Microstripe pattern substrate consisting of alternating planar and nanoprotrusive regions improved hiPSC-derived cardiomyocytes' unidirectional alignment and functional properties.

The alignment of each cell in human myocardium is considered critical for the efficient movement of cardiac tissue. We investigated 96-well microstripe-patterned plates to align human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes (CMs), which resemble fetal myocardium. The aligned CMs (ACMs) cultured on the microstripe-patterned plates exhibited pathology, motor function, gene expression, and drug response that more closely resembled those of adult cells than did unaligned CMs cultured on a flat plate (FCMs). We used these ACMs to evaluate drug side effects and efficacy, and to determine whether these were similar to adult-like responses. When CMs from patients with hypertrophic cardiomyopathy (HCMs) were seeded and cultured on the microstripe-patterned plates or layered on top of the ACMs, both sets of HCMs showed increased heart rate and synchronized contractions, indicating improved cardiac function. It is suggested that the ACMs could be used for drug screening as cells representative of adult-like CMs and be transplanted in the form of a cell sheet for regenerative treatment of heart failure.

[Acute pancreatitis triggered by an impacted minor papillary stone in a patient with pancreas divisum improved after spontaneous evacuation of the pancreatic stone:a case report].

A 56-year-old woman with epigastric pain was admitted to our hospital. Blood tests and abdominal computed tomography (CT) suggested acute pancreatitis. Abdominal CT revealed a pancreas divisum and stone in the minor papilla. Stone impaction was the most likely cause of the acute pancreatitis. The patient's abdominal pain promptly improved after admission. The patient was treated conservatively, and the pancreatitis resolved. Abdominal CT on the 10th day confirmed spontaneous evacuation of the pancreatic stone. This is a rare case of acute pancreatitis in a patient with pancreas divisum triggered by an impacted stone in the minor papilla, which improved after spontaneous evacuation of the pancreatic stone.

A Case of TAFRO Syndrome Developed after COVID-19 Vaccination.

TAFRO syndrome is a systemic inflammatory disorder, which is characterized by thrombocytopenia, anasarca, fever, reticulin myelofibrosis, renal dysfunction, and organomegaly. It often presents with progressive clinical symptoms and can be fatal. COVID-19 vaccination is important to reduce the number of COVID-19-infected populations and lower the risk of becoming severe. However, serious adverse events have been reported. TAFRO syndrome that progresses after the COVID-19 mRNA vaccination has not yet been reported. A 45-year-old man developed fever, gross hematuria, renal dysfunction, pleural effusions, and ascites immediately after vaccination. This case fulfilled three major categories (thrombocytopenia, anasarca, and systemic inflammation) and two minor categories (renal insufficiency and myelofibrosis) and was diagnosed with TAFRO syndrome. High-dose steroid treatment was initiated, followed by prednisolone administration. After treatment, renal dysfunction and fluid retention were resolved. Universal vaccination against COVID-19 is important for lowering the risk of spreading COVID-19 infection. Several complications, such as renal, hematological, and heart diseases, have been reported; however, its pathogenesis is unclear. The possibility of various complications after the COVID-19 vaccination, including TAFRO syndrome, should be considered.

[Voluntary inoculation of hepatitis B vaccine at pediatric medical institutions].

We conducted a questionnaire survey on voluntary inoculation of hepatitis B (HB) vaccine to children at 79 pediatric clinics. The voluntary vaccination rate was 65.2%, the desired vaccination target was "all infants" at 84.8% of clinics, the recommended method was "only when the patient wishes" at a rate of 80.0%, and "actively recommended" at 20.0%. If there was a request, 71.7% of clinics answered that they would like to recommend it in the future, and 38.9% said that it was difficult to recommend it because of the voluntary nature of vaccination. The requests were "expansion of the scope of regular vaccination" at 60.9% of clinics and "enlightenment activities and information provision" at 54.3%. Since it was suggested that voluntary vaccination is difficult to recommend, it is necessary to proactively provide information and public relations activities regarding its necessity to medical personnel and parents.

Monensin-Induced Increase in Intracellular Na+ Induces Changes in Na+ and Ca2+ Currents and Regulates Na+-K+ and Na+-Ca2+ Transport in Cardiomyocytes.

BACKGROUND/AIMS: Monensin, an Na ionophore, increases intracellular Na ([Na]i). Alteration of [Na]i influences ion transport through the sarcolemmal membrane. So far, the effects of monensin on ventricular myocytes have not been examined in detail. The main objective of this study was to elucidate the mechanism via which monensin-evoked increases in [Na]i affect the membrane potential and currents in ventricular myocytes of guinea pigs. METHODS: Membrane potentials and currents were measured using the whole-cell patch-clamp technique in single myocytes. The concentration of intracellular Ca ([Ca]i) was evaluated by measuring fluorescence intensity of Fluo-4. RESULTS: Monensin (10-5M) shortened the action potential duration (APD) and reduced the amplitude of the plateau phase. In addition, monensin decreased the sodium current (INa) and shifted the inactivation curve to the hyperpolarized direction. Moreover, it decreased the L-type calcium current (ICa). However, this effect was attenuated by increasing the buffering capacity of [Ca]i. The Na-Ca exchange current (INa-Ca) was activated particularly in the reverse mode. Na-K pump current (INa-K) was also activated. Notably, the inward rectifying K current (IK1) was not affected, and the change in the delayed outward K current (IK) was not evident. CONCLUSION: These results suggest that the monensin-induced shortened APD and reduced amplitude of the plateau phase are primarily due to the decrease in the ICa, the activation of the reverse mode of INa-Ca, and the increased INa-K, and second due to the decreased INa. The IK and the IK1 may not be associated with the abovementioned changes induced by monensin. The elevation of [Na]i can exert multiple influences on electrophysiological phenomena in cardiac myocytes.

[Questionnaire survey on hepatitis B vaccine in clinics].

We conducted a questionnaire survey on the status of implementation of hepatitis B vaccination and HBs antibody testing. It involved medical personnel covering 484 regional medical institutions in the Osaka Province. Results showed that the recognition rate was 30.1%, the hepatitis B vaccination implementation rate was 38.9%, and that of HBs antibody testing was 38.9%. Although 42.5% of the medical institutions had experienced needle-stick injuries, some medical institutions did not respond properly. The low implementation rate of hepatitis B vaccination and HBs antibody test could be explained by lack of recognition for hepatitis B infection control guidelines. Therefore, we can achieve a possible improvement in the control of infection in the Province, if sensitization programs on hepatitis B infection control are organized in the various regional medical institutions in order to provide adequate information and raise awareness on the importance of respecting these guidelines.

[The effect of introducing an on-call system for attending physicians].

Gastroenterologists working in hospitals that have adopted the chief physician system are often required to work overtime during the night and on holidays treating critically ill patients and ordering emergency tests. To help alleviate the attending physician's duties, our hospital initiated an on-call system in October 2019 to replace the existing system. Changes in overtime hours worked and business stress before and after the introduction of the on-call system were verified. After the introduction of the on-call system, both overtime hours and the number of holidays worked decreased and work stress was reduced. We report that the on-call system is a suitable alternative to the attending physician system because it increases the work efficiency and satisfaction of attending physicians.

Application of substrate depletion assay to evaluation of CYP isoforms responsible for stereoselective metabolism of carvedilol.

To evaluate the relative contribution of cytochrome P450 (CYP) isoforms responsible for carvedilol (CAR) oxidation, enantioselective metabolism of CAR was investigated in human liver microsomes (HLMs) and recombinant human CYPs by using the substrate depletion assay. CYP2D6 exhibited the highest contribution to the metabolism of R-CAR, followed by CYP3A4, CYP1A2, and CYP2C9, whereas the metabolism of the S-enantiomer was mainly mediated by CYP1A2, followed by CYP2D6 and CYP3A4. In HLMs, metabolism of R- and S-CAR was markedly inhibited by quinidine; R-CAR metabolism (57-61% decrease) was more inhibited than S-CAR metabolism (37-43% decrease), and furafylline and ketoconazole almost equally inhibited metabolism of both enantiomers by 25-32% and 30-50%, respectively. The absence of CYP2D6 in a mixture of five major recombinant CYP isoforms at the approximate ratio as in HLMs resulted in a 42% and 25% decrease in the metabolic activities for R- and S-CAR, respectively. Moreover, the absence of CYP1A2 in the mixture resulted in a 16% and 39% decrease in the metabolic activities for R- and S-CAR, respectively. Our results suggest the stereoselective metabolism of CAR is determined by not only the activity of CYP2D6 but also of CYP1A2 and CYP3A4.

A new species of Calicotyle Diesing, 1850 (Monogenea: Monocotylidae) from the shortspine spurdog Squalus mitsukurii Jordan & Snyder and the synonymy of Gymnocalicotyle Nybelin, 1941 with this genus.

Calicotyle japonica n. sp., collected from the uterus, rectal gland, archinephric duct and cloaca of the shortspine spurdog Squalus mitsukurii Jordan & Snyder (Squaliformes) off the Pacific coast of Japan, is described. The new species can be distinguished from C. inermis Woolcock, 1936 by the shape of the male copulatory organ; in C. japonica this is directed anteriorly, is sharply bent in the middle and then increases in width toward the tip, whereas it is long, coiled and uniform in width throughout its entire length in C. inermis. Furthermore, the intestinal caeca have many irregular diverticula on both sides and the vaginal apertures are at the level of the common genital pore in C. japonica, whereas in C. inermis the intestine is smooth and the vaginae open at the level of the oötype. A phylogeny constructed using LSU rDNA data indicates that the new species is grouped with other Calicotyle species; based on this, Gymnocalicotyle Nybelin, 1941 is synonymised with Calicotyle Diesing, 1850. Calicotyle is divided into two major clades, with the new species being grouped with Calicotyle species infecting sharks and the second clade consisting of Calicotyle spp. infecting rays. This suggests that the loss of the hamuli in C. japonica and C. inermis, both parasites of sharks, is a comparatively recent event in the evolution of the genus.

In vitro culture technique for Cryptocaryon irritans, a parasitic ciliate of marine teleosts.

A medium for the in vitro culture of Cryptocaryon irritans, which is an obligatorily parasitic ciliate of marine teleosts and causes 'white spot disease', was developed. The medium consisted of a layer of cultured fish cells (FHM), with an agarose gel layer covering the cell layer. The agarose gel contained 0.22% agarose, 10% fetal calf serum, 100 I.U. ml(-1) Penicillin G potassium and 100 microg ml(-1) streptomycin sulphate. Theronts of C. irritans transformed to trophonts and grew to 180 microm in mean length in the medium, although they gradually decreased in number. When trophonts fully developed in medium were transferred into seawater 4 d after inoculation, approximately 70% of them transformed to encysted tomonts and released theronts. When fish were challenged with theronts obtained from in vitro-raised parasites, approximately 40% of the theronts were recovered from fish, indicating comparative infectivity of in vitro-raised theronts to those of in vivo-raised theronts. This is the first report that C. irritans fully developed in vitro and its entire life cycle was completed without a host fish.

Two new species of hexabothriid monogeneans from the ginzame Chimaera phantasma and shortspine spurdog Squalus mitsukurii.

Descriptions are given for two new species of hexabothriid monogeneans, one collected from the gills of shortspine spurdog Squalus mitsukurii and the other from the gills of the ginzame Chimaera phantasma in Sagami Bay on the Pacific coast of Japan. Squalonchocotyle mitsukurii n. sp. from Squalus mitsukurii can be distinguished from Squalonchocotyle squali MacCallum, 1931, the most closely related species, by the inwardly curved vagina. Callorhynchocotyle sagamiensis n. sp. from Chimaera phantasma can be distinguished from all the known species of Callorhynchocotyle Suriano & Incorvaia, 1982 by the structure of the cirrus; in the new species, the cirrus is entirely muscular with a widened opening, while in the four known species it comprises a tubular proximal part and a bulbous distal part.

Scuticociliata infection in the weedy sea dragon Phyllopteryx taeniolatus.

The weedy sea dragon Phyllopteryx taeniolatus at an aquarium in Kanagawa Prefecture were found infected by protozoan ciliate, 2001. The infected fish in particular, showed sloughing of the epidermis. Fish with intense infections showed sloughing of the dorsal fin, depigmentation of skin, anal distension and accumulation of ascitic fluid in the body cavity. In biopsies, ciliates were detected only in fresh mounts of abdominal dropsy and in the mucus on the body surface. Histopathological studies revealed ciliates mainly infected the dermis, and induced extensive detachment of the epidermis from the skin. Based on the arrangement and shape of the buccal structure and the number of somatic ciliature, ciliates isolated from the fishes belonged to the order Scuticociliatida. We discuss the major factors that bring about the death of weedy sea dragons.

Cytochrome p450 induction and gonadal status alteration in common carp (Cyprinus carpio) associated with the discharge of dioxin contaminated effluent to the Hikiji River, Kanagawa Prefecture, Japan.

Accumulations of polychlorinated dibenzo-p-dioxins, dibenzofurans and coplanar polychlorinated biphenyls were analyzed in common carp (Cyprinus carpio) collected in the Hikiji River, Kanagawa Prefecture, Japan in which dioxin contaminated effluent was released during the period starting from November 1992 to March 2000. Higher levels of 2,3,7,8-tetrachlorinated dibenzo-p-dioxin toxic equivalents were observed in carps collected downstream to the dioxin release site (contaminated site) than the reference site. Modulations of cytochrome p450 (CYP) enzyme in liver, serum estrogen concentration and gonadal somatic index (GSI) were also measured as biomarkers for the contaminants. Total CYP content in livers was markedly higher in male and female carps from the contaminated site relative to the reference site fish. The expression level of the cytochrome p450 1A and Ethoxyresorufin O-deethylase activity were significantly higher in female carps from the contaminated site than from the reference site. A lower level of plasma estrogen was observed in carps from the contaminated site. The GSI in female carps from the contaminated site was smaller than that recorded at the reference site. The present study indicates that dioxins released to the Hikiji River might induce the CYP enzyme and inhibit the reproductive functions in common carps dwelling downstream from the release site.