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Abstract During the production of bispecific antibodies (bsAbs), nonspecific pairing results in low yields of target bsAb molecules, an issue known as the “mispairing problem.” Several antibody engineering techniques have been developed to overcome mispairing issues. Here, we introduce “bsAb by external pairing and excision” (BAPE), a novel chain pairing method that induces specific chain pairing by fusing external SpyCatcher/Tag and SnoopCatcher/Tag units. These tags are then removed via protease cleavage. In this study, we applied this method to force the correct pairings of heavy and light chains while the heavy-chain pairing was achieved by the Knobs-into-Holes mutation. We then confirmed the formation of interchain bridges with covalent isopeptide bonds. Both anti-CD3/anti-Her2 and anti-CD3/anti-EGFR bsAbs displayed satisfactory target binding activities and in vitro cell-killing activity with activated T-cells.

Abstract Skyrmion lattices (SkL) in centrosymmetric materials typically have a magnetic period on the nanometer-scale, so that the coupling between magnetic superstructures and the underlying crystal lattice cannot be neglected. We reveal the commensurate locking of a SkL to the atomic lattice in Gd3Ru4Al12 via high-resolution resonant elastic x-ray scattering (REXS). Weak easy-plane magnetic anisotropy, demonstrated here by a combination of ferromagnetic resonance and REXS, penalizes placing a skyrmion core on a site of the atomic lattice. Under these conditions, a commensurate SkL, locked to the crystal lattice, is stable at finite temperatures – but gives way to a competing incommensurate ground state upon cooling. We discuss the role of Umklapp-terms in the Hamiltonian for the formation of this lattice-locked state, its magnetic space group, and the role of slight discommensurations, or (line) defects in the magnetic texture. We also contrast our findings with the case of SkLs in noncentrosymmetric material platforms.

In older horses, basal metabolic rate decreases, and plasma metabolite and hormone concentrations related to energy metabolism change. The occurrence of age-related diseases, which increases in old animals, may enhance inflammatory reactivity (inflammaging). Finding the appropriate treatment for inflammaging at an early stage may prevent various age-related diseases. Changes in metabolite and hormone concentrations and enzyme activities involved in energy metabolism in the plasma of clinically healthy riding horses of various ages were measured to identify biomarkers of inflammaging (persistent low-grade inflammation that occurs with aging). All horses were clinically healthy, and their body condition scores (BCSs) were 4 or 5 (9-point scale). Plasma triglyceride (TG), total cholesterol (T-Cho), blood urea nitrogen (BUN), insulin concentrations, malondialdehyde (MDA), and serum amyloid A (SAA) concentrations generally increased with age. Adiponectin concentrations, plasma superoxide dismutase (SOD), and leukocyte AMP-activated protein kinase (AMPK) activities decreased, while plasma aspartate aminotransferase (AST), alanine aminotransferase (ALT), and glutathione peroxidase (GPx) remained unchanged as horses aged. Although riding horses that partake in continuous exercise seems to be less likely to develop inflammaging, horses over 17 years of age tend to show proinflammatory signs with disordered lipid metabolism. In riding horses, SAA, in combination with other markers, may be a useful biomarker for inflammaging and dysregulated lipid metabolism in aging horses.

Coastal areas often experience high population density and intense human activity owing to the considerable value of the ocean. Therefore, devices for monitoring marine disasters are crucial for ensuring the safety of human life. Herein, we develop hemispherical spring origami (SO) triboelectric nanogenerators (TENGs) (HSO-TENGs) for self-powered ocean wave monitoring. Optimization is performed using two approaches. First, swing machine experiments are conducted to investigate the monitoring performance of the HSO-TENGs regarding wave height and period with satisfactory accuracy. To increase power generation and monitoring accuracy, the internal inertia and centroid of gravity of the HSO-TENGs are optimized with respect to the structural parameters (i.e., magnet weight, hammer height, and external swing arm length). Second, numerical simulations are performed using the smoothed-particle hydrodynamics (SPH) method to determine the most suitable fixed condition for the HSO-TENGs for sensing wave changes. Subsequently, wave tank experiments are conducted on the HSO-TENGs to determine their ability to sense wave height, period, frequency, and direction. Tests related to supplying other sensors are also conducted. Eventually, the ability of the HSO-TENGs to monitor wave direction and spreading parameters is investigated in a numerical SPH circular wave tank. The results prove that the optimized HSO-TENGs can achieve powering and sensing through the same device.

Abstract Vitamin E is classified into tocopherol (Toc) and tocotrienol (T3) based on its side chains. T3 generally has higher cellular uptake than Toc, though the responsible mechanism remains unclear. To elucidate this mechanism, we hypothesized and investigated whether serum albumin is a factor that induces such a difference in the cellular uptake of Toc and T3. Adding bovine serum albumin (BSA) to serum-depleted media increased the cellular uptake of T3 and decreased that of Toc, with varying degrees among α-, β-, γ-, and δ-analogs. Such enhanced uptake of α-T3 was not observed when cells were incubated under low temperature (the uptake of α-Toc was also reduced), suggesting that Toc and T3 bind to albumin to form a complex that results in differential cellular uptake of vitamin E. Fluorescence quenching study confirmed that vitamin E certainly bound to BSA, and that T3 showed a higher affinity than Toc. Molecular docking further indicated that the differential binding energy of Toc or T3 to BSA is due to the Van der Waals interactions via their side chain. Overall, these results suggested that the affinity of Toc and T3 to albumin differs due to their side chains, causing the difference in their albumin-mediated cellular uptake. Our results give a better mechanistic insight into the physiological action of vitamin E.

Abstract Although several ribosomal protein paralogs are expressed in a tissue-specific manner, how these proteins affect translation and why they are required only in certain tissues have remained unclear. Here we show that RPL3L, a paralog of RPL3 specifically expressed in heart and skeletal muscle, influences translation elongation dynamics. Deficiency of RPL3L-containing ribosomes in RPL3L knockout male mice resulted in impaired cardiac contractility. Ribosome occupancy at mRNA codons was found to be altered in the RPL3L-deficient heart, and the changes were negatively correlated with those observed in myoblasts overexpressing RPL3L. RPL3L-containing ribosomes were less prone to collisions compared with RPL3-containing canonical ribosomes. Although the loss of RPL3L-containing ribosomes altered translation elongation dynamics for the entire transcriptome, its effects were most pronounced for transcripts related to cardiac muscle contraction and dilated cardiomyopathy, with the abundance of the encoded proteins being correspondingly decreased. Our results provide further insight into the mechanisms and physiological relevance of tissue-specific translational regulation.

The mode of cytokine receptor activation is diverse. Here, the authors find that the marine-sponge derived lectin ThC, a bivalent sugar binding protein, activates human cytokine receptor MPL. This mode of action resembles the pathogenic activation of MPL by mutant molecular chaperon calreticulin in hematologic malignancies.

Strigolactones (SLs) regulate angiosperm development and promote symbiosis with arbuscular mycorrhizae. Here the authors show that bryosymbiol, an SL present in bryophytes and angiosperms, promotes AM symbiosis in Marchantia paleacea suggesting an ancestral function of SLs as rhizosphere signals.

Abstract Understanding the ultrashort time scale structural dynamics of the FeRh metamagnetic phase transition is a key element in developing a complete explanation of the mechanism driving the evolution from an antiferromagnetic to ferromagnetic state. Using an X-ray free electron laser we determine, with sub-ps time resolution, the time evolution of the (–101) lattice diffraction peak following excitation using a 35 fs laser pulse. The dynamics at higher laser fluence indicates the existence of a transient lattice state distinct from the high temperature ferromagnetic phase. By extracting the lattice temperature and comparing it with values obtained in a quasi-static diffraction measurement, we estimate the electron–phonon coupling in FeRh thin films as a function of laser excitation fluence. A model is presented which demonstrates that the transient state is paramagnetic and can be reached by a subset of the phonon bands. A complete description of the FeRh structural dynamics requires consideration of coupling strength variation across the phonon frequencies.

Abstract Background Surgeons sometimes have difficulty determining which result to favor when preoperative results (MRI + preoperative endometrial biopsy [pre-op EB]) differ from intraoperative frozen section histology (FS) results. Investigation of how FS can complement ordinary preoperative examinations like MRI and pre-op EB in identification of patients at high risk of lymph node metastasis (high-risk patients) could provide clarity on this issue. Therefore, the aim of this study is to assess the utility of pre-op EB, MRI and FS results and determine how to combine these results in identification of high-risk patients. Methods The subjects were 172 patients with endometrial cancer. Patients with a histological high-grade tumor (HGT), namely, grade 3 endometrioid cancer, clear cell carcinoma or serous cell carcinoma, or with any type of cancer invading at least half of the uterine myometrium were considered high-risk. Tumors invading at least half of the uterine myometrium were classified as high-stage tumors (HST). We compared (a) detection of HGT using pre-op EB versus FS, (b) detection of HST using MRI versus FS, and (c) identification of high-risk patients using MRI + pre-op EB versus FS. Lastly, we determined to what degree addition of FS results improves identification of high-risk patients by routine MRI + pre-op EB. Results (a) Sensitivity, specificity, and accuracy for detecting HGT were 59.6, 98.4 and 87.8% for pre-op EB versus 55.3, 99.2 and 87.2% for FS (P = 0.44). (b) These figures for detecting HST were 74.4, 83.0 and 80.8% for MRI versus 46.5, 99.2 and 86.0% for FS (P < 0.001). (c) These figures for identifying high-risk patients were 78.3, 85.4 and 82.6% for MRI + pre-op EB versus 55.1, 99.0 and 81.2% for FS (P < 0.001). The high specificity of FS improved the sensitivity of MRI + pre-op EB from 78.3 to 81.2%, but this difference was not statistically significant (P < 0.16). Conclusion Frozen section enables identification of high-risk patients with nearly 100% specificity. This advantage can be used to improve sensitivity for identification of high-risk patients by routine MRI + pre-op EB, although this improvement is not statistically significant.

Objective To report the long-term postoperative outcomes of transsclerally sutured intraocular lenses (IOLs), in which the haptics were correctly fixated into the ciliary sulcus using an auxiliary device and endoscope.Methods and analysis Data were collected from eyes that were followed up for at least 12 months after ciliary sulcus suture fixation of an IOL using an auxiliary device for securely placing the IOL haptics to the ciliary sulcus, which was confirmed using intraoperative endoscopy in all cases. The corrected distance visual acuity (CDVA), refractive error, anterior chamber depth (ACD), IOL decentration and tilt, corneal endothelial cell density (CECD) and postoperative complications were recorded. ACD and IOL deviations were compared with those of normal controls after standard cataract surgery.Results A total of 146 eyes of 142 patients were included, with a mean follow-up period of 56.0±35.3 (range 12–174) months. Postoperative CDVA from 1 month to 8 years and final CDVA were significantly better, and the mean refraction error, ACD and CECD decline rate were −0.71±0.75 dioptre, 4.01±0.37 mm and −7.4%±16.0%, respectively. Compared with normal controls, ACD was not significantly different but the tilt and decentration were significantly different. The main postoperative complications included vitreous haemorrhage (24.0％), suture thread exposure (19.2%) and corectopia (18.5%). There were no cases of IOL dislocation due to suture breakage or postoperative endophthalmitisConclusion Long-term postoperative outcomes were favorable with good CDVA and without IOL dislocation and endophthalmitis. The significance and value of fixing haptics to the ciliary sulcus should be re-evaluated.

Abstract Antibodies have been widely used for cancer therapy owing to their ability to distinguish cancer cells by recognizing cancer-specific antigens. Epidermal growth factor receptor (EGFR) is a promising target for the cancer therapeutics, against which several antibody clones have been developed and brought into therapeutic use. Another antibody clone, 528, is an antagonistic anti-EGFR antibody, which has been the focus of our antibody engineering studies to develop cancer drugs. In this study, we explored the interaction of 528 with the extracellular region of EGFR (sEGFR) via binding analyses and structural studies. Dot blotting experiments with heat treated sEGFR and surface plasmon resonance binding experiments revealed that 528 recognizes the tertiary structure of sEGFR and exhibits competitive binding to sEGFR with EGF and cetuximab. Single particle analysis of the sEGFR–528 Fab complex via electron microscopy clearly showed the binding of 528 to domain III of sEGFR, the domain to which EGF and cetuximab bind, explaining its antagonistic activity. Comparison between the two-dimensional class average and the cetuximab/sEGFR crystal structure revealed that 528 binds to a site that is shifted from, rather than identical to, the cetuximab epitope, and may exclude known drug-resistant EGFR mutations.

A man in his 60s visited the emergency department because of epigastric pain and vomiting. Noncontrast helical abdominal CT revealed a lipomatous mass on the right side of the lower esophagus. Caudal slices showed that the mass appeared to displace the gastric antrum causing gastric outlet obstruction. Further observation revealed a linear structure from the greater curvature of the stomach into the omental vessels. Based on these findings, a diagnosis of intrathoracic omental hernia through the esophageal hiatus was made. Intraoperatively, a defect in the lesser omentum was seen, and the greater omentum herniated through the defect into the esophageal hiatus. Omental herniation through the lesser omentum into the esophageal hiatus can present as gastric outlet obstruction. Keywords: Omental herniation, Hiatus hernia, Mediastinum, CT

Due to the high incidence of mammary tumors in dogs, it is important to elucidate the pathogenesis of these tumors in veterinary medicine. Radiation therapy is often used to treat mammary tumors that target DNA lesions. RAD51 is a key molecule that repairs DNA damage via homologous recombination. We examined the relationship between RAD51 expression and radiosensitivity in mammary tumor cell lines. CHMp and CHMm from the same individual were selected based on the differences in RAD51 expression. The radiosensitivity of both cell lines was examined using MTT and scratch assays; CHMm, which has high RAD51 expression, showed higher sensitivity to radiation than CHMp. However, the nuclear focus of RAD51 during DNA repair was formed normally in CHMp, but not in most of CHMm. Since irradiation resulted in the suppression of cell cycle progression in CHMp, the expression of p21, a cell cycle regulatory factor, was detected in CHMp after 15 Gy irradiation but not in CHMm. These results indicate that functional expression is more important than the quantitative expression of RAD51 in canine mammary tumor cells in response to DNA damage.

The biological effects of lead (Pb) contamination have been reported in various species. There are no restrictions on the use of Pb products, including bullets, in the areas south of Hokkaido, Japan. Local governments have announced the presence of Pb in the soil sediments of water bodies. Previous studies have confirmed the relationship between blood Pb level (BLL) and immune cells. This study was performed with the aim of clarifying the effect of Pb contamination on immune cells. In total, 170 Black-headed Gulls (Chroicocephalus ridibundus) were captured, including a population in Tokyo Bay between November 2018 and April 2021 and a population in Mikawa Bay between January 2019 and April 2021. Linear regression analysis was performed with the white blood cell count (WBC), proportion of heterophils (Het), proportion of lymphocytes (Lym), ratio of heterophils and lymphocytes (H/L ratio), copy number of CD4 messenger RNA, and copy number of CD8α messenger RNA as the objective variables, and the BLL as the explanatory variable. The group with BLL < 1.0 µg/dL had a significantly lower Het and higher Lym than that with BLL > 3.5 µg/dL (P < 0.05). In addition, the group with BLL < 1.0 µg/dL had a significantly lower H/L ratio than that with BLL > 3.5 µg/dL. CD8α and WBC were higher in the group with BLL ranging from 1.0 to 3.5 µg/dL than in the group with BLL < 1.0 µg/dL. This study suggests that the effect of Pb pollution on the immune cells of Black-headed Gulls is lower than some previous criteria values. It is possible that gulls affected by Pb contamination suffer indirect negative effects on immune function, possibly making them more susceptible to infectious diseases. Pb is a major environmental pollutant, against which measures must be taken.

Abstract Background Isolated enteric duplication cyst is an intestinal duplication cyst found in a distant location from the intestinal tract and it is said to have its own blood supply. Meckel’s diverticulm is considered as an antimesenteric structure and has its own blood supply. However, there are some reported cases of Meckele’s diverticum in the mesenteric side. Ectopic pancreas may be found in both entities. Case presentation A 5-year-old girl presented with increasing abdominal pain around the umbilicus. On laboratory investigation serum pancreatic enzymes and C-reactive protein were elevated. Abdominal computed tomography (CT) revealed a normal pancreas but a cystic lesion in the mesentery of the ileum. A nodule with a marked enhancement was observed in the wall of the lesion. During the laparoscopy, the lesion was found at the root of the mesentery and was distant from the ileum. The lesion was resected suspecting an abscess. Pathologically, the wall of the lesion consisted of small bowel like tissue, and pancreatic tissue was seen beneath the mucosa. There were some post inflammatory changes in the pancreatic tissue. Retrospectively on thin slice enhanced CT, an independent blood supply was noted. Based on these findings, a diagnosis of ectopic pancreatitis in an iliac intestinal duplication cyst was made. Conclusion Isolated enteric duplication cyst in the root of ileal mesentery and mesenteric Meckel’s diverticulum have similarities. In the present case, the diagnosis of isolated enteric duplication cyst was made since it was found distant from the ileum. It is important to consider the possibility of ectopic pancreatitis when serum pancreatic enzymes are elevated even when the pancreas appears normal.

The oxygen transporter of molluscs, hemocyanin, consists of long pearl-necklace-like subunits of several globular domains. The subunits assemble in a complex manner to form cylindrical decamers. Typically, the first six domains of each subunit assemble together to form the cylinder wall, while the C-terminal domains form a collar that fills or caps the cylinder. During evolution, various molluscs have been able to fine-tune their oxygen binding by deleting or adding C-terminal domains and adjusting their inner-collar architecture. However, squids have duplicated one of the wall domains of their subunits instead. Here, using cryo-EM and an optimized refinement protocol implemented in SPHIRE, this work tackled the symmetry-mismatched structure of squid hemocyanin, revealing the precise effect of this duplication on its quaternary structure and providing a potential model for its structural evolution.

A 33-year-old woman, gravida two para one, at 31 weeks of gestational age experienced sudden onset of left lower quadrant pain and underwent unenhanced pelvic MRI. On fast imaging employing steady state precession (FIESTA) sequence images, a marked difference was observed in the diameters of the right and left ovarian veins. The right ovarian vein was torturous and dilated, measuring 35 mm in diameter, while the left ovarian vein was thin and linear, measuring 7 mm in diameter. The left ovary showed no apparent swelling or hemorrhage, but was suspected to have been shifted anteriorly. The patient underwent explorative laparotomy and was found to have left ovarian torsion. A difference in diameter of the ovarian veins, with thinning of the twisted side and compensatory dilatation of the contralateral side for drainage of increased uterine blood flow, may be a useful imaging sign for the diagnosis of ovarian torsion during pregnancy. Keywords: Ovarian torsion, Third trimester in pregnancy, MRI

Feline coronaviruses (FCoVs) infect cats worldwide and cause severe systemic diseases, such as feline infectious peritonitis (FIP). FIP has a high mortality rate, and drugs approved by the Food and Drug Administration have been ineffective for the treatment of FIP. Investigating host factors and the functions required for FCoV replication is necessary to develop effective drugs for the treatment of FIP. FCoV utilizes an endosomal trafficking system for cellular entry after binding between the viral spike (S) protein and its receptor. The cellular enzymes that cleave the S protein of FCoV to release the viral genome into the cytosol require an acidic pH optimized in the endosomes by regulating cellular ion concentrations. Ionophore antibiotics are compounds that form complexes with alkali ions to alter the endosomal pH conditions. This study shows that ionophore antibiotics, including valinomycin, salinomycin, and nigericin, inhibit FCoV proliferation in vitro in a dose-dependent manner. These results suggest that ionophore antibiotics should be investigated further as potential broad-spectrum anti-FCoV agents.

Purpose: We report a case of corneal keloid occurring 30 years after pterygium surgery and 3 years after cataract surgery. Observations: The case of a 72-year-old man was referred because of blurred vision and corneal opacity in the right eye. Pterygium surgery had been performed on the right eye 30 years earlier, and bilateral cataract surgery had been done uneventfully via a temporal corneal incision 3 years ago. Deterioration of vision occurred in the right eye from 2 years ago. At the initial visit, his best corrected visual acuity (BCVA) was 20/2000 on the right. A white nodule that was well demarcated from the underlying stroma was seen on the right cornea. The nodule was excised by superficial keratectomy, with BCVA being 180/200 at 1 week after surgery. Pathological examination of the resected specimen revealed proliferation of fibroblasts and haphazard arrangement of collagen bundles, leading to a diagnosis of corneal keloid. Keloid-like lesion was also later noted in temporal corneal incision site of cataract surgery. Conclusions and importance: This rare case of corneal keloid occurred as a late complication of pterygium surgery.

Understanding ultrafast magnetization dynamics on the microscopic level is of strong current interest due to the potential for applications in information storage. In recent years, spin-lattice coupling has been recognized as essential for ultrafast magnetization dynamics. Magnetoelectric multiferroics of type II possess intrinsic correlations among magnetic sublattices and electric polarization (P) through spin-lattice coupling, enabling fundamentally coupled dynamics between spins and lattice. Here, we report on ultrafast magnetization dynamics in a room-temperature multiferroic hexaferrite possessing ferrimagnetic (FM) and antiferromagnetic sublattices, revealed by time-resolved resonant x-ray diffraction. A femtosecond above-bandgap excitation triggers a coherent magnon in which the two magnetic sublattices entangle and give rise to a transient modulation of P. A microscopic mechanism for triggering the coherent magnon in this FM insulator based on the spin-lattice coupling is proposed. Our finding opens a general pathway for ultrafast control of magnetism.

Magnetically induced phase behaviour in a soft matter system is of potential interest for magneto-responsive compounds. Here the authors fabricate a discotic ionic liquid crystalline hybrid material which can be switched from orthorhombic to cubic phase in the absence or presence of a strong magnetic field.

tRNA-dependent cysteine biosynthesis is catalyzed by the transsulfursome protein complex. Here, the authors use a multidisciplinary approach to structurally characterize the archaeal transsulfursome and propose a model for tRNA channeling in the complex.

In this work, we use ultrafast pump-probe nonresonant and resonant x-ray diffraction to track the periodic lattice distortion and the electronic charge density wave in 1T-TiSe_{2} upon optical excitation. We observe a fluence regime in which the periodic lattice deformation is strongly suppressed but the charge density wave related Se 4p orbital order remains mostly intact. Complete melting of both structural and electronic order occurs four to five times faster than expected from a purely electronic charge-screening process, strongly suggesting a structurally assisted weakening of excitonic correlations. Our experimental data provide insight on the intricate coupling between structural and electronic order in stabilizing the periodic-lattice-distortion/charge-density-wave state in 1T-TiSe_{2}. The results further show that electron-phonon coupling can lead to different, energy dependent phase-transition pathways in condensed matter systems, opening different possibilities in the conception of nonequilibrium phenomena at the ultrafast scale.

Japanese morning glory (Ipomoea nil) has diverse flowering traits. Here, the authors describe the reference genome sequence of I. nil, annotations of genes and transposons, and compare evolution of the I. nilgenome to other Convolvulaceae and Solanales genomes.

Material properties can be controlled via strain, pressure, chemical composition, or dimensionality. Nickelates are particularly susceptible due to their strong variations of the electronic and magnetic properties on such external stimuli. Here, we analyze the photoinduced dynamics in a single crystalline NdNiO3 film upon excitation across the electronic gap. Using time-resolved reflectivity and resonant x-ray diffraction, we show that the pump pulse induces an insulator-to-metal transition, accompanied by the melting of the charge order. Finally, we compare our results with similar studies in manganites and show that the same model can be used to describe the dynamics in nickelates, hinting towards a unified description of these photoinduced electronic ordering phase transitions.

We investigate the structural dynamics of the incommensurately modulated phase of Sn 2 P 2 Se 6 by means of time-resolved X-ray diffraction following excitation by an optical pump. Tracking the incommensurable distortion in the time domain enables us to identify the transport effects leading to a complete disappearance of the incommensurate phase over the course of 100 ns. These observations suggest that a thin surface layer of the high-temperature phase forms quickly after photo-excitation and then propagates into the material with a constant velocity of 3.7 m/s. Complementary static structural measurements reveal previously unreported higher-order satellite reflection in the incommensurate phase. These higher-order reflections are attributed to cubic vibrational terms in the Hamiltonian.

Coronaviruses infect a variety of mammalian and avian species and cause serious diseases in humans, cats, mice, and birds in the form of severe acute respiratory syndrome (SARS), feline infectious peritonitis (FIP), mouse hepatitis, and avian infectious bronchitis, respectively. No effective vaccine or treatment has been developed for SARS-coronavirus or FIP virus, both of which cause lethal diseases. It has been reported that a cyclophilin inhibitor, cyclosporin A (CsA), could inhibit the replication of coronaviruses. CsA is a well-known immunosuppressive drug that binds to cellular cyclophilins to inhibit calcineurin, a calcium-calmodulin-activated serine/threonine-specific phosphatase. The inhibition of calcineurin blocks the translocation of nuclear factor of activated T cells from the cytosol into the nucleus, thus preventing the transcription of genes encoding cytokines such as interleukin-2. Cyclophilins are peptidyl-prolyl isomerases with physiological functions that have been described for many years to include chaperone and foldase activities. Also, many viruses require cyclophilins for replication; these include human immunodeficiency virus, vesicular stomatitis virus, and hepatitis C virus. However, the molecular mechanisms leading to the suppression of viral replication differ for different viruses. This review describes the suppressive effects of CsA on coronavirus replication.

Hypoxic regions within the tumor form due to imbalances between cell proliferation and angiogenesis; specifically, temporary closure or a reduced flow due to abnormal vasculature. They create environments where cancer cells acquire resistance to therapies. Therefore, the development of therapeutic approaches targeting the hypoxic cells is one of the most crucial challenges for cancer regression. Screening potential candidates for effective diagnostic modalities even under a hypoxic environment would be an important first step. In this study, we describe the development of a real-time imaging system to monitor hypoxic cell apoptosis for such screening. The imaging system is composed of a cyclic luciferase (luc) gene under the control of an improved hypoxic-responsive promoter. The cyclic luc gene product works as a caspase-3 (cas-3) monitor as it gains luc activity in response to cas-3 activation. The promoter composed of six hypoxic responsible elements and the CMV IE1 core promoter drives the effective expression of the cyclic luc gene in hypoxic conditions, enhancing hypoxic cell apoptosis visualization. We also confirmed real-time imaging of hypoxic cell apoptosis in the spheroid, which shares properties with the tumor. Thus, this constructed system could be a powerful tool for the development of effective anticancer diagnostic modalities.

Surface asperities were made on the surface of UHMWPE by sandblasting with sodium hydrogen carbonate abrasive in order to enhance the adhesion of hydrogen-containing carbon (HCC) coating to substrate. Sodium hydrogen carbonate abrasive was used because it easily solutes in hot water and the blasted abrasive stuck into the surface of UHMWPE can be entirely removed by washing with hot water. The HCC coating was deposited on the polished UHMWPE substrate and blasted substrate by RF-plasma CVD method. Friction test was conducted with the ball-on-disk tribometer in the distilled water. As the results of SEM observation and Raman spectroscopy analysis of the friction track on the HCC coating, it was confirmed that a substantial quantity of HCC coating deposited on the blasted substrate was remained on the friction track as compared with the polished substrate. It is concluded that the appropriate blasting treatment of UHMWPE substrate enhances the adhesiveness of the HCC coating to the substrate and improves wear resistance of HCC coating.

When human body is punched by the boxing glove, both of the body surface and the boxing glove deform in a complex shape. The purpose of this study is to develop a flexible sensor that can be used in such interface. Firstly, several mechanical phenomena, which are the cause of error signal of the sensor, are discussed. These are the influences of out-of-plane bending deformation, shear force caused by rubbing force, shear force caused by the Poisson's effect of contact material, and the transverse compressive force caused by the overhanging deformation of flexible material. Then as a sensor that can eliminate the error factors of these, a distribution type impact sensor in which sixteen sensor elements are arranged in a 4x4 matrix is developed. Punching experiments using a boxing glove are carried out by installing the sensor on the load cell, on the concrete wall and on the sandbag. From the experiment, it is found that the impact force can be measured with good accuracy by using the sensor. Despite the sensor has inadequate distribution number of sensor elements, the sensor structure includes mechanical requirements for the flexible impact sensor.

To investigate the effects of irbesartan on inflammation and apoptosis in atherosclerotic plaques by histochemical examination and molecular imaging using (14)C-FDG and (99m)Tc-annexin A5.Irbesartan has a peroxisome proliferator-activated receptor gamma (PPARγ) activation property in addition to its ability to block the AT1 receptor. Accordingly, irbesartan may exert further anti-inflammatory and anti-apoptotic effects in atherosclerotic plaques. However, such effects of irbesartan have not been fully investigated. Molecular imaging using (18)F-FDG and (99m)Tc-annexin A5 is useful for evaluating inflammation and apoptosis in atherosclerotic plaques.Female apoE(-/-) mice were treated with irbesartan-mixed (50 mg/kg/day) or irbesartan-free (control) diet for 12 weeks (n = 11/group). One week after the treatment, the mice were co-injected with (14)C-FDG and (99m)Tc-annexin A5, and cryostat sections of the aortic root were prepared. Histochemical examination with Movat's pentachrome (plaque size), Oil Red O (lipid deposition), Mac-2 (macrophage infiltration), and TUNEL (apoptosis) stainings were performed. Dual-tracer autoradiography was carried out to evaluate the levels of (14)C-FDG and (99m)Tc-annexin A5 in plaques (%ID×kg). In vitro experiments were performed to investigate the mechanism underlying the effects.Histological examination indicated that irbesartan treatment significantly reduced plaque size (to 56.4%±11.1% of control), intra-plaque lipid deposition (53.6%±20.2%) and macrophage infiltration (61.9%±20.8%) levels, and the number of apoptotic cells (14.5%±16.6%). (14)C-FDG (43.0%±18.6%) and (99m)Tc-annexin A5 levels (45.9%±16.8%) were also significantly reduced by irbesartan treatment. Irbesartan significantly suppressed MCP-1 mRNA expression in TNF-α stimulated THP-1 monocytes (64.8%±8.4% of un-treated cells). PPARγ activation was observed in cells treated with irbesartan (134%±36% at 3 µM to 3329%±218% at 81 µM) by a PPARγ reporter assay system.Remissions of inflammation and apoptosis as potential therapeutic effects of irbesartan on atherosclerosis were observed. The usefulness of molecular imaging using (18)F-FDG and (99m)Tc-annexin A5 for evaluating the therapeutic effects of irbesartan on atherosclerosis was also suggested.

Background Thrombocytopenia is a common problem in the management of patients with cancer and other conditions that affect hematopoietic cells. In previous clinical trials, the polyethylene-glycol-conjugated recombinant human megakaryocyte growth and development factor increased platelet counts in patients with idiopathic thrombocytopenic purpura and solid tumors undergoing chemotherapy. However, antibodies to polyethylene-glycol-conjugated recombinant human megakaryocyte growth and development factor develop in healthy volunteers and patients undergoing chemotherapy and cross-react with endogenous thrombopoietin. As a result, clinical development of polyethylene-glycol-conjugated recombinant human megakaryocyte growth and development factor was discontinued in 1998. The aim of this study was to identify an orally bioavailable human Mpl activator that does not develop autoantibodies against endogenous thrombopoietin.Design and Methods We screened our chemical library and created a novel non-peptidyl thrombopoietin receptor, Mpl activator named butyzamide. We evaluated the effect of butyzamide on megakaryopoiesis in vitro using Ba/F3 cells expressing Mpl and human hematopoietic stem cells. For the evaluation of its in vivo effect, we administered butyzamide orally to immunodeficient NOD/Shi-scid,IL-2Rγnull (NOG) mice transplanted with human fetal liver-derived CD34+ cells and investigated the production of human platelets.Results Butyzamide specifically reacted with human Mpl and activated the same signal transduction pathway as thrombopoietin. However, unlike thrombopoietin, butyzamide did not react with murine Mpl and was shown to require the histidine residue in the transmembrane domain of Mpl for its agonistic activity. Butyzamide induced colony-forming unit-megakaryocytes and polyploid megakaryocytes from human CD34+ hematopoietic progenitor cells, and its effects were comparable to those of thrombopoietin. When butyzamide was administered orally at the doses of 10 and 50 mg/kg for 20 days to NOG mice transplanted with human fetal liver-derived CD34+ cells, the human platelet count increased by 6.2- and 22.9-fold, respectively.Conclusions Butyzamide is an orally bioavailable human Mpl activator, and appears to have potential for clinical development as a therapeutic agent for patients with thrombocytopenia.

We synthesized a "cyborg protein," wherein a synthetic molecule partially substitutes the main peptide chain by linking 2 protein domains with a synthetic oligomer. Green fluorescent protein (GFP) served as the model for constructing the cyborg proteins. We prepared circularly permuted GFP (cpGFP) with new termini between β10 and β11, where the original N- and C-termini were linked by a cleavable peptide loop. The cyborg GFP was constructed from cpGFP by linking the β10 and β11 with oligo(ethylene glycol) (OEG) using maleimide-cysteine couplings, followed by the enzymatic cleavage of the N- and C-termini linking loop by thrombin. With the help of machine learning, we were able to obtain the cpGFP mutants that significantly alter the fluorescence intensity (53% increase) by thrombin treatment, which splits cpGFP into 2 fragments (fragmented GFP), and by heat shock. When the cyborg GFP was constructed using this mutant, the fluorescence intensity increased by 13% after heat treatment, similar to cpGFP (33% increase), and the behavior was significantly different from that of the fragmented GFP. This result suggests the possibility that the OEG chain in the cyborg protein plays a similar role to the peptide in the main chain of the protein. [ABSTRACT FROM AUTHOR]

The measurements of vector electric fields and currents were made with two plastic balloons of 5,000 m^3 launched from Syowa Station (L=8.4), Antarctica, on 28 November and 29 December, 1972. The results give the following. (1) The vertical electric field and current at the altitude of 30km are 0.12-0.21 V/m and 1.3-2.2 x10^ A/m^2 respectively, and their diurnal variations are roughly consistent with the expected variation from the average world-wide thunderstorm activity. (2) The electrical conductivities were estimated from the ratio of the electric current to the electric field giving the average value of 1 x 10^ mho/m at the altitude of 30 km. This value is about twice as large as that in Japan. This can be explained by the latitudinal effect of the cosmic ray ionization in the atmosphere. (3) Enhancements of the electric field and current were observed in both vertical and horizontal components of the magnitude of about 80 mV/m and 0.8 x 10^ A/m^2 respectively at an initial stage of a magnetic substorm. The vertical electric field on the ground was also correspondingly enhanced by about 80 V/m at the same time. (4) Comparisons of observed horizontal electric field vectors with the magnetic fields observed at Syowa Station show that the magnetic field variations are caused by the ionospheric Hall current. (5) One hour averages of the horizontal electric field show large scale convection fields consistent with S^p_q in the polar ionosphere and with DP-2 in the equatorial magnetosphere. The average ionospheric and magnetospheric equatorial electric fields were 40 mV/m and 0.98 mV/m respectively, and corresponding plasma convection velocities were estimated to be 700 m/s and 22 km/s respectively.