Total Synthesis and Biological Evaluation of 19-Hydroxysarmentogenin-3 -O-α-l-rhamnoside, Trewianin, and Their Aglycons.

Cardenolides comprise an important family of natural steroids with a wide spectrum of biological activities. Although 19-hydroxysarmentogenin-3- O-α-l-rhamnoside (1a) and trewianin (1b) were structurally determined to have cardenolide structures, their biological activities have not been evaluated. The 6/6/6/5-membered ABCD-ring systems of both 1a and 1b are decorated by a ß-oriented C17-butenolide, three C11,14,19-hydroxy groups, and a C3-O-l-rhamnoside moiety. On the other hand, 1a and 1b are epimeric at the C5-position. The structures of 1a and 1b were assembled from four simple fragments by applying a convergent and unified strategy. The AB-ring 10a/b and the D-ring 8/9 were tentatively tethered at the acetal moiety, and a subsequent stereoselective 6- exo radical reaction linked the two fragments. Next, an aldol reaction enabled simultaneous introduction of three new stereocenters of the C-rings of 5aa and 54. Attachment of the C17-butenolide led to aglycons 2a and 2b. l-Rhamnose was then installed into 2a and 2b to yield the targets 1a and 1b, respectively. Finally, the growth inhibitory activity of 1a, 1b, 2a, and 2b was assessed against MCF-7 human breast carcinoma cells. The significantly higher activities of 1a and 1b in comparison to 2a and 2b demonstrated the biological importance of the monosaccharide substructure.

Identification of candidate genes involved in the etiology of sporadic Tourette syndrome by exome sequencing.

Tourette Syndrome (TS) is a neurodevelopmental disorder characterized by chronic motor and vocal tics. Although there is a large genetic contribution, the genetic architecture of TS remains unclear. Exome sequencing has successfully revealed the contribution of de novo mutations in sporadic cases with neuropsychiatric disorders such as autism and schizophrenia. Here, using exome sequencing, we investigated de novo mutations in individuals with sporadic TS to identify novel risk loci and elucidate the genetic background of TS. Exome analysis was conducted for sporadic TS cases: nine trio families and one quartet family with concordant twins were investigated. Missense mutations were evaluated using functional prediction algorithms, and their population frequencies were calculated based on three public databases. Gene expression patterns in the brain were analyzed using the BrainSpan Developmental Transcriptome. Thirty de novo mutations, including four synonymous and four missense mutations, were identified. Among the missense mutations, one in the rapamycin-insensitive companion of mammalian target of rapamycin (RICTOR)-coding gene (rs140964083: G > A, found in one proband) was predicted to be hazardous. In the three public databases analyzed, variants in the same SNP locus were absent, and variants in the same gene were either absent or present at an extremely low frequency (3/5,008), indicating the rarity of hazardous RICTOR mutations in the general population. The de novo variant of RICTOR may be implicated in the development of sporadic TS, and RICTOR is a novel candidate factor for TS etiology.

Rotational plane-wise analysis of angular movement of neck motor tics in Tourette's syndrome.

Motor tics are sudden, rapid, recurrent, non-rhythmic movements. There is a lack of quantitative assessment methods for the motor tics despite severe neck complications. We aimed to provide an improved quantitative method for neck tic assessment in motor tic disorders. We recorded neck motor tics in patients with motor tic disorders and voluntary neck movements in healthy controls. The maximum peak angular velocities and angular accelerations were calculated. Motor tics were assessed in three orthogonal planes (yaw, pitch, and roll) separately, and compared between the patients with motor tic disorders and controls. Correlations between the maximum angular velocities/accelerations and tic counts were also assessed. In the pitch plane, motor tics of the patients showed higher angular velocities/accelerations than voluntary movements of the controls. Angular acceleration in the yaw, and roll planes showed positive correlations with tic count. Some of the observed tics were comparable to the movements experienced in contact sports. Our findings may aid in the identification of populations at a high risk for severe neck complications among motor tic disorder patients.

A 2-year longitudinal follow-up of quantitative assessment neck tics in Tourette's syndrome.

BACKGROUND: Neck motor tics in Tourette's syndrome can cause severe neck complications. Although addressed in a few longitudinal studies, the clinical course of Tourette's syndrome has not been quantitatively assessed. We had previously developed a method for quantifying the angular movements of neck tics using a compact gyroscope. Here, we present a follow-up study aimed at elucidating the clinical course of neck tics at both the group and individual levels. METHODS: Eleven patients with Tourette's syndrome from our previous study participated in the present study, and their neck tics were recorded during a 5-min observation period. The severity of neck symptoms was assessed using the Yale Global Tic Severity Scale. The peak angular velocities and accelerations, tic counts, and severity scores in our previous study (baseline) and the present study (2-year follow-up) were compared at the group and individual levels. The individual level consistency between baseline and follow-up were calculated using intra-class correlation coefficients (ICCs, one-way random, single measure). RESULTS: At the group level, no significant change was observed between baseline and follow-up. At the individual level, angular velocity (ICC 0.73) and YGTSS scores (ICC 0.75) had substantial consistency over the two time points, and angular acceleration (ICC 0.59) and tic counts (ICC 0.69) had moderate consistency. CONCLUSIONS: The intensity and frequency of neck tics did not change over time. Therefore, quantification of angular neck motor tics will aid in identifying patients with neck tics at high risk for severe neck complications.

Correction: Cows painted with zebra-like striping can avoid biting fly attack.

[This corrects the article DOI: 10.1371/journal.pone.0223447.].

Cows painted with zebra-like striping can avoid biting fly attack.

Experimental and comparative studies suggest that the striped coats of zebras can prevent biting fly attacks. Biting flies are serious pests of livestock that cause economic losses in animal production. We hypothesized that cows painted with black and white stripes on their body could avoid biting fly attacks and show fewer fly-repelling behaviors. Six Japanese Black cows were assigned to treatments using a 3 × 3 Latin-square design. The treatments were black-and-white painted stripes, black painted stripes, and no stripes (all-black body surface). Recorded fly-repelling behaviors were head throw, ear beat, leg stamp, skin twitch, and tail flick. Photo images of the right side of each cow were taken using a commercial digital camera after every observation and biting flies on the body and each leg were counted from the photo images. Here we show that the numbers of biting flies on Japanese Black cows painted with black-and-white stripes were significantly lower than those on non-painted cows and cows painted only with black stripes. The frequencies of fly-repelling behaviors in cows painted with black-and-white stripes were also lower than those in the non-painted and black-striped cows. These results thus suggest that painting black-and-white stripes on livestock such as cattle can prevent biting fly attacks and provide an alternative method of defending livestock against biting flies without using pesticides in animal production, thereby proposing a solution for the problem of pesticide resistance in the environment.

Right bundle branch block in acute myocardial infarction treated by primary coronary angioplasty and stenting.

Patients with right bundle branch block (RBBB) in acute myocardial infarction (AMI) have a significantly higher mortality rate even with the advent of thrombolytic therapy. This study was undertaken to assess the impact of primary percutaneous transluminal coronary angioplasty (PTCA) and stenting on the outcome of patients with RBBB in AMI. A total of 600 patients with AMI who underwent primary PTCA and stenting (rate: 61%) < 12 hours of onset were studied. A 12-lead ECG was obtained at least every 6 hours. Serial creatine kinase was measured, and left ventricular ejection fraction was obtained during the hospital stay. Among 600 patients with AMI, 94 patients (15.7%) had RBBB; it was persistent in 31 (33%) and transient in 63 (67%). In-hospital mortality rate was 7.3% in patients without RBBB, 7.9% in transient RBBB, and 25.8% in persistent RBBB (p < 0.02). The incidence of heart failure was 26.5% in those without RBBB, 34.9% in transient RBBB, and 58.1% in persistent RBBB (p < 0.001). There was no significant difference among these 3 groups in ventricular arrhythmias and complete atrioventricular block. Peak creatine kinase was 3,214+/-2,293 U/L in those without RBBB, 4,558+/-3,316 U/L in transient RBBB (p < 0.001), and 5,635+/-3,920 U/L in persistent RBBB (p < 0.001). Left ventricular ejection fraction was 50+/-11% in those without RBBB, 47+/-11% in transient RBBB (p < 0.05), and 42+/-13% in persistent RBBB (p < 0.001). Patients with AMI treated by primary PTCA and stenting had an increased incidence of transient RBBB, especially following reperfusion therapy, although the clinical outcome was similar to that of those without RBBB. In contrast, there was no satisfactory improvement in clinical outcomes in those with persistent RBBB.

Inhibition of Na(+),K(+)-ATPase by the extract of Stephania cephararantha HAYATA and bisbenzylisoquinoline alkaloid cycleanine, a major constituent.

The Stephania cephararantha HAYATA extract, and its constituent bisbenzylisoquinoline alkaloids, such as cycleanine, cepharanthine, isotetrandrine, berbamine, homoaromoline, and cepharanoline were studied for effects on Na(+),K(+)-ATPase activity. The S. cephararantha HAYATA extract inhibited Na(+),K(+)-ATPase activity with an apparent IC(50) value of 540 microg/mL. Cycleanine markedly inhibited Na(+),K(+)-ATPase activity with an IC(50) value of 6.2 x 10(-4)M. It slightly inhibited Mg(2+)-ATPase, H(+)-ATPase, and Ca(2+)-ATPase. No effects on alkaline and acid phosphatase activities were observed. The inhibition by isotetrandrine, homoaromoline, cepharanthine, and berbamine was less marked, and cepharanoline showed no effect. Five synthetic analogues of cepharanthine slightly inhibited the activity. The mechanism of inhibition by cycleanine on Na(+),K(+)-ATPase activity was examined in detail, and the following results were obtained in the overall reaction: (1) the mode of inhibition was noncompetitive with respect to ATP; (2) the degree of inhibition was decreased with a decrease of K(+) concentration; (3) it was not affected by Na(+) concentration; (4) the inhibition mechanism was different from that of ouabain. The activity of K(+)-dependent p-nitrophenyl phosphatase, a partial reaction of Na(+),K(+)-ATPase, did not appear to have been inhibited by cycleanine in the reaction mixture containing 15 mM K(+) (optimum condition). However, cycleanine increased the K(0.5) value for K(+) and reduced the K(i) values for Na(+) and ATP, in K(+)-dependent p-nitrophenyl phosphatase. Cycleanine might interact with the enzyme in Na.E(1)-P form and prevents the reaction step from Na.E(1)-P to E(2)-P.

Bone marrow stimulation and left ventricular function in acute myocardial infarction.

BACKGROUND: Experimental studies have demonstrated that bone marrow stem cells can migrate into peripheral blood and regenerate damaged myocardium. HYPOTHESIS: Bone marrow stimulation might improve myocardial functional recovery after acute myocardial infarction (AMI). METHODS: In all, 104 consecutive patients with anterior wall AMI treated by primary coronary angioplasty and stenting within 12 h after onset, and who underwent left ventriculography on admission and 6 months after discharge, were studied. Among these, 23 patients (Group 1) demonstrated transient appearance of immature blood cells including myelocytes, promyelocytes, and myeloblasts during hospital stay. Thirty-eight matched patients in whom no immature blood cells were detected were studied as a control group (Group 2). RESULTS: There was no significant difference in baseline characteristics between the two groups. There was no significant difference in left ventricular ejection fraction (EF) on admission between Group 1 (33 +/- 12%) and Group 2 (34 +/- 8%). In contrast, EF was significantly better in Group 1 (47 +/- 12%) than in Group 2 (40 +/- 10%, p = 0.016) 6 months after discharge. CONCLUSION: The study suggests significantly greater improvement in left ventricular function in patients with AMI with sign of bone marrow stimulation than in matched patients with no sign of bone marrow stimulation.

Prolongation of QT interval as a predictor of no-reflow induced by rotational atherectomy.

Although coronary rotational atherectomy (RA) is widely applied to clinical cases, the incidence of coronary no-reflow associated with it is higher than in percutaneous transluminal coronary angioplasty (PTCA) and stenting. This study was undertaken to predict no-reflow by using conventional electrocardiograms (ECGs). A total of 105 patients who underwent RA (group 1) and 40 who underwent PTCA (group 2) were studied. Eight patients of group 1, all of whom had long calcified coronary lesions, were complicated with no-reflow following RA. Standard 12-lead ECGs were recorded before and throughout the interventional procedures. Maximum and minimum QT intervals and QT dispersion were measured and corrected by heart rate. Corrected and uncorrected QT intervals and QT dispersion were significantly prolonged by RA in group 1 patients without no-reflow: maximum QTc, 428 +/- 28 ms --> 485 +/- 53 ms, p<0.001. The increases in QT intervals were more remarkable in group 1 patients with no-reflow: maximum QTc, 434 +/- 15 ms --> 552 +/- 39 ms, p<0.001. Of the 33 patients with maximum QTc > or = 500 ms, 8 were complicated with no-reflow. No patients with maximum QTc < 500 ms had no-reflow. There was no significant increase in QT intervals in group 2. Adsorption of calcium ions from the myocardium by pulverized calcified atheromatous debris when these pass through coronary capillaries, resulting in transient myocardial hypocalcemia, was considered as a possible mechanism of QT prolongation. Because QT prolongation appears during the initial RA trial, prolonged QT intervals could be a predictor of no-reflow. It is recommended to avoid repetitive RA if marked QT prolongation is observed at the initial RA trial.

Coronary blood flow in evolving myocardial infarction preceded by preinfarction angina: a critical reevaluation of preconditioning effects in clinical cases.

This study was undertaken to reevaluate the protective effects of preinfarction (pre-MI) angina in acute MI. The mechanisms involved in the apparent protective effects of pre-MI angina have been presumed to be preconditioning effects as defined by experimental studies. The phenomenon, has not, however, been observed in diabetic and/or elderly patients or in those treated by primary percutaneous coronary intervention (PCI). A total of 202 patients with anterior wall MI without a history of MI who underwent primary PCI with coronary balloon dilation and stenting (rate: 50%) <6 hours after onset were studied. Patients included 59 with pre-MI angina (group 1) and 143 without pre-MI angina (group 2). The infarct-related coronary artery was patent on admission in 46% of group 1 and 31% of group 2 (p=0.045). Thrombolysis in Myocardial Infarction (TIMI) 1-2 flow was significantly more frequent in group 1 (29%) than in group 2 (11%, p=0.005) on admission. Among risk factors, clinical background, coronary anatomy, and clinical outcome, the only significant predictor of pre-MI angina was a patent infarct-related coronary artery on admission (odds ratio: 2.39, p = 0.015). There was no significant difference in left ventricular ejection fraction, peak creatine kinase, or the incidences of heart failure and in-hospital/follow-up deaths between these groups. In conclusion, the findings suggest that the protective effects reported in MI with pre-MI angina treated by thrombolysis are due to more fragile thrombotic occlusion, which can be more easily recanalized by thrombolysis, whereas the beneficial effects are not evident in those treated by primary PCI.

Clinical evaluation of coronary lesion characteristics in acute myocardial infarction.

Coronary lesion instability at the onset of acute myocardial infarction (AMI) was evaluated. The mechanism of AMI has been considered to be coronary lesion instability with occlusive thrombus, although more than one half of AMI occurs in clinically stable patients. A total of 313 AMI patients treated by primary percutaneous transluminal coronary angioplasty with provisional stenting (rate, 41%) were studied. They were divided into 2 groups: group 1A (n = 211), without unstable angina before AMI onset, and group 1B (n = 102), with unstable angina before onset. Moreover, angina patients treated similarly were studied: group 2A (n = 180), with stable angina, and group 2B (n = 204), with unstable angina. Coronary lesion instability at AMI onset was also predicted by C-reactive protein (CRP) levels within 6 hours after onset, before they were affected by myocardial damage. The incidence of repeated AMI and/or target vessel revascularization was 1.9% in group 1A, 7.8% in 1B (p=0.035), 1.7% in 2A, and 5.9% in 2B (p=0.043). Event-free survival curves were consistent with each other in groups 1A and 2A and in groups 1B and 2B. CRP levels on admission were 2.0 +/- 1.7 mg/L in group 1A, 3.3 +/- 4.8 mg/L in group 1B (p<0.001), 2.1 +/- 1.7 mg/L in group 2A, and 3.4 +/- 4.7 mg/L in group 2B (p<0.001). Thus coronary lesion characteristics at AMI onset appeared to be similar in groups 1A and 2A and in groups 1B and 2B. A substantial number of patients have stable culprit lesions at the onset of AMI.

Time course of changes in antioxidant activity of milk from dairy cows fed a trehalose-supplemented diet.

This study was to investigate the time course of changes to the antioxidant activity of milk from cows fed a trehalose-supplemented diet, and to determine possible underlying mechanisms for observed changes. Six Holstein cows were used, and subjected to two experimental feeding periods consisting of a 1% trehalose-supplemented diet for 10 days, followed by a basal diet only (no trehalose) for 10 days. 1,1-Diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activities in milk were gradually increased during the trehalose supplementation period and were highest at the end of the second period. However, trehalose was not detected in the milk and plasma of dairy cows fed a diet supplemented with trehalose for 10 days, indicating that the increased antioxidant activity in the milk of trehalose-fed cows is not due to the direct transfer of trehalose to the milk. Plasma DPPH activities exhibited a similar time course to that seen for milk. Relative superoxide dismutase (SOD) activities in the rumen were higher 3 days after the end of trehalose supplementation than at any other time during the experimental periods. These results suggested that the improved antioxidant activity in milk and plasma of cows fed a trehalose-supplemented diet was due to improved ruminal relative SOD activity.

Missense mutation with/without nonsense mutation of the p53 gene is associated with large cell morphology in human malignant lymphoma.

Mutations in p53 gene exons 5-9 were studied in 44 non-Hodgkin's lymphomas (NHL) consisting of 35 B-NHL and 9 T-NHL. Missense mutations were found in two diffuse large B-cell lymphomas (DLBL) and one peripheral T-cell lymphoma (unspecified). Double transversion missense and nonsense mutations were detected in one DLBL and one adult T-cell leukemia/lymphoma. Silent mutations were found in two DLBL. Detailed histomorphological study showed that cases harboring p53 missense mutation with/without nonsense mutation tended to have larger nuclei with much more prominent nucleoli. Cytomorphometric analysis was therefore conducted by measuring the gross area of 100 lymphoma cell nuclei in 44 cases and the results were compared between lymphomas harboring p53 missense mutation with/without nonsense mutation and lymphomas harboring p53 silent mutation or lacking mutation. It was found that the lymphomas harboring p53 missense mutation with/without nonsense mutation had a highly significantly larger nuclear gross area than lymphomas with silent p53 mutation or lacking mutation (two-sample t-test, P < 0.00001; Exact Wilcoxon rank-sum test, P < 0.00001). This result suggests that p53 mutation might induce enlargement of neoplastic cell nuclei by some molecular mechanism.

Age-related urinary excretion of BK polyomavirus by nonimmunocompromised individuals.

Two polyomaviruses, BK virus (BKV) and JC virus (JCV), are ubiquitous in the human population, generally infecting children asymptomatically and then persisting in renal tissue. It is generally thought that reactivation leads to productive infection for both viruses, with progeny shed in the urine. Several studies have shown that the rate of JC viruria increases with the age of the host, but a systematic approach to examine the shedding of BKV has not been developed. To elucidate the relationship between BK viruria and host age, we obtained urine from donors (healthy volunteers or nonimmunocompromised patients) who were divided into nine age groups, each containing 50 members. A high-sensitivity PCR was used to detect BKV and JCV DNA from urinary samples, and the specificity of amplification was confirmed by sequencing or restriction analysis of the amplified fragments. The rate of BK viruria was relatively low in subjects aged <30 years but gradually increased with age in subjects aged > or =30 years. However, BK viruria was less frequent than JC viruria in adults. The detected BKV isolates were classified into subtypes, and detection rates for individual subtypes were compared among age groups; this analysis showed that viruria of subtypes I (the most prevalent subtype) and IV (the second most prevalent subtype) occurred more frequently in older subjects. Therefore, our results reveal new aspects of BK viruria in nonimmunocompromised individuals.

Maintenance and characterization of an Epstein Barr virus-infected CD56-negative T cell lymphoma.

T cell lymphoma carrying Epstein Barr virus (EBV(+) TL) is very rare among Western countries while it is much more common among Japanese. Here we report an EBV(+) TL which has been maintained for years by the use of mice with severe combined immune deficiency (SCID) mice. Lymphoma was obtained from a 55-year-old male suffering from oculomotor nerve palsy and lymphadenopathy. A small piece of biopsied tumor was transplanted into SCID mice and the lymphoma has been maintained for over 3 years with passages every 2 - 3 weeks. The maintained lymphoma, termed as TMS24, and the original lymphoma cells showed identical phenotype and genotype, including diffuse medium-sized cell morphology lacking granules, suppressor / cytotoxic immunophenotype and identical T cell receptor beta-chain gene rearrangement mode. Further, both were shown to carry an identical EBV clone in terms of the number of terminal repeats and the latency II-type restricted gene expression profile. Cytogenetically, TMS24 retained two characteristic chromosomal translocations of t(1;18)(q32;q21) and t(6;12)(p21;q24). Since only one cell line with such characters has been reported previously, TMS24 should be useful for detailed analysis of EBV(+) TL.