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Articles by Pamela C. Roehm in JoVE

 JoVE Immunology and Infection

A Primary Neuron Culture System for the Study of Herpes Simplex Virus Latency and Reactivation


JoVE 3823 4/02/2012

1Department of Microbiology, New York University School of Medicine, 2Molecular Neurobiology Program, Skirball Institute for Biomolecular Medicine, New York University School of Medicine, 3Department of Otolaryngology, New York University School of Medicine, 4Department of Cell Biology, New York University School of Medicine, 5Department of Physiology and Neuroscience, New York University School of Medicine, 6Department of Psychiatry, New York University School of Medicine, 7Center for Neural Science, New York University School of Medicine

The protocol describes an efficient and reproducible model system to study herpes simplex virus type 1 (HSV-1) latency and reactivation. The assay employs homogenous sympathetic neuron cultures and allows for the molecular dissection of virus-neuron interactions using a variety of tools including RNA interference and expression of recombinant proteins.

Other articles by Pamela C. Roehm on PubMed

Adenosquamous Carcinoma of the Upper Aerodigestive Tract: a Clinicopathologic Study of 12 Cases and Review of the Literature

Adenosquamous carcinoma is an uncommon, controversial neoplasm. To further comprehend its natural history, the clinical and pathological features of 12 new cases were reviewed and analyzed collectively with those described in the English literature.

Dynamic Voice Assessment Using Flexible Laryngoscopy--how I Do It: a Targeted Problem and Its Solution

Strategies to Preserve or Regenerate Spiral Ganglion Neurons

Degeneration of spiral ganglion neurons following hair cell loss carries critical implications for efforts to rehabilitate severe cases of hearing loss with cochlear implants or hair cell regeneration. This review considers recently identified neurotrophic factors and therapeutic strategies which promote spiral ganglion neuron survival and neurite growth. Replacement of these factors may help preserve or regenerate the auditory nerve in patients with extensive hair cell loss.

Constitutive Neuregulin-1/ErbB Signaling Contributes to Human Vestibular Schwannoma Proliferation

Vestibular schwannomas (VSs) are benign tumors that arise from the Schwann cells (SCs) lining the vestibular nerve. VS cells survive and proliferate far from neurons and axonally derived growth factors. We have previously shown that VSs produce the glial growth factor, neuregulin-1 (NRG1), and its receptors, ErbB2 and ErbB3. In the present work, we explore the contribution of constitutive NRG1:ErbB signaling to human VS cell proliferation. We confirm that human VSs, which express markers of immature and denervated SCs, also express endogenous NRG1 and activated ErbB2. We find that a blocking anti-NRG1 antibody and trastuzumab (Herceptin, HCN), a humanized anti-ErbB2 inhibitory monoclonal antibody, effectively inhibit NRG1 induced SC proliferation. Treatment of primary VS cultures with anti-NRG1 or HCN reduces cell proliferation in the absence of exogenous NRG1. Furthermore, conditioned medium from VS cell cultures contains NRG1 and stimulates SC proliferation in SC cultures, an effect that is inhibited by anti-NRG1 and HCN. These data suggest an autocrine pathway of VS growth stimulation involving NRG and ErbB receptors. Inhibition of constitutive NRG:ErbB signaling reduces VS cell proliferation in vitro and may have therapeutic potential for patients with VSs.

Cochlear Implant Explantation As a Sequela of Severe Chronic Otitis Media: Case Report and Review of the Literature

In the 1980s, intracranial and inner ear infections were feared complications in patients with recurrent or chronic otitis media (COM) who had undergone cochlear implantation. Current studies show a low incidence of such complications. We present a case of a patient who developed severe COM requiring cochlear explantation.

Large Extradural Epidermoid Tumor of the Temporal Bone and Posterior Fossa Cranium

Disseminated Histoplasmosis Presenting As a Unilateral Cranial Nerve VIII Mass: a Case Report

To report a unique presentation of disseminated histoplasmosis.

Overexpression of Bcl-2 or Bcl-xL Prevents Spiral Ganglion Neuron Death and Inhibits Neurite Growth

Spiral ganglion neurons (SGNs) provide afferent innervation to the cochlea and rely on contact with hair cells (HCs) for their survival. Following deafferentation due to hair cell loss, SGNs gradually die. In a rat culture model, we explored the ability of prosurvival members of the Bcl-2 family of proteins to support the survival and neurite outgrowth of SGNs. We found that overexpression of either Bcl-2 or Bcl-xL significantly increases SGN survival in the absence of neurotrophic factors, establishing that the Bcl-2 pathway is sufficient for SGN cell survival and that SGN deprived of trophic support die by an apoptotic mechanism. However, in contrast to observations in central neurons and PC12 cells where Bcl-2 appears to promote neurite growth, both Bcl-2 and Bcl-xL overexpression dramatically inhibit neurite outgrowth in SGNs. This inhibition of neurite growth by Bcl-2 occurs in nearly all SGNs even in the presence of multiple neurotrophic factors implying that Bcl-2 directly inhibits neurite growth rather than simply rescuing a subpopulation of neurons incapable of extending neurites without additional stimuli. Thus, although overexpression of prosurvival members of the Bcl-2 family prevents SGN loss following trophic factor deprivation, the inhibition of neurite growth by these molecules may limit their efficacy for support of auditory nerve maintenance or regeneration following hair cell loss.

Management of Acoustic Neuromas in Patients 65 Years or Older

OBJECTIVE:: To analyze an optimal management protocol for patients 65 years or older at the time of acoustic neuroma diagnosis. STUDY DESIGN:: Retrospective case review. SETTING:: Tertiary care hospital. PATIENTS:: Two hundred sixteen patients with acoustic neuroma 65 years or older at time of diagnosis. INTERVENTION:: Patients with smaller tumors (<2.5 cm) were followed with serial magnetic resonance imaging. If significant growth occurred, they were treated with surgery. Surgery was performed at initial diagnosis on patients with larger tumors or in selected patients for hearing preservation. Stereotactic radiotherapy was performed for poor surgical candidates and for patient choice. OUTCOME MEASURES:: Measurement of acoustic neuroma growth and tabulation of complications. RESULTS:: One hundred fourteen patients were initially managed by observation, 80 with surgery, and 3 with radiation therapy, with an average follow-up of 35.4 months. For patients in the observation group, average tumor growth was 1.2 mm/yr. Thirty-two patients required crossover to surgery or radiotherapy due to tumor growth (average growth, 4.1 versus 0.3 mm/yr for those remaining in the observation group). One of the patients in the observation group had a complication (0.9%). CONCLUSION:: Management of acoustic neuromas in elderly patients can be based on size and "biological age" criteria. Surgical treatment can safely be reserved for the few patients who have significant tumor growth.

Management of Acoustic Neuromas in Patients 65 Years or Older

To analyze an optimal management protocol for patients 65 years or older at the time of acoustic neuroma diagnosis.

Membrane Depolarization Inhibits Spiral Ganglion Neurite Growth Via Activation of Multiple Types of Voltage Sensitive Calcium Channels and Calpain

The effect of membrane electrical activity on spiral ganglion neuron (SGN) neurite growth remains unknown despite its relevance to cochlear implant technology. We demonstrate that membrane depolarization delays the initial formation and inhibits the subsequent extension of cultured SGN neurites. This inhibition depends directly on the level of depolarization with higher levels of depolarization causing retraction of existing neurites. Cultured SGNs express subunits for L-type, N-type, and P/Q type voltage-gated calcium channels (VGCCs) and removal of extracellular Ca(2+) or treatment with a combination of L-type, N-type, and P/Q-type VGCC antagonists rescues SGN neurite growth under depolarizing conditions. By measuring the fluorescence intensity of SGNs loaded with the fluorogenic calpain substrate t-butoxy carbonyl-Leu-Met-chloromethylaminocoumarin (20 microM), we demonstrate that depolarization activates calpains. Calpeptin (15 microM), a calpain inhibitor, prevents calpain activation by depolarization and rescues neurite growth in depolarized SGNs suggesting that calpain activation contributes to the inhibition of neurite growth by depolarization.

Petrous Apex Mucocele

Pathology Quiz Case 2. Pneumocystis Carinii (also Known As Pneumocystis Jiroveci) Infection of the Right Temporal Bone

Actinomycosis of the Temporal Bone and Brain: Case Report and Review of the Literature

Actinomycosis is a rare disease with a typically indolent course in the head and neck. During the modern era, only 12 cases within the ear and temporal bone and 75 intracranial cases have been reported. We present a case of actinomycosis of the petrous apex that led to meningitis and encephalitis.

The Effects of Cochlear Implantation on Speech Perception in Older Adults

To determine whether significant differences in cochlear implant (CI) performance exist between older and younger CI recipients.

Sudden Sensorineural Hearing Loss: a Review of Diagnosis, Treatment, and Prognosis

Sudden sensorineural hearing loss (SSNHL) is commonly encountered in audiologic and otolaryngologic practice. SSNHL is most commonly defined as sensorineural hearing loss of 30dB or greater over at least three contiguous audiometric frequencies occurring within a 72-hr period. Although the differential for SSNHL is vast, for the majority of patients an etiologic factor is not identified. Treatment for SSNHL of known etiology is directed toward that agent, with poor hearing outcomes characteristic for discoverable etiologies that cause inner ear hair cell loss. Steroid therapy is the current mainstay of treatment of idiopathic SSNHL in the United States. The prognosis for hearing recovery for idiopathic SSNHL is dependent on a number of factors including the severity of hearing loss, age, presence of vertigo, and shape of the audiogram.

Auditory Rehabilitation of Patients with Neurofibromatosis Type 2 by Using Cochlear Implants

The aim of this study was to determine whether patients with neurofibromatosis Type 2 (NF2) who have intact ipsilateral cochlear nerves can have open-set speech discrimination following cochlear implantation.

Cultured Vestibular Ganglion Neurons Demonstrate Latent HSV1 Reactivation

Vestibular neuritis is a common cause of both acute and chronic vestibular dysfunction. Multiple pathologies have been hypothesized to be the causative agent of vestibular neuritis; however, whether herpes simplex type I (HSV1) reactivation occurs within the vestibular ganglion has not been demonstrated previously by experimental evidence. We developed an in vitro system to study HSV1 infection of vestibular ganglion neurons (VGNs) using a cell culture model system.

Does Addition of Antiviral Medication to High-dose Corticosteroid Therapy Improve Hearing Recovery Following Idiopathic Sudden Sensorineural Hearing Loss?

Glossal Abscess As a Complication of Tongue-base Suspension Surgery

Approximately 60 cases of tongue abscess have been reported in the English-language literature over the past 30 years. We report what we believe is the first case of a glossal abscess that arose as a complication of tongue-base suspension surgery. The patient was a 31-year-old man who presented with a several-day history of odynophagia, tongue swelling, voice changes, and increased snoring. Two years earlier, he had undergone a tongue-base suspension procedure for the treatment of obstructive sleep apnea. Computed tomography (CT) revealed a tongue abscess. During peroral incision and drainage, a knotted 0 Prolene suture was discovered within the abscess cavity. The suture was removed, the area was thoroughly irrigated, the drain was placed in the abscess cavity, and the incision was loosely closed. On postoperative day 5, repeat CT revealed resolution of the abscess, and the patient was discharged on oral antibiotics. Although glossal abscess is very rare, physicians should consider it in the differential diagnosis of any patient who presents with lingual swelling following tongue-base suspension surgery.

A Cell Culture Model of Facial Palsy Resulting from Reactivation of Latent Herpes Simplex Type 1

Reactivation of herpes simplex virus type 1 (HSV-1) in geniculate ganglion neurons (GGNs) is an etiologic mechanism of Bell's palsy (BP) and delayed facial palsy (DFP) after otologic surgery.

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