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Abstract Cisplatin is an effective chemotherapeutic agent widely used for the treatment of various solid tumors. However, cisplatin has an important limitation in its use; currently, there is no method to ameliorate cisplatin-induced acute kidney injury (AKI). Thrombomodulin (TM) is well known not only for its role as a cofactor in the clinically important natural anticoagulation pathway but also for its anti-inflammatory properties. Here, we investigated the effects of TM in cisplatin-induced AKI. In mice intraperitoneally injected with 15 mg/kg cisplatin, TM (10 mg/kg) or PBS was administered intravenously at 24 h after cisplatin injection. TM significantly attenuated cisplatin-induced nephrotoxicity with the suppressed elevation of blood urea nitrogen and serum creatinine, and reduced histological damages. Actually, TM treatment significantly alleviated oxidative stress-induced apoptosis by reducing reactive oxygen species (ROS) levels in cisplatin-treated renal proximal tubular epithelial cells (RPTECs) in vitro. Furthermore, TM clarified cisplatin-induced apoptosis by reducing caspase-3 levels. In addition, TM attenuated the endoplasmic reticulum (ER) stress signaling pathway in both renal tissues and RPTECs to protect the kidneys from cisplatin-induced AKI. These findings suggest that TM is a potential protectant against cisplatin-induced nephrotoxicity through suppressing ROS generation and ER stress in response to cisplatin.

Abstract When exposed to oxidative and electrophilic stress, a protective antioxidant response is initiated by nuclear factor erythroid 2-related factor 2 (Nrf2). However, the extent of its importance in the forensic diagnosis of acute ischemic heart diseases (AIHD), such as myocardial infarction (MI), remains uncertain. On the other hand, immunohistochemical analyses of fibronectin (FN) and the terminal complement complex (C5b-9) prove valuable in identifying myocardial ischemia that precedes necrosis during the postmortem diagnosis of sudden cardiac death (SCD). In this study, we investigated the immunohistochemical levels of Nrf2, FN, and C5b-9 in human cardiac samples to explore their forensic relevance for the identification of acute cardiac ischemia. Heart samples were obtained from 25 AIHD cases and 39 non-AIHD cases as controls. Nrf2 was localized in the nuclei of cardiomyocytes, while FN and C5b-9 were detected in the myocardial cytoplasm. The number of intranuclear Nrf2 positive signals in cardiomyocytes increased in AIHD cases compared to control cases. Additionally, the grading of positive portions of cardiac FN and C5b-9 in the myocardium was also significantly enhanced in AIHD, compared to controls. Collectively, these results indicate that the immunohistochemical investigation of Nrf2 combined with FN, and/or C5b-9 holds the potential for identifying early-stage myocardial ischemic lesions in cases of SCD.

Abstract Heat shock proteins (HSPs) are molecular chaperones whose primary function is cytoprotection, supporting cell survival under (sub) lethal conditions. They have been implicated in various diseases such as inflammatory diseases and cancer due to their cytoprotective and immunomodulatory effects, and their biological mechanisms have been studied. Central family members include, HSP27, which is induced by various stimuli such as heat shock, hypoxia, hyperoxia, ultraviolet exposure, and nutritional deficiency, and HSP70, which is homeostatically expressed in many organs such as the gastrointestinal tract and has anti-cell death and anti-inflammatory effects. In this study, HSP27 and HSP70 were investigated during thrombus formation and dissolution in a deep vein thrombosis model by immunohistochemistry to determine their involvement in this process and whether their expression could be used as a forensic marker. In the process of thrombus formation and lysis, HSP27 and HSP70 were found to be expressed by immunohistochemical analysis. The role of inhibitors of HSP27 and HSP70 in the pathogenesis of thrombosis in mice was also investigated. When HSP27 or HSP70 inhibitors were administered, thrombi were significantly smaller than in the control group on day 5 after inferior vena cava ligation, indicating pro-thrombotic effects HSP27 and HSP70. If HSP27- or HSP70-positive cells were clearly visible and easily identifiable in the thrombus sections, the thrombus was presumed to be more than 10 days old. Thus, the detection of intrathrombotic HSP27 and HSP70 could forensically provide useful information for the estimation of thrombus ages. Collectively, our study implied that both HSP27 and HSP70 might be molecular targets for thrombus therapy and that the detection of HSP-related molecules such as HSP27 and HSP70 could be useful for the determination of thrombus ages.

Inborn errors of bile acid metabolism (IEBAM) cause cholestasis during the neonatal period, and 8 types of IEBAM have been reported to date. IEBAM accounts for approximately 2% of cases of cholestasis of unknown cause. As only 10 patients have been identified in Japan, IEBAM presents diagnostic challenges due to the similarity of clinical symptoms with biliary atresia, thus necessitating precise differentiation to avoid unnecessary invasive procedures. Laboratory tests in IEBAM are characterized by normal γ-glutamyltransferase (GGT) and serum total bile acid (STBA) levels despite the presence of cholestasis; therefore, measuring STBA and GGT is essential to distinguishing biliary atresia from IEBAM. With suspected IEBAM, liquid chromatography–mass spectrometry (LC/MS) analysis of urinary bile acids is needed to optimize diagnostic and therapeutic efficacy and avoid open cholangiography and initiate treatment for primary bile acids such as cholic acid or chenodeoxycholic acid. This prospective report aims to increase awareness of IEBAM by highlighting the characteristics of general blood test and bile acid profiles from LC/MS analyses of blood, urine, and stool samples.

FeCrAl ODS steel is the promising candidate cladding material for the accident tolerant fuel (ATF) of the light water reactors (LWR), due to the formation of Al2O3 in alloys which could suppress the oxidation reaction with hot steam. In the present work, change in microstructures and the corresponding irradiation hardening of two kinds of FeCrAl ODS alloys (15.5Cr-4Al), with 0.6 wt% Zr (denoted as P1) or 0.8 wt% Hf addition (denoted as P2), were investigated. Electron backscattered diffraction (EBSD) and transmitted electron microscopy (TEM) were used to characterize grains and dispersion morphology of oxides, respectively. Compared to P1, P2 has finer and denser oxides. 56Fe17+ ions with kinetic energy of 352.8 MeV were used to produce a quasi-uniformly distributed damage region from the near-surface to a depth of 24 μm in the specimens by using an energy degrader. A damage level of 0.8 dpa (displacement per atom) was attained in the specimens of both alloys simultaneously at 200℃. The Vickers hardness test shows that both specimens exhibited an apparent irradiation hardening. The extent of hardening of P2 (6 %) is significantly lower than P1 (12 %). TEM results show that irradiation-induced dislocation loops occur in both specimens but with a relatively smaller average size in P2. The results demonstrate that compared to the 0.6 wt% Zr addition, the 0.8 wt% Hf addition improves more significantly the hardening resistance, which was discussed in terms of the calculated sink strength originating from the oxides/matrix interface.

Abstract Estimating the age and vitality of human skin wounds is essential in forensic practice, and the use of immunohistochemical parameters in this regard remains a challenge. Heat shock proteins (HSPs) are evolutionarily conserved universal proteins that protect biological systems from various types of stress. However, its importance in forensic pathology for determining wound activation in neck compression skin remains unclear. The expression of HSP27 and HSP70 in neck skin samples was immunohistochemically examined to understand its forensic applicability in determining wound vitality. Skin samples were obtained from 45 cases of neck compression (hanging, 32 cases; strangulation, 10 cases; manual strangulation, 2 cases; other, 1 case) during forensic autopsies; intact skin from the same individual was used as a control. HSP27 expression was detected in 17.4% of keratinocytes in the intact skin samples. In the compressed region, the frequency of HSP27 expression in keratinocytes was 75.8%, which was significantly higher than that in intact skin. Similarly, HSP70 expression was 24.8% in intact skin samples and 81.9% in compressed skin samples, significantly higher in compressed skin than in intact skin samples. This increase in case compression cases may be due to the cell defence role of HSPs. From a forensic pathology perspective, the immunohistochemical examination of HSP27 and HSP70 expression in neck skin could be considered a valuable marker for diagnosing traces of antemortem compression.

Abstract Estimating time of death is one of the most important problems in forensics. Here, we evaluated the applicability, limitations and reliability of the developed biological clock-based method. We analyzed the expression of the clock genes, BMAL1 and NR1D1, in 318 dead hearts with defined time of death by real-time RT-PCR. For estimating the time of death, we chose two parameters, the NR1D1/BMAL1 ratio and BMAL1/NR1D1 ratio for morning and evening deaths, respectively. The NR1D1/BMAL1 ratio was significantly higher in morning deaths and the BMAL1/NR1D1 ratio was significantly higher in evening deaths. Sex, age, postmortem interval, and most causes of death had no significant effect on the two parameters, except for infants and the elderly, and severe brain injury. Although our method may not work in all cases, our method is useful for forensic practice in that it complements classical methods that are strongly influenced by the environment in which the corpse is placed. However, this method should be applied with caution in infants, the elderly, and patients with severe brain injury.

Abstract We investigated the dynamics of the gene expression of M1 and M2 macrophage markers during skin wound healing in mice. Expression of M1-macrophage markers, such as Il12a, Tnf, Il6, Il1b, and Nos2 was upregulated after wounding and peaked at 1 or 3 days after injury, and that of M2-macrophage markers such as Mrc1, Cd163, Ccl17, Arg, and Tgfb1, peaked at 6 days after injury. Consistent with these findings, using triple-color immunofluorescence analysis revealed that F4/80+CD80+ M1 macrophages were more abundant than F4/80+CD206+ M2 macrophages on day 3 in mouse wound specimens, and that M2 macrophages were prominently detected in day 6 wounds. For application in forensic practice, we examined macrophage polarization using human wound specimens. The average ratios of CD68+iNOS+ M1 macrophages to CD68+CD163+ M2 macrophages (M1/M2 ratios) were greater than 2.5 for the wounds aged 2–5 days. Out of 11 wounds aged 1–5 days, five samples had the M1/M2 ratios of > 3.0. These observations propose that the M1/M2 ratios of 3.0 would indicate a wound age of 1–5 days as the forensic opinion. This study showed that M1 and M2 macrophages in human skin wound might be a promising marker for wound age determination.

Abstract Heme oxygenase-1 (HO-1), an inducible stress-response protein, exerts anti-oxidant and anti-apoptotic effects. However, its significance in forensic diagnosis of acute ischemic heart diseases (AIHD) such as myocardial infarction (MI) is still unknown. We examined the immunohistochemical expression of HO-1 in the heart samples to discuss their forensic significance to determine acute cardiac ischemia. The heart samples were obtained from 23 AIHD cases and 33 non-AIHD cases as controls. HO-1 positive signals in cardiomyocyte nuclear were detected in 78.2% of AIHD cases, however, that were detected in only 24.2% control cases with statistical difference between AIHD and non-AIHD groups. In contrast to HO-1 protein expression, there was no significant difference in the appearance of myoglobin pallor regions and leukocyte infiltration in the hearts between AIHD and non-AIHD groups. From the viewpoints of forensic pathology, intracardiac HO-1 expression would be considered a valuable marker to diagnose AIHD as the cause of death.

Organic anion transporting polypeptide (OATP) 1B1 (gene, solute carrier organic anion transporter family member 1B1 [SLCO1B1]) and OATP1B3 (SLCO1B3) serve as transporters for hepatic uptake of important endogenous substances and several commonly prescribed drugs. Inactivation of both proteins together causes Rotor syndrome. How this OATP1B1/1B3 defect disturbs bile acid (BA) metabolism is largely unknown. In this study, we performed detailed BA analysis in 3 patients with genetically diagnosed Rotor syndrome. We found that BAs glucuronidated at the C‐3 position (BA‐3G) accounted for 50% or more of total BAs in these patients. In contrast but similarly to healthy controls, only trace amounts of BA‐3G were detected in patients with constitutional indocyanine green excretory defect (OATP1B3 deficiency) or sodium‐taurocholate cotransporting polypeptide (NTCP; gene, solute carrier family 10 member 1 [SLC10A1]) deficiency. Therefore, substantial amounts of BA‐3G are synthesized in hepatocytes. The cycling pathway of BA‐3G, consisting of excretion from upstream hepatocytes and uptake by downstream hepatocytes by OATP1B1/1B3 may exist to reduce the burden on upstream hepatocytes. Conclusion: Detailed BA analysis revealed glucuronidated bile acidemia in patients with Rotor syndrome. Further exploration of the physiologic role of glucuronidated BAs is necessary.

Abstract Diagnosis of biliary atresia (BA) can involve uncertainties. In the present prospective multicenter study, we considered whether urinary oxysterols represent a useful marker for diagnosis of BA in Japanese children. Subjects under 6 months old at 7 pediatric centers in Japan were prospectively enrolled, including patients with cholestasis and healthy controls (HC) without liver disease. Patients with cholestasis constituted 2 groups representing BA patients and others with cholestasis from other causes (non-BA). We quantitatively analyzed 7 oxysterols including 4β-, 20(S)-, 22(S)-, 22(R)-, 24(S)-, 25-, and 27-hydroxycholesterol by liquid chromatography/electrospray ionization-tandem mass spectrometry. Enrolled subjects included 14 with BA (median age 68 days; range 26–170) and 10 non-BA cholestatic controls (59; 14–162), as well as 10 HC (57; 25–120). Total urinary oxysterols were significantly greater in BA (median, 153.0 μmol/mol creatinine; range 24.1–486.7; P

Intense neutrophil infiltration into the liver is a characteristic of acetaminophen-induced acute liver injury. Neutrophil elastase is released by neutrophils during inflammation. To elucidate the involvement of neutrophil elastase in acetaminophen-induced liver injury, we investigated the efficacy of a potent and specific neutrophil elastase inhibitor, sivelestat, in mice with acetaminophen-induced acute liver injury. Intraperitoneal administration of 750 mg/kg of acetaminophen caused severe liver damage, such as elevated serum transaminase levels, centrilobular hepatic necrosis, and neutrophil infiltration, with approximately 50% mortality in BALB/c mice within 48 h of administration. However, in mice treated with sivelestat 30 min after the acetaminophen challenge, all mice survived, with reduced serum transaminase elevation and diminished hepatic necrosis. In addition, mice treated with sivelestat had reduced NOS-II expression and hepatic neutrophil infiltration after the acetaminophen challenge. Furthermore, treatment with sivelestat at 3 h after the acetaminophen challenge significantly improved survival. These findings indicate a new clinical application for sivelestat in the treatment of acetaminophen-induced liver failure through mechanisms involving the regulation of neutrophil migration and NO production.

Ubiquitin is a member of the heat shock protein family and is rapidly induced by various types of stimuli, including ischemic and mechanical stress. However, its significance in determining wound vitality of neck compression skin in forensic pathology remains unclear. We immunohistochemically examined the expression of ubiquitin in the neck skin samples to understand its forensic applicability in determining wound vitality. Skin samples were obtained from 53 cases of neck compression (hanging, 42 cases; strangulation, 11 cases) during forensic autopsies. Intact skin from the same individual was used as the control. Ubiquitin expression was detected in 73.9% of keratinocytes in intact skin samples, but only in 21.2% of keratinocytes in the compression regions, with statistical differences between the control and compression groups. This depletion in the case of neck compression may be caused by the impaired conversion of conjugated to free ubiquitin and failure of de novo ubiquitin synthesis. From a forensic pathological perspective, immunohistochemical examination of ubiquitin expression in the skin of the neck can be regarded as a valuable marker for diagnosing traces of antemortem compression.

Inflammatory cytokines and chemokines are involved in the development of abdominal aortic aneurysm (AAA). Here the authors show that CCL3 prevents the development of CaCl2-induced AAA by suppressing MMP-9 expression.

Background: Inborn errors of bile acid metabolism (IEBAM) cause rare but treatable genetic disorders that can present as neonatal cholestasis or neurological diseases. Without timely primary bile acid treatment, patients may develop liver failure early in life. This study aimed to analyze the types and treatment outcomes of IEBAM in Taiwanese infants and document the allele frequency of CYP7B1 hot spot mutations in the population. Methods: Urine samples from patients with infantile intrahepatic cholestasis and suspected IEBAM were subjected to urinary bile acid analysis by gas chromatography-mass spectrometry (GC/MS). Genetic diagnoses were made using direct sequencing or next-generation sequencing. We also tested healthy control subjects for a probable hot spot point mutation of CYP7B1. Results: Among the 75 patients with infantile intrahepatic cholestasis tested during 2000 –2016, three had ∆4-3-oxosteroid 5β-reductase deficiency with AKR1D1 mutations, and three had oxysterol-7α-hydroxylase deficiency with CYP7B1 mutation. Two patients with ∆4-3-oxosteroid 5β-reductase deficiency were successfully treated with cholic acid. The three unrelated infants with oxysterol 7α-hydroxylase deficiencies had the same p.R112X homozygous CYP7B1 mutation. Two had mild renal or neurological involvement. Among 608 healthy control subjects, the allele frequency of the heterozygous mutation for p.R112X was 2/1216 (0.16%). The only surviving patient with oxysterol 7α-hydroxylase deficiency recovered from liver failure after chenodeoxycholic acid (CDCA) treatment beginning at 3 months of age. Conclusion: Distinct types of IEBAM disease were found in the Taiwanese population. Patients with early diagnosis and early treatment had a favorable outcome. IEBAM prevalence rates may be higher than expected due to the presence of heterozygous mutations in the general population. Key Words: chenodeoxycholic acid, cholic acid, inborn errors of bile acid metabolism, neonatal cholestasis, oxysterol 7α-hydroxylase deficiency

After the ligation of the inferior vena cava (IVC) of wild-type (WT) mice, venous thrombi formed and grew progressively until 5 days and resolved thereafter. Concomitantly, intrathrombotic gene expression of Il6 was enhanced later than 5 days after IVC ligation. IL-6 protein expression was detected mainly in F4/80-positive macrophages in thrombus. When Il6-deficient (Il6−/−) mice were treated in the same manner, thrombus mass was significantly larger than in WT mice. Moreover, the recovery of thrombosed IVC blood flow was markedly delayed in Il6−/− compared with WT mice. F4/80-positive macrophages in thrombus expressed proteolytic enzymes such as matrix metalloproteinase (Mmp) 2, Mmp9, and urokinase-type plasminogen activator (Plau); and their mRNA expression was significantly reduced in Il6−/− mice. Consistently, the administration of anti-IL-6 antibody delayed the thrombus resolution in WT mice, whereas IL-6 administration accelerated thrombus resolution in WT and Il6−/− mice. Moreover, IL-6 in vitro enhanced Mmp2, Mmp9, and Plau mRNA expression in WT-derived peritoneal macrophages in a dose-dependent manner; and the enhancement was abrogated by a specific Stat3 inhibitor, Stattic. Thus, IL-6/Stat3 signaling pathway can promote thrombus resolution by enhancing Mmp2, Mmp9, and Plau expression in macrophages.

A mechanically alloyed austenitic stainless steel (MA304LZ) was produced from pre-alloyed SUS304L powder with a small amount of Zr addition. The yield stress of MA304LZ was more than 3 times larger than that of SUS304L or 316L, while total elongation was reduced to about one third of the conventional steels. Microstructure analysis revealed an average grain size of 0.42 µm in MA304LZ and about 34/30 µm in SUS304L/316 L. In MA304LZ, two types of precipitates were observed; inhomogeneously distributed fine precipitates with an average size of 6.0 nm and homogeneously distributed coarse precipitates (d > 20 nm) with an average size of 47 nm. The strengthening mechanism of MA304LZ was discussed on the bases of Hall-Petch and Orowan equations, and the strengthening of MA304LZ was attributed mostly to refined grains. The dislocation barrier strength factor, α, is estimated to be 0.277 for the Zr-rich precipitates in MA304LZ. Keywords: Powder metallurgy, Grain size effect, Precipitation hardening, Zr-oxides

15Cr-ODS ferritic steel was implanted with 3500 appm He (0.2 dpa) at 300, 550 and 700 °C. The post-implantation annealing (PIA) at 800 °C/100 h was conducted on the specimen implanted at 300 °C. Nanoindentation (NI) tests were carried out to investigate the temperature dependent hardening, and transmission electron microscopy (TEM) observation was performed to characterize the cavity evolution. A limited hardening was found in the as-implanted ODS specimens at all implantation temperatures, which tended to reduce with increasing the temperature. The PIA caused no further hardening, while TEM images revealed that there was an evident change in the cavity distribution morphology. The cavity diameter increased from 2.5 ± 0.4 nm to 5.0 ± 1.7 nm and the corresponding number density decreased from (22.2 ± 1.6) × 1022 m−3 to (6.7 ± 0.5) × 1022 m−3 before and after the PIA. The negligible hardness changes induced by the PIA were interpreted in terms of the Orowan-type dislocation barrier model with the corresponding barrier strength factor of cavities to be less than 0.1. Keywords: Cavity evolution, Nanoindentation hardness, ODS steel, He implantation, Annealing

The stress corrosion cracking (SCC) susceptibility of solution-annealed 310S stainless steel (SS) was examined by means of slow-strain rate test (SSRT) method in hot water (288 °C) with dissolved-hydrogen (DH), and the results were compared with those of 316L SS. Almost no SCC was observed in 310S SS, while a remarkable SCC was found in 316L SS showing brittle fracture surfaces and cracks on specimen side surfaces. The SCC observed in 316L SS started as IGSCC while necking, and transferred to TGSCC. It was also shown that sensitization resulted in the suppression of IGSCC of 316L SS. On the assumption that the SCC in hydrogenated hot water can be interpreted in terms of hydrogen assisted SCC, the mechanism of the suppression of SCC in 310S SS is related with a higher nickel content which increases stacking fault energy and facilitates cross slip of dislocations, resulting in the reduction of hydrogen concentration on a unit area of slip plane. Keywords: SSRT, Hydrogen assisted SCC, Cross slip, Stacking fault energy

Tetrahydroxy bile acids (THBAs) are hydrophilic and are present at minimal or undetectable levels in healthy human adults, but are present at high levels in bile salt export pump (abcb11)-knockout mice. The roles of THBAs in human cholestatic diseases are unclear. We aimed to investigate the presence of THBAs in patients with infantile intrahepatic cholestasis and its correlation with outcome. Urinary bile acids (BAs) were analyzed by GC-MS. Data were compared between good (n = 21) (disease-free before 1 year old) and poor prognosis groups (n = 19). Good prognosis patients had a higher urinary THBA proportion than poor prognosis patients [25.89% (3.45–76.73%) vs. 1.93% (0.05–48.90%)]. A urinary THBA proportion >7.23% predicted good prognosis with high sensitivity (95.24%), specificity (84.21%), and area under the curve (0.91) (P < 0.0001). A THBA proportion ≤7.23% was an independent factor for decreased transplant-free survival (hazard ratio = 7.16, confidence interval: 1.24–41.31, P = 0.028). Patients with a confirmed ABCB11 or tight junction protein 2 gene mutation (n = 7) had a minimally detectable THBA proportion (0.23–2.99% of total BAs). Three patients with an ATP8B1 mutation had an elevated THBA proportion (7.51–37.26%). In conclusion, in addition to disease entity as a major determinant of outcome, a high THBA level was associated with good outcome in the infantile intrahepatic cholestasis patients.

As-received and recrystallized pure tungsten (W) were irradiated with 6.4MeV Fe3+ up to 2dpa with or without He+ at 300°C, 500°C, 700°C and 1000°C respectively. Irradiation hardening was measured by the nano-indentation method. An equation to evaluate the bulk equivalent hardness was derived on the assumption that the geometrically necessary dislocation (GND) densities at an indentation depth were the same before and after irradiation. Ion-irradiation always induces hardening in both as-received and recrystallized W at all the experiment temperatures. In the case of single-beam irradiation, the recrystallized W exhibited higher hardening than as-received one. The effect of helium on the irradiation hardening is dependent on the material condition: as-received W showed an additional hardening by helium at all the irradiation temperatures, while in recrystallized W the hardening was not affected by helium below 700°C.

The age-hardening in oxide dispersion strengthened (ODS) ferritic steels with various additions of Cr (12, 15 and 18wt.%) and Al (0, 5, 7 and 9wt.%) were investigated. After 5000h aging at 475ºC, the hardness increases in all these ODS steels, while the increased level depends on the Cr/Al contents. In 12Cr-ODS steels, the more the Al, the higher the increased hardness is. However, in 18Cr-ODS steels, higher Al addition suppresses the age-hardening. TEM observations of 18Cr-ODS steels reveal that 9Al suppresses the formation of Cr-enriched α' phase, while the 18Cr-5Al-ODS steel comprises a plenty of α′ phases. Adding Zr in ODS steels appears to increase the age-hardening. The susceptibility to age-hardening is remarkably lower in the ODS ferritic steels than in the non-ODS ferritic steel with the similar concentration of Cr.

Aortic dissection and aneurysm are associated with abnormal hemodynamic loads originating from hypertension. Our previous study demonstrated that cyclic mechanical stretch (CMS, mimicked hypertension) caused the death of rat aortic smooth muscle cells (RASMCs) in a mitogen activated-protein kinases (MAPKs)-dependent manner. The current study investigated the effects of inducible nitric oxide synthase (iNOS) on CMS-induced RASMC death. cDNA microarrays for CMS-treated RASMCs showed that iNOS expression levels were increased in response to CMS. Real-time polymerase chain reaction (PCR) analysis demonstrated that this increase was p38 MAPK (p38)-dependent. NO production was also increased. This increase could be inhibited by p38 and iNOS inhibitors. Thus, CMS-induced iNOS synthesized NO. CMS-induced cell death in RASMCs was increased by the iNOS inhibitor but abrogated by the long-acting NO donor DETA-NONOate. Increased iNOS expression was confirmed in the abdominal aortic constriction mouse model. Signal transducers and activators of transcription 1 (STAT1) was activated in stretched RASMCs, and iNOS expression and NO production were inhibited by the STAT1 inhibitor nifuroxazide. Our findings suggest that RASMCs were protected by iNOS from CMS-stimulated cell death through the STAT1 and p38 signal pathways independently.

BackgroundA clear understanding of the molecular mechanisms underlying hemodynamic stress‐initiated cardiac hypertrophy is important for preventing heart failure. Interferon‐γ (IFN‐γ) has been suggested to play crucial roles in various diseases other than immunological disorders by modulating the expression of myriad genes. However, the involvement of IFN‐γ in the pathogenesis of cardiac hypertrophy still remains unclear. Methods and ResultsIn order to elucidate the roles of IFN‐γ in pressure overload–induced cardiac pathology, we subjected Balb/c wild‐type (WT) or IFN‐γ‐deficient (Ifng−/−) mice to transverse aortic constriction (TAC). Three weeks after TAC, Ifng−/− mice developed more severe cardiac hypertrophy, fibrosis, and dysfunction than WT mice. Bone marrow–derived immune cells including macrophages were a source of IFN‐γ in hearts after TAC. The activation of PI3K/Akt signaling, a key signaling pathway in compensatory hypertrophy, was detected 3 days after TAC in the left ventricles of WT mice and was markedly attenuated in Ifng−/− mice. The administration of a neutralizing anti‐IFN‐γ antibody abrogated PI3K/Akt signal activation in WT mice during compensatory hypertrophy, while that of IFN‐γ activated PI3K/Akt signaling in Ifng−/− mice. TAC also induced the phosphorylation of Stat5, but not Stat1 in the left ventricles of WT mice 3 days after TAC. Furthermore, IFN‐γ induced Stat5 and Akt phosphorylation in rat cardiomyocytes cultured under stretch conditions. A Stat5 inhibitor significantly suppressed PI3K/Akt signaling activation in the left ventricles of WT mice, and aggravated pressure overload–induced cardiac hypertrophy. ConclusionsThe IFN‐γ/Stat5 axis may be protective against persistent pressure overload–induced cardiac hypertrophy by activating the PI3K/Akt pathway.

Preterm labor (PTL) is the most common cause of neonatal death and long-term adverse outcome. The pharmacological agents for PTL prevention are palliative and frequently fail to prevent PTL and improve neonatal outcome. It is essential to fully understand the molecular mechanisms of PTL in order to develop novel therapeutic methods against PTL. Several lines of evidence indicate some chemokines are expressed in gestational tissues during labor or PTL. To reveal the pathophysiological roles of the CX3CL1-CX3CR1 axis in PTL, we performed present study using LPS-induced PTL mice model in CX3CR1-deficient (Cx3cr1-/-) mice. We indicated that PTL was suppressed in Cx3cr1-/- mice and immunoneutralization of CX3CL1 in WT mice. From immunohistochemical and the gene expression analyses, the CX3CL1-CX3CR1 axis has detrimental roles in PTL through intrauterine recruitment of macrophages and the enhancement of macrophage-derived inflammatory mediators. Thus, the CX3CL1-CX3CR1 axis may be a good molecular target for preventing PTL.

The effect of cold rolling on the recrystallization behavior of Al-free and Al-added 15Cr-oxide dispersion strengthened (ODS) ferritic steels was investigated. The recrystallization of both steels are enhanced by cold rolling. The Al-free ODS steel with finer oxide particles is harder to recrystallize than Al-added ODS steel with coarser oxide particles. The effect of Al addition on the recrystallization behavior is evident. It is estimated that the recrystallization temperature of Al-free and Al-added 15Cr-ODS ferritic steel is 900 °C and 1250 °C with the annealing period of 1 h. In Al-free ODS steel, a small hardening was observed in the temperature range between 850 °C and 1200 °C, while no such phenomenon was observed in Al-added ODS steel, which is indicative of retardation of recovery by finely dispersed oxide particles. Oxide particle growth is mostly dependent on annealing temperature, while recrystallization and grain growth are controlled by not only the temperature, but the cold rolling ratio, which alters the multiple factors such as dislocation density, initial grain shape and oxide particle dispersion morphology. The cold rolling direction also influences the grain morphology and grain orientation in Al-added ODS steel, and the second rolling in a perpendicular direction to the first cold rolling direction induces the rotation of the grains from to . The recrystallization temperature is not significantly changed by the cold rolling direction. Recrystallization after cold rolling appears to increase the {111} grain orientation on the cold rolled specimen surface.

A narrow-band dual camera system demonstrated a new close-range sensing technique to seasonally track trends in leaf greenness in rice paddies. A weatherproof digital imaging system for the visible red (RED, 620−650 nm) and near infrared band (NIR, 820−900 nm) was positioned 12 m above a 600-m2 rice field. During the 2009 and 2010 paddyrice seasons, the system automatically logged images at 10-min intervals throughout the day. Radiometric corrections for the images utilized solar irradiance sensors and prior calibration to calculate 0900-1500 JST daily-averaged reflectance factors (DARF). The DARF in RED (DARF-RED) and NIR (DARF-NIR) values were transformed to provide a daily-averaged normalizeddifference vegetation index (DA-NDVI). The DA-NDVI increased more rapidly in the vegetative growth period, and reached an asymptotic plateau earlier than the DARF-NIR. From transplanting to harvest, leaf greenness values (measured by the SPAD index) were measured for the central part of the uppermost leaves of targeted canopies weekly with a chlorophyll meter. We developed a leaf greenness index (LGI), the ratio of DA-NDVI to DARF-NIR, and a simple calculation method for area means to reduce the background effect. The modified area means of LGI followed the seasonal trend in SPAD value well; its patternwas inherently different from the patterns of any of the original three parameters: DARF-RED, DARF-NIR or DA-NDVI. Throughout the paddy seasons in the two years, a regression equation for estimating SPAD values using the LGI, daily solar radiation, the cosine of angle between the view and the meridian directions and the cosine of culmination solar zenith angle performed favorably (R2=0.815). The nitrogen concentration per dry plant hill (g kg-1) had a close relation to the SPAD values estimated using the equation.

A two-band digital imaging system—one band for visible red (RED, 630–670 nm) and the other for near infrared (NIR, 820–900 nm)—was positioned 12 m above a 600-m2 rice field. The imaging system automatically logged bird’s-eye-view images at 10-min intervals from 0700 to 1700 JST daily during the 2008 paddy rice season. Radiometric corrections and midterm field validations for the pairs of two-band images utilized solar irradiance sensors and prior calibrations to calculate 0900-1500 JST daily-averaged reflectance factor (DARF). The DARF values in the RED and the NIR bands agreed with observations made with a portable spectroradiometer and showed smooth seasonal changes in terms of plant growth. During the before-heading period, NIR DARF values had a correlation coefficient (r) of 0.91 with the leaf area index (LAI). An empirically derived equation for LAI using NIR DARF values and the cosines of angles between the view and the planting row directions, and the view and the meridian directions, showed R2 values of 0.93, in estimations of LAI from the dataset (number of observations=52) acquired at four sample points centrally located within the viewing field. Validation indicated that the equation also worked well for the other six observation points spread across the viewing field, the data of which were not used in deriving the equation parameters. The DARF values observed in the NIR band calibrated with solar spectral irradiance sensors were useful for assessing rice LAI during the before-heading periods.

A weatherproof digital imaging system for the near infrared band (NIR, 820–900 nm) was positioned 12 m above a 600-m2 rice field. During the 2008 and 2009 paddy rice seasons, the system automatically logged images at 10-min intervals throughout the day. Radiometric corrections for the NIR images utilized a solar irradiance sensor and prior calibrations to calculate 0900–1500 JST daily-averaged reflectance factors (DARF). Prior to heading, empirically derived equations for predicting leaf area index (LAI) using the 2008 DARF values in NIR, the cosines of angles between the view and the planting row directions, and between the view and the meridian directions were verified with the 2009 data set. Transformation of a model variable by arcsine square root function improved the performance of the LAI prediction by reducing the errors and bias at low LAI values. Adding variables to incorporate lateral angular components to the horizontal viewing angular parameters hardly affected the overall performance of the models and did not reduce variation. This was probably because the height and position of the camera system were the same in successive years. In-plot means of two or four predicted values in each plot reduced the root-mean square error 30%. These results indicate that radiometric NIR images derived using a fixed-point observation system can accurately predict LAI and the simple multiple linear regression equations developed for a given year can be used the following year without in-situ recalibration.

A spectral and polarization image observation technique for detecting multiband polarimetric characteristics of reflected light from field-growing plants under daylight conditions was developed and the potential application of the method to in-situ assessments of wheat-leaf orientation at the heading stage was assessed. The developed digital imaging system corresponded to wavelength bands centered at 470, 550, and 647 nm, each with bandwidths of 10 nm. The instrument was fitted with a glass polarizer, which rotated from 0 to 360º in 15º steps, and polarized images of 1360 ×1024 pixels were captured at heading in wheat plots subjected to different fertilizer regimes at the jointing stage. Degree of polarization (DP) and mean brightness (MB) of the three bands were calculated from images of several pairs of top-dressed and non-top-dressed (non-dressed) plots, with a camera depression angle of 15–20º on two clear days. The relative azimuth angles between the view and insolation were approximately 135º (oblique front) and 180º (right in front), respectively. The mean DP for each plot area in the images varied between 0.3 and 1.4%. Although most of the top-dressed plots had significantly higher DPs than the non-dressed plots in the 550 nm band, few of the MB images in any band showed a clear difference between the top-dressed and non-dressed plots.

Here we describe the configuration and operation of a field-use digital imaging system (multi-band spectrum camera, MBSC) that captures images in two spectral bands in the visible and near infrared wavelength ranges, and automatically converts and stores data as reflectance factor (RF) images. The instrument consists of a weatherproof camera unit, a skylight sensor and a PC and hard disc for a control unit. The camera unit has two industrial use digital camera boards attached with interference band-pass filters and objective lenses. The skylight sensor has light detectors equipped with band-pass filters identical to those used in the camera unit. The system is designed to collect images in each band and convert data into RF using the skylight intensity at prescheduled times. The MBSC has potential to automatically monitor agronomic variables in fields up to several hundred square meters.

A two-band digital imaging system —one band for the visible red band (RED, 630−670 nm) and the other for the near infrared band (NIR, 820−900 nm)— was devised and positioned at a height of 12 m above a rice field of 300 m2 in area during the 2007 growing season. The imaging system automatically logged bird’seye view images at 10-min intervals from 0800−1600 every day. Radiometric corrections for the pairs of two-band images were done using solar irradiance sensors and preceding calibrations to calculate daily band-reflectance and the normalized difference vegetation index (NDVI) values for 9 plots of rice plants, with 3 levels of planting density and basal fertilization. The daily- averaged reflectance values in the RED and the NIR bands showed different but smooth seasonal changing patterns according to the growth of plants. At the maximum tiller number and the panicle formation stages, the RED and NIR reflectance values had correlation coefficients (r) of 0.79 and 0.81 with above-ground nitrogen absorption per unit land area (NA, g m-2), respectively, whereas the NDVI using the two band reflectance values showed r-value of -0.13. An empirically derived equation for the NA using two band reflectance values showed r-value of 0.96 and a root mean square of error (RMSE) 0.5 g m–2 (10% of the mean observed NA) in the estimation for the original (not validated) data set acquired at the maximum tiller number and the panicle formation stages. The results indicated that reflectance observation in the RED and NIR bands using the digital imaging system was potentially effective for assessing rice growth.