The first record of Brachylaima ezohelicis (Trematoda: Brachylaimidae) in the red fox (Vulpes vulpes schrencki).

Brachylaima spp. are trematodes that have a unique life cycle as they exclusively use land snails as the intermediate host. Although their intermediate host has been well studied, very little information is available about their definitive host, partly as isolation of its adult stage from wild animals is rare. We found three trematodes in the small intestine of a red fox (Vulpes vulpes schrencki) in Hokkaido, the northernmost island of Japan. The trematodes were identified as Brachylaima ezohelicis based on morphological features and genetic analysis, which is believed to have a definitive avian host. The morphological features of the isolated trematodes were consistent with B. ezohelicis samples grown in the definitive host except for body length. Our study suggests that B. ezohelicis uses mammals as definitive hosts as well as birds.

Quantifying the direct and indirect effects of sika deer (Cervus nippon) on the prevalence of infection with Rickettsia in questing Haemaphysalis megaspinosa: A field experimental study.

Sika deer (Cervus nippon) are important hosts for all life stages of Haemaphysalis megaspinosa, a suspected Rickettsia vector. Because some Rickettsia are unlikely to be amplified by deer in Japan, the presence of deer may decrease the prevalence of Rickettsia infection in questing H. megaspinosa. As sika deer decrease vegetation cover and height and thereby indirectly cause changes in the abundance of other hosts, including reservoirs of Rickettsia, the prevalence of Rickettsia infection in questing ticks can also change. We investigated these possible effects of deer on the prevalence of infection with Rickettsia in questing ticks in a field experiment in which deer density was manipulated at three fenced sites: a deer enclosure (Deer-enclosed site); a deer enclosure where deer had been present until 2015 and only indirect effects remained (Indirect effect site); and a deer exclosure in place since 2004 (Deer-exclosed site). Density of questing nymphs and the prevalence of infection with Rickettsia sp. 1 in questing nymphs at each site were compared from 2018 to 2020. The nymph density at the Deer-exclosed site did not significantly differ from that at the Indirect effect site, suggesting that the deer herbivory did not affect the nymph density by reducing vegetation and increasing the abundance of other host mammals. However, the prevalence of infection with Rickettsia sp. 1 in questing nymphs was higher at the Deer-exclosed site than at the Deer-enclosed site, possibly because ticks utilized alternative hosts when deer were absent. The difference in Rickettsia sp. 1 prevalence between the Indirect effect and Deer-exclosed sites was comparable to that between the Indirect effect and Deer-enclosed sites, indicating that the indirect effects of deer were as strong as the direct effects. Examining the indirect effects of ecosystem engineers in the study of tick-borne diseases may be more important than previously recognized.

Killing Two Birds with One Stone: Discovery of Dual Inhibitors of Oxygen and Fumarate Respiration in Zoonotic Parasite, Echinococcus multilocularis.

Ascofuranone (AF), a meroterpenoid isolated from various filamentous fungi, including Acremonium egyptiacum, has been reported as a potential lead candidate for drug development against parasites and cancer. In this study, we demonstrated that AF and its derivatives are potent anthelminthic agents, particularly against Echinococcus multilocularis, which is the causative agent of alveolar echinococcosis. We measured the inhibitory activities of AF and its derivatives on the mitochondrial aerobic and anaerobic respiratory systems of E. multilocularis larvae. Several derivatives inhibited complex II (succinate:quinone reductase [SQR]; IC50 = 0.037 to 0.135 µM) and also complex I to III (NADH:cytochrome c reductase; IC50 = 0.008 to 0.401 µM), but not complex I (NADH:quinone reductase), indicating that mitochondrial complexes II and III are the targets. In particular, complex II inhibition in the anaerobic pathway was notable because E. multilocularis employs NADH:fumarate reductase (fumarate respiration), in addition to NADH oxidase (oxygen respiration), resulting in complete shutdown of ATP synthesis by oxidative phosphorylation. A structure-activity relationship study of E. multilocularis complex II revealed that the functional groups of AF are essential for inhibition. Binding mode prediction of AF derivatives to complex II indicated potential hydrophobic and hydrogen bond interactions between AF derivatives and amino acid residues within the quinone binding site. Ex vivo culture assays revealed that AF derivatives progressively reduced the viability of protoscoleces under both aerobic and anaerobic conditions. These findings confirm that AF and its derivatives are the first dual inhibitors of fumarate and oxygen respiration in E. multilocularis and are potential lead compounds in the development of anti-echinococcal drugs.

Data on the combined effect of atovaquone, mefloquine, and 3-bromopyruvic acid against Echinococcus multilocularis protoscoleces.

The dataset presented here is related to a previous research article titled "Mitochondrial Complex III in Larval Stage of Echinococcus multilocularis as a Potential Chemotherapeutic Target and in vivo Efficacy of Atovaquone Against Primary Hydatid Cysts"[1]. In this report, data were collected from aerobic and anaerobic culture assays of E. multilocularis protoscoleces in the presence of three anti-echinococcal drug candidates (atovaquone, mefloquine, and 3-bromopyruvic acid). The data were analyzed for viability of the protoscoleces between day 0 and day 7 upon adding drug candidates. In aerobic condition, all drug candidates caused damage to the protoscoleces, as described previously [1], [2], [3], [4], [5], [6]. Mefloquine, alone as well as in combination with atovaquone, immediately eliminated the protoscoleces, whereas combination of atovaquone with 3-bromopyruvic acid did not show clear synergy. In anaerobic condition, mefloquine, alone as well as in combination with atovaquone, eliminated protoscoleces immediately. 3-Bromopyruvic acid showed stronger efficacy in anaerobic condition than in aerobic condition. Combination of atovaquone with 3-bromopyruvic acid eliminated the protoscoleces, indicating that synergy occurred only under anaerobic condition. The data clarified that combined use of the three drugs eliminated protoscoleces in both aerobic and anaerobic conditions, hence suggesting that these could inhibit aerobic and anaerobic respiration pathways of Echinococcus multilocularis in vivo. The obtained data would be useful for the development of new drug dosing method for alveolar echinococcosis.

Associations between Japanese spotted fever (JSF) cases and wildlife distribution on the Boso Peninsula, Central Japan (2006-2017).

Populations of large mammals have been dramatically increasing in Japan, resulting in damage to agriculture, forestry, and ecosystems. However, their effects on tick-borne diseases have been poorly studied. Here, we focused on the relationship between Japanese spotted fever (JSF), a tick-borne disease caused by Rickettsia japonica, and populations of large mammals. To explore factors that affected the area in which JSF cases occur, we used generalized linear mixed models (GLMMs). We demonstrated that the expansion of the area of JSF occurrence can be predicted by deer density and geographical factors, which is likely due to differences in landscape structure. However, the associated models have limitations because of the lack of information about the distribution of vectors and reservoirs. To reduce the risk of humans contracting JSF, potential reservoirs should be confirmed.

Morphologically abnormal beaks observed in chickens that were beak-trimmed at young ages.

A survey of beak morphological abnormalities was performed on 6,201 chickens (egg-laying hens and chickens for meat production belonging to 25 flocks) brought to a poultry processing plant. The observed abnormalities varied among flocks with occurrence rates ranging from 0.48 to 46.67%. The occurrence was high in flocks subjected to beak trimming and varied significantly according to chicken breed, with the highest rates of abnormalities in a certain chicken breed. The most widely observed abnormalities were: 1) uneven growth of the upper and lower mandibles, mostly with elongation of the lower mandible (accounting for 64.8% of all abnormalities); 2) misalignment of the upper and lower mandibles, causing lateral deviation or crossing (16.3%); 3) sharp or jagged deformities of the mandible tips (10.1%); 4) permanent open beak, a deformity in which the beak did not close completely even when closed (5.8%); and 5) formation of tubercular swellings at the tips of the upper or lower mandibles (3.1%). This is the first report on the occurrence of beak abnormalities in beak-trimmed poultry in Japan.

Febuxostat (Feburic tablet) in the management of hyperuricemia in a general practice cohort of Japanese patients with a high prevalence of cardiovascular problems.

Hyperuricemia is increasing in prevalence and this is paralleled by an increased incidence of acute gout. In addition, there is growing evidence of an association between high serum levels of uric acid (sUA) and cardiovascular disease (CVD). In this preliminary report, we present 12-16 week results from a multicenter, general practice study in which we evaluated the usefulness of febuxostat in a cohort of untreated patients with hyperuricemia with a high prevalence of CVD. Febuxostat titrated from 10 mg/day up to 40 mg/day resulted in statistically significant and clinically relevant reductions in sUA after 12-16 weeks. A "responder" level of 6.0 mg/dL or lower was achieved in 95 of 100 (95%) patients. Significant reductions in sUA were achieved regardless of the presence/absence of coexisting diseases (e.g. CVD, renal insufficiency, diabetes and obesity) or the class of antihypertensive agent being used by the patient. No serious adverse reactions were noted with febuxostat. Although allopurinol has been used generally for hyperuricemia/gout, it is excreted fully via the kidneys, restricting its use in patients with reduced renal function, and its three-times-daily administration leads to poor adherence. Based on the results of this study, febuxostat may provide an easier option than allopurinol for clinicians specializing in CVDs.

Significance of stroke volume variation during hepatic resection under infrahepatic inferior vena cava and portal triad clamping.

PURPOSE: Stroke volume variation (SVV), which is measured by analyzing arterial blood pressure waveform characteristics, is a simple and sensitive indicator of fluid responsiveness. The current retrospective study was to investigate SVV and central venous pressure (CVP) during hepatic resection under clamping of both the infrahepatic inferior vena cava (IVC) and the portal triad. METHODS: All hepatic resections performed from December 2009 to February 2010 at the Department of Surgery at Iizuka Hospital in Japan were included in this study. Invasive hemodynamic monitoring including CVP and SVV were performed in 14 patients. RESULTS: CVP was significantly lower in patients with blood loss < or = 486 g than in those with blood loss > 486 g. SVV was significantly higher in patients with blood loss < or = 486 g than those with blood loss > 486 g during both IVC clamping and IVC + portal triad clamping. Estimated blood loss was significantly less in the group with SVV values > 18% compared to the group with values < or = 18%. There was a significant correlation between SVV and CVP (R2 = 0.714; P < .01). CONCLUSION: SVV is a useful indicator of intraoperative blood loss without the monitoring of CVP during hepatic resection under clamping of both the infrahepatic IVC and the portal triad.

[Clinical evaluation of the new creatine kinase MB reagent kit "L-System CK-MB MtO"].

The creatine kinase MB (CK-MB) activity assay, which is based on the immunoinhibition method, has long been used in the diagnosis of acute myocardial infarction (AMI) because of its good cost-performance ratio and simplicity. However, the immunoinhibition method can not differentiate between CK-MB and MtCK, and therefore, CK-MB activity determined using this method is higher than the actual value in the sample which MtCK appears; this may lead to the misdiagnosis of AMI. We, therefore, evaluated the analytical and clinical performance of a new CK-MB reagent kit "L-System CK-MB MtO," which can inhibit MtCK. The kit yielded good precision and linearity and no interference from hemolysis, bilirubin or chyle. A good correlation was observed between the values determined using this kit and those determined using the conventional kit for samples of patients with acute coronary syndromes. However, differences were observed in the CK-MB values determined for samples from patients with malignancy. CK isoenzyme analysis indicated that MtCK was present in all these samples. The new method permits the accurate estimation of CK-MB activity in samples of patients with high serum levels of MtCK activity and indicates that the conventional method has a high false-positive rate for CK-MB activity. CK-MB activity in the serum of healthy individuals measured using the new and the conventional kits was 1.9-9.5 U/l and 4.5-15.3 U/l, respectively. The new kit, enables accurate estimation of CK-MB activity and is, therefore, more useful than the conventional kit in the diagnosis of acute coronary syndromes.

Dietary sphingolipids ameliorate disorders of lipid metabolism in Zucker fatty rats.

Dietary sphingolipids (SL) inhibit colon carcinogenesis, reduce serum cholesterol, and improve skin barrier function and are considered to be "functional lipids". For comparative determination of the effects of SL with different chemical compositions on lipid metabolism and its related hepatic gene expression, Zucker fatty rats were fed pure sphingomyelin (SM) of animal origin and glucosylceramide (GC) of plant origin. After 45 days, the SM and GC diets led to significant reductions in hepatic lipid and plasma non-HDL cholesterol. Both SM and GC diets decreased plasma insulin levels, whereas only the GC diet increased the plasma adiponectin level. Hepatic gene expression analysis revealed increased expression of adiponectin receptor 2 (Adipor2), peroxisome proliferator-activated receptor alpha (PPARalpha), and pyruvate dehydrogenase kinase 4 (Pdk4). However, expression of stearoyl CoA desaturase (Scd1) was significantly decreased. These results suggest that dietary SL, even of different origins and chemical compositions, may prevent fatty liver and hypercholesterolemia through improvement of adiponectin signaling and consequent increases in insulin sensitivity.

The effects of ezetimibe on surrogate markers of cholesterol absorption and synthesis in Japanese patients with dyslipidemia.

AIM: To demonstrate the clinical benefit of inhibiting intestinal cholesterol absorption, we evaluated the effects of ezetimibe on surrogate markers of cholesterol absorption and synthesis, lipid and glucose metabolism, and markers of obesity and inflammation. METHODS: A total of 120 patients with dyslipidemia (46 men; mean age 66.5 years), who had not achieved the low density lipoprotein cholesterol (LDL-C) goal recommended by the Japan Atherosclerosis Society Guideline despite diet and exercise or any statin therapy, were enrolled and additionally treated with ezetimibe (10 mg/day) for 12 weeks. RESULTS: Compared to the baseline, LDL-C was reduced by 19.2% (p<0.001) after ezetimibe monotherapy and by 24.7% (p<0.001) after co-administration with ezetimibe and any statin. Ezetimibe therapy decreased cholesterol absorption markers and increased a cholesterol synthesis marker. Treatment with ezetimibe reduced the fasting serum insulin level (p<0.05) and HbA1c (p<0.05), increased serum adiponectin (p<0.01), and showed a significant decrease of high-sensitive C-reactive protein (hsCRP, p<0.01). No adverse events occurred during the study. CONCLUSION: Thus, cholesterol absorption inhibition by ezetimibe is an important therapeutic strategy since LDL-C and cholesterol absorption markers had a positive correlation. Ezetimibe not only reduced the serum LDL-C level but also improved glucose metabolism as well as obesity and inflammation markers. These findings support the benefit of ezetimibe as a new option for the treatment of dyslipidemia.

Improved specificity of a new homogeneous assay for LDL-cholesterol in serum with abnormal lipoproteins.

BACKGROUND: Although a homogeneous assay for serum LDL-cholesterol (LDL-C) has become a routine clinical procedure, problems remain in assay performance characteristics. METHODS: We examined the performance of a recently developed automated homogeneous assay (New-Daiichi assay) for serum LDL-C and compared the results with those obtained by the current homogeneous method (Denka-Seiken assay) or by ultracentrifugation as a control. RESULTS: The New-Daiichi assay showed satisfactory basic performance characteristics such as reproducibility, linearity, and stability. There was no interference in the assay by various substances examined. The LDL-C values obtained with this method correlated well with those obtained by ultracentrifugation. In samples from patients with obstructive jaundice, both methods detected cholesterol from abnormal lipoproteins (such as lipoprotein-X and -Y), but the New-Daiichi assay was less reactive and more specific for LDL-C. CONCLUSION: The new method has improved performance for the accurate measurement of LDL-C in clinical practice.

Questionnaire survey on the state of routine echocardiographic examinations in Japan: second report.

We mailed questionnaires to 748 registered medical sonographers [RMSs (cardiology)] to gather information for a large-scale survey of RMSs in May 2000. We wanted to evaluate the current state of routine echocardiography in Japan. Altogether, 530 (70.9%) of these sonographers responded; 18 respondents employed by equipment manufacturers were excluded from the start of the study, and 20 others later found to be employed by equipment manufacturers were also excluded. Responses from all personnel at a single institution were treated as a single response; 436 institutions were thus included in the survey. Most or all examinations were carried out by sonographers at 77.3% of the responding institutions but were performed mainly by physicians at 11.1% of the institutions. At least 80% of sonographer or physician working hours were used for echocardiographic examination at 18.9% of the surveyed institutions, whereas up to half the working hours were devoted to echocardiographic examinations at 67.0% of the institutions. The most frequently reported examination time was 30-40 min [35.6% (n = 116) of the institutions], whereas 32.8% (n = 107) of the institutions indicated durations of 10-20 min per examination. Left ventricular (LV) dimensions and wall thickness were measured on M-mode images in all patients at 14.1% (n = 59) of the institutions and mainly on M-mode images but from two-dimensional echocardiography in problematic patients at 74.2% (n = 311) of the institutions. The LV ejection fraction was calculated from LV dimensions at 55.7% (n = 205) of the institutions and from the LV cross-sectional area at 44.3% (n = 163) of the institutions. The LV ejection fraction was estimated visually at 57.0% of the institutions. Only 5.5% (n = 22) of the facilities always scored the wall motion. Pulsed Doppler echocardiography was used to assess LV inflow in all patients at 65.9% of the institutions; regurgitation and shunting were always assessed quantitatively at only 2% (n = 8). Comments concerning image quality were reported in all cases or in cases of poor image quality at 98% of the institutions, and the sonographer was also involved in writing the diagnostic report at 94% of institutions. Echocardiography is less expensive than other diagnostic imaging methods, and its importance is thus likely to continue to increase. Echocardiographic examinations should be carried out by skilled RMSs and physician echocardiographers; and an accurate, readily comprehensible report of the findings should be provided promptly to the referring physician. More such surveys are required to ensure that these practices are adopted.

[Intravascular ultrasound and angioscopy].

We observed pulmonary artery thrombi and parietal lesions in patients with acute pulmonary thromboembolism (APTE), using intravascular ultrasound (IVUS) and angioscopy (AS). In APTE without underlying disease mainly non-echorich intraluminal mass was noted, with a pulsatile and thin intima. On AS red thrombi with white fibrin coating could be directly observed, and no parietal lesions were found. The findings of the pulmonary arterial intima and thrombus were different between APTE and chronic pulmonary thromboembolism (CPTE), and even among CPTE cases. IVUS and AS are useful in characterizing the thrombi and related pulmonary artery lesions in PTE.