-AFM in an electrolyte solution. The surface charge density of the top surface of the SA molecule was estimated by fitting the experimental force curves to the Derjaguin-Landau-Verwey-Overbeck theory. We found that the surface charge density of the top surface of the SA molecule is comparable to those reported earlier for the biotin-binding sides of the molecule. We expect that, by using the DNA origami technology, one can control the orientation of a biomolecule attached to the substrate and measure the surface charge density of the specific surface areas of the biomolecule to obtain information that will help us to understand the relationship between their structures and functions.

Late-onset pneumothorax in a COVID-19 patient treated with ventilation and ECMO: A case report and literature review.

Coronavirus disease 2019 (COVID-19) has become a major threat to public health since the outbreak in Wuhan in 2019. Chest computed tomography is recommended for COVID-19 cases for evaluation and follow up of pneumonia and related complication. We report the case of a 66-year-old man with underlying hypertension and a history of smoking 76 packs a year; he was frequently monitored by computed tomography for pulmonary changes during the period from early symptom onset to death. Furthermore, he developed a pneumothorax during the course. The occurrence of pneumothorax in COVID-19 patients is not common, and there have been only a few previous reports. This is a valuable case of pneumothorax in a patient with COVID-19 treated with a ventilator and extracorporeal membrane oxygenation. This case and previous reports suggest that pneumothorax occurs in COVID-19 with a relatively late onset (3-8 weeks). Long-term pneumonia morbidity, steroid therapy, positive pressure ventilation, and extracorporeal membrane oxygenation can cause pneumothorax, leading to capillary and alveolar damage.

Molecular-Scale Solvation Structures of Ionic Liquids on a Heterogeneously Charged Surface.

Understanding the sub-nanoscale solvation structures of ionic liquids is crucial for the development of innovative functional "devices" across numerous fields. We previously demonstrated the atomic-scale solvation measurements using an ultralow noise 3D frequency-modulation atomic force microscopy combined with molecular dynamics simulations. However, to facilitate practical applications, the molecular distribution on a heterosurface must be verified. Here, we unveil the local solvation structures on a heterogeneously charged phyllosilicate surface in an ionic liquid solution and pure liquid. By identifying adsorbed ion species from the molecular sizes and orientations, we experimentally demonstrate that anions and cations preferentially adsorbed onto the positive and negative surfaces exhibit different orientations and water miscibility. Moreover, we reveal that neutral intermediate regions are formed at the boundary region in ionic liquid media as well as a KCl solution. In the future, this technique will be essential for the evolution of ionic-liquid functional "devices".

Evaluation of the Usefulness of CO-RADS for Chest CT in Patients Suspected of Having COVID-19.

The purpose of this study was to use the Coronavirus Disease 2019 (COVID-19) Reporting and Data System (CO-RADS) to evaluate the chest computed tomography (CT) images of patients suspected of having COVID-19, and to investigate its diagnostic performance and interobserver agreement. The Dutch Radiological Society developed CO-RADS as a diagnostic indicator for assessing suspicion of lung involvement of COVID-19 on a scale of 1 (very low) to 5 (very high). We investigated retrospectively 154 adult patients with clinically suspected COVID-19, between April and June 2020, who underwent chest CT and reverse transcription-polymerase chain reaction (RT-PCR). The patients' average age was 61.3 years (range, 21-93), 101 were male, and 76 were RT-PCR positive. Using CO-RADS, four radiologists evaluated the chest CT images. Sensitivity, specificity, and area under the receiver operating characteristic curve (AUC) were calculated. Interobserver agreement was calculated using the intraclass correlation coefficient (ICC) by comparing the individual reader's score to the median of the remaining three radiologists. The average sensitivity was 87.8% (range, 80.2-93.4%), specificity was 66.4% (range, 51.3-84.5%), and AUC was 0.859 (range, 0.847-0.881); there was no significant difference between the readers (p > 0.200). In 325 (52.8%) of 616 observations, there was absolute agreement among observers. The average ICC of readers was 0.840 (range, 0.800-0.874; p < 0.001). CO-RADS is a categorical taxonomic evaluation scheme for COVID-19 pneumonia, using chest CT images, that provides outstanding performance and from substantial to almost perfect interobserver agreement for predicting COVID-19.

Atomic-Scale Three-Dimensional Local Solvation Structures of Ionic Liquids.

Room-temperature ionic liquids are promising media for next-generation energy devices because of their various superior characteristics. Because device performance is often dictated by the solvation structures at the solid-liquid interfaces, particularly at the local reactive sites, their atomistic pictures are in great demand. However, there has been no experimental technique for their three-dimensional solvation structures. Here, we first demonstrate the measurement of the atomic-scale ionic liquids using a recently established ultralow-noise three-dimensional frequency-modulation atomic force microscopy technique supported by molecular dynamics simulations. We conducted the experiments in protic and aprotic aqueous solutions and reveal that the aprotic solvation structure exhibits the higher site specificity, which resolves atomic-scale surface charge distribution on mica because of the absence of the H-bonding network. Our methodology is also applicable to pure liquids and would be a breakthrough for expanding their future applications.

TS-1 add-on therapy in Japanese patients with triple-negative breast cancer after neoadjuvant or adjuvant chemotherapy: a feasibility study.

Purpose We examined the feasibility, efficacy, and safety of TS-1 add-on therapy (TAT) in Japanese patients with triple-negative breast caner (TNBC). Methods TAT (TS-1, 80 mg/m2/day, BID, PO), consisting of the 21-day cycles of 14-day consecutive administration followed by 7-day drug holiday, was conducted for 365 days. The median follow-up was 75.2 months (range, 7.3-103.3 months). The primary endpoint was the feasibility of TAT. The secondary endpoints included relapse-free survival (RFS), overall survival (OS), and safety. Results 63 Japanese patients with TNBC (median age, 52.5 years; range, 23.7-68.6 years) were examined. Among them, 34 (54.0%) were postmenopausal, 54 (93.7%) had TNBC of common histological type, 51 (81.0%) had T1 to 3 tumors, 63 (100%) had undergone standardized surgery, and 44 (69.8%) and 19 (30.2%) had undergone neoadjuvant chemotherapy and adjuvant chemotherapy, respectively. The 365-day cumulative rate of TS-1 administration was 68.3% (95% confidence interval, 55.3-79.4), being comparable to 65.8% previously reported for gastric cancer. The 5-year RFS rates were 52.3% and 84.2% in the neoadjuvant and adjuvant chemotherapy groups, respectively, and the 5-year OS rates were 68.0% and 89.5%, respectively. The most common adverse events (AEs) were leucocyte count decreased (50.8%), total bilirubin decreased (44.4%), and pigmentation (42.9%). AEs were manageable clinically, and any grade 4 AEs did not develop. Conclusions The 365-day cumulative rate of TS-1 administration in TNBC patients was comparable to that in gastric cancer patients despite previous chemotherapy with anthracyclines and/or taxanes. TAT was feasible for TNBC patients after standard primary therapy.

Multicenter Observational Study of Fulvestrant 500 mg in Postmenopausal Japanese Women with Estrogen Receptor-Positive Advanced or Recurrent Breast Cancer after Prior Endocrine Treatment (SBCCSG29 Study).

BACKGROUND: Fulvestrant 500 mg has been an option for endocrine therapy for advanced or recurrent breast cancer after prior endocrine treatment since November 2011 in Japan. This study aimed to clarify the effectiveness and safety of fulvestrant 500 mg in clinical settings. METHODS: This was a multicenter, both prospective and retrospective, observational study of 132 postmenopausal women (median age 66) with locally advanced or metastatic breast cancer, who had been treated with fulvestrant. Information from medical records was retrospectively obtained from 9 hospitals (Saitama Breast Cancer Clinical Study Group: SBCCSG) in Saitama prefecture, Japan, from October 2012 to April 2014. The primary end point was time to treatment failure (TTF). The secondary end points were overall survival (OS), objective response rate (ORR), clinical benefit rate (CBR), and adverse events (AE) (CTCAE ver. 4). The choice of subsequent therapy after fulvestrant was also evaluated. RESULTS: The median TTF was 6.1 months. Median OS was 28.5 months (the starting date was the first day of fulvestrant). ORR was 12.9% and CBR was 45.5%. The most common AEs were injection site reactions (9.1%). The rate of grade 3 AE was only 2.3% (3/132). The number of patients who underwent subsequent therapy after fulvestrant were 54 (55.7%) receiving chemotherapy, 42 (43.3%) receiving non-fulvestrant endocrine therapy, and 1 (1%) receiving mammalian target of rapamycin inhibitor (mTORi) + endocrine therapy (ET). CONCLUSION: Fulvestrant 500 mg is an effective and safe treatment for patients with advanced or recurrent breast cancer after prior endocrine treatment.

Molecular-scale visualization and surface charge density measurement of Z-DNA in aqueous solution.

The DNA in the left-handed conformation (Z-conformation) was first discovered by A. Rich, who revealed the crystalline structure of a DNA oligomer d(GC)3 by X-ray diffraction method. Later it was also found that DNA molecules change their conformations from typical right-handed form (B-DNA) to the left-handed form (Z-DNA) under specific conditions (B-Z transition). Furthermore, the detailed structures of the interface between B- and Z-DNAs, B-Z junction, was also determined with an atomic resolution. Recently it was found that some proteins have the Z-DNA binding domains, but the biological functions of Z-DNA are not well understood yet. Therefore the investigation of Z-DNA under physiological conditions is highly essential. In this study, we demonstrated the high-resolution real-space imaging of DNA molecules having the Z- and B-form conformations by frequency-modulation atomic force microscopy (FM-AFM), that has made a great progress in recent years, in an aqueous solution. The major and minor grooves of both DNA conformations were clearly visualized. Furthermore, the surface charge density was measured by three-dimensional (3D) force mapping method. We found that Z-form region was less negatively charged than the B-form region.

Atomic-Level Viscosity Distribution in the Hydration Layer.

The viscosity of solvation structures is crucial for the development of energy-efficient biofunctional and electrochemical devices. Elucidating their subnanoscale distributions can cause the formation of a sustainable energy society. Here, we visualize the site-specific three-dimensional damping distribution on a CaCO_{3} surface composed of binary ion species using ultra-low-noise frequency modulation atomic force microscopy. With the support from molecular dynamics simulation, we found a strikingly large damping at the calcium sites, which demonstrates the capability of this methodology to visualize atomic-scale viscosity in the hydration layers. Our finding will expedite the evolutions of various functional devices.

Correction to: Eribulin, trastuzumab, and pertuzumab as first-line therapy for patients with HER2-positive metastatic breast cancer: a phase II, multicenter, collaborative, open-label, single-arm clinical trial.

The authors would like to note the replacement of Fig. 2b, for which Fig. 2a was placed erringly, with appropriate Fig. 2b.

Eribulin, trastuzumab, and pertuzumab as first-line therapy for patients with HER2-positive metastatic breast cancer: a phase II, multicenter, collaborative, open-label, single-arm clinical trial.

Purpose To examine the efficacy and safety of triple therapy with eribulin, trastuzumab, and pertuzumab in patients with HER2-positive metastatic breast cancer (MBC) who never received any prior therapy in the first-line metastatic/advanced setting. Methods Eribulin 1.4 mg/m2 (days 1 and 8), trastuzumab 8 mg/kg over 90 min and 6 mg/kg over 30 min, and pertuzumab 840 mg/body over 60 min and 420 mg/body over 30 min were administered intravenously in 21-day cycles. Results 25 women (median age, 57 years [range, 41-75 years]) received a median of 10 cycles (range, 0-34 cycles); 24 had performance status (PS) 0, 1 PS 1, 8 stage IV breast cancer, and 17 recurrence. Lung and liver metastases occurred in 9 and 9 patients, respectively. Median time to treatment failure with eribulin was 9.1 months (95% confidence interval [CI], 4.3-13.9 months), and median progression-free survival was 23.1 months (95% CI, 14.4-31.8 months). The overall response rate (complete response [CR] + partial response [PR]) was 80.0% (95% CI, 59.3-93.2%), and the clinical benefit rate (CR + PR + stable disease ≥24 weeks) was 84.0% (95% CI, 63.9-95.5%). The most common treatment-emergent adverse events (TEAEs) were alopecia (92.0%), fatigue (68.0%), and sensory peripheral neuropathy (60.0%). Grade 3/4 TEAEs occurred in 11 patients (44.0%). The only grade 4 TEAE was neutrophil count decreased (16.0%). Neither grade 4 peripheral neuropathy nor febrile neutropenia occurred. Conclusions ETP therapy showed acceptable efficacy and safety and is a potential first-line therapy for patients with HER2-positive MBC.

Investigation of Local Hydration Structures of Alkanethiol Self-Assembled Monolayers with Different Molecular Structures by FM-AFM.

Hydration structures play crucial roles in a wide variety of chemical and biological phenomena. However, the key factors that determine a hydration structure remain an open question. Most recent studies have focused on the electrostatic interactions between the surface charges and dipoles of water molecules, which are determined by the atomic/ionic species of the outermost solid surface, as the dominating factor. The number of studies on the correlation between the hydration structure and the atomic-scale surface corrugation has been limited. In this study, we investigated the hydration structures of alkanethiol self-assembled monolayers terminated with a hydroxyl group using frequency-modulated atomic force microscopy. We observed two molecular structures, namely, the (√3 × âˆš3) R30° structure and the c(4 × 2) superlattice structure, and found that their hydration structures are different mainly because of the slight differences in their molecular arrangements. This result suggests that a slight difference in the molecular/atomic arrangements as well as the atomic/ionic species in the outermost solid surface strongly influences the local hydration structures.