Preoperative prognostic predictors and treatment strategies for surgical procedure focused on the sequential organ failure assessment score in nonocclusive mesenteric ischemia: a multicenter retrospective cohort study.

BACKGROUND: Several large-scale studies have assessed endovascular and surgical treatment methods for nonocclusive mesenteric ischemia (NOMI); however, the prognostic factors for NOMI remain unclear. Therefore, this study aimed to evaluate risk factors for in-hospital mortality among patients with NOMI who underwent laparotomy and to examine therapeutic strategies that may improve the prognosis. MATERIALS AND METHODS: In this multicenter retrospective study, the authors reviewed the electronic medical records retrieved from the inpatient database of patients with NOMI at eight district general hospitals between January 2011 and January 2021. A total of 88 patients who underwent laparotomies were divided into survivor and nonsurvivor groups, and statistical analysis was performed to determine clinical and physiological factors. RESULTS: Exploratory laparotomy based on second-look surgery was the first treatment choice. The overall mortality rate was 48.8%, with a male-to-female ratio of 1.1:1. The median Sequential Organ Failure Assessment (SOFA) score was 8 [interquartile range: 3.75-14.2], and the median SOFA scores were 5 [3-7] in the survivor group and 13 [9-17.5] in the nonsurvivor group. Univariate analysis revealed a significant difference in BMI ( P <0.001), hypoglycemia ( P =0.0012), previous cardiovascular surgery ( P =0.0019), catecholamine use ( P <0.001), SOFA score ( P <0.001), platelet count ( P =0.0023), and lactate level ( P <0.001). Logistic regression analysis using the factors with significant differences revealed that SOFA score ≥10 (odds ratio 23.3; 95% CI: 1.94-280.00; P =0.013) was an independent prognostic factor. In addition, catecholamine use was suggested as a factor with a SOFA score greater than or equal to 10. CONCLUSION: This study confirmed that a SOFA score of greater than or equal to 10 may be associated with increased mortality. While closely monitoring low blood pressure and renal dysfunction, survival rates may be improved if surgical intervention is performed before the SOFA score reaches greater than or equal to 10.

Intrahelical Interactions in an α-Helical Coiled Coil Determine the Structural Stability of Tropomyosin.

Tropomyosin (Tpm) is a two-stranded parallel α-helical coiled-coil protein, and studying its structure is crucial for understanding the nature of coiled coils. Previously, we found that the N-terminal half of the human skeletal muscle α-Tpm (α-Tpm 140) was less structurally stable in the presence of phosphate ions than the coiled-coil protein carrier (CCPC) 140 variant with 18 mutated residues, in which all amino acid residues located at the interface between the two α-helices were completely conserved. A classical hypothesis explains that interhelical interactions stabilize the coiled-coil structure. In this study, we tested the hypothesis that the structural stability of Tpm and its variant is governed by the binding of multivalent ions that form a bridge between charged side chains located at positions b, c, and f of the heptad repeat on a single α-helical chain. We found that the structural stability of α-Tpm 140 and CCPC 140 markedly increased upon addition of divalent cations and divalent anions, respectively. We also clarified that the structural stability of the α-Tpm 140/CCPC 140 heteromeric coiled-coil molecule was governed by the stability of a less stable α-helical chain. These results demonstrated that the entire structural stability of Tpm is determined by the stability of a single α-helix. Our findings provide new insights into the study of the structure of coiled-coil proteins.

Effect of the Aspect Ratio of Coiled-Coil Protein Carriers on Cellular Uptake.

We showed previously that a rigid and fibrous-structured cationic coiled-coil artificial protein had cell-penetrating activity that was significantly greater when compared with a less-structured cell-penetrating peptide. Nanomaterials with anisotropic structures often show aspect-ratio-dependent unique physicochemical properties, as well as cell-penetrating activities. In this report, we have designed and demonstrated the cell-penetrating activity of a shorter cationic coiled-coil protein. An aspect ratio at 4.5:1 was found to be critical for ensuring that the cationic coiled-coil protein showed strong cell-penetrating activity. At an aspect ratio of 3.5:1, the cationic coiled-coil protein showed cell-penetrating activity that was similar to a less-structured short cationic cell-penetrating peptide. Interestingly, at an aspect ratio of 4:1, the cationic coiled-coil protein exhibited intermediate cell-penetrating activity. These findings should aid in the principle design of intracellular drug delivery carriers including coiled-coil artificial proteins, their derivatives, and α-helical cell-penetrating peptides as well as provide a framework for developing synthetic nanomaterials, such as metal nanorods and synthetic polymers.

Draft Genome Sequence of Brevibacillusreuszeri Strain NIT02, Isolated from a Laundered Rental Cloth Hot Towel.

Brevibacillus reuszeri is a Gram-positive spore-forming bacterium. Here, we present the draft genome sequence of Brevibacillus reuszeri strain NIT02, which was isolated from a laundered rental cloth hot towel.

Noncationic Rigid and Anisotropic Coiled-Coil Proteins Exhibit Cell-Penetration Activity.

Numerous cationic peptides that penetrate cells have been studied intensively as drug delivery system carriers for cellular delivery. However, cationic molecules tend to be cytotoxic and cause inflammation, and their stability in the blood is usually low. We have previously demonstrated that a rigid and fibrous cationic coiled-coil protein exhibited cell-penetrating ability superior to that of previously reported cell-penetrating peptides. Making use of structural properties, here we describe the cell-penetrating activity of a rigid and fibrous coiled-coil protein with a noncationic surface. A fibrous coiled-coil protein of pI 6.5 penetrated 100% of the cells tested in vitro at a concentration of 500 nM, which is comparable to that of previously reported cell-penetrating peptides. We also investigated the effect of cell-strain dependency and short-term cytotoxicity.

A novel missense mutation of RYR1 in familial idiopathic hyper CK-emia.

Persistent elevation of serum creatine kinase (CK) without any symptoms has been called idiopathic hyper CK-emia (IHCK). We examined a four-generation Japanese pedigree of familial IHCK. The multipoint linkage analysis of the pedigree showed seven clear peaks of logarithm of odds (LOD) scores (>1.4). By the exome sequencing followed by multiple filtering processes, we identified one novel heterozygous nonsynonymous single nucleotide variant (SNV), c.7034G>C, p.S2345T in the ryanodine receptor 1 gene, RYR1 cosegregated with IHCK in the pedigree. Mutation Taster predicted this substitution as "disease causing" (p=0.999). The PolyPhen-2 and PANTHER subPSEC scores for the substitution are 0.911 (possibly damaging) and -3.56 (probably damaging), respectively. We confirmed the absence of the SNV in 511 healthy Japanese individuals excluding the possibility of a normal variant with a very low frequency. Immunohistochemistry and Western blotting of biopsy samples consistently showed the expression level of RYR1 reduced in the patient. In real-time RT-PCR, the mRNA expression level of RYR1 was also significantly reduced in the patient (p=0.009). These results suggest that the novel nonsynonymous SNV contribute to the vulnerability of the RYR1 protein through the dominant negative effect. We conclude that the SNV in the RYR1 gene is one of the responsible genes of IHCK.

Development of a rapid and simple method for detection of protein contaminants in carmine.

Protein contaminants in carmine can cause dyspnea and anaphylactic reactions in users and consumers of products containing this pigment. The method generally used for detection of proteins in carmine has low reproducibility and is time-consuming. In this study, a rapid, simple, and highly reproducible method was developed for the detection of protein contaminants in carmine. This method incorporates acidic protein denaturation conditions and ultrafiltration. To prevent protein aggregation, sodium dodecyl sulfate containing gel electrophoresis running buffer was used for dispersing the carmine before filtration. An ultrafiltration device was used to separate the protein contaminants from carminic acid in the carmine solution. Two ultrafiltration devices were compared, and a cylindrical device containing a modified polyethersulfone membrane gave the best results. The method had high reproducibility.

Superior cell penetration by a rigid and anisotropic synthetic protein.

Molecules with structural anisotropy and rigidity, such as asbestos, demonstrate high cell-penetrating activity but also high toxicity. Here we synthesize a biodegradable, rigid, and fibrous artificial protein, CCPC 140, as a potential vehicle for cellular delivery. CCPC 140 penetrated 100% of cells tested in vitro, even at a concentration of 3.1 nM-superior to previously reported cell-penetrating peptides. The effects of cell-strain-dependency and aspect ratio on the cell-penetrating activity of CCPC 140 were also investigated.

Nonvolatile flash memory based on biologically integrated hierarchical nanostructures.

The first six peptides of multifunctional titanium binding peptide-1 bestowed recombinant L-ferritin, minT1-LF, was genetically engineered and used to fabricate multilayered nanoparticle architecture. The multifunctionality of minT1-LF enables specific binding of nanoparticle-accommodated minT1-LF to the silicon substrate surface and wet biochemical fabrication of gate oxide layer by its biomineralization activity. Three-dimensional (3D) nanoparticle architecture with multilayered structure was fabricated by the biological layer-by-layer method and embedded in a metal oxide-semiconductor device structure as a charge storage node of a flash memory device. The 3D-integrated multilayered nanoparticle architecture successfully worked as a charge storage node in flash memory devices that exhibited improved charge storage capacity compared with that of a conventional monolayer structure device.

Hydrophilic Double-Network Polymers that Sustain High Mechanical Modulus under 80% Humidity.

The effects of water on the mechanical properties of synthetic hydrophilic polymers with double-network (DN) structures were studied under different relative humidities (RH). It was found that they could sustain nearly the same high Young's modulus as dry DN polymers in the RH range 10-80% (water content 3-17 wt %), that is, more than 102 MPa. However, when the RH exceeds 80%, DN polymers abruptly absorb large amounts of water (water content 90 wt %) and transform to a highly water-swollen "gel state" with a decrease in the Young's modulus of 3 orders of magnitude. Spectroscopic analyses revealed that water molecules below RH 80% are strongly bound to hydrophilic moieties with highly restricted mobility; water under such states improves rather than reduces mechanical properties by behaving as a plasticizer. DN polymers capable of sustaining high mechanical properties, even under RH 80%, have potential uses as hydrophilic materials.

Self-repairing filamentous actin hydrogel with hierarchical structure.

A chemically cross-linked filamentous actin (F-actin) gel consisting of globular actin (G-actin) as repeating units was prepared. The F-actin gel was cross-linked by covalent bonds, and the main chain is represented by the self-assembly of G-actin with a high-ordered hierarchical structure. The gel exhibited good mechanical performance with a storage modulus >1 kPa and undergoes reversible sol-gel transitions in response to changes in the salt concentration (chemical-induced sol-gel transition) as well as to shear strain (mechanical-induced sol-gel transition). Therefore, the gel exhibits self-repairing ability through dynamic polymerization and depolymerization across the structure hierarchies under repeated shear stress.

Thermoresponsive microtubule hydrogel with high hierarchical structure.

A thermoresponsive 3D microtubule hydrogel (MT gel) was prepared by simultaneous polymerization and chemical cross-linking of tubulins. The main chain of this gel is composed of cross-linked MTs, which consists of a cylindrical assembly of tubulin covalently connected by polyethylene glycol. This gel, which contains 10 mg/mL of tubulin, exhibits a storage modulus G' as high as 1 × 10(3), which is 10 times higher than the loss modulus G'' over a wide range of frequencies. The MT gel exhibits a reversible sol-gel transition by temperature changes at 4-37 °C via depolymerization and polymerization of the MT network. Notable effects of the presence of the cross-linkage on the process of polymerization and depolymerization of tubulin were experimentally observed, and the role of the cross-linkage was discussed.

Autonomous silica encapsulation and sustained release of anticancer protein.

We present a novel method for preparing a silica carrier for the sustained release of a proteinaceous pharmaceutical. This method makes use of the silicification activity of the protein itself, which autonomously formed a protein-silica composite upon simple incubation with a silica precursor. The composite was dissolved, and the encapsulated protein was released into a culture medium, thereby sustaining the protein's activity for a long period of time.

Critical amino acid residues for the specific binding of the Ti-recognizing recombinant ferritin with oxide surfaces of titanium and silicon.

The interactions of ferritins fused with a Ti-recognizing peptide (RKLPDA) and their mutants with titanium oxide substrates were explored with an atomic force microscope (AFM). The amino acid sequence of the peptide was systematically modified to elucidate the role of each amino acid residue in the specific interaction. Force measurements revealed a clear correlation among the sequences in the N-terminal domain of ferritin, surface potentials, and long-range electrostatic interactions. Measurements of adhesion forces clearly revealed that hydrogen bonds take part in the specific binding as well as the electrostatic interaction between charged residues and surface charges of Ti oxides. Moreover, our results indicated that not only the charged and polar residues but also a neutral residue (proline) govern the strength of the specific binding, with the order of the residues also being significant. These results demonstrate that the local structure of the peptide governs the special arrangement of charged residues and strongly affects the strength of the bindings.

In aqua structuralization of a three-dimensional configuration using biomolecules.

Ferritin nanoparticles ornamented with a Ti-binding peptide are versatile nanoscaled building blocks. Their specific binding ability is strong enough to position them on nanopatterned Ti regions on a Pt substrate. Furthermore, the peptides mineralization activity enables the formation of titania on the outer side of the particle, and the particle's inner nanospaces can serve as a carrier for inorganic nanodots. Making use of all these properties, here we show controlled in aqua fabrication of three-dimensional nanoscale structures. The X-Y positioning obeyed the specific binding of the peptide, while fabrication in the Z-dimension entailed stepwise formation of titania and ferritin layers by alternately applying the binding and mineralization abilities of the Ti-binding peptide. This method paves the way for in aqua fabrication of nanodevices having complicated structures and functions.

Realizing a two-dimensional ordered array of ferritin molecules directly on a solid surface utilizing carbonaceous material affinity peptides.

A two-dimensional hexagonally close-packed (2D-HCP) array of ferritin molecules with a nanoparticle core was fabricated directly on a carbonaceous solid substrate by genetically modifying the outer surface of the ferritin with carbonaceous materials-specific binding peptides. The displayed peptides endowed ferritins with a specific protein-substrate interaction and masked the strong nonspecific interaction. The specific interaction was weak enough to allow ferritins to be rearranged as well as an attractive protein-protein interaction that could be adjusted by selecting the buffer conditions. This method not only produced 2D-HCP arrays of ferritin but also 2D-ordered arrays of independent inorganic nanoparticles after protein elimination that can be applied to floating gate memories.

Conversion of a monodispersed globular protein into an amyloid-like filament by appending an artificial peptide at the N-terminal.

The soluble, globular, alpha-helix-rich peptide SipA(446-684) is a domain of a bacterial protein that binds to mammalian filamentous-actin and re-arranges the host cell's cytoskeleton. We show that adding two copies of NHBP-1, a carbon nanomaterial binding peptide, to its N-terminal can induce SipA(446-684) to polymerize and assume a fibrillar structure under physiological conditions. The fibrils formed showed thioflavine T and Congo red staining profiles that are characteristic of and specific for amyloid-like structures. The alpha-helical structure of the globular protein was retained in the fibrils, suggesting the appended NHBP-1 sequence plays a key role in the formation of cross-beta spines within the fibrils. Consistent with that idea, we observed that a synthetic NHBP-1 peptide can form an amyloid-like structure under appropriate conditions. Thus, our findings add a new subtype of amyloid-like structure formation and suggest this method of assembly could be exploited in nano-biotechnology.

Mechanism underlying specificity of proteins targeting inorganic materials.

Adhesion force analysis using atomic force microscopy clearly revealed for the first time the mechanism underlying the specific binding between a titanium surface and ferritin possessing the sequence of Ti-binding peptide in its N-terminal domain. Our results proved that the specific binding is due to double electrostatic bonds between charged residue and surface groups of the substrate. Furthermore, it is also demonstrated that the accretion of surfactant reduces nonspecific interactions, dramatically enhancing the selectivity and specificity of Ti-binding peptide.

Utilization of the pleiotropy of a peptidic aptamer to fabricate heterogeneous nanodot-containing multilayer nanostructures.

Peptide aptamers (=binders) against inorganic materials often show a capacity for mineralization of their target atoms; thus they are able to function both as binding molecules and as mediators for mineralization. Although the mechanisms underlying these two properties of peptide aptamers are not yet fully understood, they have been used separately to fabricate various nanostructures. Here, we present a novel method of nanofabrication, in which binding and mineralization by a peptide aptamer are alternately utilized to assemble multilayered nanostructures comprised of metal loaded cage proteins ornamented with Ti-binding peptides.