Consistent Negative Correlations between Parasite Infection and Host Body Condition Across Seasons Suggest Potential Harmful Impacts of Salmincola markewitschi on Wild White-Spotted Charr, Salvelinus leucomaenis.

Assessing the impacts of parasites on wild fish populations is a fundamental and challenging aspect of the study of host-parasite relationships. Salmincola, a genus of ectoparasitic copepods, mainly infects salmonid species. This genus, which is notorious in aquaculture, damages host fishes, but its impacts under natural conditions remain largely unknown or are often considered negligible. In this study, we investigated the potential impacts of mouth-attaching Salmincola markewitschi on white-spotted charr (Salvelinus leucomaenis) through intensive field surveys across four seasons using host body condition as an indicator of harmful effects. The prevalence and parasite abundance were highest in winter and gradually decreased in summer and autumn, which might be due to host breeding and/or wintering aggregations that help parasite transmissions. Despite seasonal differences in prevalence and parasite abundance, consistent negative correlations between parasite abundance and host body condition were observed across all seasons, indicating that the mouth-attaching copepods could reduce the body condition of the host fish. This provides field evidence suggesting that S. markewitschi has a potential negative impact on wild white-spotted charr.

Parasites either reduce or increase host vulnerability to fishing: a case study of a parasitic copepod and its salmonid host.

Parasites generally increase host vulnerability to predators via host manipulation for trophic transmission and reduction of host activities. Predators also select prey depending on the parasite infection status. Despite such parasites' roles in prey-predator interactions in wild animals, how parasites affect human hunting probability and resource consumption remains unknown. We examined the effects of the ectoparasitic copepod Salmincola cf. markewitschi on fish vulnerability to angling. We found that infected fish were less vulnerable compared with non-infected fish when the fish body condition was low, which was probably due to reduced foraging activity. On the contrary, infected fish were more vulnerable when the host body condition was high, probably due to the compensation of parasites' negative effects. A Twitter analysis also suggested that people avoided eating fish with parasites and that anglers' satisfaction decreased when captured fish were parasitized. Thus, we should consider how animal hunting is affected by parasites not only for catchability but also for avoiding parasite infection sources in many local regions.

Salmincola markewitschi or S. carpionis (Copepoda: Lernaeopodidae)? A requirement for taxonomic revision due to their high morphological variations.

Salmincola markewitschi Shedko et Shedko, 2002 (Copepoda: Lernaeopodidae) is an ectoparasitic copepod mainly infecting the buccal cavities of white-spotted charr Salvelinus leucomaenis (Pallas) (Salmonidae). This species has only been recorded from Northeast Asia, where a morphologically similar congener Salmincola carpionis (Krøyer, 1837) is also distributed, using the same host species. These copepods are hard to distinguish from each other because of their similarities. We thus examined the newly collected specimens morphologically and genetically from five populations of white-spotted charr in Japan. Most of the specimens were morphologically consistent with S. markewitschi but showed great variations in the numbers of spines on the exopods of the antennae, shape of the maxilliped myxal palps, and the bulla diameter. Consequently, some specimens shared characteristics with S. carpionis. In addition to the mophological continuities, genetic analyses of 28S rDNA and COI mitochondrial DNA confirmed that all specimens belong to a single species. Further taxonomic revisions are required to draw conclusions of whether S. markewitschi is a valid species different from S. carpionis, by collecting samples from across their wide distributional ranges, such as Europe, North America, and Northeast Asia. A key to identification of species of Salmincola Wilson, 1915 occurring in Japan is also provided.

Potential negative effects and heterogeneous distribution of a parasitic copepod Salmincola edwardsii (Copepoda: Lernaeopodidae) on Southern Asian Dolly Varden Salvelinus curilus in Hokkaido, Japan.

The genus Salmincola is an ectoparasitic copepod group commonly infesting the branchial and buccal cavities of salmonids. While negative impacts on hatchery fishes have been reported, their impacts on wild fish populations and distribution patterns are critically understudied. In the Shiretoko Peninsula, Hokkaido, Japan, we found parasites belonging to this genus on the branchial cavity of a stream salmonid, Southern Asian Dolly Varden Salvelinus curilus. All parasites recovered were identified as Salmincola edwardsii based on morphological characteristics and partial 28S rDNA sequences. Prevalence was highly heterogeneous even among neighboring streams (0-54.8%, < 10 km) with the mean intensity among streams being generally low (2.19 parasites/infeted fish). Despite the low intensity, quantile regression analysis showed negative trends between parasite intensity and host condition, suggesting that the infestation of S. edwardsii has a potential negative impact on the host salmonid. In addition, a single copepod was found from an anadromous fish, which could indicate some salinity tolerance of the copepods. It is important to evaluate the effects of Salmincola spp. on host species and determine the limiting factors on the parasite's distribution for proper management.

[Multicenter Evaluation of Cine Angiography and Digital Subtraction Angiography Dose Rate for Various Angiography Programs: Comparison with Fluoroscopic Dose Rate].

We conducted a nationwide multicenter survey of various interventional radiology (IVR) procedures. Data were collected from 385 X-ray systems in 126 institutions, including 432 cine programs and 380 digital subtraction angiography (DSA) programs for diagnostic catheterization, percutaneous coronary intervention (PCI), ablation, transcatheter aortic valve implantation (TAVI), neurologic IVR, thorax IVR, abdominal IVR, and endovascular therapy (EVT). Fluoroscopic and cine dose rates were 10.1 mGy/min and 110.7 mGy/min, respectively, whereas for DSA programs, the median fluoroscopic and DSA dose rates were 8.0 mGy/min and 224.8 mGy/min, respectively. The DSA dose rate was more than twice the cine dose rate. The largest difference between dose rates was for diagnostic catheterization, which had a cine dose rate of 142.6 mGy/min and a fluoroscopic dose rate of 12.6 mGy/min (by a factor of 12.5), followed by EVT, which had a DSA dose rate of 216.0 mGy/min and a fluoroscopic dose rate of 7.7 mGy/min (by a factor of 29.6). The smallest difference between dose rates was for TAVI, which had a cine dose rate of 96.8 mGy/min and a fluoroscopic dose rate of 12.0 mGy/min (by a factor of 8.9), followed by neurologic IVR, which had a DSA dose rate of 227.9 mGy/min and a fluoroscopic dose rate of 9.6 mGy/min (by a factor of 22.6). Compared with the fluoroscopic dose rates, the cine dose rates were 9-13 times higher and the DSA dose rates were 22-30 times higher; the DSA dose rates were more than double the cine dose rates.

[Multicenter Survey on Radiation Dose of Cardiac Intervention].

We conducted a nationwide survey of multiple institutions and collected data of various interventional procedures in the field of cardiology. Included in the analysis were 126 institutions, 381 X-ray systems, and 805 protocols. The dose values were compared with the Japanese diagnostic reference levels (DRLs) 2015. Fluoroscopy time, air kerma at the patient entrance reference point (Ka, r), and air kerma-area product (PKA ) were analyzed for various interventional procedures in 5,734 cardiology patients. The fluoroscopic dose rate (FDR) for pulmonary vein isolation (PVI) was less than half that of the 75th percentile of the Japanese DRLs 2015. The 75th percentiles of fluoroscopy time, Ka, r, and PKA for the respective interventional procedures were as follows: 11.0 min, 735 mGy, and 64 Gyï½¥cm2 for diagnostic coronary angiography (CA); 13.2 min, 839 mGy, and 75 Gyï½¥cm2 for CA + left ventriculography; 34.4 min, 1,810 mGy, and 148 Gyï½¥cm2 for percutaneous coronary intervention (PCI) excluding chronic total occlusion; 80.1 min, 4,338 mGy, and 312 Gyï½¥cm2 for PCI for chronic total occlusion; 74.4 min, 833 mGy, and 90 Gyï½¥cm2 for PVI; and 34.0 min, 795 mGy, and 94 Gyï½¥cm2 for transcatheter aortic valve implantation, respectively. In assessing dose values in interventional radiology, the difficulty of the technique needs to be considered, and the DRL values for FDR, fluoroscopic time, Ka, r, and PKA for each interventional procedure are considered necessary when reassessing or updating DRLs.

Structural dynamics and stability of corticocortical and thalamocortical axon terminals during motor learning.

Synaptic plasticity is the cellular basis of learning and memory. When animals learn a novel motor skill, synaptic modifications are induced in the primary motor cortex (M1), and new postsynaptic dendritic spines relevant to motor memory are formed in the early stage of learning. However, it is poorly understood how presynaptic axonal boutons are formed, eliminated, and maintained during motor learning, and whether long-range corticocortical and thalamocortical axonal boutons show distinct structural changes during learning. In this study, we conducted two-photon imaging of presynaptic boutons of long-range axons in layer 1 (L1) of the mouse M1 during the 7-day learning of an accelerating rotarod task. The training-period-averaged rate of formation of boutons on axons projecting from the secondary motor cortical area increased, while the average rate of elimination of those from the motor thalamus (thalamic boutons) decreased. In particular, the elimination rate of thalamic boutons during days 4-7 was lower than that in untrained mice, and the fraction of pre-existing thalamic boutons that survived until day 7 was higher than that in untrained mice. Our results suggest that the late stabilization of thalamic boutons in M1 contributes to motor skill learning.

Exploration of the Key Factors for Optimizing the in Vivo Oral Delivery of Insulin by Using a Noncovalent Strategy with Cell-Penetrating Peptides.

This present study aimed to determine the optimal oral insulin delivery conditions that would maximize the utility of cell-penetrating peptides (CPPs) by using a noncovalent strategy. We first compared the effectiveness of two potential CPPs, penetratin and its analog PenetraMax, as absorption enhancers for insulin. The combined effect was evaluated under in vivo oral administration conditions. Both D-forms of CPPs were highly effective for increasing the oral absorption of insulin, and D-PenetraMax showed a more rapid onset of absorption enhancement effects compared with those of D-penetratin. However, synergistic absorption enhancement effects after combination treatment were not observed. Next, we tried a theoretical approach to establish optimized oral insulin delivery conditions. A surface plasmon resonance (SPR)-based analysis demonstrated that binding between insulin and penetratin (2 mM) might be saturated at 100-500 µM penetratin, while the bound concentration of penetratin could increase in accordance with an increased concentration of mixed insulin. To test this hypothesis, we investigated the effectiveness of different insulin doses in the gastric pH-neutralized mice. The results showed that the dissociation of noncovalent complexes of insulin and CPPs at the low gastric pH was prevented in these mice. Our findings clearly suggested that a noncovalent strategy with CPPs represents an effective approach for the L-form of CPP to increase the concentration of CPP-bound insulin to attain greater absorption of insulin, although this approach may not be appropriate for the D-form of CPP. Our findings will contribute to the development of oral dosage forms of insulin for noncovalent strategies involving CPP.

A transient disruption of fibroblastic transcriptional regulatory network facilitates trans-differentiation.

Transcriptional Regulatory Networks (TRNs) coordinate multiple transcription factors (TFs) in concert to maintain tissue homeostasis and cellular function. The re-establishment of target cell TRNs has been previously implicated in direct trans-differentiation studies where the newly introduced TFs switch on a set of key regulatory factors to induce de novo expression and function. However, the extent to which TRNs in starting cell types, such as dermal fibroblasts, protect cells from undergoing cellular reprogramming remains largely unexplored. In order to identify TFs specific to maintaining the fibroblast state, we performed systematic knockdown of 18 fibroblast-enriched TFs and analyzed differential mRNA expression against the same 18 genes, building a Matrix-RNAi. The resulting expression matrix revealed seven highly interconnected TFs. Interestingly, suppressing four out of seven TFs generated lipid droplets and induced PPARG and CEBPA expression in the presence of adipocyte-inducing medium only, while negative control knockdown cells maintained fibroblastic character in the same induction regime. Global gene expression analyses further revealed that the knockdown-induced adipocytes expressed genes associated with lipid metabolism and significantly suppressed fibroblast genes. Overall, this study reveals the critical role of the TRN in protecting cells against aberrant reprogramming, and demonstrates the vulnerability of donor cell's TRNs, offering a novel strategy to induce transgene-free trans-differentiations.

Slow reactant-water exchange and high catalytic performance of water-tolerant Lewis acids.

(31)P nuclear magnetic resonance (NMR) spectroscopic measurement with trimethylphosphine oxide (TMPO) was applied to evaluate the Lewis acid catalysis of various metal triflates in water. The original (31)P NMR chemical shift and line width of TMPO is changed by the direct interaction of TMPO molecules with the Lewis acid sites of metal triflates. [Sc(OTf)3] and [In(OTf)3] had larger changes in (31)P chemical shift and line width by formation of the Lewis acid-TMPO complex than other metal triflates. It originates from the strong interaction between the Lewis acid and TMPO, which results in higher stability of [Sc(OTf)3TMPO] and [In(OTf)3TMPO] complexes than other metal triflate-TMPO complexes. The catalytic activities of [Sc(OTf)3] and [In(OTf)3] for Lewis acid-catalyzed reactions with carbonyl compounds in water were far superior to the other metal triflates, which indicates that the high stability of metal triflate-carbonyl compound complexes cause high catalytic performance for these reactions. Density functional theory (DFT) calculation suggests that low LUMO levels of [Sc(OTf)3] and [In(OTf)3] would be responsible for the formation of stable coordination intermediate with nucleophilic reactant in water.

Involvement of the N-methyl-D-aspartate receptor GluN2D subunit in phencyclidine-induced motor impairment, gene expression, and increased Fos immunoreactivity.

BACKGROUND: Noncompetitive N-methyl-d-aspartate (NMDA) receptor antagonists evoke a behavioral and neurobiological syndrome in experimental animals. We previously reported that phencyclidine (PCP), an NMDA receptor antagonist, increased locomotor activity in wildtype (WT) mice but not GluN2D subunit knockout mice. Thus, the aim of the present study was to determine whether the GluN2D subunit is involved in PCP-induced motor impairment. RESULTS: PCP or UBP141 (a GluN2D antagonist) induced potent motor impairment in WT mice but not GluN2D KO mice. By contrast, CIQ, a GluN2C/2D potentiator, induced severe motor impairment in GluN2D KO mice but not WT mice, suggesting that the GluN2D subunit plays an essential role in the effects of PCP and UBP141, and an appropriate balance between GluN2C and GluN2D subunits might be needed for appropriate motor performance. The level of the GluN2D subunit in the mature mouse brain is very low and restricted. GluN2D subunits exist in brainstem structures, the globus pallidus, thalamus, and subthalamic nucleus. We found that the expression of the c-fos gene increased the most among PCP-dependent differentially expressed genes between WT and GluN2D KO mice, and the number of Fos-positive cells increased after PCP administration in the basal ganglia motor circuit in WT mice but not GluN2D KO mice. CONCLUSION: These results suggest that the GluN2D subunit within the motor circuitry is a key subunit for PCP-induced motor impairment, which requires an intricate balance between GluN2C- and GluN2D-mediated excitatory outputs.

A new reference point for patient dose estimation in neurovascular interventional radiology.

In interventional radiology, dose estimation using the interventional reference point (IRP) is a practical method for obtaining the real-time skin dose of a patient. However, the IRP is defined in terms of adult cardiovascular radiology and is not suitable for dosimetry of the head. In the present study, we defined a new reference point (neuro-IRP) for neuro-interventional procedures. The neuro-IRP was located on the central ray of the X-ray beam, 9 cm from the isocenter, toward the focal spot. To verify whether the neuro-IRP was accurate in dose estimation, we compared calculated doses at the neuro-IRP and actual measured doses at the surface of the head phantom for various directions of the X-ray projection. The resulting calculated doses were fairly consistent with actual measured doses, with the error in this estimation within approximately 15%. These data suggest that dose estimation using the neuro-IRP for the head is valid.

Identification of ZNF395 as a novel modulator of adipogenesis.

Adipogenesis is the process of cell differentiation by which mesenchymal stem cells (MSC) become adipocytes. Investigating the transcriptional regulatory process during adipogenesis may provide strategies to prevent obesity and other metabolic disorders. In recent years, numerous zinc finger proteins (ZFPs) have been implicated in regulating differentiation and cell fate determination. To investigate the regulatory role of ZFPs involved in adipogenesis, we performed genome-wide microarray expression profiling of an adipogenesis time series. Particularly focusing on the transiently responsive ZFPs, we identified and characterized the functional role of ZNF395 in adipogenesis. A systematic ablation of the ZNF395 transcript during adipogenesis revealed 40% reduction of adipocytes when compared to control. Furthermore, the number of adipocytes as well as the expression of key adipocyte markers were greatly induced when MSC were co-transduced with ZNF395 and PPARG2. To further elucidate the functional role of ZNF395 during adipogenesis, we attempted to trans-differentiate human dermal fibroblasts with PPARG2. The test remarkably revealed that ZNF395 in conjunction with PPARG2 greatly induced adipogenesis from dermal fibroblasts when compared to PPARG2 alone. These loss and gain of function experiments firmly establish that ZNF395 coordinate the transcriptional regulatory pathway with PPARG2, which may be necessary for the genesis of adipocytes.

CpG site-specific alteration of hydroxymethylcytosine to methylcytosine beyond DNA replication.

Hydroxymethylcytosines (hmC), one of several reported cytosine modifications, was recently found to be enriched in embryonic stem cells and neuronal cells, and thought to play an important role in regulating gene expression and cell specification. However, unlike methylcytosines (mC), the fate of hmC beyond DNA replication is not well understood. Here, to monitor the status of hmC during DNA replication, we prepared a stable episomal vector-based monitoring system called MoCEV in 293T cells. The MoCEV system containing fully hydroxymethylated-cytosine fragments revealed a significant modification towards mC after several rounds of DNA replication. Strikingly this modification was specifically observed at the CpG sites (71.9% of cytosines), whereas only 1.1% of modified cytosines were detected at the non-CpG sites. Since the unmodified MoCEV did not undergo any DNA methylation during cell division, the results strongly suggest that somatic cells undergo hmC to mC specifically at the CpG sites during cell division.

Inhibitory effect of selective serotonin reuptake inhibitors on the vesicular monoamine transporter 2.

The neuronal vesicular monoamine transporter (VMAT2) is the target molecule of action of some psychostimulants, such as methamphetamine and 3,4-methylenedioxymethamphetamine (MDMA). The present study examined the effect of antidepressants, such as selective serotonin reuptake inhibitors (SSRIs), on VMAT2 activity by measuring adenosine triphosphate-dependent [(3)H]dopamine uptake into synaptic vesicles prepared from rat striatum. SSRIs, fluoxetine, paroxetine, and fluvoxamine, inhibited vesicular [(3)H]dopamine uptake in vitro. The rank order of potency was reserpine>>fluoxetine, paroxetine>fluvoxamine, methamphetamine>MDMA. Moreover, kinetic analysis revealed that inhibition by reserpine, a typical VMAT2 inhibitor, was uncompetitive, decreasing maximum velocity and affinity for dopamine. Inhibition by fluoxetine was noncompetitive, only decreasing maximum velocity for dopamine. These results suggest that fluoxetine inhibited the activity of VMAT2 by a mechanism different from that of reserpine and did not directly interact with the active site of VMAT2.