Safety profile of vonoprazan compared with proton pump inhibitors: insight from a pharmacovigilance study.

Proton pump inhibitors (PPIs) are used to treat acid-related disorders such as peptic ulcer and gastroesophageal reflux disease. Recently, vonoprazan, a novel potassium-competitive acid blocker (P-CAB), has been introduced as more effective treatment option. The purpose of this study was to clarify the adverse events associated with vonoprazan compared to PPIs using a spontaneous reporting system database. We performed a retrospective pharmacovigilance disproportionality analysis using the Japanese Adverse Drug Event Report (JADER) database. Adverse event reports submitted to the Pharmaceuticals and Medical Devices Agency between 2004 and 2017 were analyzed, and the reporting odds ratio (ROR) and 95% confidence interval (CI) for each adverse event were calculated. The database comprised 11,433 reports associated with PPIs, and 636 reports with vonoprazan. Hepatic and skin disorders were commonly detected in both PPIs and vonoprazan. There was a significant association of interstitial lung disease with PPIs as a class (ROR: 1.61, 95%CI: 1.47-1.77), but not with vonoprazan. Vonoprazan was strongly associated with haemorrhagic enterocolitis (ROR, 86.5; 95%CI, 59.7125). Among the PPIs, the signal score of microscopic colitis was noteworthy in the case of lansoprazole (ROR, 405; 95%CI, 348-472). It is suggested that there is a diversity in the strength of the association between PPIs and vonoprazan with adverse events. Our results may provide useful information for the treatment of acid-related disorders, but further research with more data is needed to finally clarify this.

Emerging applications of single-cell diagnostics.

The performance of DNA sequencers (next generation sequencing) is rapidly enhanced these days, being used for genetic diagnostics. Although many phenomena could be elucidated with such massive genome data, it is still a big challenge to obtain comprehensive understanding of diseases and the relevant biology at the cellular level. In general terms, the data obtained to date are averages of ensembles of cells, but it is not certain whether the same features are the same inside an individual cell. Accordingly, important information may be masked by the averaging process. As the technologies for analyzing bio-molecular components in single cells are being developed, single cell analysis seems promising to address the current limitations due to averaging problems. Although the technologies for single cell analysis are still at the infant stage, the single cell approach has the potential to improve the accuracy of diagnosis based on knowledge of intra- and inter-cellular networks. In this review several technologies and applications (especially medical applications) of genome and transcriptome analysis or single cells are described.

Development of capillary array DNA sequencers for genome analysis.

Massive analysis of biological molecules is being successfully applied to elucidate various biological phenomena using techniques of molecular science. This new era was initiated by the Human Genome Project, which was supported by the development of various automated tools including DNA sequencers and DNA chips. A small project led by Akiyoshi Wada in Japan triggered the development of automated DNA analysis instruments. As one of the members of the project, the author has been engaged in developing DNA analyzers including capillary array DNA sequencers. Here, the principles and development of the main technologies related to capillary array DNA sequencers, which contributed to the completion of the Human Genome Project, are reviewed.

Endpoint stiffness magnitude increases linearly with a stronger power grasp.

Humans can increase the endpoint stiffness of their arm to reduce self-generated movement variability and to reject unpredictable perturbations from the environment, like during handheld drilling, thereby increasing movement precision. Existing methods to estimate changes in the endpoint stiffness use robotic interfaces to apply position or force perturbations to measure the arm's dynamic response. We propose an alternative method of measuring changes in the power grasp force to estimate adaptations in the magnitude of the arm's endpoint stiffness. To validate our method, we examined how the strength of the power grasp, when holding onto a robotic manipulandum, affected the arm's endpoint stiffness in three different locations of the workspace. The endpoint stiffness magnitude increased linearly with the grasp force, and this linear relationship did not depend on the arm's posture or position in the workspace. The endpoint stiffness may have increased as a combination of greater grasp stiffness and greater arm stiffness, since larger co-contraction was observed in the elbow and shoulder with a stronger grasp. Changes in the grasp force could serve as a metric in assessing how humans adapt their endpoint stiffness magnitude.

Increase in Grasp Force Reflects a Desire to Improve Movement Precision.

Grasping is an action engraved in the human genome, enabling newborn infants to hang from a monkey-bar immediately after birth. The grasp force provides rich information about the brain's control of arm movements. In this study, we tested the hypothesis that the grasp force increases to improve the hand's movement precision during reaching. In two reaching experiments, subjects increased grasp force to suppress movement imprecision that arose from both self-generated motor noise and from an unpredictable environment. Furthermore, the grasp force did not increase constantly, but increased specifically along the movement where the hand's deviation was greatest. The increased grasp was premeditated and was not a reaction to environmental forces, suggesting that the central nervous system has a predictive, state-dependent model of movement precision during reaching. The grasp force provides a high temporal resolution and calibration-less estimate of movement precision adaptation.

Construction of a versatile ultralow temperature scanning tunneling microscope.

We constructed a dilution-refrigerator (DR)-based ultralow temperature scanning tunneling microscope (ULT-STM) which works at temperatures down to 30 mK, in magnetic fields up to 6 T and in ultrahigh vacuum (UHV). Besides these extreme operation conditions, this STM has several unique features not available in other DR-based ULT-STMs. One can load STM tips as well as samples with clean surfaces prepared in an UHV environment to a STM head keeping low temperature and UHV conditions. After then, the system can be cooled back to near the base temperature within 3 h. Due to these capabilities, it has a variety of applications not only for cleavable materials but also for almost all conducting materials. The present ULT-STM has also an exceptionally high stability in the presence of magnetic field and even during field sweep. We describe details of its design, performance, and applications for low temperature physics.

Quantitative detection of single nucleotide polymorphisms for a pooled sample by a bioluminometric assay coupled with modified primer extension reactions (BAMPER).

A new method for SNP analysis based on the detection of pyrophosphate (PPi) is demonstrated, which is capable of detecting small allele frequency differences between two DNA pools for genetic association studies other than SNP typing. The method is based on specific primer extension reactions coupled with PPi detection. As the specificity of the primer-directed extension is not enough for quantitative SNP analysis, artificial mismatched bases are introduced into the 3'-terminal regions of the specific primers as a way of improving the switching characteristics of the primer extension reactions. The best position in the primer for such artificial mismatched bases is the third position from the primer 3'-terminus. Contamination with endogenous PPi, which produces a large background signal level in SNP analysis, was removed using PPase to degrade the PPi during the sample preparation process. It is possible to accurately and quantitatively analyze SNPs using a set of primers that correspond to the wild-type and mutant DNA segments. The termini of these primers are at the mutation positions. Various types of SNPs were successfully analyzed. It was possible to very accurately determine SNPs with frequencies as low 0.02. It is very reproducible and the allele frequency difference can be determined. It is accurate enough to detect meaningful genetic differences among pooled DNA samples. The method is sensitive enough to detect 14 amol ssM13 DNA. The proposed method seems very promising in terms of realizing a cost-effective, large-scale human genetic testing system.

Multiplex polymerase chain reaction (PCR) with color-tagged module-shuffling primers for comparing gene expression levels in various cells.

A method based on the multiplex polymerase chain reaction (PCR) and gel electrophoresis for the comparative analysis of gene expression levels was developed. Using the method many cDNA fragments from different sources can be compared simultaneously. Competitive PCR amplification of expressed genes from different sources was performed by using 'module-shuffling primers' (MPSs). The MPSs (labeled with different fluorophores) consist of sequence modules of 3 or 4 nt. The modules are arranged in different orders in each primer; therefore, the base sequences of the primers are different but their melting temperatures are identical. The genes expressed in different sources are ligated with tags complementary with the MPSs. Tag-ligated fragments are mixed in one tube and amplified at the same amplification efficiency by the MPSs. Amplified fragments are detected separately by multiple-color gel electrophoresis. This method can detect different amounts of each expressed gene, up to a difference in amounts of 30%, and its detection limit is 0.1 amol per assay.

Individualistic weight perception from motion on a slope.

Perception of an object's weight is linked to its form and motion. Studies have shown the relationship between weight perception and motion in horizontal and vertical environments to be universally identical across subjects during passive observation. Here we show a contradicting finding in that not all humans share the same motion-weight pairing. A virtual environment where participants control the steepness of a slope was used to investigate the relationship between sliding motion and weight perception. Our findings showed that distinct, albeit subjective, motion-weight relationships in perception could be identified for slope environments. These individualistic perceptions were found when changes in environmental parameters governing motion were introduced, specifically inclination and surface texture. Differences in environmental parameters, combined with individual factors such as experience, affected participants' weight perception. This phenomenon may offer evidence of the central nervous system's ability to choose and combine internal models based on information from the sensory system. The results also point toward the possibility of controlling human perception by presenting strong sensory cues to manipulate the mechanisms managing internal models.

Stress thallium-201 transaxial emission computed tomography: quantitative versus qualitative analysis for evaluation of coronary artery disease.

Stress thallium-201 myocardial distribution was quantitatively evaluated by emission transaxial tomography in 104 patients who underwent coronary arteriography. The initial uptake and percent washout of thallium were assessed by the circumferential profile curves of the three short-axis sections and one middle right anterior oblique long-axis section. This quantitative tomographic analysis showed abnormal distribution in all but two patients (98%) with coronary artery disease, whereas qualitative analysis showed abnormality in 76 of the patients (93%). Quantitative analysis showed better sensitivity (91%) for detecting involved coronary vessels than qualitative analysis (80%, p less than 0.01), especially in three vessel disease (82 versus 67%, p less than 0.05). For localization of individual vessel involvement, quantitative analysis showed high sensitivity (right coronary artery: 96%, left anterior descending artery: 90% and left circumflex artery: 88%) as compared with qualitative analysis (88, 83 and 63%, respectively, p less than 0.05), while similar specificity was observed (92% for quantitative and 93% for qualitative analyses). Furthermore, in the study of patients without infarction, myocardial segments supplied by coronary vessels with moderate stenosis (51 to 75%) revealed abnormality more often with quantitative (81%) than with qualitative (56%) analysis. Thus, quantitative analysis of stress thallium emission tomography provides improved sensitivity for the detection of diseased coronary vessels in patients with three vessel disease and those with moderate stenosis. It is a valuable technique for the evaluation of coronary artery disease.

Non-Q wave versus Q wave myocardial infarction: regional myocardial metabolism and blood flow assessed by positron emission tomography.

This study compared regional myocardial blood flow at rest and during supine exercise as well as regional myocardial glucose utilization in the fasting condition in 22 patients, 11 with antecedent non-Q wave and 11 with antecedent Q wave infarction. With use of N-13 (nitrogen-13) ammonia and F-18 (fluorine-18) deoxyglucose as tracers of blood flow and exogenous glucose utilization and positron emission tomography, hypoperfused areas were noted at rest and during exercise in all 11 patients (100%) with Q wave infarction. Among the 11 patients with non-Q wave infarction such areas were noted in only 5 (45%) at rest and in 8 (73%) during exercise. Furthermore, segmentally enhanced F-18 deoxyglucose uptake corresponding to the infarcted areas (identified electrocardiographically) was seen in 10 (91%) of the 11 patients with non-Q wave infarction but in only 4 (36%) of the 11 patients with Q wave infarction (p less than 0.01). In conclusion, segmental F-18 deoxyglucose uptake as a possible sign of myocardial viability was seen more frequently in non-Q wave than in Q wave infarction and, importantly, regionally enhanced F-18 deoxyglucose uptake occurred even in the absence of segmental rest or exercise blood flow abnormalities, or both, in 5 (45%) of 11 patients with non-Q wave infarction.

Effects of coronary artery reperfusion on relation between creatine kinase-MB release and infarct size estimated by myocardial emission tomography with thallium-201 in man.

The quantitative relations between serum creatine kinase-MB isoenzyme (CK-MB) release and the final infarct size estimated by myocardial emission computed tomography with thallium-201 was assessed in 37 patients with a first acute transmural myocardial infarction who underwent intracoronary thrombolysis using urokinase 4.6 +/- 1.9 hours after the onset of symptoms. Serial CK-MB determinations were used to calculate the accumulated release of CK-MB (sigma CK-MB). Myocardial emission tomography with thallium-201 was performed 4 weeks after the onset, and infarct volume was measured from reconstructed tomographic images by computerized planimetry. The results are presented for two groups of patients: 11 patients with unsuccessful thrombolysis (group A) and 26 patients with successful thrombolysis (group B). An excellent linear relation was found for group A (sigma CK-MB = 6.4 X infarct volume + 47.7, r = 0.91), whereas a different linear relation was observed for group B (sigma CK-MB = 10.5 X infarct volume + 89.1, r = 0.80). Moreover, serum CK-MB activity reached a peak at 21.1 +/- 2.2 hours after the onset in group A and reached an earlier peak at 12.5 +/- 2.9 hours in group B (p less than 0.001). These data suggest that acute coronary recanalization alters the kinetics of CK-MB release, resulting in greater CK-MB release into the serum for equivalent infarct volume estimated by myocardial emission tomography with thallium-201. Thus, serum CK-MB time-activity curves after acute myocardial infarction may be influenced considerably by acute reperfusion, which is an important factor that should be incorporated in the interpretation of enzymatic estimates of infarct size in human patients.

Prognostic value of an increase in fluorine-18 deoxyglucose uptake in patients with myocardial infarction: comparison with stress thallium imaging.

OBJECTIVES: This study was undertaken to evaluate the prognostic value of an increase in fluorine (F)-18 deoxyglucose uptake compared with clinical, angiographic and stress thallium findings in patients with myocardial infarction. BACKGROUND: Positron emission tomography (PET) imaging using F-18 deoxyglucose has been applied to assess tissue viability in patients with coronary artery disease. We hypothesized that patients with a myocardial segment with augmented F-18 deoxyglucose uptake are at high risk for a future cardiac event. METHODS: One hundred fifty-eight consecutive patients with myocardial infarction referred for F-18 deoxyglucose PET and stress thallium scans were studied. Follow-up was obtained in 84 patients at a mean interval of 23 months to investigate prognostic implications of radionuclide studies. RESULTS: Seventeen patients had a cardiac event during the follow-up interval. Univariate analysis showed that an increase in F-18 deoxyglucose uptake was the best predictor of a future cardiac event (p = 0.0006), followed by the number of stenosed vessels (p = 0.008). In the multivariate analysis, when an increase in F-18 deoxyglucose uptake was entered into the model, only angiographic variables had an independent prognostic value, whereas no other radionuclide variables showed significant prognostic value. Among patients who did not show redistribution, a future cardiac event was observed more often in patients with than in those without an increase in F-18 deoxyglucose uptake (p < 0.05). CONCLUSIONS: Thus, an increase in F-18 deoxyglucose uptake seemed to be the best predictor of a future cardiac event among all clinical, angiographic and radionuclide variables in this study of stable patients with myocardial infarction. Even when a stress thallium-201 scan does not show redistribution, those patients who have an increase in F-18 deoxyglucose uptake in a PET study may be at risk for a future cardiac event, and these patients may need aggressive treatment to prevent a future cardiac event.

DNA sequencing using ultra small amounts of reagents and template.

One of the key points in the genome project is finding ways to reduce the running cost in DNA sequencing. One way is to use a highly-sensitive fluorescent DNA sequencer, where only trace amounts of template DNA and reagents are needed. An experimental protocol optimized for the trace amounts of DNA analysis was established by using the hybridization reaction rate coefficient of primers on template DNA, which was estimated to be 7.5 x 10(5) M-1sec-1 at 37 degrees C. One femtomole of template DNA with 0.001 unit of modified T7 DNA polymerase (Sequenase Ver. 2.0) and also 0.45 fmol of M13 template DNA with 0.01 unit of Taq DNA polymerase were enough to sequence DNA of up to 400 bases.

DNA sequencing directly from a mixture using terminal-base-selective primers.

DNA cloning is often used to select and amplify one DNA species from a mixture. However, the cloning process is complex and labor-intensive. We have developed a new two-step method for DNA sequencing directly from a mixture. The first is the introduction of a known oligonucleotide (common part) into the terminus of unknown DNA by ligation. The second is selective DNA sequencing using primers with two additional nucleotides at the 3' terminus in addition to the common part (terminal-base-selective primers). The primers work only for templates on which the primers perfectly hybridized. This method was found to be effective for the HindIII digestion products of lambda phage.

Fragment walking for long DNA sequencing by using a library as small as 16 primers.

A DNA sequence can be rapidly and efficiently determined by digesting it into segments small enough to be sequenced at one time, then assembling them into contigs by searching for overlaps. Fragment walking achieves this without subcloning or preparing many kinds of primers; fragments obtained by digesting a template DNA are sequenced in parallel directly from the fragment mixture by using a set of 16 primers. Since the sequence adjacent to each cutting site is determined together with the fragment sequence, the contiguous fragment can be easily determined. The complete template DNA sequence can thus efficiently determined. The sequencing of pUC19 (2.7 kb) from both sides was done in one step by using only 15 fragments. The redundancy was only about 1.3, greatly reducing base-reading redundancy. Automating this strategy would increase the speed and efficiency of large-scale DNA sequencing.

Heterogeneity within the nonstructural protein 5-encoding region of hepatitis C viruses from a single patient.

Nucleotide (nt) sequence heterogeneity of the hepatitis C virus (HCV) genome derived from a single carrier was investigated. A polymerase chain reaction (PCR) product of 311 bp in the putative nonstructural protein 5-encoding region was directly sequenced, while part of a PCR product was cloned, and sequence analyses were carried out for 27 independent clones. Although 14 of the 27 clones were conserved, ten other types of nt sequences were found. The difference was at most 3 nt (1.1%). A directly determined sequence showed the major sequence of the cloned products. Since most of the nt changes occurred in the third letter of a codon, these nt changes might not have originated from random misincorporation during the PCR. These results of natural divergence of genome population in a single carrier suggest that HCV is a typical RNA virus with a quasi-species nature due to high mutation rates.

Chemical robot for enzymatic reactions and extraction processes of DNA in DNA sequence analysis.

A chemical robot capable of performing enzymatic reactions and extraction processes of DNA has been developed. The basic functions of this robot include handling of plastic tubes with caps, micropipetting, mixing, microcentrifuging and incubating. As a result, almost all of the pre-electrophoresis steps can be carried out. In addition, because these processes are automated, the working time of each process can be reduced. The reproducibility of the automated operation is equivalent to that of a skilled operator.

Myocardial uptake of technetium-99m stannous pyrophosphate in experimental viral myopericarditis.

Distribution of technetium-99m stannous pyrophosphate was studied in mice with experimentally induced viral myopericarditis. Myocardial and bone uptakes of Tc-PPi were compared in 55 mice inoculated with coxsackievirus B3 (Nancy strain). The myocardium-to-bone uptake ratio in 33 mice with myopericarditis was increased to a greater extent than that seen in 22 mice without myopericarditis (p less than 0.001). In the severely involved heart, the uptake per gram exceeded that in the bone. Myocardial uptake in myopericarditis can be visualized on a whole-body image using a pinhole collimator and a left lateral view. Our experimental studies suggest the potential clinical usefulness of myocardial scintigraphy in viral myopericarditis.

Clinical evaluation of thallium-201 emission myocardial tomography using a rotating gamma camera: comparison with seven-pinhole tomography.

Single-photon emission computed tomography (SPECT) for thallium-201 myocardial imaging with a rotating gamma camera was evaluated in comparison with planar imaging and seven-pinhole tomography (7P). Cardiac phantom studies indicated that defects 2 cm in diameter can be visualized by both tomographic methods, but the 7P method showed propagation of the image into nearby planes, with lower image contrast. In a clinical study of 47 patients with myocardial infarction, both sensitivity and specificity for the SPECT system were high (96 and 89%, respectively); the 7P system, on the other hand, showed good sensitivity (93%) but poor specificity (68%), while planar imaging performed conversely (75 against 89%). The overall accuracy was not significantly improved in the 7P method (planar: 81%, 7P: 83%, and SPECT: 94%). Our study indicates that SPECT, which can reconstruct reliable tomographic sections in either the transaxial, frontal, or sagittal planes, will result in a remarkable improvement in the clinical evaluation of ischemic heart disease.