ABSTRACT
To cope with seasonal environmental changes, organisms have evolved approximately 1-y endogenous circannual clocks. These circannual clocks regulate various physiological properties and behaviors such as reproduction, hibernation, migration, and molting, thus providing organisms with adaptive advantages. Although several hypotheses have been proposed, the genes that regulate circannual rhythms and the underlying mechanisms controlling long-term circannual clocks remain unknown in any organism. Here, we show a transcriptional program underlying the circannual clock in medaka fish (Oryzias latipes). We monitored the seasonal reproductive rhythms of medaka kept under natural outdoor conditions for 2 y. Linear regression analysis suggested that seasonal changes in reproductive activity were predominantly determined by an endogenous program. Medaka hypothalamic and pituitary transcriptomes were obtained monthly over 2 y and daily on all equinoxes and solstices. Analysis identified 3,341 seasonally oscillating genes and 1,381 daily oscillating genes. We then examined the existence of circannual rhythms in medaka via maintaining them under constant photoperiodic conditions. Medaka exhibited approximately 6-mo free-running circannual rhythms under constant conditions, and monthly transcriptomes under constant conditions identified 518 circannual genes. Gene ontology analysis of circannual genes highlighted the enrichment of genes related to cell proliferation and differentiation. Altogether, our findings support the "histogenesis hypothesis" that postulates the involvement of tissue remodeling in circannual time-keeping.
Subject(s)
Oryzias , Animals , Oryzias/genetics , Seasons , Circadian Rhythm/physiology , Gonads , PhotoperiodABSTRACT
BACKGROUND: Radiofrequency (RF) heating during MRI theoretically increases with magnetic field strength. In addition, implanted metallic devices are reported to further increase RF heating. However, a detailed evaluation of this type of heating remains scarce in clinical practice. PURPOSE: To assess possible risks and discomfort related to RF heating during MRI examinations of patients with and without metallic implantable devices. STUDY TYPE: A retrospective study of previous questionnaire results on the heating sensation during MRI examinations of the lumbar spine. SUBJECTS: In all, 715 patients, of whom 101 had implanted lumbar spine fixation devices. FIELD STRENGTH/SEQUENCE: 1T and 3T/T1 - and T2 -weighted imaging. ASSESSMENT: The number of patients who perceived heating around the lumbar spine or other regions during the MRI examination. STATISTICAL TESTS: A chi-square test with respect to static field strength B0 , presence of lumbar spine fixation devices, and duration of the MRI examination. RESULTS: The number of patients who perceived heating around the lumbar spine during the MRI examination significantly increased from 5.0% at 1T to 47.5% at 3T (P < 0.001), without a significant difference between patients with and without lumbar spine fixation devices (P = 0.23 at 1T, P = 0.48 at 3T), and regardless of the duration of the MRI examination (P = 0.88 at 1T, P = 0.15 at 3T). DATA CONCLUSION: Sensation of RF heating increased by around 10 times from 1T to 3T MRI examination, but the influence of implanted lumbar spine fixation devices on the RF heating sensation has not been observed. LEVEL OF EVIDENCE: 4 Technical Efficacy: Stage 1 J. Magn. Reson. Imaging 2019;49:525-533.
Subject(s)
Hot Temperature , Magnetic Fields , Magnetic Resonance Imaging , Spinal Fusion/methods , Adult , Aged , Female , Humans , Lumbar Vertebrae/surgery , Male , Metals , Middle Aged , Prostheses and Implants , Radio Waves , Retrospective Studies , Surveys and Questionnaires , ThermosensingABSTRACT
Glycation, the nonenzymatic reaction between proteins and excess blood sugar, is implicated in multiple disorders and occurs via the formation and accumulation of advanced glycation end products (AGEs). In our previous studies, we demonstrated that the red-leaf variant of the Persicaria hydropiper sprout (Japanese red water pepper, Benitade) is one of the potent plants that inhibit formation of AGEs. In this study, we aimed to identify antiglycative compounds in Benitade. Benitade extracts were prepared with hot water, then fractionated by using high-performance liquid chromatography (HPLC). The antiglycative efficacy of each fraction was evaluated by measuring the formation of fluorescent AGEs (Ex 370 nm/Em 440 nm). Two fractions, which contained peaks at 26.4 min and 31.8 min, showed potent antiglycative efficacy. When we hydrolyzed these peaks, they shifted to 32.5 and 41.4 min, which are the same retention times as cyanidin and quercetin, respectively. Based on thin-layer chromatography, both compounds contained galactose. Finally, ultrahigh-performance liquid chromatography/quadrupole-time of flight mass spectrometry (UHPLC-QqTOF-MS) analyses were performed to determine the structure of those compounds. Overall, we identified two glycosides, cyanidin 3-O-galactoside (idaein) and quercetin 3-O-galactoside (hyperin), as representative antiglycative compounds in Benitade.
Subject(s)
Glycation End Products, Advanced/drug effects , Glycosides/pharmacology , Polygonaceae/chemistry , Anthocyanins/chemistry , Anthocyanins/pharmacology , Chromatography, High Pressure Liquid , Chromatography, Thin Layer , Glycosides/chemistry , Glycosides/isolation & purification , Plant Extracts/chemistry , Quercetin/analogs & derivativesABSTRACT
In the past few years, the face mask has been recommended for the prevention of exposing others to COVID-19. Wearing a face mask may have the potential to increase dyspnea and discomfort during exercise; however, controversy exists on whether wearing face masks during exercise affects exercise performance, perception, and mood in runners. We investigated the physiological and perceptual responses of healthy male adults who had experienced long-distance running while exercising at different intensities. Nine healthy young adults who were long-distance runners wearing surgical face mask conducted an incremental treadmill protocol. The protocol was three 6-min stages (20%, 40%, and 60% of maximal heart rate, respectively). The rating of perceived exertion (RPE) and the feeling scale (FS) were measured. RPE was higher in mask condition than in unmask condition (No mask vs. Face mask, light; 8.22 vs. 8.78, p = 0.615, middle; 10.00 vs. 10.78, p = 0.345, high; 12.33 vs. 13.67, p = 0.044.), while FS was not different between conditions. The present study shows that wearing a mask may increase rating of perceived exertion and discomfort when the exercise intensity exceeds a certain threshold in healthy male adults who have experienced long-distance running.
Subject(s)
Affect , COVID-19 , Masks , Running , Humans , Male , Masks/adverse effects , Running/physiology , Affect/physiology , Pilot Projects , Adult , COVID-19/prevention & control , Young Adult , Exercise Test/methods , Physical Exertion/physiology , Perception/physiology , Heart Rate/physiology , SARS-CoV-2ABSTRACT
Blackcurrant is available as a traditional medicine in Europe. However, the detailed effects of blackcurrant on the human gut microbiota remain unknown. In this study, we investigated the prebiotic effects of a blackcurrant extract using a human fecal culture model in six healthy subjects. Feces were individually inoculated into a medium with or without the blackcurrant extract and then fermented for 48 hr under anaerobic conditions. The results obtained from analysis of samples from the fermented medium demonstrated that after 48 hr of fermentation, the pH of the medium with the blackcurrant extract was significantly decreased (control, 6.62 ± 0.20; blackcurrant extract, 6.41 ± 0.33; p=0.0312). A 16S rRNA gene sequencing analysis of the microbiota of the fermented medium showed a significant increase in the relative abundance of Bifidobacteriaceae. In measuring the concentrations of putrefactive components in the fermented medium, we found that the blackcurrant extract significantly reduced ammonia levels and displayed a tendency toward reduced indole levels. Our results suggest that blackcurrant extract could be a potential ingredient for relief of putrefactive components in the gut.
ABSTRACT
Freezing of gait (FOG) is a debilitating problem that markedly impairs the mobility and independence of 38-65% of people with Parkinson's disease. During a FOG episode, patients report that their feet are suddenly and inexplicably "glued" to the floor. The lack of a widely applicable, objective FOG detection method obstructs research and treatment. To address this problem, we organized a 3-month machine-learning contest, inviting experts from around the world to develop wearable sensor-based FOG detection algorithms. 1,379 teams from 83 countries submitted 24,862 solutions. The winning solutions demonstrated high accuracy, high specificity, and good precision in FOG detection, with strong correlations to gold-standard references. When applied to continuous 24/7 data, the solutions revealed previously unobserved patterns in daily living FOG occurrences. This successful endeavor underscores the potential of machine learning contests to rapidly engage AI experts in addressing critical medical challenges and provides a promising means for objective FOG quantification.
Subject(s)
Algorithms , Gait , Machine Learning , Parkinson Disease , Humans , Gait/physiology , Parkinson Disease/diagnosis , Parkinson Disease/physiopathology , Wearable Electronic Devices , Gait Disorders, Neurologic/diagnosis , Gait Disorders, Neurologic/physiopathology , Male , FemaleABSTRACT
Three thienopyrrole-fused thiadiazole (TPT) fluorescent dyes featuring a common amide linker and different alkoxy substituents on peripheral trialkoxybenzene moieties were synthesized, and their self-assembly behavior in solution was investigated. The obtained results revealed a substantial steric effect of the alkoxy substituents on the supramolecular polymerization mechanism, which results from a combination of π-stacking and hydrogen (H)-bonding interactions. Detailed spectroscopic measurements revealed that with increasing steric demand of the substituents, the supramolecular polymerization processes in pure methylcyclohexane (MCH) or a mixture of MCH and toluene become temperature-sensitive and enthalpically favorable, resulting in a change from the isodesmic assembly mechanism to the cooperative mechanism. Theoretical calculations suggested that in TPTs with bulky substituents, steric hindrance causes the H-bonding array of the amide moieties to be aligned along the stacking axis of the π-systems; thus, the H-bonding interactions are strengthened compared to those in TPTs with less bulky substituents, compensating for the weakened π-stacking interactions. A chiral TPT derivative with (S) stereogenic centers was found to form homochiral helical supramolecular assemblies that generate discernible circularly polarized luminescence. Achiral TPTs also generate helical assemblies to which preferential helicity can be imparted through the external chiral bias of the solvents (R)- and (S)-limonene.
ABSTRACT
We have proposed a new method for the exploration of organic functional molecules, using an exhaustive molecular generator combined without combinatorial explosion and electronic state predicted by machine learning and adapted for developing n-type organic semiconductor molecules for field-effect transistors. Our method first enumerates skeletal structures as much as possible and next generates fused ring structures using substitution operations for atomic nodes and bond edges. We have succeeded in generating more than 4.8 million molecules. We calculated the electron affinity (EA) of about 51 thousand molecules with DFT calculation and trained the graph neural networks to estimate EA values of generated molecules. Finally, we obtained the 727 thousand molecules as candidates that satisfy EA values over 3â eV. The number of these possible candidate molecules is far beyond what we have been able to propose based on our knowledge and experience in synthetic chemistry, indicating a wide diversity of organic molecules.
ABSTRACT
Background: Thrombosis is a unique complication of coronavirus disease 2019 (COVID-19). Although antiphospholipid antibodies (aPL) are detected in COVID-19 patients, their clinical significance remains elusive. We evaluated the prevalence of aPL and serum concentrations of beta-2 glycoprotein I (ß2GPI), a major self-antigen for aPL, in Japanese COVID-19 patients with and without thrombosis. Methods: This retrospective single-center nested case-control study included 594 hospitalized patients with COVID-19 between January 2020 and August 2021. Thrombotic complications were collected from medical records. Propensity score-matching method (PSM) (1:2 matching including age, sex, severity on admission, and prior history of thrombosis) was performed to compare the prevalence and titer of aPL (anti-cardiolipin (aCL) IgG/IgM, anti-ß2GPI IgG/IgM/IgA, and anti-phosphatidylserine/prothrombin antibody (aPS/PT) IgG/IgM) and serum ß2GPI concentration. In addition, PSM (1:1 matching including age and sex) was performed to compare the serum ß2GPI concentration between COVID-19 patients and healthy donors. Results: Among the patients, 31 patients with thrombosis and 62 patients without were compared. The prevalence of any aPLs was indifferent regardless of the thrombosis (41.9% in those with thrombosis vs. 38.7% in those without, p =0.82). The positive rates of individual aPL were as follows: anti-CL IgG (9.7% vs. 1.6%, p =0.11)/IgM (0% vs. 3.2%, p =0.55), anti-ß2GP1 IgG (22.6% vs. 9.7%, p =0.12)/IgA (9.7% vs. 9.7%, p =1.0)/IgM (0% vs. 0%, p =1.0), and anti-PS/PT IgG (0% vs. 1.6%, p =1.0)/IgM (12.9% vs. 21.0%, p =0.41), respectively. The aPL titers were also similar regardless of thrombosis. The levels of ß2GPI in COVID-19 patients were lower than those in the healthy donors. Conclusion: Although aPLs were frequently detected in Japanese COVID-19 patients, their prevalence and titer were irrelevant to thrombotic complications. While COVID-19 patients have lower levels of serum ß2GPI than healthy blood donors, ß2GPI levels were indifferent regardless of thrombosis. Although most of the titers were below cut-offs, positive correlations were observed among aPLs, suggesting that the immune reactions against aPL antigens were induced by COVID-19. We should focus on the long-term thromboembolic risk and the development of APS in the aPL-positive patients with high titer or multiple aPLs.
Subject(s)
COVID-19 , Thrombosis , Humans , Retrospective Studies , Case-Control Studies , East Asian People , Propensity Score , Antibodies, Antiphospholipid , Antibodies, Anticardiolipin , beta 2-Glycoprotein I , Immunoglobulin M , Immunoglobulin A , Phosphatidylserines , Immunoglobulin GABSTRACT
Human body awareness is adaptive to context changes. The illusory sense of body ownership has been studied since the publication of the rubber hand illusion, where ambiguous body ownership feeling was first defined. Phenomenologically, the ambiguous body ownership is attributed to a conflict between feeling and judgement: it characterises a discrepancy between first- and third-person processes. Although Bayesian inference can explain this malleability of body image, it still fails to relate the subjective feeling to physiological data. This study attempts to explain subjective experience during rubber hand illusions by using integrated information theory (IIT). The integrated information [Formula: see text] in IIT measures the difference between the whole system and its subsystems. By analysing seven different time-series of physiological data representing a small body-brain system, we demonstrate that the integrity of the whole system during the illusion decreases, while the integrity of its subsystems increases. These general tendencies agree with many brain-image analyses and subjective reports; furthermore, we found that subjective ratings as ambiguous body ownership were associated with [Formula: see text]. Our result suggests that IIT can explain the general tendency of the sense of ownership illusions and individual differences in subjective experience during the illusions.
Subject(s)
Illusions , Touch Perception , Humans , Illusions/physiology , Information Theory , Bayes Theorem , Proprioception/physiology , Hand/physiology , Body Image , Touch Perception/physiology , Visual Perception/physiologyABSTRACT
Many organisms living along the coastlines synchronize their reproduction with the lunar cycle. At the time of spring tide, thousands of grass puffers (Takifugu alboplumbeus) aggregate and vigorously tremble their bodies at the water's edge to spawn. To understand the mechanisms underlying this spectacular semilunar beach spawning, we collected the hypothalamus and pituitary from male grass puffers every week for 2 months. RNA sequencing (RNA-seq) analysis identified 125 semilunar genes, including genes crucial for reproduction (e.g., gonadotropin-releasing hormone 1 [gnrh1], luteinizing hormone ß subunit [lhb]) and receptors for pheromone prostaglandin E (PGE). PGE2 is secreted into the seawater during the spawning, and its administration activates olfactory sensory neurons and triggers trembling behavior of surrounding individuals. These results suggest that PGE2 synchronizes lunar-regulated beach-spawning behavior in grass puffers. To further explore the mechanism that regulates the lunar-synchronized transcription of semilunar genes, we searched for semilunar transcription factors. Spatial transcriptomics and multiplex fluorescent in situ hybridization showed co-localization of the semilunar transcription factor CCAAT/enhancer-binding protein δ (cebpd) and gnrh1, and cebpd induced the promoter activity of gnrh1. Taken together, our study demonstrates semilunar genes that mediate lunar-synchronized beach-spawning behavior. VIDEO ABSTRACT.
Subject(s)
Moon , Takifugu , Humans , Animals , Male , Takifugu/genetics , Takifugu/metabolism , In Situ Hybridization, Fluorescence , Reproduction/physiology , Prostaglandins E/metabolism , Prostaglandins/metabolismABSTRACT
Computational material discovery is under intense study owing to its ability to explore the vast space of chemical systems. Neural network potentials (NNPs) have been shown to be particularly effective in conducting atomistic simulations for such purposes. However, existing NNPs are generally designed for narrow target materials, making them unsuitable for broader applications in material discovery. Here we report a development of universal NNP called PreFerred Potential (PFP), which is able to handle any combination of 45 elements. Particular emphasis is placed on the datasets, which include a diverse set of virtual structures used to attain the universality. We demonstrated the applicability of PFP in selected domains: lithium diffusion in LiFeSO4F, molecular adsorption in metal-organic frameworks, an order-disorder transition of Cu-Au alloys, and material discovery for a Fischer-Tropsch catalyst. They showcase the power of PFP, and this technology provides a highly useful tool for material discovery.
Subject(s)
Metal-Organic Frameworks , Neural Networks, Computer , Adsorption , CatalysisABSTRACT
BACKGROUND: There are some clinical reports on dysphagia in patients with chronic obstructive pulmonary disease (COPD); however, its pathophysiology remains largely unknown.Changes in respiratory function occur in patients with COPD causing a decrease in tidal volume and an increase in respiratory rate (tachypnea). In addition, it leads to lack of coordination between respiration and swallowing.A new treatment called nasal high flow (NHF) has been introduced for patients with COPD, replacing the traditional non-invasive ventilation (NIV) procedure. The NHF therapy involves inhalation of high flow of humidified air, which reduces respiratory effort in patients with COPD. Furthermore, NHF therapy facilitates swallowing of saliva even during respiratory management. A recent clinical study reported that high-flow nasal cannula oxygen therapy for 6 weeks improved the health-related quality of life and reduced hypercapnia in patients with stable COPD. Taken together, NHF therapy is gaining attention in the clinical management of patients with COPD.Therefore, in this study, we aim to examine the efficacy of NHF therapy on the coordination between breathing and swallowing of saliva during daytime nap in patients with COPD. METHODS/DESIGN: This open-label, investigator-initiated, single center study will evaluate the efficacy of NHF therapy on the coordination between breathing and swallowing of saliva during the daytime nap in COPD patients with forced expiratory volume in 1âsecond (FEV1%) of <70% during treatment at the Nagasaki University Hospital Respiratory Rehabilitation Center. Evaluations will be performed during the 90 to 180âminute "daytime nap" in the measurement room of the hospital. The primary endpoint will be the rate of appearance of the expiratory phase after swallowing of saliva and the frequency of swallowing during the measurement period. DISCUSSION: The purpose of this study is to obtain evidence regarding the utility of NHF as a potential therapeutic device for COPD patients to prevent aspiration of saliva during the sleep stage of daytime nap. The utility will be assessed by comparing the decrease in incidence rates of the expiratory phase after swallowing of saliva in the NHF device group and the control group, wherein this device was not used.
Subject(s)
Deglutition/physiology , Noninvasive Ventilation/methods , Pulmonary Disease, Chronic Obstructive/physiopathology , Pulmonary Disease, Chronic Obstructive/therapy , Respiratory Mechanics/physiology , Cannula , Humans , Noninvasive Ventilation/adverse effects , Research Design , SalivaABSTRACT
BACKGROUND: Body position might affect the coordination between respiration and swallowing. This study was carried out to test the hypothesis that during swallowing, coordinated movements of muscle groups such as the diaphragm and rectus abdominis muscles are important to control normal swallowing apnea. OBJECTIVE: To investigate this hypothesis, respiratory parameters, swallowing apnea and muscle activity were measured in each of four body positions: sitting position with feet on the floor, 30° reclining position, lateral position, and standing position. METHODS: All measurements were performed in nine healthy subjects. Nasal airflow was measured using a pneumotachometer and muscle activity was measured using an electromyograph. All lung volume fraction parameters were measured using spirometer and swallowing apnea time was calculated. RESULTS: The maximum inspiratory volume was 2.76 ± 0.83 L in the 30° reclining position, which was significantly larger than that in the other positions (p = .0001). The preliminary expiratory volume was 1.05 ± 0.42 L in the 30° reclining position, which was significantly smaller than that in the other positions (p < .0001). The swallowing apnea time during water swallowing was 1.17 ± 0.35 sec in the lateral position and 0.87 ± 0.28 sec in the 30° reclining position, which tended to be longer than the 0.78 sec in the sitting position. CONCLUSION: We conclude that both lateral and reclining positions require a longer period of swallowing apnea compared to the sitting and standing positions. Differences in body position may significantly influence the coordination between respiration and swallowing.
Subject(s)
Abdominal Muscles/physiology , Apnea/physiopathology , Deglutition , Posture , Reflex/physiology , Respiration , Adult , Female , Healthy Volunteers , Humans , Lung Volume Measurements , Male , Pilot Projects , Young AdultABSTRACT
Background: Several studies have reported a strong association between the presence of oropharyngeal secretions in the laryngeal vestibule and the likelihood of aspiration of food or liquid. However, no previous studies have evaluated the accumulation of saliva and swallowing dynamics. Objective: The objective of this study was to examine the factors related to decreased function that result in saliva accumulation based on images from videofluoroscopic examination of swallowing (VF) performed on the same day as videoendoscopic examination of swallowing (VE). Methods: This retrospective study investigated 47 patients with dysphagia who underwent VF and VE on the same day. Saliva accumulation in the pharynx was assessed on VE and classified by the Murray secretion scale. Pharyngeal residue was assessed on VF. In addition, displacement of the hyoid bone and larynx on swallowing and the opening size of the esophageal orifice were measured, and contact between the base of the tongue and the posterior pharyngeal wall was examined on VF. Results: Moderate correlations were found between saliva accumulation and perpendicular displacement of the larynx and upper esophageal sphincter opening. The percentage of patients showing contact between the base of the tongue and the posterior pharyngeal wall was significantly greater in those with a saliva accumulation score of 0 or 1. Conclusion: Less laryngeal elevation and upper esophageal sphincter opening and absence of contact between the base of the tongue and the posterior pharyngeal wall when swallowing tended to result in accumulation of saliva in the pharynx.
Subject(s)
Deglutition/physiology , Oropharyngeal Neoplasms/physiopathology , Saliva/physiology , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Retrospective StudiesABSTRACT
Lim15/Dmc1 is a meiosis specific RecA-like protein. Here we propose its participation in meiotic chromosome pairing-related events along with DNA topoisomerase II. Analysis of protein-protein interactions using in vitro binding assays provided evidence that Coprinus cinereus DNA topoisomerase II (CcTopII) specifically interacts with C.cinereus Lim15/Dmc1 (CcLim15). Co-immunoprecipitation experiments also indicated that the CcLim15 protein interacts with CcTopII in vivo. Furthermore, a significant proportion of CcLim15 and CcTopII could be shown to co-localize on chromosomes from the leptotene to the zygotene stage. Interestingly, CcLim15 can potently activate the relaxation/catenation activity of CcTopII in vitro, and CcTopII suppresses CcLim15-dependent strand transfer activity. On the other hand, while enhancement of CcLim15's DNA-dependent ATPase activity by CcTopII was found in vitro, the same enzyme activity of CcTopII was inhibited by adding CcLim15. The interaction of CcLim15 and CcTopII may facilitate pairing of homologous chromosomes.
Subject(s)
Cell Cycle Proteins/metabolism , DNA Topoisomerases, Type II/metabolism , DNA-Binding Proteins/metabolism , Meiosis , Base Sequence , Cell Cycle Proteins/analysis , Cell Cycle Proteins/chemistry , Cell Nucleus/chemistry , Cell Nucleus/enzymology , Coprinus/enzymology , DNA Topoisomerases, Type II/analysis , DNA Topoisomerases, Type II/chemistry , DNA-Binding Proteins/analysis , DNA-Binding Proteins/chemistry , Immunoprecipitation , Molecular Sequence Data , Sequence Deletion , Two-Hybrid System TechniquesABSTRACT
DNA ligase I is thought to be essential for DNA replication, repair and recombination, at least in the mitotic cell cycle, but whether this is also the case during the meiotic cell cycle is still obscure. To investigate the role of DNA ligase I during the meiotic cell cycle, we cloned the Coprinus cinereus DNA ligase I cDNA (CcLIG1). Northern blotting analysis indicated that CcLIG1 is expressed not only in the premeiotic S-phase but also during the meiotic cell cycle itself. Especially, intense signals were observed in the leptotene and zygotene stages. Western blotting analysis indicated that CcLIG1 is expressed through the meiotic cell cycle and immunofluorescence also showed CcLIG1 protein staining in meiotic cells. Interestingly, the patterns was similar to that for the C. cinereus proliferating cell nuclear antigen gene (CcPCNA) and immunoprecipitation analysis suggested that CcPCNA binds to CcLIG1 in crude extracts of meiotic prophase I tissues. Based on these observations, relationships and roles during the meiotic cell cycle are discussed.
Subject(s)
Coprinus/enzymology , Coprinus/genetics , DNA Ligases/biosynthesis , Meiosis/physiology , Amino Acid Sequence , Blotting, Northern , Blotting, Western , DNA Ligase ATP , DNA Ligases/genetics , DNA, Complementary , Fungal Proteins/genetics , Fungal Proteins/metabolism , Gene Dosage , In Situ Hybridization, Fluorescence , Molecular Sequence Data , Prophase/physiologyABSTRACT
In the basidiomycete Coprinus cinereus (C. cinereus), which shows a highly synchronous meiotic cell cycle, the meiotic prophase I cells demonstrate flap endonuclease-1 activity. To investigate its role during meiosis, we isolated a C. cinereus cDNA homolog of flap endonuclease-1 (CcFEN-1), 1377bp in length with the open reading frame (ORF) encoding a predicted molecular mass of 51 kDa. At amino-acid residues Glu276-Pro345, a specific inserted sequence composed of 70 amino acids rich in polar forms was found to exist, without sequence identity to other eukaryotic FEN-1 or the polar amino acid rich sequences found in C. cinereus PCNA and C. cinereus DNA ligase IV, although the lengths and percentages of polar amino acids were similar. Northern hybridization analysis indicated CcFEN-1 to be expressed not only in the pre-meiotic S phase but also in meiotic prophase I. The roles of CcFEN-1 during meiosis are discussed.