Cochlear implant surgery facilitates intracochlear distribution of perioperative systemic steroids.

BACKGROUND: Systemically administered steroids are widely utilised for hearing preservation therapies. More recently, steroids have been administered to achieve hearing protection after cochlear implant surgery. Currently there is a lack of understanding as to which administration route offers most therapeutic efficacy, local or systemic administration. Paramount to this are observations in animal studies that systemic administration following implantation offers hearing protection and reduced cochlear fibrosis, despite observations that perilymphatic levels are up to 10-fold higher after local administration in non-implanted cochleae. AIMS/OBJECTIVES: This paper explores the impact that cochlear implantation and associated acute inflammation has on steroid distribution and uptake following systemic administration of dexamethasone. MATERIAL AND METHODS: Eight guinea pigs received systemic dexamethasone 60 min prior to cochlear implantation. Implanted and contralateral non-implanted cochlea were harvested for tissue immunohistochemistry and detection of dexamethasone. RESULTS: Cochleostomy with scala tympani implantation resulted in a significant increase in cochlear dexamethasone signal. This was most notable at the organ of Corti, stria vascularis, and blood product in the scala tympani. CONCLUSIONS AND SIGNIFICANCE: This study demonstrates that the inner ear distribution of systemically administered steroids is enhanced following surgery for cochlear implantation and provides rationale for systemic perioperative steroids in hearing preservation surgery.

Spironolactone Ameliorates Cochlear Implant Induced Endolymphatic Hydrops.

BACKGROUND: Endolymphatic hydrops (EH) has been observed in both animal and human cochleae following cochlear implant (CI) surgery. We tested whether EH could be eliminated by administration of mineralocorticoid steroid antagonist spironolactone and explored the electrophysiological consequences of this. METHODS: Sixty-four adult guinea pigs underwent cochlear implantation with a dummy electrode. Animals then survived either 2, 7, or 28 days. Auditory function was monitored by recording electrocochleography from the round window membrane preimplantation, and on the last day of the experiment. Spironolactone or control solution was added to animals' feed for 7 days (if they survived that long) beginning immediately prior to surgery. The presence of EH was determined using thin-sheet laser imaging microscopy. RESULTS: Treatment with spironolactone resulted in significant reduction in EH in the second cochlear turn 7 days postimplantation. In all animals, the compound action potential (CAP) threshold was elevated 2 days postimplantation, but for most frequencies had recovered substantially by 28 days. There was no treatment effect on CAP thresholds. SP/AP ratios were elevated at day 2. The amplitude growth of the CAP did not differ between test and control groups at any time after implantation. CONCLUSIONS: EH can be suppressed by antagonism of mineralocorticoid receptors in the week after cochlear implantation. Reduction in EH did not lead to any change in hearing, and there was no indication of synaptopathy signalled by reduced CAP amplitude at high sound intensities. We found no electrophysiological evidence that EH early after implantation impacts negatively upon preservation of residual hearing.

Four-point impedance as a biomarker for bleeding during cochlear implantation.

Cochlear implantation has successfully restored the perception of hearing for nearly 200 thousand profoundly deaf adults and children. More recently, implant candidature has expanded to include those with considerable natural hearing which, when preserved, provides an improved hearing experience in noisy environments. But more than half of these patients lose this natural hearing soon after implantation. To reduce this burden, biosensing technologies are emerging that provide feedback on the quality of surgery. Here we report clinical findings on a new intra-operative measurement of electrical impedance (4-point impedance) which, when elevated, is associated with high rates of post-operative hearing loss and vestibular dysfunction. In vivo and in vitro data presented suggest that elevated 4-point impedance is likely due to the presence of blood within the cochlea rather than its geometry. Four-point impedance is a new marker for the detection of cochlear injury causing bleeding, that may be incorporated into intraoperative monitoring protocols during CI surgery.

Glucocorticoid for Hearing Preservation After Cochlear Implantation: A Systemic Review and Meta-analysis of Animal Studies.

OBJECTIVES: To conduct systematic review and meta-analyses of preclinical studies describing the efficacy of glucocorticoids administered via different routes for hearing preservation after cochlear implantation. DATA SOURCES: A literature search was performed in PubMed to identify peer-reviewed articles published before December 31, 2017, with no language restrictions. Search components were "Cochlear implant," "Glucocorticoids," and "Hearing preservation." The results were specified for animal studies. STUDY SELECTION: Original studies in which glucocorticoids were administered before or during cochlear implantation in animal models and hearing threshold shifts were measured using auditory brainstem response. DATA EXTRACTION: Quality of included studies was assessed using the SYstematic Review Centre for Laboratory animal Experimentation protocol. Threshold Shift reduction between the "study" and "control" groups at 1-month postimplantation was the parameter used to evaluate hearing preservation. DATA SYNTHESIS: The random-effects models were used to combine the results of selected studies. Separate meta-analyses were performed for drug-eluting electrodes, systemic, and local administration. CONCLUSIONS: Administering either systemic or topical glucocorticosteroids had a significant effect on preserving low and high-frequency hearing. Topical administration was equally effective across a range of concentration levels and provided maximal hearing preservation when applied 120 minutes before implantation. The effect of systemic treatment was achieved with high doses, equivalent to 26 mg of dexamethasone per day in humans. No significant effect was found with the use of drug-eluting electrodes and more studies are needed to characterise the utility and efficacy of this administration method.

Electrical Impedance as a Biomarker for Inner Ear Pathology Following Lateral Wall and Peri-modiolar Cochlear Implantation.

OBJECTIVES/HYPOTHESIS: Spikes in cochlear implant impedance are associated with inner ear pathology after implantation. Here, we correlate these spikes with episodes of hearing loss and/or vertigo, with a comparison between lateral wall and peri-modiolar electrode arrays. METHODS: Seven hundred seventy recipients of Cochlear's slim-straight, lateral wall electrode (CI422), or peri-modiolar (CI512) electrode were investigated for impedance spikes. Impedance fluctuations were defined as a median rise of ≥ 4 kΩ across all intracochlear electrodes from baseline measurements taken 2 weeks after switch-on. Medical records were analyzed from 189 of the 770 patients. RESULTS: The slim straight, lateral wall electrode was found to spike in impedance at a significantly higher rate than the peri-modiolar array (17% vs 12%). The peri-modiolar electrode tended to spike in impedance earlier than the slim-straight electrode. Impedance spikes were found to significantly correlate with medical events (hearing loss, vertigo, or tinnitus). Overall, in the "spike" group, 42 of 75 patients (56%) demonstrated a clinical event during the impedance spike, whereas 26 of 114 patients (22%) of the "non-spike" group had a clinical event. This significant difference existed with both implant types. CONCLUSION: These results demonstrate a small, but significant increase in impedance spikes in lateral wall electrodes, and support the relationship between spikes in cochlear implant impedances and postoperative inner-ear events, including the loss of residual hearing and vertigo. Monitoring cochlear implant impedance may be a method for early detection, and so the prevention, of these events in the future.

Intracochlear tPA infusion may reduce fibrosis caused by cochlear implantation surgery.

BACKGROUND: Experiments show that the extent of ongoing fibrotic change within the cochlea can be determined by the volume and pattern of bleeding within the first 24 h following cochlear implantation. Tissue-type plasminogen activator (tPA) is effective at reducing thrombus volume when administered both within and external to the systemic circulation. AIMS/OBJECTIVES: To determine if tPA delivered into the scala tympani immediately following implantation will reduce thrombus volume within the lower basal turn of the cochlea. MATERIALS AND METHODS: Guinea pigs were implanted with either 'soft' or 'hard' arrays and administered tPA or saline via an intra-cochlear infusion immediately after implantation. Hearing was checked prior to, and 2 weeks after implantation. Cochleae were then harvested and imaged. RESULTS: Animals implanted with 'soft' arrays had 4.2% less tissue response compared with animals implanted with 'hard' arrays. In animals receiving 'soft' arrays, tPA reduced the volume of tissue response (measured by the percentage of the lower basal turn of the scala tympani occupied by tissue response) compared with saline. CONCLUSIONS AND SIGNIFICANCE: tPA may be effective in reducing the overall volume of tissue response in routine 'soft' cochlear implantation and may have a greater effect in the event of significant surgical trauma.

Adjuvant agents enhance round window membrane permeability to dexamethasone and modulate basal to apical cochlear gradients.

Glucocorticoids have direct anti-inflammatory, anti-oxidant and anti-apoptotic effects on cochlear hair cells. Cochlear glucocorticoid therapy has gained particular attention for its ability to enhance the protection of residual hearing following hearing preservation cochlear implantation. Local drug delivery methods achieve high drug concentrations within the inner ear fluids but are reliant upon diffusion across the round window membrane. Diffusion has been shown to demonstrate large individual variability. This study explores the role of "adjuvant agents", which when administered with glucocorticoids, enhance inner ear absorption and distribution. Guinea pig cochleae were administered either dexamethasone alone or in combination with hyaluronic acid, histamine, or combination histamine and hyaluronic acid, targeted at the round window membrane. Control subjects received saline. Perilymph was sampled from the cochlear apex, and basal to apical dexamethasone concentrations recorded with mass spectroscopy. Cochleae were harvested, and immunohistochemistry employed to explore dexamethasone tissue penetration and distribution. Basal to apical gradients were observed along the scala tympani, with higher dexamethasone concentrations observed at the cochlear base. Gradients were more pronounced and uniform when administered on a hyaluronic acid sponge, while histamine increased absolute concentrations reaching the inner ear. Tissue penetration correlated with perilymph concentration. Our results demonstrate that adjuvant agents can be employed to enhance dexamethasone absorption and distribution in the inner ear, thus proposing therapeutic strategies that may enhance steroid facilitated hearing protection.

A comparison of cochlear distribution and glucocorticoid receptor activation in local and systemic dexamethasone drug delivery regimes.

Local and systemically delivered glucocorticoids are commonly administered to protect the cochlea against damage associated with a variety of insults. There is reason to believe that dexamethasone administered by these routes may arrive at cochlear target sites via different pathways. Clinically, there is a lack of clarity as to which route is more effective in any specific circumstance. This study explores dexamethasone distribution within the guinea pig cochlea following local and systemic delivery methods. A combination of mass spectroscopy and immunohistochemistry were employed to compare both perilymph distribution, tissue uptake and receptor activation. Local administration of dexamethasone to the round window membrane resulted in greater perilymph concentrations, with a basal to apical gradient that favours the cochlear base. Tissue immunofluorescence was intimately related to perilymph concentration following local administration. Systemic administration resulted in much lower perilymph concentrations, with an inverse basal to apical gradient favouring the cochlear apex. Lower perilymph concentrations following systemic administration were associated with minimal tissue immunofluorescence. Despite this, GR activation of the SGNs was equivalent in both administration regimes. These results bring into question the efficacy of measuring perilymph concentrations alone as a surrogacy for dexamethasone distribution and activity in the cochlea, suggesting that the steroid ligand may arrive at its target receptor via alternative pathways. Our results suggest an equivalence in efficacy between local and systemic administration routes early after drug delivery, when the ultimate outcome of GR activation is the goal.

Electrode Impedance Fluctuations as a Biomarker for Inner Ear Pathology After Cochlear Implantation.

OBJECTIVES/HYPOTHESIS: Cochlear implant surgery now aims to preserve residual low frequency hearing. The current research explores whether fluctuations in the electrical impedance of cochlear implant electrodes may act as a biomarker for pathological changes that lead to the delayed loss of residual hearing. STUDY DESIGN: Secondary analysis of a double-blinded randomized trial, where methylprednisolone was administered intravenously before cochlear implantation with a view to preserving residual hearing. METHODS: Seventy-four patients with residual hearing after cochlear implant surgery were investigated for an impedance "spike," defined as a median rise of ≥4 kΩ across all electrodes from the baseline measurements. Spikes were related to objective and subjective hearing loss, dizziness, and tinnitus. RESULTS: An impedance spike occurred in 14% (10/74) of enrolled patients. Three months after surgery, five patients exhibited spikes and three of these patients had a total loss of their residual hearing. 4.3% of the 69 patients without spikes lost residual hearing. At 1 year, 9 of 10 patients who exhibited spikes had lost all their residual hearing. 8.1% of the 37 patients who did not experience a spike lost their residual hearing. Seventy percent of patients exhibiting a spike also experienced vertigo. The administration of steroids at the time of surgery did not influence the occurrence of spikes. CONCLUSION: Our results suggest that there is a relationship between a spike and the loss of residual hearing. It seems that rises in impedance can reflect pathology within the inner ear and predict the future loss of residual hearing.

The Role of Preoperative Steroids in Atraumatic Cochlear Implantation Surgery.

HYPOTHESIS: Depth of insertion is related to the extent of tissue response and low frequency hearing loss. Intravenous steroids have greatest effect in reducing postimplantation fibrosis and hearing loss in the presence of significant electrode insertion trauma, when compared with saline treatment. BACKGROUND: Experiments exploring the enhancement of cochlear implantation (CI) outcomes with glucocorticosteroids have produced mixed results, possibly due to lack of standardization of the CI model. METHODS: Forty-eight normal-hearing guinea pigs were randomly implanted with a highly flexible electrode to a depth of 1.5, 3.0, or 5.0 mm. For each insertion depth, sub-cohorts received either intravenous saline ("saline") or dexamethasone ("steroid") 60 minutes before implantation. Shifts in electrocochleography thresholds at 2 to 32 kHz were determined before and 4 weeks after implantation. Cochleae were harvested and imaged. RESULTS: Low-frequency hearing loss was greatest with 5.0 mm insertions. Fracture of the osseous spiral lamina and/or fibrotic involvement of the round window membrane exacerbated hearing loss. The extent of intracochlear fibrosis was directly related to the depth of insertion. Steroids reduced the intracochlear tissue response for deepest insertions and in apical regions of the cochlea where basilar membrane contact was prevalent. Steroids preserved no more hearing than saline at all insertion depths. CONCLUSION: Cochlear trauma influenced postimplantation hearing loss and steroid effect on fibrosis. Fibrosis, and to a lesser extent, postimplantation hearing loss increased proportionally to the depth of insertion. Steroids did not influence fibrosis relating to the cochleostomy, but could reduce scarring as the electrode negotiated the hook region or near the electrode tip.

Defining the Hook Region Anatomy of the Guinea Pig Cochlea for Modeling of Inner Ear Surgery.

HYPOTHESIS: The aim of this study was to describe the hook region anatomy of the guinea pig cochlea to identify the optimal surgical approach for cochlear implantation and to determine what anatomical structures are at risk. BACKGROUND: Animal studies investigating hearing loss after cochlear implantation surgery are currently constrained by the lack of a reproducible implantation model. METHODS: Guinea pig cochleae were imaged using thin-sheet laser imaging microscopy. Images were stitched, reconstructed, and segmented for analysis. Insertion vectors were determined by tracing their paths to the outer wall and converting to Cartesian coordinates. Spherical surface and multiplane views were generated to analyze outer wall and radial forces of the insertion vector. RESULTS: Thin-sheet laser imaging microscopy enabled quantitative, whole specimen analysis of the soft and bony tissue relationships of the complex cochlear hook region in any desired plane without loss of image quality. Round window or cochleostomy approaches in the anteroinferior plane avoided direct damage to cochlear structures. Cochleostomy approach had large interindividual variability of angular depth and outer wall forces but predictable radial force. CONCLUSION: The guinea pig hook region and lower basal turn have similar structural relationships to humans. Careful cochleostomy placement is essentially for minimizing cochlear trauma and for ensuring a straight insertion vector that successfully advances around the outer wall. Experiments with guinea pigs that control for the surgical approach are likely to provide useful insights into the aetiology and the development of therapies directed at postimplantation hearing loss.

The Effect of Systemic Steroid on Hearing Preservation After Cochlear Implantation via Round Window Approach: A Guinea Pig Model.

HYPOTHESIS: When administered perioperatively, systemic dexamethasone will reduce the hearing loss associated with cochlear implantation (CI) performed via the round window approach. BACKGROUND: The benefits of electroacoustic stimulation have led to interest in pharmacological interventions to preserve hearing after CI. METHODS: Thirty guinea pigs were randomly divided into three experimental groups: a control group; a 3-day infusion group; and a 7-day infusion group. Dexamethasone was delivered via a mini-osmotic pump for either 3 or 7 days after CI via the round window. Pure tone-evoked auditory brainstem response (ABR) thresholds were monitored for a period of 12 weeks after CI. The cochleae were then collected for histology. RESULTS: At 4 and 12 weeks after CI, ABR threshold shifts were significantly reduced in both 7-day and 3-day infusion groups compared with the control group. Furthermore, the 7-day infusion group has significantly reduced ABR threshold shifts compared with the 3-day infusion group. The total tissue response, including fibrosis and ossification, was significantly reduced in the 7-day infusion group compared with the control group. On multiple regression the extent of fibrosis predicted hearing loss across most frequencies, while hair cell counts predicted ABR thresholds at 32 kHz. CONCLUSION: Hearing protection after systemic administration of steroids is more effective when continued for at least a week after CI. Similarly, this treatment approach was more effective in reducing the fibrosis that encapsulates the CI electrode. Reduced fibrosis seemed to be the most likely explanation for the hearing protection.

The Effect of Round Window Sealants on Delayed Hearing Loss in a Guinea Pig Model of Cochlear Implantation.

AIM: To determine whether the type of material used to seal the cochlea after round window cochlear implantation influences delayed hearing loss. BACKGROUND: Cochlear implants are now prescribed to patients with residual, low-frequency hearing. This hearing-which provides perceptual benefits for the implanted ear-is frequently lost for unknown reasons weeks to months after surgery in a proportion of patients. A post-surgical change in cochlear mechanics, related to the material used to seal the cochlea after round window implantation, may contribute to this loss. METHODS: An electrode array was implanted in guinea pigs via the round window, which was then sealed with muscle, periosteum, or fibrin glue. Auditory brainstem responses (ABRs) to pure tones (2, 8, 16, 24, and 32 kHz) were recorded before surgery and 1, 4, and 12 weeks after surgery, with subjects then euthanized and their cochleae harvested for histological analysis. RESULTS: Muscle and periosteum, but not fibrin glue, exhibited delayed threshold rises at 2 kHz. Twelve weeks after implantation, 2 kHz threshold shifts differed significantly between muscle (mean, 27.1 dB) and fibrin glue (9.3 dB), but not between these groups and periosteum (19.3 dB). Muscle was sometimes associated with much greater tissue reactions than the other sealants. Most cochleae had injuries to the basilar membrane and/or osseous spiral lamina, regardless of sealant. Hair cell counts did not differ significantly among sealants. CONCLUSION: Delayed, low-frequency hearing loss was observed when cochleae were sealed with muscle or periosteum, but not when cochleae were sealed with fibrin glue.

Extended use of systemic steroid is beneficial in preserving hearing in guinea pigs after cochlear implant.

CONCLUSION: Seven-day administration of systemic steroids was more effective in preserving hearing for 12 weeks after cochlear implantation (CI) than a 3-day delivery. OBJECTIVES: To determine the effectiveness of extended delivery of systemic steroids to preserve hearing in guinea pigs after CI. METHODS: Dexamethasone (4 mg/ml) was delivered parenterally via a mini-osmotic pump for either 3 or 7 days. A dummy CI electrode was inserted via cochleostomy approach in 8-week-old guinea pigs. Auditory thresholds were assessed from tone burst auditory brainstem responses (2, 8, 16, 24, and 32 kHz) at 1 day prior to CI, and 1, 4, and 12 weeks after implantation. Histologic evaluation of the cochleae was carried out. RESULTS: No differences were observed in hearing thresholds among groups before CI. Significant hearing preservation was achieved at 8, 16, 24, and 32 kHz only in the 7-day infusion group compared with the control group at 1 week after CI. The same trend was maintained at 4 weeks (16, 24 kHz) and 12 weeks (16, 24, and 32 kHz). Histologic review of the 7-day infusion group revealed less fibrosis and ossification in the scala tympani and the preservation of more spiral ganglion cells, compared with the control group.

Delayed low frequency hearing loss caused by cochlear implantation interventions via the round window but not cochleostomy.

Cochlear implant recipients show improved speech perception and music appreciation when residual acoustic hearing is combined with the cochlear implant. However, up to one third of patients lose their pre-operative residual hearing weeks to months after implantation, for reasons that are not well understood. This study tested whether this "delayed" hearing loss was influenced by the route of electrode array insertion and/or position of the electrode array within scala tympani in a guinea pig model of cochlear implantation. Five treatment groups were monitored over 12 weeks: (1) round window implant; (2) round window incised with no implant; (3) cochleostomy with medially-oriented implant; (4) cochleostomy with laterally-oriented implant; and (5) cochleostomy with no implant. Hearing was measured at selected time points by the auditory brainstem response. Cochlear condition was assessed histologically, with cochleae three-dimensionally reconstructed to plot electrode paths and estimate tissue response. Electrode array trajectories matched their intended paths. Arrays inserted via the round window were situated nearer to the basilar membrane and organ of Corti over the majority of their intrascalar path compared with arrays inserted via cochleostomy. Round window interventions exhibited delayed, low frequency hearing loss that was not seen after cochleostomy. This hearing loss appeared unrelated to the extent of tissue reaction or injury within scala tympani, although round window insertion was histologically the most traumatic mode of implantation. We speculate that delayed hearing loss was related not to the electrode position as postulated, but rather to the muscle graft used to seal the round window post-intervention, by altering cochlear mechanics via round window fibrosis.

Effect of both local and systemically administered dexamethasone on long-term hearing and tissue response in a Guinea pig model of cochlear implantation.

Dexamethasone administered prior to cochlear implantation has been shown to reduce the loss of residual hearing in experimental settings. However, its effect on the tissue response around the implant has not been extensively studied. In this study dexamethasone sodium phosphate was administered to guinea pigs via local delivery to the round window (2% dexamethasone for 120 min prior to surgery, 'local 2/120', or 20% dexamethasone for 30 min prior to surgery) or intravenously (2 mg/kg dexamethasone for 60 min) prior to implantation. Auditory brainstem responses (ABR) were monitored for 3 months, after which the cochleae were embedded in Spurr's resin and sectioned. The extent of the tissue response and the survival of the neurosensory structures were analysed. Both local 2/120 and systemically delivered dexamethasone improved ABR thresholds when compared with control animals. Systemic dexamethasone also reduced the tissue response around the electrode. This suggests that whilst both locally and systemically administered dexamethasone can protect residual hearing after cochlear implantation, their effects upon the tissue response to implantation may differ.

Early cochlear response and ICAM-1 expression to cochlear implantation.

AIM: To examine the early cochlear response and intercellular cell adhesion molecule-1 (ICAM-1) expression to implantation of a cochlear electrode into the scala tympani. BACKGROUND: Understanding the early response of the cochlea to implantation may inform the duration which drug therapies should be delivered to protect hearing. METHODS: Guinea pigs were implanted with a cochlear electrode and survived 1, 2, or 7 days before they were euthanized, cochleae harvested, processed, and cryosectioned for light microscopy or ICAM-1 immunohistochemistry. RESULTS: On hematoxylin and eosin staining, scala tympani was characterized by the presence of fibrin and blood clot at 1 to 2 days after surgery, with a leukocytic infiltrate, primarily of neutrophils and macrophage-like cells. By 7 days after surgery, fibroblasts had infiltrated the clot, and the numbers of red blood cells (RBCs) and neutrophils had diminished. ICAM-1 expression was greatest in the lateral cochlear wall with highest expression found in the basal turn in the region of the electrode at 24 hours postimplantation. CONCLUSION: The cochlear vasculature is maximally primed to recruit cells from the circulation, as evidenced by ICAM-1 expression levels, at 24 hours after cochlear implantation. This response is similar to that seen after other types of injury. Where cochlear implantation differs is the predominance of fibrin and clot early after electrode insertion before infiltration by fibroblasts by the end of the first postoperative week. These results suggest that anti-inflammatory drugs aimed at reducing the extravasation of immunecompetent cells into the cochlea must be effective over the first few days after surgery. Whether this can be achieved through preoperative treatment alone, or whether therapy will need to continue postoperatively, awaits further experimentation.

How should we best estimate the mean recency duration for the BED method?

BED estimates of HIV incidence from cross-sectional surveys are obtained by restricting, to fixed time T, the period over which incidence is estimated. The appropriate mean recency duration (Ω(T)) then refers to the time where BED optical density (OD) is less than a pre-set cut-off C, given the patient has been HIV positive for at most time T. Five methods, tested using data for postpartum women in Zimbabwe, provided similar estimates of Ω(T) for C = 0.8: i) The ratio (r/s) of the number of BED-recent infections to all seroconversions over T = 365 days: 192 days [95% CI 168-216]. ii) Linear mixed modeling (LMM): 191 days [95% CI 174-208]. iii) Non-linear mixed modeling (NLMM): 196 days [95% CrI 188-204]. iv) Survival analysis (SA): 192 days [95% CI 168-216]. Graphical analysis: 193 days. NLMM estimates of Ω(T)--based on a biologically more appropriate functional relationship than LMM--resulted in best fits to OD data, the smallest variance in estimates of VT, and best correspondence between BED and follow-up estimates of HIV incidence, for the same subjects over the same time period. SA and NLMM produced very similar estimates of Ω(T) but the coefficient of variation of the former was .3 times as high. The r/s method requires uniformly distributed seroconversion events but is useful if data are available only from a single follow-up. The graphical method produces the most variable results, involves unsound methodology and should not be used to provide estimates of Ω(T). False-recent rates increased as a quadratic function of C: for incidence estimation C should thus be chosen as small as possible, consistent with an adequate resultant number of recent cases, and accurate estimation of Ω(T). Inaccuracies in the estimation of Ω(T) should not now provide an impediment to incidence estimation.

An enzyme-initiated Smiles rearrangement enables the development of an assay of MshB, the GlcNAc-Ins deacetylase of mycothiol biosynthesis.

MshB is the N-acetyl-1-D-myo-inosityl-2-amino-2-deoxy-D-glucopyranoside (GlcNAc-Ins) deacetylase active as one of the enzymes involved in the biosynthesis of mycothiol (MSH), a protective low molecular weight thiol present only in Mycobacterium tuberculosis and other actinomycetes. In this study, structural analogues of GlcNAc-Ins in which the inosityl moiety is replaced by a chromophore were synthesized and evaluated as alternate substrates of MshB, with the goal of identifying a compound that would be useful in high-throughput assays of the enzyme. In an unexpected and surprising finding one of the GlcNAc-Ins analogues is shown to undergo a Smiles rearrangement upon MshB-mediated deacetylation, uncovering a free thiol group. We demonstrate that this chemistry can be exploited for the development of the first continuous assay of MshB activity based on the detection of thiol formation by DTNB (Ellman's reagent); such an assay should be ideally suited for the identification of MshB inhibitors by means of high-throughput screens in microplates.

Systemic immunity influences hearing preservation in cochlear implantation.

HYPOTHESIS: To determine whether a systemic immune response influences hearing thresholds and tissue response after cochlear implantation of hearing guinea pigs. METHODS: Guinea pigs were inoculated with sterile antigen (Keyhole limpet hemocyanin) 3 weeks before cochlear implantation. Pure-tone auditory brainstem response thresholds were performed before implantation and 1 and 4 weeks later. Dexamethasone phosphate 20% was adsorbed onto a hyaluronic acid carboxymethylcellulose sponge and was applied to the round window for 30 minutes before electrode insertion. Normal saline was used for controls. Cochlear histology was performed at 4 weeks after implantation to assess the tissue response to implantation. To control for the effect of keyhole limpet hemocyanin priming, a group of unprimed animals underwent cochlear implantation with a saline-soaked pledget applied to the round window. RESULTS: Keyhole limpet hemocyanin priming had no significant detrimental effect on thresholds without implantation. Thresholds were elevated after implantation across all frequencies tested (2-32 kHz) in primed animals but only at higher frequencies (4-32 kHz) in unprimed controls. In primed animals, dexamethasone treatment significantly reduced threshold shifts at 2 and 8 kHz. Keyhole limpet hemocyanin led to the more frequent observation of lymphocytes in the tissue response to the implant. CONCLUSION: Systemic immune activation at the time of cochlear implantation broadened the range of frequencies experiencing elevated thresholds after implantation. Local dexamethasone provides partial protection against this hearing loss, but the degree and extent of protection are less compared to previous studies with unprimed animals.