C2H2-zinc-finger transcription factors bind RNA and function in diverse post-transcriptional regulatory processes.

Cys2-His2 zinc-finger proteins (C2H2-ZNFs) constitute the largest class of DNA-binding transcription factors (TFs) yet remain largely uncharacterized. Although certain family members, e.g., GTF3A, have been shown to bind both DNA and RNA, the extent to which C2H2-ZNFs interact with-and regulate-RNA-associated processes is not known. Using UV crosslinking and immunoprecipitation (CLIP), we observe that 148 of 150 analyzed C2H2-ZNFs bind directly to RNA in human cells. By integrating CLIP sequencing (CLIP-seq) RNA-binding maps for 50 of these C2H2-ZNFs with data from chromatin immunoprecipitation sequencing (ChIP-seq), protein-protein interaction assays, and transcriptome profiling experiments, we observe that the RNA-binding profiles of C2H2-ZNFs are generally distinct from their DNA-binding preferences and that they regulate a variety of post-transcriptional processes, including pre-mRNA splicing, cleavage and polyadenylation, and m6A modification of mRNA. Our results thus define a substantially expanded repertoire of C2H2-ZNFs that bind RNA and provide an important resource for elucidating post-transcriptional regulatory programs.

A central chaperone-like role for 14-3-3 proteins in human cells.

14-3-3 proteins are highly conserved regulatory proteins that interact with hundreds of structurally diverse clients and act as central hubs of signaling networks. However, how 14-3-3 paralogs differ in specificity and how they regulate client protein function are not known for most clients. Here, we map the interactomes of all human 14-3-3 paralogs and systematically characterize the effect of disrupting these interactions on client localization. The loss of 14-3-3 binding leads to the coalescence of a large fraction of clients into discrete foci in a client-specific manner, suggesting a central chaperone-like function for 14-3-3 proteins. Congruently, the engraftment of 14-3-3 binding motifs to nonclients can suppress their aggregation or phase separation. Finally, we show that 14-3-3s negatively regulate the localization of the RNA-binding protein SAMD4A to cytoplasmic granules and inhibit its activity as a translational repressor. Our work suggests that 14-3-3s have a more prominent role as chaperone-like molecules than previously thought.

Regulation of alternative polyadenylation by the C2H2-zinc-finger protein Sp1.

Alternative polyadenylation (APA) enhances gene regulatory potential by increasing the diversity of mRNA transcripts. 3' UTR shortening through APA correlates with enhanced cellular proliferation and is a widespread phenomenon in tumor cells. Here, we show that the ubiquitously expressed transcription factor Sp1 binds RNA in vivo and is a common repressor of distal poly(A) site usage. RNA sequencing identified 2,344 genes (36% of the total mapped mRNA transcripts) with lengthened 3' UTRs upon Sp1 depletion. Sp1 preferentially binds the 3' UTRs of such lengthened transcripts and inhibits cleavage at distal sites by interacting with the subunits of the core cleavage and polyadenylation (CPA) machinery. The 3' UTR lengths of Sp1 target genes in breast cancer patient RNA-seq data correlate with Sp1 expression levels, implicating Sp1-mediated APA regulation in modulating tumorigenic properties. Taken together, our findings provide insights into the mechanism for dynamic APA regulation by unraveling a previously unknown function of the DNA-binding transcription factor Sp1.

Transcanal endoscopic ear surgery for glomus tympanicum removal: A case series on the necessity of preoperative embolization.

BACKGROUND: Transcanal endoscopic ear surgery (TEES) has become popular in recent years in the treatment of glomus tympanicum tumors (GTT). The most significant risk for TEES is bleeding. In some cases, preoperative vascular embolization is performed to mitigate bleeding during TEES. However, guidelines regarding the necessity and efficacy of preoperative vascular embolization have not been established yet. CASE PRESENTATION: This report aimed to assess the necessity and usefulness of preoperative vascular embolization in TEES for GTT by comparing the surgical findings of TEES without preoperative vascular embolization (Case 1) and TEES with preoperative vascular embolization (Case 2). Compared to Case 1, Case 2 included less bleeding and a more convenient procedure. However, no significant difference was observed. CONCLUSIONS: For GTT confined to the middle ear cavity (Glasscock-Jackson Grade II or less), when performed by a proficient otolaryngologist, TEES alone is sufficient without preoperative vascular embolization.

Functional characterization of RebL1 highlights the evolutionary conservation of oncogenic activities of the RBBP4/7 orthologue in Tetrahymena thermophila.

Retinoblastoma-binding proteins 4 and 7 (RBBP4 and RBBP7) are two highly homologous human histone chaperones. They function in epigenetic regulation as subunits of multiple chromatin-related complexes and have been implicated in numerous cancers. Due to their overlapping functions, our understanding of RBBP4 and 7, particularly outside of Opisthokonts, has remained limited. Here, we report that in the ciliate protozoan Tetrahymena thermophila a single orthologue of human RBBP4 and 7 proteins, RebL1, physically interacts with histone H4 and functions in multiple epigenetic regulatory pathways. Functional proteomics identified conserved functional links for Tetrahymena RebL1 protein as well as human RBBP4 and 7. We found that putative subunits of multiple chromatin-related complexes including CAF1, Hat1, Rpd3, and MuvB, co-purified with RebL1 during Tetrahymena growth and conjugation. Iterative proteomics analyses revealed that the cell cycle regulatory MuvB-complex in Tetrahymena is composed of at least five subunits including evolutionarily conserved Lin54, Lin9 and RebL1 proteins. Genome-wide analyses indicated that RebL1 and Lin54 (Anqa1) bind within genic and intergenic regions. Moreover, Anqa1 targets primarily promoter regions suggesting a role for Tetrahymena MuvB in transcription regulation. RebL1 depletion inhibited cellular growth and reduced the expression levels of Anqa1 and Lin9. Consistent with observations in glioblastoma tumors, RebL1 depletion suppressed DNA repair protein Rad51 in Tetrahymena, thus underscoring the evolutionarily conserved functions of RBBP4/7 proteins. Our results suggest the essentiality of RebL1 functions in multiple epigenetic regulatory complexes in which it impacts transcription regulation and cellular viability.

Long Noncoding RNAs and Repetitive Elements: Junk or Intimate Evolutionary Partners?

Our recent ability to sequence entire genomes, along with all of their transcribed RNAs, has led to the surprising finding that only ∼1% of the human genome is used to encode proteins. This finding has led to vigorous debate over the functional importance of the transcribed but untranslated portions of the genome. Currently, scientists tend to assume coding genes are functional until proven not to be, while the opposite is true for noncoding genes. This review takes a new look at the evidence for and against widespread noncoding gene functionality. We focus in particular on long noncoding RNA (noncoding RNAs longer than 200 nucleotides) genes and their 'junk' associates, transposable elements, and satellite repeats. Taken together, the suggestion put forward is that more of this junk DNA may be functional than nonfunctional and that noncoding RNAs and transposable elements act symbiotically to drive evolution.

CRISPR/Cas9-Mediated Editing of AGAMOUS-like Genes Results in a Late-Bolting Phenotype in Chinese Cabbage (Brassica rapa ssp. pekinensis).

Due to the sudden change in temperature in spring, Chinese cabbage, a leafy vegetable cultivated for consumption, loses its commercial value due to the onset of bolting­the phenomenon of switching from vegetative to reproductive growth. In this study, we applied clustered regularly interspaced short palindromic repeats/(CRISPR)-associated system 9 (CRISPR/Cas9) technology to analyze AGAMOUS-like genes. We performed functional analysis of AGL19 and AGL24 genes related to bolting and flowering using CRISPR/Cas9-mediated Chinese cabbage transformation. Single-guide RNA (sgRNA) sequences were created with a low off-targeting probability to construct gene-editing vectors. Agrobacterium-mediated transformation was conducted, and tentative E0 AGL-edited lines were analyzed using molecular biotechnological methods. Two AGL19-edited lines with nucleotide sequence mutations in the target sequence of the AGL19 genes and four AGL24-edited lines with nucleotide sequence mutations in the target sequence of the AGL24 genes showed particularly late bolting compared to the inbred line 'CT001.' Generational progression using bud pollination obtained T-DNA-free E1 AGL-edited lines, which also showed late bolting. The loss of function of the AGL protein was caused by the occurrence of an indel mutation in the AGL19 and AGL24 genes, which results in an early stop codon. Furthermore, frameshift mutations led to structural changes and the introduction of an early stop codon in the AGL19 and AGL24 proteins. Our results indicate that CRISPR/Cas9-mediated editing of AGAMOUS-like genes results in a late-bolting phenotype and that CRISPR/Cas9 is a useful technology for analyzing gene function in Chinese cabbage (Brassica rapa ssp. pekinensis).

Navigating the COVID-19 pandemic in the contingency framework: Antecedents and consequences of public's stance toward the CDC.

This study applied the contingency theory of conflict management to examine how contingency factors influence the public's perceptual and behavioral responses to COVID-19 and stance toward the Centers for Disease Control and Prevention (CDC). In particular, we tested political ideology as an important individual characteristic variable to examine its roles in the contingency theory framework. The findings revealed that two situational variables (i.e., threat appraisal and attitudes toward CDC) positively influenced the public's contingency accommodation stance toward the CDC. Furthermore, greater conservatism was significantly associated with lower levels of threat appraisal and more negative attitudes toward the CDC, however it did not influence the stance toward the CDC. Theoretical and practical implications of the findings and directions for future research are discussed.

Mercury Chloride but Not Lead Acetate Causes Apoptotic Cell Death in Human Lung Fibroblast MRC5 Cells via Regulation of Cell Cycle Progression.

Heavy metals are important for various biological systems, but, in excess, they pose a serious risk to human health. Heavy metals are commonly used in consumer and industrial products. Despite the increasing evidence on the adverse effects of heavy metals, the detailed mechanisms underlying their action on lung cancer progression are still poorly understood. In the present study, we investigated whether heavy metals (mercury chloride and lead acetate) affect cell viability, cell cycle, and apoptotic cell death in human lung fibroblast MRC5 cells. The results showed that mercury chloride arrested the sub-G1 and G2/M phases by inducing cyclin B1 expression. In addition, the exposure to mercury chloride increased apoptosis through the activation of caspase-3. However, lead had no cytotoxic effects on human lung fibroblast MRC5 cells at low concentration. These findings demonstrated that mercury chloride affects the cytotoxicity of MRC5 cells by increasing cell cycle progression and apoptotic cell death.

Transcriptome Analysis Reveals HgCl2 Induces Apoptotic Cell Death in Human Lung Carcinoma H1299 Cells through Caspase-3-Independent Pathway.

Mercury is one of the detrimental toxicants that can be found in the environment and exists naturally in different forms; inorganic and organic. Human exposure to inorganic mercury, such as mercury chloride, occurs through air pollution, absorption of food or water, and personal care products. This study aimed to investigate the effect of HgCl2 on cell viability, cell cycle, apoptotic pathway, and alters of the transcriptome profiles in human non-small cell lung cancer cells, H1299. Our data show that HgCl2 treatment causes inhibition of cell growth via cell cycle arrest at G0/G1- and S-phase. In addition, HgCl2 induces apoptotic cell death through the caspase-3-independent pathway. Comprehensive transcriptome analysis using RNA-seq indicated that cellular nitrogen compound metabolic process, cellular metabolism, and translation for biological processes-related gene sets were significantly up- and downregulated by HgCl2 treatment. Interestingly, comparative gene expression patterns by RNA-seq indicated that mitochondrial ribosomal proteins were markedly altered by low-dose of HgCl2 treatment. Altogether, these data show that HgCl2 induces apoptotic cell death through the dysfunction of mitochondria.

EEG-Based Evidence of Mirror Neuron Activity from App-Mediated Stroke Patient Observation.

Background and Objectives: The mirror neuron system in the sensorimotor region of the cerebral cortex is equally activated during both action observation and execution. Action observation training mimics the functioning of the mirror neuron system, requiring patients to watch and imitate the actions necessary to perform activities of daily living. StrokeCare is a user-friendly application based on the principles of action observation training, designed to assist people recovering from stroke. Therefore, when observing the daily life behavior provided in the StrokeCare app, whether the MNS is activated and mu inhibition appears. Materials and Methods: We performed electroencephalography (EEG) on 24 patients with chronic stroke (infarction: 11, hemorrhage: 13) during tasks closely related to daily activities, such as dressing, undressing, and walking. The StrokeCare app provided action videos for patients to watch. Landscape imagery observation facilitated comparison among tasks. We analyzed the mu rhythm from the C3, CZ, and C4 regions and calculated the mean log ratios for comparison of mu suppression values. Results: The EEG mu power log ratios were significantly suppressed during action observation in dressing, undressing, walking, and landscape conditions, in decreasing order. However, there were no significant activity differences in the C3, C4 and CZ regions. The dressing task showed maximum suppression after a color spectrum was used to map the relative power values of the mu rhythm for each task. Conclusions: These findings reveal that the human mirror neuron system was more strongly activated during observation of actions closely related to daily life activities than landscape images.

The usefulness of Valsalva computed tomography as an assessment tool for the Eustachian tube.

PURPOSE: To determine the usefulness of 'computed tomography (CT) with Valsalva maneuver (VM)' (Valsalva CT) for localizing lesions of the eustachian tube (ET) and identifying characteristics of ET dysfunction (ETD). MATERIALS AND METHODS: In this case-control study, 12, 17, and 25 patients with obstructive ETD (OETD), patulous eustachian tube (PET), and conductive hearing loss (control), respectively, underwent Valsalva CT in the supine position. The visualized length to total length ratio of the cartilaginous ET (VTRET) with and without VM were compared in the three groups. In the OETD group, obstructed areas of the ET were identified on multiplanar reconstructed images. RESULTS: VTRET without VM in the PET group was significantly higher than that in the control and OETD groups (p-values, 0.003 and 0.007, respectively). However, there was no significant difference between the control and OETD groups (p = 0.053). VTRET with VM in the PET group was significantly higher than that in the other two groups (p < 0.001), whereas that in the control group was significantly higher than that in the OETD group (p < 0.001). In the OETD group, the obstructed area was clearly identified, and there were one, one, six, and eight patients in the isthmus; bony portion and isthmus; bony portion, isthmus, and cartilaginous portion; and isthmus and cartilaginous portion subgroups, respectively. CONCLUSION: Valsalva CT is particularly valuable as a visualization assessment tool for identifying obstructed areas in the OETD. It may help future research of disease-specific mechanical characteristics of the ET on controlling pressure variables.

Audiologic, surgical and subjective outcomes of active transcutaneous bone conduction implant system (Bonebridge).

Objective: Our objective was to evaluate the surgical restrictions, audiologic benefits and satisfaction from using an active transcutaneous bone conduction device (BCD), Bonebridge® (BB) in patients with mixed/conductive hearing loss (MCHL) or single-sided deafness (SSD).Design: A retrospective review from all patients who underwent BB surgery at the Pusan National University Hospital from 2015 to 2017 for SSD or MCHL was performed.Study sample: Twenty-two patients with SSD and five with MCHL had a BB implanted and analysed.Results: Complete transmastoid implantation of the device was possible for all patients with an intact canal wall (ICW), using lifts if necessary. The overall functional hearing gain (FHG) in SSD and MCHL was 31.4 and 37.6 dB, respectively. The mean percentage of speech recognition in a quiet was 81% (vs. 11% unaided) for MCHL group and 82% (vs. 29% unaided) for SSD group. Mean speech recognition scores in noise improved significantly under various signal-to-noise ratio (SNR) for both groups. Questionnaires showed overall improvement, and there was no significant difference between the two groups.Conclusions: The BB provides improved functional gain, and there were no limitations during surgery despite the large device. Both MCHL and SSD group had benefit and improved quality of life with BB.

Comparison of Two Different Treatment Protocols Using Systemic and Intratympanic Steroids with and without Hyperbaric Oxygen Therapy in Patients with Severe to Profound Idiopathic Sudden Sensorineural Hearing Loss: A Randomized Controlled Trial.

OBJECTIVES: This study aimed to investigate the efficacy of simultaneous steroid and hyperbaric oxygen therapy (HBOT) in patients with severe to profound idiopathic sudden sensorineural hearing loss (ISSNHL), which has a poor prognosis. METHODS: Sixty patients diagnosed with severe to profound ISSNHL (≥70 dB HL) were randomly divided into two groups in a prospective controlled trial: an oral steroid + intratympanic steroid injection (ITSI) group (control group) and an oral steroid + ITSI + HBOT group (study group). Pure-tone audiometry (PTA) results and word discrimination scores (WDS) were compared between the two groups before treatment and 10 days and 1, 2, and 3 months after treatment. Hearing improvement was assessed using the modified American Academy of Otolaryngology-Head and Neck Surgery criteria. Analyses were by both intention to treat and per protocol. RESULTS: A total of 58 patients completed the 3-month follow-up, and 2 patients in the study group were excluded due to follow-up loss in the per-protocol analysis. In the intention-to-treat and per-protocol analyses, the study group showed significantly better hearing levels than did the control group at 500 Hz (p < 0.05) 1 month after treatment and at 1 kHz (p < 0.05) 3 months after treatment. However, the average PTA values and PTA at 2, 4, and 8 kHz showed no significant difference. WDS improvement was significantly higher in the study group compared to the control group 3 months after treatment by both per-protocol (66.4 ± 13.3 and 56.7 ± 19.1%, respectively; p = 0.029) and intention-to-treat analyses (65.9 ± 14.1 and 56.7 ± 19.1%, respectively; p = 0.035). The sum of complete and partial hearing recovery for the study group was significantly higher than that for the control group by per-protocol analysis (60.7 vs. 33.3%; p = 0.037) and intention-to-treat analysis (60.0 vs. 33.3%; p = 0.038). CONCLUSION: These results demonstrate that the addition of HBOT to steroid combination therapy does not improve the average PTA values in severe to profound ISSNHL; however, it was associated with a better outcome at 500 Hz 1 month after treatment and, at 1 kHz, WDS 3 months after treatment. The sum of complete and partial hearing recovery was significantly higher for the study group than for the control group.

Application of endoscopy in otology: Changes over the last 8years in Korean Otological Society.

OBJECTIVES: Despite the worldwide availability of endoscope in otology, it still has not replaced the microscopic surgery as it has for nose surgery. In 2008, we administered a questionnaire on the preference of using otoscopes in Korea. In light of the worldwide availability of endoscopic ear surgery, we have now conducted a more detailed survey to determine if this preference has changed over 8years. SUBJECTS AND METHODS: A questionnaire consisting of 10 questions was used to survey members of the Korean Otological Society who were actively performing middle ear surgery. The responses to this questionnaire were compared to those from 2008. The study also determined the preference for endoscope use based on the surgeon's experience. RESULTS: The mean surgical experience of the otologists was 12.7years. Endoscopy for tympanoplasty and tympanomastoidectomy increased over a period of 8years. In the outpatient clinic, the use of endoscopy also increased; over 8-year period, it surpassed the microscope as the most preferred diagnostic tool in the outpatient clinic. Greater than half of the operating rooms were equipped with endoscopic sets; however, only 4.5% of otologists acknowledged having endoscopic instruments. CONCLUSION: Endoscopy for surgery and in outpatient clinic assessment increased in otology in Korea; however, endoscopes have consistently been used as an adjuvant to the microscope to improve visualization of the tympanic cavity rather than as a tool for totally endoscopic ear surgery.

Utilizing Audiovisual and Gain-Framed Messages to Attenuate Psychological Reactance Toward Weight Management Health Messages.

Guided by the psychological reactance theory, this study predicted that gain-framed messages and audiovisual content could counteract state reactance and increase the persuasiveness of weight management health messages. Data from a 2 (message frame: gain/loss) × 2 (modality: audiovisual/text) × 2 (message repetition) within-subjects experiment (N = 82) indicated that in the context of weight management messages for college students, gain-framed messages indeed mitigate psychological reactance. Furthermore, the modality and the frame of the health message interacted in such a way that gain-framed messages in an audiovisual modality generated the highest motivations to comply with the recommendations in the persuasive health messages.

Hearing aid silicone impression material as a foreign body in the middle ear.

We report an extremely rare case of hearing aid silicone impression material as a foreign body in the middle ear. Symptoms of the patient were otorrhea and vertigo after taking of a mold impression on his only hearing ear, and the symptoms mimicked chronic otitis media. A temporal bone CT scan revealed foreign body material in the middle ear and Eustachian tube. An intact canal wall mastoidectomy with a facial recess approach and type IV tympanoplasty was performed to remove the silicone impression material. In addition to the case report, we review the literature regarding impression material foreign bodies.

Clinical and audiological characteristics of 1000Hz audiometric notch patients.

PURPOSE: Among the hearing loss patients, we can confirm that the hearing loss of the specific frequency decreases, such as the 2000Hz notch in otosclerosis and the 4000Hz notch (c5-dip) in noise-induced hearing loss. The 1000Hz notch (c3-dip), however, is rarely studied. We fortuitously encountered a group of patients with a 1kHz hearing loss and report it with a review of the literature. METHODS: Otological history, audiogram, diagnosis, occupation, and history of noise exposure were reviewed from charts and telephone interview, and compared between c3-dip and c5-dip patients (n=98). RESULTS: Thirty-one patients (mean age: 46.2years) demonstrated 1kHz hearing loss; these included 11 males. The pure-tone threshold was 37.97dB at 1kHz and the average threshold was 22.38dB at other frequencies. In the c3-dip group, tinnitus was the most common complaint, while sudden sensorineural hearing loss and idiopathic tinnitus (n=8 each) were the most common diagnoses. Female patients and unilateral cases were more common in the c3-dip than in the c5-dip group, and ear fullness was more common in the c3-dip group than in the c5-dip group. The duration of occupation-related noise exposure was longer in the c5 group, and head or ear trauma was more frequent in the c3-dip group. CONCLUSION: We have defined a new clinical entity of 1kHz hearing loss in patients, defined as the c3-dip, which was clinically and audiologically distinct from the c5-dip. Further study is needed to clarify this new entity of hearing loss.

Third-Person Effect and Pandemic Flu: The Role of Severity, Self-Efficacy Method Mentions, and Message Source.

Within the context of a pandemic flu, this experiment investigated whether source (government officials or physicians), severity condition (high or low), and mention of self-efficacy method (mention present or absent) in H1N1 health news affected participants' (a) perception of media influence on self and others and (b) intentions to get vaccinated. Results found support for third-person effects, and the magnitude of the effects grew with social distance. Main effect of source, as well as interaction effects among the independent variables on third-person effect and vaccination intentions, were also found.

Trimethylation enhancement using diazomethane (TrEnDi) II: rapid in-solution concomitant quaternization of glycerophospholipid amino groups and methylation of phosphate groups via reaction with diazomethane significantly enhances sensitivity in mass spectrometry analyses via a fixed, permanent positive charge.

A novel mass spectrometry (MS)-based lipidomics strategy that exposes glycerophospholipids to an ethereal solution of diazomethane and acid, derivatizing them to contain a net fixed, permanent positive charge, is described. The sensitivity of modified lipids to MS detection is enhanced via improved ionization characteristics as well as consolidation of ion dissociation to form one or two strong, characteristic polar headgroup fragments. Our strategy has been optimized to enable a priori prediction of ion fragmentation patterns for four subclasses of modified glycerophospholipid species. Our method enables analyte ionization regardless of proton affinity, thereby decreasing ion suppression and permitting predictable precursor ion-based quantitation with improved sensitivity in comparison to MS-based methods that are currently used on unmodified lipid precursors.