Comparative analysis of sorafenib and lenvatinib on HepG2 cells and human umbilical vein endothelial cells: Involvement of transforming growth factor-ß signaling in their molecular effects.

AIM: This study aimed to compare the effects of the molecular targeted drugs, sorafenib and lenvatinib, on the survival, invasion, and angiogenesis of hepatocellular carcinoma cells. Additionally, we investigated the involvement of transforming growth factor beta (TGF-ß) signaling in their molecular mechanisms. METHODS: To investigate the effects of sorafenib and lenvatinib, we conducted cell viability, invasion, and angiogenesis assays, as well as western blotting analyses. RESULTS: In human hepatocellular carcinoma cells (HepG2), sorafenib demonstrated potent inhibitory effects on cell proliferation, but induced cell invasion similar to TGF-ß. In contrast, lenvatinib showed weaker cytotoxicity compared with sorafenib, but suppressed cell invasion induced by TGF-ß. The actions of these two molecular targeted drugs were suggested to involve the regulation of the TGFßR2/ERK pathway. Moreover, in human umbilical vein endothelial cells, Sorafenib showed weaker cytotoxicity and enhanced the effects of TGF-ß on angiogenesis. Conversely, lenvatinib showed potent cytotoxic abilities and suppressed angiogenesis induced by TGF-ß. The actions of these two molecular targeted drugs were suggested to involve the regulation of the crosstalk between TGF-ß signaling and vascular endothelial growth factor signaling. CONCLUSIONS: Our findings indicate that both sorafenib and lenvatinib possess anticancer abilities by inducing the cytotoxicity of hepatocellular carcinoma cells. Furthermore, they show opposing effects on TGF-ß-induced cell invasion and angiogenesis, thereby enhancing the understanding of the multifaceted functions of molecular targeted drugs in treating hepatocellular carcinoma.

Optimal clinical dose of medetomidine for sedation of young cows during dehorning surgery.

The present paper reports on the clinical efficacy and optimal clinical dose of medetomidine for sedation of young cows during dehorning surgery. Medical records of 24 female Holstein cows that underwent dehorning surgery were used in this study. In four groups, the sedation of animals was carried out by one of the four intravenous treatments: 0.1 mg kg-1 xylazine (Xyl group, n = 6), 5.0 µg kg-1 medetomidine (5.0 Med group, n = 6), 10.0 µg kg-1 medetomidine (10.0 Med group, n = 6) or 20.0 µg kg-1 medetomidine (20.0 Med group, n = 6). The clinical sedation score (CSS) and heart rate (HR) were recorded. The CSSs after intravenous administration of each α2-adrenergic receptor agonist increased rapidly and peaked at 12.5 (10.0-16.0) at t = 20 min in the Xyl group, 11.5 (10.0-15.0) at t = 10 min in the 5.0 Med group, 16.0 (14.0-16.0) at t = 20 min in the 10.0 Med group and 16.0 (14.0 - 16.0) at t = 20 min in the 20.0 Med group. A similar degree of bradycardia was observed after every sedative treatment. We conclude that the intravenous administration of 10.0-20.0 µg kg-1 medetomidine is appropriate for sedation of young cows without severe side effects.

Long-term outcomes of lupus nephritis with low-level proteinuria: a multicenter, retrospective study.

OBJECTIVES: Reportedly, patients with lupus nephritis (LN) and low-level proteinuria have favorable short-term renal outcomes. We aimed to clarify the long-term renal outcomes and overall survival of them, and the significance of renal biopsy in the early phase with low-level proteinuria. METHODS: We included 144 Japanese patients with biopsy-proven LN from ten hospitals. Low-level proteinuria was defined by a urine protein: creatinine ratio (UPCR) of ≤ 1 g/gCr based on previous reports. The outcomes were end-stage renal disease (ESRD) and death. RESULTS: Compared with patients with high-level proteinuria (UPCR > 1), those with low-level proteinuria (n = 67 [46.5%]) had significantly improved renal function at the time of renal biopsy, and low activity index and chronicity index (CI) while the frequency of class III/IV was similar (79.1% vs 84.4%, p = 0.409). In patients with low-level proteinuria, cyclophosphamide usage was less, and the incidence of ESRD (3.0% vs 13.0%, p = 0.036) or death (3.0% vs 16.9%, p = 0.006) during the total observation period (median, 72 months) were low. Kaplan-Meier analysis showed significant differences in the incidence of ESRD and death between the groups. Multivariate Cox regression analysis revealed that the significant risk factors for ESRD were high CI and hypertension, whereas those for death were increased age and high-level proteinuria. CONCLUSION: Patients with LN and low-level proteinuria had favorable long-term renal and life outcomes. As these patients have substantial active pathological lesions, renal biopsy in the early phase with low-level proteinuria could enable early diagnosis and treatment and thus improve prognosis.

Evaluation of blood serum iron concentration as an alternative biomarker for inflammation in dairy cows.

This study aimed to clarify the relationship between acute phase protein (APP) concentrations and serum Fe concentrations to determine whether serum iron (Fe) can be clinically applied as a substitute for APPs in cows. One hundred five Holstein-Friesian breed lactating dairy cows were enrolled in this study. Cows with inflammatory diseases were 16 subclinical, and 15 severe mastitis cows, in addition to 15 mild and 16 severe sole ulcer cows. The plasma haptoglobin (HPT), alpha-1 acid glycoprotein (AGP), SAA, serum Fe levels, and other biochemical parameters in the cows were measured. The two-sample t-tests and multiple logistic regression analysis were used to compare the control and inflammatory disease groups. ROC analysis was used to evaluate the ability to diagnose inflammation disease. From the results, the proposed diagnostic cutoff value for plasma SAA and serum Fe concentrations to identify dairy cows with inflammatory diseases based on analyses of ROC curves were set at > 3.65 mg/l and < 120.50 µg/dl, respectively. Therefore, instead of using expensive inflammatory markers to evaluate the inflammatory state at the first treatment day for inflammatory diseases in cow, it shows the useful for screening with serum Fe concentration that can be measured easily and inexpensively as alternative inflammatory biomarkers.

Mastitis causes negative reproduction performance similar to genital diseases.

The economic efficiency of dairy farms is directly influenced by reproductive performance. Reproductive performance is affected by postpartum health; however, it is unknown which diseases reduce reproductive performance or how often these diseases occur in tie-stall dairy farms. In this study, the 15 most frequent diseases in tie-stalls and their impact on reproduction were determined by logistic analysis. Data from reproductive and veterinary records of 814 cows (473 Holstein) calving between April 2016 and March 2018 reared in 9 tie-stall commercial dairy farms were used. The 15 most frequent diseases were identified by medical records. The odds ratio (OR) of the diseases reducing reproductive performance (days open, number of artificial insemination, and first artificial insemination days in milk) was measured by logistic regression analysis. We focused on mastitis, and factors (inflammation grade, causative bacteria) having a negative effect on reproduction were measured similarly. Of the 15 most frequent diseases, endometritis (OR: 5.45, p < 0.001), ovarian quiescence (OR: 3.95, p < 0.001), mastitis (OR: 2.49, p < 0.001), and follicular cyst (OR: 2.33, p = 0.006) significantly prolonged days open after adjusting for environmental factors (farm, parity, and calving season). Focusing on mastitis, mastitis with mild inflammation significantly prolonged days open. Mastitis and genital diseases were frequent diseases that reduced reproductive performance. Strategies to reduce the incidence of these diseases are considered effective for the economics of tie-stall dairy farms.

Comparison of the alkalizing abilities between 1.35% sodium bicarbonate solutions with and without dextrose in healthy calves.

The present study aimed to clarify the alkalizing ability of 1.35% isotonic sodium bicarbonate solution (ISBS), which did not contain dextrose, compared with that of 1.35% isotonic bicarbonate sodium solution containing 4.03% dextrose (ISBD) in healthy calves. The calves were intravenously administered with 20.7 mL/kg of the solutions for 30 min as the volume required to correct base deficit of 10 mM. ISBS increased the blood pH, HCO3-, and base excess from 7.44 ± 0.02, 29.6 ± 1.9 mM, and 5.3 ± 2.1 mM to 7.49 ± 0.02, 36.9 ± 2.3 mM, and 13.5 ± 2.6 mM respectively (P<0.05). These factors for the ISBD group increased from 7.41 ± 0.02, 29.0 ± 1.1 mM, and 4.5 ± 1.3 mM to 7.43 ± 0.02, 33.5 ± 1.9 mM, and 9.5 ± 1.7 mM (P<0.05), respectively. Furthermore, in the ISBD group, the relative plasma volume and blood glucose level increased while the K+ level decreased, which did not occur in the ISBS group. Therefore, the results revealed that ISBS had better alkalizing ability in calves than ISBD.

Massive transformation in FeNi nanopowders with nanotwin-assisted nitridation.

L10-ordered FeNi alloy (tetrataenite), a promising candidate for rare-earth-free and low-cost permanent magnet applications, is attracting increasing attention from academic and industrial communities. Highly ordered single-phase L10-FeNi is difficult to synthesis efficiently because of its low chemical order-disorder transition temperature (200-320 °C). A non-equilibrium synthetic route utilizing a nitrogen topotactic reaction has been considered a valid approach, although the phase transformation mechanism is currently unknown. Herein, we investigated the basis of this reaction, namely the formation mechanism of the tetragonal FeNiN precursor phase during the nitridation of FeNi nanopowders. Detailed microstructure analysis revealed that the FeNiN precursor phase could preferentially nucleate at the nanotwinned region during nitridation and subsequently grow following a massive transformation, with high-index irrational orientation relationships and ledgewise growth motion detected at the migrating phase interface. This is the first report of a massive phase transformation detected in an Fe-Ni-N system and provides new insights into the phase transformation during the nitriding process. This work is expected to promote the synthetic optimization of fully ordered FeNi alloys for various magnetic applications.

Identification and phylogenetic analysis of Babesia parasites in domestic dogs in Nigeria.

The present study examined the presence of Babesia parasites in 104 domestic dogs in Nigeria. Sequentially, Babesia parasites infecting domestic dogs underwent genetic and phylogenetic analyses. The results of nested PCR based on the Piroplasmida 18S rRNA gene illustrated that 13.5% (14/104) of the samples were positive. The obtained positive samples determined the nucleotide sequences of the 18S rRNA genes. In the genetic and phylogenetic analyses, four of five nucleotide sequences were similar to Babesia canis rossi, and one sample exhibited a close similarity to a Babesia sp. isolated from a raccoon in Hokkaido, Japan. The present study revealed the widespread presence of B. canis rossi among domestic dogs in Nigeria.

Over 3-Years Follow-Up of a Case of Primary Biliary Cholangitis With Autoimmune Hyperthyroidism: Factors Associated With Liver Dysfunction.

We present a rare case of a 50-year-old woman with simultaneous primary biliary cholangitis (PBC) and autoimmune hyperthyroidism in which the factors associated with liver dysfunction were evaluated over a 3-year follow-up period. Although serum thyroid hormone levels improved after the administration of thiamazole, thyroid dysfunction was not directly associated with liver dysfunction. During the follow-up period, anti-hyperlipidemic drug therapy and eradication therapy for Helicobacter pylori (HP) infection caused transient elevation of serum transaminase levels. It should be recognized that serum liver enzyme levels might be affected by various factors, including the therapies for the many complications of PBC.

The effect of sodium butyrate supplementation on ruminal and fecal pH and serum lipopolysaccharide-binding protein after ruminal acidosis challenge in nonlactating cows.

The objective of this study was to evaluate effects of sodium-butyrate supplementation on gastrointestinal function and the inflammatory response to ruminal acidosis (RA) challenge in cows. Four nonlactating cows with a rumen cannula were assigned to two treatments in a crossover design. Treatments were ruminal administration of sodium-butyrate (BUT) or control (CON). Sodium-butyrate was provided as Gustor BP70 and administered at a butyrate dose of 0.04% per kg body weight. The CON premix was made by replacing sodium-butyrate with wheat bran. Experimental periods were 28 days long with 21-day washout period separating the treatments. On Day 25 of each period, corn starch was ruminally administered at 0.7% per kg body weight as RA challenge. After RA challenge, ruminal pH was lower, and endotoxin concentration was higher for cows provided with BUT than those with CON, but the increase in fecal starch and the decrease in fecal pH were attenuated by BUT. The effect of butyrate supplementation on serum lipopolysaccharide-binding protein after RA challenge was not found. From these findings, butyrate supplementation mitigated rectal acidosis by reducing the flux of fermentable carbohydrate into the large intestine. An anti-inflammatory effect of butyrate was not observed, possibly due to lower pH and higher endotoxin concentration in the rumen.

Supplementation with Branched-Chain Amino Acids Induces Unexpected Deleterious Effects on Astrocyte Survival and Intracellular Metabolism with or without Hyperammonemia: A Preliminary In Vitro Study.

INTRODUCTION: Ammonia is a key component in the pathogenesis of hepatic encephalopathy. Branched-chain amino acids (BCAA) have been reported to improve the symptoms of HE induced by hyperammonemia; however, we recently reported that ammonia increases intracellular levels of BCAA and exerts toxic effects on astrocytes. OBJECTIVES: This follow-up study was designed to confirm the direct effects of BCAA on human astrocytes and clarify their underlying mechanisms using metabolome analysis and evaluation of associated signaling. METHODS: We performed cytotoxicity and cell proliferation tests on astrocytes following BCAA treatment with and without ammonium chloride (NH4Cl) and then compared the results with the effects of BCAA on hepatocytes and neurons. Subsequently, we used metabolomic analysis to investigate intracellular metabolite levels in astrocytes with and without BCAA treatment. RESULTS: The astrocytes showed increased leakage of intracellular lactate dehydrogenase and reduced proliferation rate upon BCAA treatment. Interestingly, our analysis showed a BCAA-induced impairment of intracellular glycolysis/glyconeogenesis as well as amino acid and butyric acid metabolism. Furthermore, BCAA treatment was found to cause decreased levels of Glut-1 and phosphorylated GSK-3ß and mTOR in astrocytes. CONCLUSIONS: Although further investigations of the effect of BCAA on human astrocytes with hyperammonemia are needed, our work demonstrates that BCAA supplementation has direct negative effects on astrocyte survival and intracellular metabolism.

Clistobothrium sp. (Cestoda: Tetraphyllidea) in oarfish (Regalecus russelii) stranded on the coast of Akita Prefecture, Japan.

Oarfish (Regalecus russelii Cuvier) are mesopelagic fish with little known about their life history. Oarfish live in deep water, making it difficult for researchers to collect specimens; thus, records of their parasitic helminths are limited. Two plerocercoids were found for the first time in an oarfish stranded on the coast of Akita Prefecture, Japan. These plerocercoids were identified as Clistobothrium sp. RR-1 using morphological and molecular analyses. It was revealed that oarfish represent one of the intermediate hosts of the genus Clistobothrium, and large sharks are the definitive hosts for these parasites.

Retrospective study on the outcomes and risk factors of right paramedian abomasopexy for right abomasal disorders in 47 dairy cows.

Very few epidemiologic studies have verified the utility of the right paramedian abomasopexy (RPA) technique in cows with right abomasal disorders. This study aimed to investigate the outcomes and risk factors for non-survival in the herd within 30 days of surgery in cows with right abomasal disorders who underwent the RPA technique. Forty-seven Holstein cows with right abomasal disorders (25 with right abomasal displacement [RDA] and 22 with right abomasal volvulus [RAV]) were included. Twenty-two cows with RDA (22/25, 88.0%) and 10 cows with RAV (10/22, 45.5%) survived at 30 days post-surgery. Multivariate logistic regression analysis indicated that hyponatremia, hypokalemia, and the presence of abomasal volvulus were the major risk factors associated with non-survival.

Serum IgG4 levels at diagnosis can predict unfavorable outcomes of untreated patients with IgG4-related disease.

The outcomes of patients with immunoglobulin G4 (IgG4)-related disease (IgG4-RD) who are not treated are unclear. This study aimed to clarify these outcomes and identify the factors related to them. We retrospectively evaluated various clinical features including laboratory data and involved organs at diagnosis in 107 patients with IgG4-RD, who were followed up for more than 6 months, at a single center in Japan. We compared the clinical features of the 27 untreated patients with those of the 80 patients treated with glucocorticoid. The patient outcomes were investigated, and logistic regression analysis was performed to identify factors related to them. The patients comprised 73 men and 34 women (median age 67 years). The untreated patients had significantly lower IgG4-RD responder index (9 vs. 12) and fewer affected organs (1 vs. 3) than did those treated with glucocorticoid. Of these 27 patients, 8 experienced deterioration of IgG4-RD after the diagnosis. In the age- and sex-adjusted logistic regression analysis, serum IgG4 elevation (per 100 mg/dL, odds ratio 1.194, 95% confidence interval 1.017-1.402) was the only significant factor related to disease deterioration in untreated patients with IgG4-RD, whereas not serum IgG4 levels (per 100 mg/dL, odds ratio 0.995, 95% confidence interval 0.921-1.075) but history of allergy (OR 3.134, 95% confidence interval 1.094-8.977, P = 0.033) related to deterioration in patients who underwent treatment. Serum IgG4 levels may be a useful predictor of unfavorable outcomes in untreated patients with IgG4-RD, who tend to have fewer affected organs and lower IgG4-RD responder index.

Evaluation of the Long-term Administration of Rifaximin for More than Three Years in the Treatment of Repeated and Recurrent Overt Hepatic Encephalopathy.

The patient was a 65-year-old man with alcoholic liver cirrhosis who had been admitted to hospital 5 times for repeated and recurrent overt hepatic encephalopathy (HE) despite numerous therapies, including disaccharide, branched-chain amino acid (BCAA) formula, L-carnitine and zinc. After the additional administration of rifaximin (1,200 mg/day orally), his consciousness level was well controlled for 3 years without any adverse effects. The long-term administration of rifaximin may be useful and safe for managing recurrent overt HE, although the maintenance dosage and duration of rifaximin and safety should be evaluated in patients with ameliorated HE.

Effects of temporary cessation of milking following intramammary cefazolin sodium infusion on residual antibiotic in lactating dairy cows.

The aims of studies were to estimate the withdrawal period of antibiotic from milk after the intramammary infusion of cefazolin sodium (CEZ) in cows with difficulties in frequent milk discharge due to disease such as teat injury. The period was compared among cows milked twice a day after 150 or 450 mg of CEZ were administered to all quarters (Study 1, 2) and the cows in which milking of front-right quarter was ceased for five days after administration of these infusions to only that quarter (Study 3). In Studies 1 and 2, the median of 17.66 µg/ml and 83.18 µg/ml of CEZ were detected in the samples of first milking after intramammary administration, respectively; however, there was no residual antibiotic by 72 hr in all cows. In Study 3, the median of 1.96 µg/ml of CEZ was detected in the sample after the resumption of milking at 120 hr, and the residual was eliminated by 174 hr. The withdrawal period may be prolonged by the cessation of milking after administration, and the period is the total time from cessation to 72 hr after the resumption of milking.

Validation of the bovine blood calcium checker as a rapid and simple measuring tool for the ionized calcium concentration in cattle.

Point-of-care (POC) devices that veterinary practitioners can use to easily and rapidly measure blood ionized calcium (iCa) levels in cows immediately after withdrawing a blood sample on the dairy farm are needed. Aims of present studies was to compare the commercially available ion-selective electrode handheld iCa meter (bovine blood iCa checker) with the benchtop blood gas analyzer GEM premier 3500 and handheld analyzer i-STAT 1. Sixty-two paired-point whole blood samples were obtained from three cows with hypocalcemia experimentally induced by Na2-EDTA infusion. Whole blood samples were also obtained from the 36 cows kept on a farm in field conditions. The results using the bovine blood iCa checker correlated with those using the GEM premier 3500 and i-STAT 1. Bovine blood iCa checker was "compatible" with the GEM premier 3500 and i-STAT 1 because the frequency of differences between the measurements within ± 20% of the mean were 100% (65/65, >75%) and 90.8% (59/65, >75%), respectively. In the field trial, the blood iCa concentration measured by the bovine blood Ca checker was significantly positively correlated with that measured by the i-STAT 1 portable analyzer. Bovine blood iCa checker was "compatible" with the i-STAT 1 because the frequency of differences between the measurements within ± 20% of the mean was 100% (36/36, >75%). Results from these findings, the bovine blood iCa checker may be applied as a simplified system to measure the iCa concentration in bovine whole blood.

Serum iron concentration is candidate inflammatory marker for respiratory diseases in beef cows.

We hypothesized that the serum iron (Fe) concentration in cows with respiratory diseases is a satisfactory substitute for major inflammatory markers such as haptoglobin (HPT) and serum amyloid A (SAA). Twenty Japanese Black cows aged 279.6 ± 120.0 days were enrolled, and divided into respiratory diseases and control groups based on the presence of clinical findings of respiratory diseases. As a result, area under the receiver operating characteristic curves for plasma HPT, SAA and serum Fe concentrations for respiratory disease-associated systemic inflammation were excellent, at 1.00, 0.96 and 0.97, respectively. Therefore we confirmed that the serum Fe concentration is a satisfactory substitute for HPT and SAA in beef cows with respiratory diseases.

Association between serum albumin and cognitive dysfunction in hepatic encephalopathy: An exploratory data analysis.

BACKGROUND AND AIM: Because covert hepatic encephalopathy (CHE) has been shown to affect the prognosis of cirrhotic patients, early diagnosis of hepatic encephalopathy (HE) is a prerequisite for the preservation of patients' quality of life and for prophylaxis of overt HE. The aim of this study was to identify a clinical parameter to predict impairment of cognitive function in cirrhotic patients with early-stage HE. METHODS: We investigated the data from 172 patients with cirrhotic or idiopathic portosystemic shunt (PSS) in phase II/III trials of rifaximin in Japan. Classification and regression trees (CARTs) were constructed to identify clinical profiles related to cognitive dysfunction as indicated by the prolongation of time required for the Number Connection Test (NCT-B). RESULTS: CART analysis detected age 65 years as the variable for the initial split, and serum albumin level was selected as the variable for the second split among patients aged ≤65 years. In 27 cirrhotic patients aged ≤65 years without PSS, receiver operating characteristic curve analysis revealed that the optimal albumin level cutoff point was 3.05 g/dL, and the area under the curve was 0.80 for the prolongation of NCT-B time, which was higher than that of the branched-chain amino acids-to-tyrosine ratio (0.46), the prothrombin time-international normalized ratio (PT-INR) (0.68), serum ammonia (0.61), and total bilirubin (0.69). CONCLUSIONS: Lower serum albumin level as a clinical biomarker associated with impaired cognitive function may be available as a screening examination for early-stage HE in cirrhotic patients aged ≤65 years without PSS before undergoing neuropsychological tests.

Antiphospholipid antibody syndrome-associated renal thrombotic microangiopathy improved not with rivaroxaban but with warfarin in a systemic lupus erythematosus patient without lupus nephritis.

A 34-year-old Japanese woman with systemic lupus erythematosus (SLE) was admitted to our hospital for exacerbation of renal dysfunction, hemolytic anemia and thrombocytopenia. Twenty-two years before admission, she was diagnosed with SLE. Eight years before, lupus anticoagulant (LAC) positivity was detected without any thrombotic findings. Fourteen months before, renal function started to worsen. Three months before, unprovoked left leg swelling appeared. She was diagnosed with deep vein thrombosis (DVT) by ultrasonography. Blood examination revealed mild anemia, thrombocytopenia, and renal dysfunction. Rivaroxaban was started after which the left leg swelling subsided. When she was referred to our hospital, LAC was positive, but hypocomplementemia nor elevation of serum anti-double-stranded DNA antibodies was detected. Renal biopsy showed acute and chronic thrombotic microangiopathy (TMA) without concurrent lupus nephritis. Brain magnetic resonance imaging showed new small multiple cerebral infarcts. Antiphospholipid antibody syndrome (APS), causing renal TMA, new cerebral infarction, and DVT was diagnosed. Rivaroxaban was changed to warfarin. Two months after admission, renal impairment improved, and the complete disappearance of DVT and brain infarcts was confirmed. This case suggests that warfarin may be more effective than direct oral anticoagulants in the treatment of APS-associated renal TMA.