Effect of reducing electrical stimulation rate on hearing performance of Nucleus® cochlear implant recipients.

OBJECTIVE: To evaluate whether a 500 pulses per second per channel (pps/ch) rate would provide non-inferior hearing performance compared to the 900 pps/ch rate in the Advanced Combination Encoder (ACE™) sound coding strategy. DESIGN: A repeated measures single-subject design was employed, wherein each subject served as their own control. All except one subject used 900 pps/ch at enrolment. After three weeks of using the alternative rate program, both programs were loaded into the sound processor for two more weeks of take-home use. Subjective performance, preference, words in quiet, sentences in babble, music quality, and fundamental frequency (F0) discrimination were assessed using a balanced design. STUDY SAMPLE: Data from 18 subjects were analysed, with complete datasets available for 17 subjects. RESULTS: Non-inferior performance on all clinical measures was shown for the lower rate program. Subjects' preference ratings were comparable for the programs, with 53% reporting no difference overall. When a preference was expressed, the 900 pps/ch condition was preferred more often. CONCLUSION: Reducing the stimulation rate from 900 pps/ch to 500 pps/ch did not compromise the hearing outcomes evaluated in this study. A lower pulse rate in future cochlear implants could reduce power consumption, allowing for smaller batteries and processors.

Collaborative Cohort of Cohorts for COVID-19 Research (C4R) Study: Study Design.

The Collaborative Cohort of Cohorts for COVID-19 Research (C4R) is a national prospective study of adults comprising 14 established US prospective cohort studies. Starting as early as 1971, investigators in the C4R cohort studies have collected data on clinical and subclinical diseases and their risk factors, including behavior, cognition, biomarkers, and social determinants of health. C4R links this pre-coronavirus disease 2019 (COVID-19) phenotyping to information on severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and acute and postacute COVID-related illness. C4R is largely population-based, has an age range of 18-108 years, and reflects the racial, ethnic, socioeconomic, and geographic diversity of the United States. C4R ascertains SARS-CoV-2 infection and COVID-19 illness using standardized questionnaires, ascertainment of COVID-related hospitalizations and deaths, and a SARS-CoV-2 serosurvey conducted via dried blood spots. Master protocols leverage existing robust retention rates for telephone and in-person examinations and high-quality event surveillance. Extensive prepandemic data minimize referral, survival, and recall bias. Data are harmonized with research-quality phenotyping unmatched by clinical and survey-based studies; these data will be pooled and shared widely to expedite collaboration and scientific findings. This resource will allow evaluation of risk and resilience factors for COVID-19 severity and outcomes, including postacute sequelae, and assessment of the social and behavioral impact of the pandemic on long-term health trajectories.

Two Interferon-Stimulated Response Elements Cooperatively Regulate Interferon-Stimulated Gene Expression in West Nile Virus-Infected IFNAR-/- Mouse Embryo Fibroblasts.

We previously identified a subset of interferon-stimulated genes (ISGs) upregulated by West Nile virus (WNV) infection in wild-type mouse embryo fibroblasts (MEFs) after viral proteins had inhibited type I interferon (IFN)-mediated JAK-STAT signaling and also in WNV-infected RIG-I-/-, MDA5-/-, STAT1-/-, STAT2-/-, IFNAR-/-, IRF3-/-, IRF7-/-, and IRF3/7-/- MEFs. In this study, ISG upregulation by WNV infection in IFNAR-/- MEFs was confirmed by transcriptome sequencing (RNA-seq). ISG upregulation by WNV infection was inhibited in RIG-I/MDA5-/- MEFs. ISGs were upregulated in IRF1-/- and IRF5-/- MEFs but only minimally upregulated in IRF3/5/7-/- MEFs, suggesting redundant IRF involvement. We previously showed that a single proximal interferon-stimulated response element (ISRE) in the Oas1a and Oas1b promoters bound the ISGF3 complex after type I IFN treatment. In this study, we used wild-type and mutant promoter luciferase reporter constructs to identify critical regions in the Oas1b and Ifit1 promoters for gene activation in infected IFNAR-/- MEFs. Two ISREs were required in both promoters. Mutation of these ISREs in an Ifit1 promoter DNA probe reduced in vitro complex formation with infected nuclear extracts. An NF-κB inhibitor decreased Ifit1 promoter activity in cells and in vitro complex formation. IRF3 and p50 promoter binding was detected by chromatin immunoprecipitation (ChIP) for upregulated ISGs with two proximal ISREs. The data indicate that ISREs function cooperatively to upregulate the expression of some ISGs when type I IFN signaling is absent, with the binding complex consisting of IRF3, IRF5, and/or IRF7 and an NF-κB component(s) as well as other, as-yet-unknown factors. IMPORTANCE Type I IFN signaling in mammalian cells induces formation of the ISGF3 transcription factor complex, which binds to interferon stimulated response elements (ISREs) in the promoters of interferon-stimulated genes (ISGs) in the cell nucleus. Flavivirus proteins counteract type I IFN signaling by preventing either the formation or nuclear localization of ISGF3. A subset of ISRE-regulated ISGs was still induced in West Nile virus (WNV)-infected mouse embryo fibroblasts (MEFs), indicating that cells have an alternative mechanism for activating these ISGs. In this study, cellular components involved in this ISG upregulation mechanism were identified using gene knockout MEFs and ChIP, and critical promoter regions for gene activation were mapped using reporter assays. The data indicate a cooperative function between two ISREs and required binding of IRF3, IRF5, and/or IRF7 and an NF-κB component(s). Moreover, type I IFN signaling-independent ISG activation requires different additional promoter activation regions than type I IFN-dependent activation.

Remote check test battery for cochlear implant recipients: proof of concept study.

OBJECTIVES: (1) To investigate the remote check test battery, designed for self-administration by cochlear implant (CI) recipients, parents/caregivers, to determine if the results give adequate information for clinicians to decide the necessity of an appointment and to capture suggestions for improvement. (2) To gauge acceptance of remote monitoring by CI-recipients and their parents/caregivers. DESIGN: Prospective, multicentre, un-blinded, non-randomized, single-subject, repeated-measures evaluation. The test battery includes an implant-site photograph, impedance measurements, datalogs, questionnaires, speech perception and aided threshold tests. Clinicians reviewed test battery results, followed by a clinical appointment with each CI-recipient, and reported if the battery identified all the issues. Study sample: n = 93 CI-recipients (73 adults, 20 children) and 28 clinicians. RESULTS: The test battery identified 94% (615/656) of all issues. The test battery and clinician observations agreed in 99% (92/93) of cases on the need for a clinic visit. For 68% (63/93) of cases, the test battery identified all clinician observed issues. The majority (77%, 72/93) of recipients would be satisfied if clinic visits were based on their test battery results. A significantly high proportion agreed that remote monitoring was more convenient than clinic visits and could result in travel, time and cost reductions. CONCLUSION: This is the first comprehensive test battery designed for CI-recipient remote monitoring.

Lesions of anogenital mammary-like glands: Four cases including novel pathologic and immunohistochemical observations.

Anogenital mammary-like glands, formerly described as ectopic breast tissue, are currently considered to be normal histologic components of the anogenital region. Anogenital mammary-like glands can give rise to many lesions identical to counterparts in the native female breast. We describe four cases of such lesions, including fibroadenoma, gynecomastia-like hyperplasia, and ectopic mammary-type tissue with a spectrum of usual ductal hyperplasia, apocrine metaplasia, adenosis, and pseudolactational change. All four cases occurred in young women (ages 29-38) who presented with vulvar or perianal masses. Similar to previously reported cases, these lesions shared histologic and immunohistochemical characteristics identical to native female breast lesions. Novel findings in our cases included (1) the first case of gynecomastia-like change to be reported in the perianal area of a female, (2) Immunohistochemical staining identifying a 3-layered epithelium characterized by a population of CK14 and CK5/6 positive and hormone receptor negative superficial luminal cells, and (3) diffuse, strong positivity for GATA3 in all cases. Our study adds to the literature on these rare lesions and highlights findings which may be useful in understanding the pathogenesis and improving the diagnosis of anogenital mammary-like gland lesions.

Protection against ß-amyloid neurotoxicity by a non-toxic endogenous N-terminal ß-amyloid fragment and its active hexapeptide core sequence.

High levels (µM) of beta amyloid (Aß) oligomers are known to trigger neurotoxic effects, leading to synaptic impairment, behavioral deficits, and apoptotic cell death. The hydrophobic C-terminal domain of Aß, together with sequences critical for oligomer formation, is essential for this neurotoxicity. However, Aß at low levels (pM-nM) has been shown to function as a positive neuromodulator and this activity resides in the hydrophilic N-terminal domain of Aß. An N-terminal Aß fragment (1-15/16), found in cerebrospinal fluid, was also shown to be a highly active neuromodulator and to reverse Aß-induced impairments of long-term potentiation. Here, we show the impact of this N-terminal Aß fragment and a shorter hexapeptide core sequence in the Aß fragment (Aßcore: 10-15) to protect or reverse Aß-induced neuronal toxicity, fear memory deficits and apoptotic death. The neuroprotective effects of the N-terminal Aß fragment and Aßcore on Aß-induced changes in mitochondrial function, oxidative stress, and apoptotic neuronal death were demonstrated via mitochondrial membrane potential, live reactive oxygen species, DNA fragmentation and cell survival assays using a model neuroblastoma cell line (differentiated NG108-15) and mouse hippocampal neuron cultures. The protective action of the N-terminal Aß fragment and Aßcore against spatial memory processing deficits in amyloid precursor protein/PSEN1 (5XFAD) mice was demonstrated in contextual fear conditioning. Stabilized derivatives of the N-terminal Aßcore were also shown to be fully protective against Aß-triggered oxidative stress. Together, these findings indicate an endogenous neuroprotective role for the N-terminal Aß fragment, while active stabilized N-terminal Aßcore derivatives offer the potential for therapeutic application.

Preoperative autologous blood collection before bone marrow harvests in haploidentical related donors: is it justified?

BACKGROUND: With the increasing safety of allogeneic blood supply and declining need for transfusion due to patient blood management, the practice of preoperative autologous donation (PAD) continues to decline. The practice gained popularity during the 1980s and 1990s with the emergence of transfusion-transmitted human immunodeficiency virus and hepatitis C. At the peak of this public concern, the National Marrow Donor Program recommended that marrow donors have 1 to 3 autologous units of blood collected before their marrow harvest to minimize the likelihood of allogeneic transfusion. After three decades, the practice remains prevalent in marrow donors. We aimed to study the efficacy of PAD in healthy marrow donors. STUDY DESIGN AND METHODS: PADs performed before marrow harvest in healthy donors at our center between January 2013 and July 2015 were reviewed. The utilization of autologous units and decrease in hemoglobin levels due to PAD and marrow harvest were studied. Similar practices were assessed in the rest of the United States through a brief survey. RESULTS: Of a total of 262 autologous units collected from 136 donors, 25.2% were wasted. Ninety-nine percent of the marrow donors received at least 1 unit of blood irrespective of the need. PAD contributed to preoperative anemia, exposing three donors to allogeneic blood transfusion. The survey results showed a mixed response with some institutions continuing and others not practicing PAD. CONCLUSION: PADs are not justified in healthy marrow donors as they expose them to a risk of preoperative anemia and hence a greater risk of transfusion.

Revisiting Cardiac Cellular Composition.

RATIONALE: Accurate knowledge of the cellular composition of the heart is essential to fully understand the changes that occur during pathogenesis and to devise strategies for tissue engineering and regeneration. OBJECTIVE: To examine the relative frequency of cardiac endothelial cells, hematopoietic-derived cells, and fibroblasts in the mouse and human heart. METHODS AND RESULTS: Using a combination of genetic tools and cellular markers, we examined the occurrence of the most prominent cell types in the adult mouse heart. Immunohistochemistry revealed that endothelial cells constitute >60%, hematopoietic-derived cells 5% to 10%, and fibroblasts <20% of the nonmyocytes in the heart. A refined cell isolation protocol and an improved flow cytometry approach provided an independent means of determining the relative abundance of nonmyocytes. High-dimensional analysis and unsupervised clustering of cell populations confirmed that endothelial cells are the most abundant cell population. Interestingly, fibroblast numbers are smaller than previously estimated, and 2 commonly assigned fibroblast markers, Sca-1 and CD90, under-represent fibroblast numbers. We also describe an alternative fibroblast surface marker that more accurately identifies the resident cardiac fibroblast population. CONCLUSIONS: This new perspective on the abundance of different cell types in the heart demonstrates that fibroblasts comprise a relatively minor population. By contrast, endothelial cells constitute the majority of noncardiomyocytes and are likely to play a greater role in physiological function and response to injury than previously appreciated.

Coronary Artery Disease Associated Transcription Factor TCF21 Regulates Smooth Muscle Precursor Cells That Contribute to the Fibrous Cap.

Recent genome wide association studies have identified a number of genes that contribute to the risk for coronary heart disease. One such gene, TCF21, encodes a basic-helix-loop-helix transcription factor believed to serve a critical role in the development of epicardial progenitor cells that give rise to coronary artery smooth muscle cells (SMC) and cardiac fibroblasts. Using reporter gene and immunolocalization studies with mouse and human tissues we have found that vascular TCF21 expression in the adult is restricted primarily to adventitial cells associated with coronary arteries and also medial SMC in the proximal aorta of mouse. Genome wide RNA-Seq studies in human coronary artery SMC (HCASMC) with siRNA knockdown found a number of putative TCF21 downstream pathways identified by enrichment of terms related to CAD, including "vascular disease," "disorder of artery," and "occlusion of artery," as well as disease-related cellular functions including "cellular movement" and "cellular growth and proliferation." In vitro studies in HCASMC demonstrated that TCF21 expression promotes proliferation and migration and inhibits SMC lineage marker expression. Detailed in situ expression studies with reporter gene and lineage tracing revealed that vascular wall cells expressing Tcf21 before disease initiation migrate into vascular lesions of ApoE-/- and Ldlr-/- mice. While Tcf21 lineage traced cells are distributed throughout the early lesions, in mature lesions they contribute to the formation of a subcapsular layer of cells, and others become associated with the fibrous cap. The lineage traced fibrous cap cells activate expression of SMC markers and growth factor receptor genes. Taken together, these data suggest that TCF21 may have a role regulating the differentiation state of SMC precursor cells that migrate into vascular lesions and contribute to the fibrous cap and more broadly, in view of the association of this gene with human CAD, provide evidence that these processes may be a mechanism for CAD risk attributable to the vascular wall.

Cancer type predicts alloimmunization following RhD-incompatible RBC transfusions.

BACKGROUND: Immunosuppressed, RhD-negative oncology patients tend to have lower rates of sensitization to the D antigen when they receive transfusion with RhD-positive blood components. Clinical factors associated with alloimmunization to the D antigen in RhD-negative oncology patients when they receive transfusion with RhD-positive red blood cells (RBCs) have not been well defined. STUDY DESIGN AND METHODS: This was a 4-year, retrospective analysis identifying RhD-negative oncology patients who received RhD-positive RBCs and were not previously alloimmunized to the D antigen. Age, sex, race, ABO group, primary oncology diagnosis, and numbers of RhD-incompatible RBC transfusions were recorded. The association between antibody formation and clinical factors was studied. The incidence of alloanti-D was calculated from a subsequent antibody-detection test performed at least 28 days after receipt of the first transfusion of RhD-positive RBCs. RESULTS: In total, 545 RhD-negative oncology patients received 4295 RhD-positive RBC transfusions. Of these, 76 (14%) became alloimmunized to the D antigen. Diagnosis type was the only factor significantly associated with responder status. The logistic regression model indicated that patients who had myelodysplastic syndrome or solid malignancies were more likely to be responders than those who had acute leukemia. CONCLUSION: We measured a 14% sensitization rate to the D antigen in our RhD-negative oncology population. The rate of alloimmunization was higher in patients who had solid cancers (22.6%) or myelodysplastic syndrome (23%) compared with those who had other hematologic malignancies (7%). Knowledge of diagnoses that predispose to RhD alloimmunization enables better utilization of RhD-negative RBCs during times of shortage.

Sarcoid-like granulomas in renal cell carcinoma: The Houston Methodist Hospital experience.

Sarcoid-like (SL) granulomas have been previously described in association with malignant tumors. These granulomas appear to be tumor-related but are not indicative of systemic sarcoidosis, and hence are referred to as SL reactions. These SL reactions can be seen within the primary tumor, its vicinity, or in uninvolved sites such as the spleen, bone marrow, skin, and/or regional lymph nodes draining the tumor. It is a widely held view that SL granulomas are caused by soluble antigenic factors, shed by tumor cells or released due to tumor necrosis. SL reactions reported in Hodgkin lymphoma have been associated with a better prognosis. SL granulomas are thought to play an important role in the host's defenses against metastatic extension. SL granulomas have been reported in approximately 4.4% of carcinomas. Isolated cases of renal cell carcinoma (RCC) with SL granulomas have been reported with questionable prognostic significance. We identified 11 cases of RCCs with SL granulomas. Interestingly, all cases had abundant clear cell cytoplasm (10 clear cell RCC cases and 1 clear cell papillary RCC). We propose that this clear, abundant cytoplasm of the tumor cells with high content of glycogen and lipids may trigger granuloma formation akin to that seen in seminomas with SL granulomas. To date, this is the largest case series of RCCs with SL granulomas.

Results using the OPAL strategy in Mandarin speaking cochlear implant recipients.

OBJECTIVE: To evaluate the effectiveness of an experimental pitch-coding strategy for improving recognition of Mandarin lexical tone in cochlear implant (CI) recipients. DESIGN: Adult CI recipients were tested on recognition of Mandarin tones in quiet and speech-shaped noise at a signal-to-noise ratio of +10 dB; Mandarin sentence speech-reception threshold (SRT) in speech-shaped noise; and pitch discrimination of synthetic complex-harmonic tones in quiet. Two versions of the experimental strategy were examined: (OPAL) linear (1:1) mapping of fundamental frequency (F0) to the coded modulation rate; and (OPAL+) transposed mapping of high F0s to a lower coded rate. Outcomes were compared to results using the clinical ACE™ strategy. STUDY SAMPLE: Five Mandarin speaking users of Nucleus® cochlear implants. RESULTS: A small but significant benefit in recognition of lexical tones was observed using OPAL compared to ACE in noise, but not in quiet, and not for OPAL+ compared to ACE or OPAL in quiet or noise. Sentence SRTs were significantly better using OPAL+ and comparable using OPAL to those using ACE. No differences in pitch discrimination thresholds were observed across strategies. CONCLUSIONS: OPAL can provide benefits to Mandarin lexical tone recognition in moderately noisy conditions and preserve perception of Mandarin sentences in challenging noise conditions.

Nicotinic Acetylcholine Receptors Sensitize a MAPK-linked Toxicity Pathway on Prolonged Exposure to ß-Amyloid.

Among putative downstream synaptic targets of ß-amyloid (Aß) are signaling molecules involved in synaptic function, memory formation and cognition, such as the MAP kinases, MKPs, CaMKII, CREB, Fyn, and Tau. Here, we assessed the activation and interaction of signaling pathways upon prolonged exposure to Aß in model nerve cells expressing nicotinic acetylcholine receptors (nAChRs). Our goal was to characterize the steps underlying sensitization of the nerve cells to neurotoxicity when Aß-target receptors are present. Of particular focus was the connection of the activated signaling molecules to oxidative stress. Differentiated neuroblastoma cells expressing mouse α4ß2-nAChRs were exposed to Aß1-42 for intervals from 30 min to 3 days. The cells and cell-derived protein extracts were then probed for activation of signaling pathway molecules (ERK, JNK, CaMKII, CREB, MARCKS, Fyn, tau). Our results show substantial, progressive activation of ERK in response to nanomolar Aß exposure, starting at the earliest time point. Increased ERK activation was followed by JNK activation as well as an increased expression of PHF-tau, paralleled by increased levels of reactive oxygen species (ROS). The impact of prolonged Aß on the levels of pERK, pJNK, and ROS was attenuated by MEK-selective and JNK-selective inhibitors. In addition, the MEK inhibitor as well as a JNK inhibitor attenuated Aß-induced nuclear fragmentation, which followed the changes in ROS levels. These results demonstrate that the presence of nAChRs sensitizes neurons to the neurotoxic action of Aß through the timed activation of discrete intracellular signaling molecules, suggesting pathways involved in the early stages of Alzheimer disease.

Effects of Threshold Adjustment on Speech Perception in Nucleus Cochlear Implant Recipients.

OBJECTIVE: The objective of this study was to investigate the impact of using smaller and larger electric dynamic ranges on speech perception, aided thresholds, and subjective preference in cochlear implant (CI) subjects with the Nucleus device. DESIGN: Data were collected from 19 adults using the Nucleus CI system. Current levels (CLs) used to set threshold stimulation levels (T-levels) were set above or below the measured hearing thresholds to create smaller or larger electric output dynamic ranges, respectively, whereas the upper stimulation level (C-level) was fixed. The base (unadjusted) condition was compared against two conditions with higher T-levels (compression), by 30% and 60% of the measured hearing dynamic range, and three conditions with lower T-levels (expansion), by 30%, 60%, and 90% of the measured hearing dynamic range. For each subject, the clinical CL units were adjusted on each electrode to achieve these conditions. The slow-acting dynamic acoustic gains of ADRO and Autosensitivity™ were enabled. Consonant-nucleus-consonant (CNC) word scores were measured in quiet at 50 dB and 60 dB SPL presentation levels. The signal-to-noise ratios (SNRs) for 50% understanding of sentences in noise were measured for sentences presented at 55 dB and 65 dB SPL in 4-talker babble noise. Free-field aided thresholds were measured at octave frequencies using frequency-modulated (warble) tones. Thirteen of the 19 subjects had take-home experience with the base and experimental conditions and provided subjective feedback via a questionnaire. RESULTS: There were no significant effects of 30% expansion and 30% compression of the electric dynamic range on scores for words in quiet and SNRs for sentences in noise, at the two presentation levels. There was a significant decrement in scores for words in quiet for 60% and 90% expansion compared with the base condition at the 50 dB and 60 dB SPL presentation levels. The score decrement was much less at 60 dB SPL. For the 50 dB SPL presentation level, the decrements in word scores at 60% and 90% expansion were linearly related to the reduction in CL units required to achieve these experimental conditions, with a greater decrement in scores for a larger CL change. There was a significant increase in SNR for sentences in noise for 60% compression compared with the base condition at the 55 dB and 65 dB SPL presentation levels. There was also a significant increase in SNR for sentences at the 55 dB SPL presentation level for 90% expansion. Aided thresholds were significantly elevated for the three expansion conditions compared with the base condition, although the mean elevation at 30% expansion was only 4 dB. The questionnaire results showed no clear preference for any condition; however, subjects reported a reduced preference for the extreme compression (60%) and expansion (90%) conditions. CONCLUSIONS: The results showed that CI subjects using the Nucleus sound processor had no significant change in performance or preference for adjustments in T-levels by ±30% of the hearing dynamic range. In quiet, speech perception scores were reduced for the more marked expansion (60% and 90%) conditions, whereas in noise, performance was poorer for the highest compression (60%) condition. Across subjects, the decrement in scores for words at 50 dB SPL for the 60% and 90% expansion conditions was related to the changes in CL units required for these conditions, with greater decrements for larger changes in levels.

Impact of sustained exposure to ß-amyloid on calcium homeostasis and neuronal integrity in model nerve cell system expressing α4ß2 nicotinic acetylcholine receptors.

Although the interaction between ß-amyloid (Aß) and nicotinic acetylcholine receptors has been widely studied, the impact of prolonged exposure to Aß on nAChR expression and signaling is not known. In this study, we employed a neuronal culture model to better understand the impact of sustained exposure of Aß on the regulation of cellular and synaptic function. The differentiated rodent neuroblastoma cell line NG108-15 expressing exogenous high-affinity α4ß2 nAChRs was exposed to soluble oligomeric Aß for several days. Ca(2+) responses, expression levels of α4ß2 nAChRs, rate of mitochondrial movement, mitochondrial fission, levels of reactive oxygen species, and nuclear integrity were compared between Aß-treated and untreated cells, transfected or not (mock-transfected) with α4ß2 nAChRs. Sustained exposure of Aß(1-42) to α4ß2 nAChR-transfected cells for several days led to increased Ca(2+) responses on subsequent acute stimulation with Aß(1-42) or nicotine, paralleled by increased expression levels of α4ß2 nAChRs, likely the result of enhanced receptor recycling. The rate of mitochondrial movement was sharply reduced, whereas the mitochondrial fission protein pDrp-1 was increased in α4ß2 nAChR-transfected cells treated with Aß(1-42). In addition, the presence of α4ß2 nAChRs dramatically enhanced Aß(1-42)-mediated increases in reactive oxygen species and nuclear fragmentation, eventually leading to apoptosis. Our data thus show disturbed calcium homeostasis coupled with mitochondrial dysfunction and loss of neuronal integrity on prolonged exposure of Aß in cells transfected with α4ß2 nAChRs. Together, the results suggest that the presence of nAChRs sensitizes neurons to the toxic actions of soluble oligomeric Aß, perhaps contributing to the cholinergic deficit in Alzheimer disease.

The critical role of interleukin-6 in protection against neurotropic flavivirus infection.

West Nile virus (WNV) and Japanese encephalitis virus (JEV) are emerging mosquito-borne flaviviruses causing encephalitis globally. No specific drug or therapy exists to treat flavivirus-induced neurological diseases. The lack of specific therapeutics underscores an urgent need to determine the function of important host factors involved in flavivirus replication and disease progression. Interleukin-6 (IL-6) upregulation has been observed during viral infections in both mice and humans, implying that it may influence the disease outcome significantly. Herein, we investigated the function of IL-6 in the pathogenesis of neurotropic flavivirus infections. First, we examined the role of IL-6 in flavivirus-infected human neuroblastoma cells, SK-N-SH, and found that IL-6 neutralization increased the WNV or JEV replication and inhibited the expression of key cytokines. We further evaluated the role of IL-6 by infecting primary mouse cells derived from IL-6 knockout (IL-6-/-) mice and wild-type (WT) mice with WNV or JEV. The results exhibited increased virus yields in the cells lacking the IL-6 gene. Next, our in vivo approach revealed that IL-6-/- mice had significantly higher morbidity and mortality after subcutaneous infection with the pathogenic WNV NY99 or JEV Nakayama strain compared to WT mice. The non-pathogenic WNV Eg101 strain did not cause mortality in WT mice but resulted in 60% mortality in IL-6-/- mice, indicating that IL-6 is required for the survival of mice after the peripheral inoculation of WNV or JEV. We also observed significantly higher viremia and brain viral load in IL-6-/- mice than in WT mice. Subsequently, we explored innate immune responses in WT and IL-6-/- mice after WNV NY99 infection. Our data demonstrated that the IL-6-/- mice had reduced levels of key cytokines in the serum during early infection but elevated levels of proinflammatory cytokines in the brain later, along with suppressed anti-inflammatory cytokines. In addition, mRNA expression of IFN-α and IFN-ß was significantly lower in the infected IL-6-/- mice. In conclusion, these data suggest that the lack of IL-6 exacerbates WNV or JEV infection in vitro and in vivo by causing an increase in virus replication and dysregulating host immune response.

Role of key aromatic residues in the ligand-binding domain of alpha7 nicotinic receptors in the agonist action of beta-amyloid.

Soluble ß-amyloid (Aß) resides in certain regions of the brain at or near picomolar concentration, rising in level during the prodromic stage of Alzheimer disease. Recently, we identified the homomeric α7 nicotinic acetylcholine receptor (α7-nAChR) as one possible functional target for picomolar Aß. This study was aimed at addressing which residues in α7-nAChRs potentially interact with Aß to regulate the presynaptic function of this receptor. Site-directed mutagenesis was carried out to study the key aromatic residues in the mouse α7-nAChR agonist-binding pocket. Mutations of tyrosine188 resulted in a decrease in activation of presynaptic α7-nAChRs by ACh and Aß but with no change in response to nicotine, indicating the critical role of Tyr-188 in presynaptic regulation by Aß. Coimmunoprecipitation additionally revealed direct binding of Aß to α7-nAChRs and to the Tyr-188 mutant receptor. In contrast, mutations of Tyr-195 in α7-nAChR led to decreased activation by nicotine without apparent effects on ACh- or Aß-induced responses. Agonist-induced responses of Tyr-93 mutant α7-nAChRs indicated possible interactions of nicotine and Aß with its hydroxyl group, but there was no change in presynaptic responses after mutation of Trp-149. All of the mutants were shown to be expressed on the plasma membrane using cell surface labeling. Together, these results directly demonstrate an essential role for the aromatic residue Tyr-188 as a key component in the agonist binding domain for the activation of α7-nAChRs by Aß.

Combining directional microphone and single-channel noise reduction algorithms: a clinical evaluation in difficult listening conditions with cochlear implant users.

OBJECTIVES: This study tested a combination of algorithms designed to improve cochlear implant performance in noise. A noise reduction (NR) algorithm, based on signal to noise ratio estimation was evaluated in combination with several directional microphone algorithms available in the Cochlear CP810 sound processor. DESIGN: Fourteen adult unilateral cochlear implant users participated in the study. Evaluation was conducted using word recognition in quiet, sentence recognition in noise, and subjective feedback via questionnaire after a period of take-home use. Music appreciation was also evaluated in a controlled listening task. The sentence recognition task measured speech reception threshold for 50% morphemes correct. The interfering maskers were speech-weighted noise and competing talkers, which were spatially separated from the target speech. In addition, the locations of the noise maskers changed during the test in an effort to replicate relevant real-world listening conditions. SmartSound directionality settings Standard, Zoom, and Beam (used in the SmartSound programs Everyday, Noise, and Focus, respectively) were all evaluated with and without NR. RESULTS: Microphone directionality demonstrated a consistent benefit in sentence recognition in all noise conditions tested. The group average speech reception threshold benefit over the Standard setting was 3.7 dB for Zoom and 5.3 dB for Beam. Addition of the NR algorithm further improved sentence recognition by 1.3 dB when the noise maskers were speech-weighted noise. There was an overall group preference for the NR algorithm in noisy environments. Group mean word recognition in quiet, preference in quiet conditions, and music appreciation were all unaffected by the NR algorithm. CONCLUSIONS: Multimicrophone directionality was effective in improving speech understanding in spatially separated noisy conditions. The single-channel NR algorithm further enhanced speech intelligibility in speech-weighted noise for cochlear implant users while maintaining equivalent performance in quiet situations and when listening to music.

Differential Pathogenesis of SARS-CoV-2 Variants of Concern in Human ACE2-Expressing Mice.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused the current pandemic, resulting in millions of deaths worldwide. Increasingly contagious variants of concern (VoC) have fueled recurring global infection waves. A major question is the relative severity of the disease caused by previous and currently circulating variants of SARS-CoV-2. In this study, we evaluated the pathogenesis of SARS-CoV-2 variants in human ACE-2-expressing (K18-hACE2) mice. Eight-week-old K18-hACE2 mice were inoculated intranasally with a representative virus from the original B.1 lineage or from the emerging B.1.1.7 (alpha), B.1.351 (beta), B.1.617.2 (delta), or B.1.1.529 (omicron) lineages. We also infected a group of mice with the mouse-adapted SARS-CoV-2 (MA10). Our results demonstrate that B.1.1.7, B.1.351 and B.1.617.2 viruses are significantly more lethal than the B.1 strain in K18-hACE2 mice. Infection with the B.1.1.7, B.1.351, and B.1.617.2 variants resulted in significantly higher virus titers in the lungs and brain of mice compared with the B.1 virus. Interestingly, mice infected with the B.1.1.529 variant exhibited less severe clinical signs and a high survival rate. We found that B.1.1.529 replication was significantly lower in the lungs and brain of infected mice in comparison with other VoC. The transcription levels of cytokines and chemokines in the lungs of B.1- and B.1.1.529-infected mice were significantly less when compared with those challenged with other VoC. Together, our data provide insights into the pathogenesis of previous and circulating SARS-CoV-2 VoC in mice.

SARS-CoV-2 Infects Primary Neurons from Human ACE2 Expressing Mice and Upregulates Genes Involved in the Inflammatory and Necroptotic Pathways.

Transgenic mice expressing human angiotensin-converting enzyme 2 under the cytokeratin 18 promoter (K18-hACE2) have been extensively used to investigate the pathogenesis and tissue tropism of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection. Neuroinvasion and the replication of SARS-CoV-2 within the central nervous system (CNS) of K18-hACE2 mice is associated with increased mortality; although, the mechanisms by which this occurs remain unclear. In this study, we generated primary neuronal cultures from K18-hACE2 mice to investigate the effects of a SARS-CoV-2 infection. We also evaluated the immunological response to SARS-CoV-2 infection in the CNS of K18-hACE2 mice and mouse neuronal cultures. Our data show that neuronal cultures obtained from K18-hACE2 mice are permissive to SARS-CoV-2 infection and support productive virus replication. Furthermore, SARS-CoV-2 infection upregulated the expression of genes involved in innate immunity and inflammation, including IFN-α, ISG-15, CXCL10, CCL2, IL-6 and TNF-α, in the neurons and mouse brains. In addition, we found that SARS-CoV-2 infection of neurons and mouse brains activates the ZBP1/pMLKL-regulated necroptosis pathway. Together, our data provide insights into the neuropathogenesis of SARS-CoV-2 infection in K18-hACE2 mice.