Post-contraction potentiation can react inversely to post-activation potentiation depending on the test contraction force.

The surface electromyographic (EMG) activity of the biceps brachii during weak elbow flexion reportedly increases immediately after strong elbow flexion, even during the exertion of a given force. This phenomenon is called post-contraction potentiation (EMG-PCP). However, the effects of test contraction intensity (TCI) on EMG-PCP remain unclear. This study evaluated PCP levels at various TCI values. Sixteen healthy participants were asked to perform a force matching task (2%, 10%, or 20% of the maximum voluntary contraction [MVC]) before (Test 1) and after (Test 2) a conditioning contraction (50% of MVC). With a 2% TCI, the EMG amplitude was higher in Test 2 than in Test 1. With a 20% TCI, the EMG amplitude was lower in Test 2 than in Test 1. Furthermore, EMG spectral analyses showed that the α- and ß-band power ratios in Test 2 were enhanced by 2% TCI compared with Test 1. These findings suggest that TCI is crucial in determining the EMG-force relationship immediately after a brief intensive contraction.

Effects of Genetic Mutation Sites in ADR Genes on Modern Chickens Produced and Domesticated by Artificial Selection.

Associations between neurotransmitters, adrenergic receptor (ADR) mutations, and behaviors in chickens produced and domesticated by artificial selection remain unclear. This study investigates the association of neurotransmitters and ADR mutations with egg laying and cockfighting-behaviors associated with significantly different breeding backgrounds-in Shaver Brown and Shamo chickens. Accordingly, the whole sequences of nine ADR genes were determined, and nine amino acid-specific mutation sites from five genes (ADRα1A: S365G, ADRα1D: T440N, ADRα2A: D273E, ADRß1: N443S, S445N, ADRß3: R342C, Q404L, and P406S) were extracted. Evolutionary analysis showed that these mutations were not ancestrally derived. These results confirm that the mutations at these sites were artificially selected for domestication and are breed specific. NST population analysis confirmed a difference in the degree of genetic differentiation between the two populations in seven genes. The results further confirm differences in the degree of genetic differentiation between the two populations in Shaver Brown (ADRA1B and ADRA1D) and Shamo (ADRA1A and ADRA2B) chickens, indicating that the ADR gene differs between the two breeds. The effects of artificial selection, guided by the human-driven selection of desirable traits, are reflected in adrenaline gene mutations. Furthermore, certain gene mutations may affect domestication, while others may affect other traits in populations or individuals.

Correction: Komiyama et al. RNA-seq and Mitochondrial DNA Analysis of Adrenal Gland Metastatic Tissue in a Patient with Renal Cell Carcinoma. Biology 2022, 11, 589.

The authors would like to make the following correction to the published paper [...].

Misencoding of ankle joint angle control system via cutaneous afferents reflex pathway in chronic ankle instability.

This study aimed to investigate how the cutaneous reflexes in the peroneus longus (PL) muscle are affected by changing the ankle joint position in patients with chronic ankle instability (CAI). We also investigated the correlation between the degree of reflex modulation and angle position sense of the ankle joint. The participants were 19 patients with CAI and 20 age-matched controls. Cutaneous reflexes were elicited by applying non-noxious electrical stimulation to the sural nerve at the ankle joint in the neutral standing and eversion/inversion standing positions. The suppressive middle latency cutaneous reflex (MLR; ~ 70-120 ms) and angle position sense of the ankle joint were assessed. During neutral standing, the gain of the suppressive MLR was more prominent in the CAI patients than in controls, although no significant difference was seen during 30° inversion standing. In addition, the ratios of the suppressive MLR and background electromyography in a neutral position were significantly larger than those at the 15°, 25°, and 30° inversion positions in CAI patients. No such difference was seen in control individuals. Furthermore, the correlations between reflex modulation degree and position sense error were quite different in CAI patients compared to controls. These findings suggest that the sensory-motor system was deteriorated in CAI patients due to changes in the PL cutaneous reflex pathway excitability and position sense of the ankle joint.

Gender Differences in Takotsubo Syndrome.

Most patients with Takotsubo Syndrome (TTS) are postmenopausal females. TTS in males is rare and gender differences have not been sufficiently investigated. Therefore, we investigated gender differences in TTS. TTS in males and females is often triggered by physical and emotional stress, respectively. Heart failure, a severe in-hospital complication, requires greater mechanical respiratory support in males. Fatal arrhythmias such as ventricular tachycardia and ventricular fibrillation and in-hospital mortality rates are higher in males. The white blood cell (WBC) count has been shown to be higher in males than in females with cardiovascular death compared with non-cardiovascular death. Therefore, the WBC count, a simple marker, may reflect severe TTS. Decreased estrogen levels, common in postmenopausal females, are a pathogenic mechanism of TTS. Females have a more significant increase in the extracellular matrix-receptor interaction than males. Moreover, the pathological findings after hematoxylin-eosin staining were different in males and females. Males had more severe complications than females in the acute phase of TTS; thus, more careful observations and interventions are likely required. From these results, it can be considered that the mechanism of the onset of TTS may be different between males and females. Therefore, it is necessary to fully understand the gender differences in order to more effectively manage TTS.

RNA-seq and Mitochondrial DNA Analysis of Adrenal Gland Metastatic Tissue in a Patient with Renal Cell Carcinoma.

This study aimed to clarify whether genetic mutations participate in renal cell carcinoma (RCC) metastasis to the adrenal gland (AG). Our study analyzed whole mitochondrial gene and ribonucleic acid sequencing (RNA-seq) data from a male patient in his 60s with metastatic RCC. We confirmed common mutation sites in the mitochondrial gene and carried out Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis using RNA-seq data for RCC and adrenal carcinoma. Furthermore, we confirmed the common mutation sites of mitochondrial genes in which the T3394Y (p.H30Y) site transitioned from histidine (His.; H) to tyrosine (Tyr.; Y) in the NADH dehydrogenase subunit 1 (ND1) gene. The R11,807G (p.T350A) site transitioned from threonine (Thr.; T) to alanine (Ala.; A). Additionally, the G15,438R or A (p.G231D) site transitioned from glycine (Gly.; G) to aspartic acid (Asp.; D) in cytochrome b (CYTB). Furthermore, pathway analysis, using RNA-seq, confirmed the common mutant pathway between RCC and adrenal carcinoma as cytokine-cytokine receptor (CCR) interaction. Confirmation of the original mutation sites suggests that transfer to AG may be related to the CCR interaction. Thus, during metastasis to the AG, mitochondria DNA mutation may represent the initial origin of the metastasis, followed by the likely mutation of the nuclear genes.

Execution of natural manipulation in the air enhances the beta-rhythm intermuscular coherences of the human arm depending on muscle pairs.

Natural manipulation tasks in air consist of two kinematic components: a grasping component, with activation of the hand muscles, and a lifting component, with activation of the proximal muscles. However, it remains unclear whether the synchronized motor commands to the hand/proximal arm muscles are divergently controlled during the task. Therefore, we examined how intermuscular coherence was modulated depending on the muscle combinations during grip and lift (G&L) tasks. Electromyograms (EMGs) were recorded from the biceps brachii (BB), triceps brachii (TB), flexor digitorum superficialis (FDS), and extensor digitorum communis (EDC) muscles. The participants were required to maintain G&L tasks involving a small cubical box with the thumb and index and middle fingers. Consequently, we found that the beta-rhythm coherence (15-35 Hz) in BB-TB, BB-FDS, and TB-EDC pairs during G&L was significantly larger than that during the isolated task with cocontraction of the two target muscles but not BB-EDC, TB-FDS, and FDS-EDC (task and muscle pair specificities). These increases in beta-rhythm coherence were also observed in intramuscular EMG recordings. Furthermore, the results from the execution of several mimic G&L tasks revealed that the separated task-related motor signals and combinations between the motor signals/sensations of the fingertips or object load had minor contributions to the increase in the coherence. These results suggest that during G&L the central nervous system regulates synchronous drive onto motoneurons depending on the muscle pairs and that the multiple combination effect of the sensations of touch/object load and motor signals in the task promotes the synchrony of these pairs.NEW & NOTEWORTHY Natural manipulation in air consists of two kinematic components: grasping, with activation of hand muscles, and lifting, with activation of proximal muscles. We show that during the maintenance of object manipulation in air the central nervous system regulates the synchronous drive onto human motoneuron pools depending on the hand/proximal muscle pairs and that the multiple combination effect of the sensations of touch/object load and motor signals in the task promotes the synchrony of these pairs.

Subcortical Contribution of Corticospinal Transmission during Visually Guided Switching Movements of the Arm.

In animal experiments, the indirect corticospinal tract (CST) system via cervical interneurons has been shown to mediate motor commands for online adjustment of visuomotor behaviors, such as target-reaching. However, it is still unclear whether the similar CST system functions to perform similar motor behaviors in humans. To clarify this, we investigated changes in motor-evoked potentials (MEPs) in the elbow muscles following transcranial magnetic stimulation, transcranial electrical stimulation, or cervicomedullary stimulation while participants executed target-reaching and switching movements. We found that the MEP, whether elicited cortically or subcortically, was modulated depending on the direction of the switching movements. MEP facilitation began around the onset of the switching activities in an agonist muscle. Furthermore, ulnar nerve-induced MEP facilitation, which could be mediated by presumed cervical interneuronal systems, also increased at the onset of MEP facilitation. In a patient with cortical hemianopsia who showed switching movements in the scotoma, the MEPs were facilitated just before the switching activities. Our findings suggested that CST excitation was flexibly tuned with the switching movement initiation, which could partly take place in the subcortical networks, including the presumed cervical interneuronal systems.

Clinical Significance of the Head-Up Tilt Test in Improving Prognosis in Patients with Possible Neurally Mediated Syncope.

Syncope is commonly encountered in daily clinical practice. Depending on its etiology (benign or life-threatening conditions or environmental triggers), syncope can be neurally mediated (reflex), cardiac, or orthostatic. Furthermore, neurologic disease can cause symptoms that mimic syncope. However, there is limited research on neurally mediated syncope (NMS), which is considered a benign disorder, and close follow-ups are rarely performed. NMS can cause serious clinical events, including severe trauma and car accidents. The head-up tilt test (HUTT) is the gold standard for diagnosing NMS; however, its clinical significance remains unknown, and its relevance to NMS prognosis requires further research. This retrospective study aimed to assess the clinical significance of the HUTT for NMS. We reviewed the charts of 101 patients who underwent HUTT at Tokai University Hospital in Japan between January 2016 and March 2019. During the HUTT, 72 patients (69.2%) experienced syncope. Patients were followed up for 886.1 ± 457.7 days (interquartile range: 518-1293 days). The syncope recurrence rate was 16.9%; however, no significant difference was observed between the two groups (HUTT positive vs. negative) (13.8% vs. 18.1%, p = 0.772). Four of 29 (13.9%) and two of 72 (2.8%) patients in the negative and positive HUTT groups, respectively, experienced cardiac events (p = 0.019). Negative HUTT results may assist in anticipating unexpected clinical events within a few years. A negative HUTT result may allow us to reconsider the NMS diagnosis based on clinical information. Close outpatient follow-up of patients with negative HUTT results is warranted.

Diagnosis and prevention of the vasodepressor type of neurally mediated syncope in Japanese patients.

We investigated circulatory dynamics in patients with vasodepressor type neurally mediated syncope (VT-NMS) by performing high-resolution Holter electrocardiography and a correlation analysis of changes in adenylate cyclase activity, blood pressure, and pulse during the head-up tilt test. Holter electrocardiography was performed for 30 patients. Adenylate cyclase activity was evaluated in lymphocytes from blood samples taken at rest and during the head-up tilt test. There was no change in autonomic nerve fluctuation during electrocardiography in VT-NMS patients, but our results showed a significant difference in blood pressure and adenylate cyclase activity between VT-NMS patients and healthy volunteers; the systolic blood pressure of VT-NMS patients decreased after 5 min, while at 10 min, the adenylate cyclase activity was the highest (0.53%) and the systolic blood pressure was the lowest (111.8 mm Hg). Pulse rates increased after 10 min. VT-NMS patients showed higher blood pressure, pulse rate, and adenylate cyclase activity during the tilt test than did healthy volunteers. In patients with syncope, standing for longer than 10 minutes may increase the risk of VT-NMS. From our results, we consider it likely that high systolic blood pressure and adenylate cyclase activity at rest cause fainting in VT-NMS patients. Our findings may be helpful for identifying individuals with a high risk of developing NMS in the healthy population.

Examination of gender differences in patients with takotsubo syndrome according to left ventricular biopsy: two case reports.

BACKGROUND: Takotsubo syndrome is a stress-induced disease that makes up 2-3% of acute coronary syndrome cases. However, its onset mechanism remains unclear. Although females are overwhelmingly affected, males end up having more cardiac complications. CASE PRESENTATION: We examined the differences in stress responses in the myocardium between sexes in patients with takotsubo syndrome. We biopsied samples from an over 70-year-old Japanese male and an over 80-year-old Japanese female. Tissues from the left ventricle apex in the acute phase and the apical ballooning-type were examined using histopathology and deoxyribonucleic acid (DNA) microarray analysis. Our data showed that left ventricular ejection fractions were 38% and 56%, and peak creatinine kinase concentrations during hospitalization were 629 U/L and 361 U/L, for the male and female patient, respectively. The pulmonary capillary wedge pressure was 26 mmHg and 11 mmHg for the male and female patient, respectively. Negative T did not return to normal in the male subject after 6 months. Histopathology results indicated that contraction band necrosis and lymphocyte infiltration were more common in the male subject. CONCLUSIONS: We noticed that possible differences may exist between male and female patients using pathological examination and some DNA analyses. In particular, it may help treat acute severity in males. We will elucidate the mechanism of takotsubo syndrome development by increasing the number of samples to support the reliability of the data in the future.

Visual information increases the indirect corticospinal excitation via cervical interneurons in humans.

Modulatory actions of inputs from the visual system to cervical interneurons (IN) for arm muscle control are poorly understood in humans. In the present study, we examined whether visual stimulation modulates the excitation of cervical IN systems mediating corticospinal tract (CST) inputs to biceps brachii (BB). Twenty-eight healthy volunteers were seated, and electromyogram recordings from the BB were performed across six experiments, each with discrete objectives. A flash stimulator for visual stimulation (50-µs duration) was placed 60 cm from the participant's eye. The CST was stimulated with transcranial magnetic/electrical stimulation (TMS/TES, respectively) contralateral to the recording site. Visual stimulation with TMS/TES was randomly delivered during weak tonic BB contractions. Single TMS/TES-induced motor-evoked potentials (MEPs) were markedly enhanced from 60-100 ms after visual stimulation compared with the control condition. The MEPs were significantly increased by combining the electrical stimulation of the ulnar nerve at the wrist [7.5-12 ms of nerve stimulation (NERVE)/TMS interval] with and without visual stimulation compared with the algebraic summation of responses obtained with either TMS or NERVE. Interestingly, the combined stimulation-induced MEP facilitation was significantly increased after visual stimulation compared with the control. Single motor unit (MU) recording also revealed the further enhancement of combined stimulation effects on the firing probabilities of MU during visual stimulation, which was observed in the peaks of the peristimulus time histogram, 1-2 ms later than the onset latency. The present findings suggest that visual stimulation facilitates the oligosynaptic CST excitation of arm motoneurons mediated by the cervical IN system.NEW & NOTEWORTHY To date, little is known about how visual information modulates the human cervical motor systems, including the presumed interneuron (IN) circuitry. This study demonstrates that photic visual stimulation influences presumed oligosynaptic corticospinal transmission to arm motoneurons, which are mediated by cervical INs. In animals, these systems are known to be crucial for visually guided switching movements, and similar visual input systems to INs may exist in humans.

Long-lasting changes in muscle activation and step cycle variables induced by repetitive sensory stimulation to discrete areas of the foot sole during walking.

We examined whether repetitive electrical stimulation to discrete foot sole regions that are phase-locked to the step cycle modulates activity patterns of ankle muscles and induces neuronal adaptation during human walking. Nonnoxious repetitive foot sole stimulation (STIM; 67 pulses at 333 Hz) was given to the medial forefoot (f-M) or heel (HL) regions at 1) the stance-to-swing transition, 2) swing-to-stance transition, or 3) midstance, during every step cycle for 10 min. Stance, but not swing, durations were prolonged with f-M STIM delivered at stance-to-swing transition, and these changes remained for up to 20-30 min after the intervention. Electromyographic (EMG) burst durations and amplitudes in the ankle extensors were also prolonged and persisted for 20 min after the intervention. Interestingly, STIM to HL was ineffective at inducing modulation, suggesting stimulation location-specific adaptation. In contrast, STIM to HL (but not f-M), at the swing-to-stance phase transition, shortened the step cycle by premature termination of swing. Furthermore, the onset of EMG bursts in the ankle extensors appeared earlier than in the control condition. STIM delivered during the midstance phase was ineffective at modulating the step cycle, highlighting phase-dependent adaptation. These effects were absent when STIM was applied while mimicking static postures for each walking phase during standing. Our findings suggest that the combination of walking-related neuronal activity with repetitive sensory inputs from the foot can generate short-term adaptation that is phase-dependent and localized to the site of STIM.NEW & NOTEWORTHY Repetitive (∼10 min) long (200 ms) trains of sensory stimulation to discrete areas of the foot sole produce persistent changes in muscle activity and cycle timing during walking. Interactions between the delivery phase and stimulus location determine the expression of the adaptations. These observations bear striking similarities to those in decerebrate cat experiments and may be usefully translated to improving locomotor function after neurotrauma.

Arm cycling increases the short-latency reflex from ankle dorsiflexor afferents to knee extensor muscles.

Low-intensity electrical stimulation of the common peroneal nerve (CPN) evokes a short latency reflex in the heteronymous knee extensor muscles (referred to as the CPN reflex). The CPN reflex is facilitated at a heel strike during walking, contributing to body weight support. However, the origin of the CPN reflex increase during walking remains unclear. We speculate that this increase originates from multiple sources due to a body of evidence suggesting the presence of neural coupling between the arms and legs. Therefore, we investigated the extent to which the CPN reflex is modulated during rhythmic arm cycling. Twenty-eight subjects sat in an armchair and were asked to perform arm cycling at a moderate cadence using a stationary ergometer while performing isometric contraction of the knee extensors, such that the CPN reflex was evoked. The CPN reflex was evoked by stimulating the CPN [0.9-2.0× the motor threshold (MT) in the tibialis anterior muscle] at the level of the neck of the fibula. The CPN-reflex amplitude was measured from the vastus lateralis (VL). The biphasic reflex response in the VL was evoked within 27-45 ms following CPN stimulation. The amplitude of the CPN reflex increased during arm cycling compared with that before cycling. The modulation of the CPN reflex during arm cycling was detected only for CPN stimulation intensity around 1.2× MT. Furthermore, CPN-reflex modulation was not observed during the isometric contraction of the arm or passive arm cycling. Our results suggest the presence of neural coupling between the CPN-reflex pathways and neural systems generating locomotive arm movement.NEW & NOTEWORTHY Whether locomotive arm movements contribute to the control of the reflex pathway from ankle dorsiflexor afferents to knee extensor muscles [common peroneal nerve (CPN)-reflex] is an unresolved issue. The CPN reflex in the stationary leg was facilitated only by arm cycling, and not by passive or isometric motor tasks. Our results suggest that the arm locomotor system modulates the reflex pathway from ankle dorsiflexor afferents to the knee extensor muscles.

Motor imagery enhances corticospinal transmission mediated by cervical premotoneurons in humans.

Motor imagery is known to affect the reacquisition of motor functioning after damage to the central nervous system. However, it remains unclear whether motor imagery influences corticospinal (CST) excitation mediated via cervical premotoneurons, which may be important for functional motor recovery in animals and humans. To investigate this, we examined the spatial facilitation of motor-evoked potentials (MEPs) induced by combined stimulation (CS) of CST and peripheral nerves. Thirty-two healthy volunteers were included and electromyograms from the biceps brachii (BB) were recorded. Transcranial magnetic stimulation (TMS) to motor cortex and electrical stimulation of ulnar nerve at wrist (NERVE) were delivered separately or in combination with 6-15 ms of interstimulus intervals (ISIs). Subjects were instructed to imagine performing an elbow flexion at rest and during tonic BB contraction. During both motor imagery and control tasks, CS (7.5-12 ms of ISIs) facilitated MEPs, compared with the mathematical summation of responses obtained with either only TMS or NERVE (P < 0.01). Interestingly, the CS-induced facilitation was significantly increased by motor imagery compared with control (P < 0.01). Single-motor unit recording also revealed increased facilitation during motor imagery, which was observed in peaks of the peristimulus time histogram 1-2 ms later than the onset latency (P < 0.01). The present findings suggest that motor imagery facilitates oligosynaptic CST excitation of arm motoneurons, mediated by cervical premotoneurons. Thus motor imagery may be a useful tool for activating the premotoneuron systems, which may contribute to motor reacquisition.NEW & NOTEWORTHY Imaging movement has positive effects on the reacquisition of motor functions after damage to the central nervous system. This study shows that motor imagery facilitates oligosynaptic corticospinal excitation that is mediated via cervical premotoneurons, which may be important for motor recovery in monkeys and humans. Current findings highlight how this imagery might be a beneficial tool for movement disorders through effects on premotoneuron circuitry.

Alopecia areata susceptibility variant in MHC region impacts expressions of genes contributing to hair keratinization and is involved in hair loss.

BACKGROUND: Alopecia areata (AA) is considered a highly heritable, T-cell-mediated autoimmune disease of the hair follicle. However, no convincing susceptibility gene has yet been pinpointed in the major histocompatibility complex (MHC), a genome region known to be associated with AA as compared to other regions. METHODS: We engineered mice carrying AA risk allele identified by haplotype sequencing for the MHC region using allele-specific genome editing with the CRISPR/Cas9 system. Finally, we performed functional evaluations in the mice and AA patients with and without the risk allele. FINDINGS: We identified a variant (rs142986308, p.Arg587Trp) in the coiled-coil alpha-helical rod protein 1 (CCHCR1) gene as the only non-synonymous variant in the AA risk haplotype. Furthermore, mice engineered to carry the risk allele displayed a hair loss phenotype. Transcriptomics further identified CCHCR1 as a novel component interacting with hair cortex keratin in hair shafts. Both, these alopecic mice and AA patients with the risk allele displayed morphologically impaired hair and comparable differential expression of hair-related genes, including hair keratin and keratin-associated proteins (KRTAPs). INTERPRETATION: Our results implicate CCHCR1 with the risk allele in a previously unidentified subtype of AA based on aberrant keratinization in addition to autoimmune events. FUNDING: This work was supported by JSPS KAKENHI (JP16K10177) and the NIHR UCLH Biomedical Research center (BRC84/CN/SB/5984).

Analysis of the source of aggressiveness in gamecocks.

Although the fighting behaviour in gamecocks has evolved because of artificial selection, it is unknown whether the selection for aggressiveness affects neurotransmitter levels in the avian central nervous system. We sought to identify the source and origin of this trait. We collected the brain samples from 6 female Shamo gamecocks and 5 Shaver Brown chickens (control; bred for egg production). The midbrain levels of norepinephrine (NE) were significantly higher in Shamo gamecocks (P = 0.0087) than in the controls. Moreover, alleles encoding adrenergic receptors differed between the breeds in terms of response to NE. Gene mutations specific to Shamo and potentially associated with fighting behaviour were in sites T440N of ADRα1D; V296I of ADRα2A; and T44I, Q232R, and T277M of ADRß2. The evolutionary analysis indicated that the ADRß2 (T44I and Q232R) mutations were heritable in all Galliformes, whereas the T440N mutation of ADRα1D and V296I mutations of ADRα2A were unique to Shamo and originated by artificial selection. A high NE level may confer a selective advantage by enabling gamecocks to be aggressive and pain tolerant. Therefore, the strong fighting behaviour of Shamo has resulted from a combination of naturally inherited and mutant genes derived by artificial selection.

Whole genome sequencing of Entamoeba nuttalli reveals mammalian host-related molecular signatures and a novel octapeptide-repeat surface protein.

The enteric protozoa Entamoeba histolytica is the causative agent of amebiasis, which is one of the most common parasitic diseases in developed and developing countries. Entamoeba nuttalli is the genetically closest species to E. histolytica in current phylogenetic analyses of Entamoeba species, and is prevalent in wild macaques. Therefore, E. nuttalli may be a key organism in which to investigate molecules required for infection of human or non-human primates. To explore the molecular signatures of host-parasite interactions, we conducted de novo assembly of the E. nuttalli genome, utilizing self-correction of PacBio long reads and polishing corrected reads using Illumina short reads, followed by comparative genomic analysis with two other mammalian and a reptilian Entamoeba species. The final draft assembly of E. nuttalli included 395 contigs with a total length of approximately 23 Mb, and 9,647 predicted genes, of which 6,940 were conserved with E. histolytica. In addition, we found an E. histolytica-specific repeat known as ERE2 in the E. nuttalli genome. GO-term enrichment analysis of mammalian host-related molecules indicated diversification of transmembrane proteins, including AIG1 family and BspA-like proteins that may be involved in the host-parasite interaction. Furthermore, we identified an E. nuttalli-specific protein that contained 42 repeats of an octapeptide ([G,E]KPTDTPS). This protein was shown to be localized on the cell surface using immunofluorescence. Since many repeat-containing proteins in parasites play important roles in interactions with host cells, this unique octapeptide repeat-containing protein may be involved in colonization of E. nuttalli in the intestine of macaques. Overall, our draft assembly provides a valuable resource for studying Entamoeba evolution and host-parasite selection.

D-loop Mutations in Renal Cell Carcinoma Improve Predictive Accuracy for Cancer-Related Death by Integrating with Mutations in the NADH Dehydrogenase Subunit 1 Gene.

Renal cell carcinoma (RCC) is associated with various genetic alterations. Although whole-genome/exome sequencing analysis has revealed that nuclear genome alterations are associated with clinical outcomes, the association between nucleotide alterations in the mitochondrial genome and RCC clinical outcomes remains unclear. In this study, we analyzed somatic mutations in the mitochondrial D-loop region, using RCC samples from 61 consecutive patients with localized RCC. Moreover, we analyzed the relationship between D-loop mutations and NADH dehydrogenase subunit 1 (MT-ND1) mutations, which we previously found to be associated with clinical outcomes in localized RCC. Among the 61 localized RCCs, 34 patients (55.7%) had at least one mitochondrial D-loop mutation. The number of D-loop mutations was associated with larger tumor diameter (> 32 mm) and higher nuclear grade (≥ ISUP grade 3). Moreover, patients with D-loop mutations showed no differences in cancer-specific survival when compared with patients without D-loop mutations. However, the co-occurrence of D-loop and MT-ND1 mutations improved the predictive accuracy of cancer-related deaths among our cohort, increasing the concordance index (C-index) from 0.757 to 0.810. Thus, we found that D-loop mutations are associated with adverse pathological features in localized RCC and may improve predictive accuracy for cancer-specific deaths when combined with MT-ND1 mutations.

Impact of rhabdoid differentiation on postoperative outcome for patients with N0M0 renal cell carcinoma.

BACKGROUND AND OBJECTIVE: We assessed the aggressiveness of localized renal cell carcinoma (N0M0 RCC) with rhabdoid differentiation (RD) after partial or radical nephrectomy. METHODS: A total of 604 patients with N0M0 RCC who had undergone partial or radical nephrectomy at a single institution were included in this study. Clinicopathological and outcome data on recurrence-free survival (RFS), cancer-specific survival (CSS), and time to recurrence (TTR) were analyzed using Kaplan-Meier methods, log-rank test, univariate and multivariable Cox proportional hazard models, and concordance index. We also evaluated the RFS and CSS in a propensity score-matched cohort to reduce inherent differences. Among the 604 patients, RD was identified in RCC specimens from 24 patients. RESULTS: At the median postoperative follow-up period of 53 months, 58 patients (12 with RD) showed recurrence and 26 patients (7 with RD) had died from RCC. Multivariate analyses showed that RD was an independent risk factor of RFS (hazard ratio 2.81; P = 0.0266) and CSS (hazard ratio 5.18; P = 0.00182). By RD adding to standard risk factors, the concordance indices for RFS and CSS increased 0.77 to 0.79, and 0.76 to 0.79, respectively. Subgroup analysis showed that the presence of RD in RCC specimens was more important for predicting poor RFS and CSS in the early pathological tumor category (≤pT2) subgroup compared to in the advanced tumor category (≥pT3) subgroup. Patients with RD showed a significantly shorter TTR than patients with RCC without RD (7.5 vs. 18 months: P = 0.0150). The propensity score-matched cohort included 24 patients with RD and 24 without RD, of which patients RD showed significantly shorter RFS than those without RD (P = 0.0026). CONCLUSIONS: In summary, the aggressiveness of N0M0 RCC with RD increased the risk of postoperative recurrence, particularly in the early pathological stage. The short TTR also demonstrated the aggressiveness of RCC with RD.