SYSTEMATIC REVIEW PROTOCOL FOR ANIMAL INTERVENTION STUDIES FORMAT BY SYRCLE (WWW.SYRCLE.NL) VERSION 2.0 (DECEMBER 2014) Item Section/Subsection/Item # A. General 1.

Title of the review

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Authors (names, affiliations, contributions)

Description The effect of the intermittent sound conditioning before a traumatic acoustic exposure on the permanent auditory threshold shift and on the loss of hair cells in rodents: a systematic review and meta-analysis Felipe C. S. Veloso1 Kelly C. L. de Andrade1 Danielle S. da S. G. Ferreira1 Gabriella O. Peixoto1 Nassib B. Bueno2 Pedro de L. Menezes1

Check for approval

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1.Hearing and Technology Lab, State University of Health Sciences of Alagoas, Brazil 2.Federal University of Alagoas, Brazil 3. 4. 5. 6. 7. 8. 9.

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Other contributors (names, affiliations, contributions) Contact person + e-mail address Funding sources/sponsors Conflicts of interest Date and location of protocol registration Registration number (if applicable) Stage of review at time of registration B. Objectives Background

Felipe C. S. Veloso ([email protected]) CNPq; UNCISAL We have no conflicts of interest to declare 05-Feb-2016. CAMARADES repository. Preliminary searches

The sound conditioning is still a little-know subject, very complex and very interesting from the point of view of preventive hearing protection. The sound conditioning comprehends a non-traumatic sound exposure before a traumatic acoustic exposure. This non-traumatic sound exposure does not cause injury to auditory system. What is already known about this This sound conditioning can be intermittent, continuous or disease/model/intervention? Why is it toughening. A conditioning can be called intermittent when important to do this review? there is a sound exposure followed by a period of silence. A continuous sound conditioning comprehends a sound exposure without interruptions. Lastly, a conditioning can be called toughening when there is a peak sound exposure followed by a period of silence. Though clearly defined here, there is still many disagreements in the literature

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regarding the definitions of the types of conditioning, encouraging further studies. This review aims to show if the intermittent sound conditioning, one of three types of sound conditioning, serves as a preventive hearing protection in rodents. For this, a meta-analysis will be performed to show, statistically, a positive or negative effect of the sound conditioning. Research question 11.

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The problem to be analysed is the permanent hearing loss, Specify the disease/health problem of in other words, the permanent auditory threshold shift and interest the loss of hair cells after an intermittent sound conditioning before a traumatic acoustic exposure. Specify the population/species Rodents studied The intervention comprehends the presence of the intermittent sound conditioning before a traumatic acoustic exposure. This intermittent sound conditioning consists on an intense sound, more than or equal to 80 dB and less than 100 dB, intermittent, 6 hours per day, during Specify the intervention/exposure 10 days on a narrow or octave band between 0,5 kHz and 16 kHz. The traumatic acoustic exposure consists on a traumatic sound, more than or equal to 100 dB, 24 hours per day, during a time more than or equal to 1 day, on a narrow or octave band between 0,5 kHz and 16 kHz. The control population consists only in a traumatic acoustic exposure. This traumatic acoustic exposure comprehends a Specify the control population traumatic sound, more than or equal to 100 dB, 24 hours per day, during a time more than or equal to 1 day, on a narrow or octave band between 0,5 kHz and 16 kHz. Permanent auditory threshold shift in decibel (dB) and loss Specify the outcome measures of the outer and inner hair cells in percentage (%) What the effect of the intermittent sound conditioning State your research question (based before a traumatic acoustic exposure on the permanent on items 11-15) auditory threshold shift and on the loss of hair cells in rodents? C. Methods Search and study identification [X] MEDLINE via PubMed [X] Web of Science

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[X]SCOPUS EMBASE [X] Other, namely: CENTRAL via Cochrane Library, ELSEVIER Identify literature databases to search via ScienceDirect, LILACS via Bireme, LILACS via SciELO, (e.g. Pubmed, Embase, Web of CiNii, OpenGrey, CiNii Dissertations, Open Access Theses science) and Dissertations, Biblioteca Digital Brasileira de Teses e Dissertações and TripDataBase.

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Define electronic search strategies (e.g. use the step by step search guide15 and animal search filters20, 21)

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□Specific journal(s), namely:

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When available, please add a supplementary file containing your search strategy: [SEARCH STRATEGIES.pdf]

[X] Reference lists of included studies 19.

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Identify other sources for study identification

□Books

□Reference lists of relevant reviews □Conference proceedings, namely: □Contacting authors/ organisations, namely: □Other, namely:

Define search strategy for these other sources Study selection 1-Selection based on title Define screening phases (e.g. pre2-Selection based on abstract screening based on title/abstract, full 3-Selection based on full text text screening, both) 4-Screening references of included studies All phases will be performed by two independent Specify (a) the number of reviewers reviewers. Discrepancies will be resolved by discussion. If per screening phase and (b) how the conflict persists, a third author will be called to assist in discrepancies will be resolved the final decision. Define all inclusion and exclusion criteria based on: Inclusion criteria: Original experimental studies Exclusion criteria: Case-control studies, cohort studies, case Type of study (design) reports, conference proceedings and all types of literature reviews Inclusion criteria: All species of rodents without weight, age Type of animals/population (e.g. age, or gender restriction. gender, disease model) Exclusion criteria: Other species of animals as well as humans. Inclusion criteria: Articles that did report an intermittent sound conditioning that comprehends an intense sound, more than or equal to 80 dB and less than 100 dB, intermittent, 6 hours per day, during 10 days on a narrow or octave band between 0,5 kHz and 16 kHz. Furthermore, after this intermittent sound conditioning, articles must report a traumatic acoustic exposure that consists on a traumatic sound, more than or equal to 100 dB, 24 hours per day, during a time more than or equal to 1 day, on a Type of intervention (e.g. dosage, narrow or octave band between 0,5 kHz and 16 kHz. timing, frequency) Exclusion criteria: Articles that did not report an intermittent sound conditioning that comprehends an intense sound, more than or equal to 80 dB and less than 100 dB, intermittent, 6 hours per day, during 10 days on a narrow or octave band between 0,5 kHz and 16 kHz, nor a traumatic acoustic exposure that consists on a traumatic sound, more than or equal to 100 dB, 24 hours per day, during a time more than or equal to 1 day, on a narrow or octave band between 0,5 kHz and 16 kHz. Inclusion criteria: Articles that did report both the Outcome measures permanent auditory threshold shift nor the percentage of the outer and inner hair cells losses.

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Language restrictions

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Publication date restrictions Other

Exclusion criteria: Articles that did not report both the permanent auditory threshold shift nor the percentage of the outer and inner hair cells losses. Inclusion criteria: Portuguese, English, Spanish, Japanese, Italian, French and Germany articles. Exclusion criteria: Other languages. No publication date restrictions Selection phase: title 1.The article is not an original experimental study. 2.The animal of the study is not a rodent. 3.The article does not report a sound conditioning before a traumatic acoustic exposure. 4.The article does not report a threshold shift or a hair cells losses.

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Selection phase: abstract 1.The article does not report an intermittent sound conditioning that comprehends an intense sound, more than or equal to 80 dB and less than 100 dB, intermittent, 6 hours per day, during 10 days on a narrow or octave band between 0,5 kHz and 16 kHz nor after that, a traumatic sound, more than or equal to 100 dB, 24 hours per day, during a time more than or equal to 1 day, on a narrow or octave band between 0,5 kHz and 16 kHz. 2.The article does not report a threshold shift or a hair cells losses. 30.

Sort and prioritize your exclusion criteria per selection phase

Selection phase: full text 1.The article does not report both the permanent auditory threshold shift nor the percentage of the outer and inner hair cells losses. Selection phase: references of included studies -Title 1.The article is not an original experimental study. 2.The animal of the study is not a rodent. 3.The article does not report a sound conditioning before a traumatic acoustic exposure. 4.The article does not report a threshold shift or a hair cells losses. -Abstract 5.The article does not report an intermittent sound conditioning that comprehends an intense sound, more than or equal to 80 dB and less than 100 dB, intermittent, 6 hours per day, during 10 days on a narrow or octave band between 0,5 kHz and 16 kHz nor after that, a traumatic sound, more than or equal to 100 dB, 24 hours per day, during a time more than or equal to 1 day, on a narrow or octave band between 0,5 kHz and 16 kHz.

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6.The article does not report a threshold shift or a hair cells losses. -Full-text 7.The article does not report both the permanent auditory threshold shift nor the percentage of the outer and inner hair cells losses. Study characteristics to be extracted (for assessment of external validity, reporting quality) Study ID (e.g. authors, year) First author, year and country. Experimental groups (number of animals of each group, intensity of sound exposure for each group, during of the exposure per hours and per days for each group), pre-test exposure (auditory threshold at 500 Hz, 1000 Hz, 2000 Hz, 4000 Hz, 5600 Hz, 8000 Hz and 16000 Hz for each group). Sound Conditioning exposure (intensity of sound exposure for each group, during of the exposure per hours and per Study design characteristics (e.g. days for each group). Post-Sound Conditioning exposure experimental groups, number of recovery time (recovery time for each group). Traumatic animals) acoustic exposure (intensity of sound exposure for each group, during of the exposure per hours and per days for each group). Permanent auditory threshold shift (time after traumatic acoustic exposure, auditory threshold at 500 Hz, 1000 Hz, 2000 Hz, 4000 Hz, 5600 Hz, 8000 Hz and 16000 Hz for each group). Histological findings (anaesthesia, percentage of outer and inner hair cells losses measure from the base to the apex of the cochlea for each group). Animal model characteristics (e.g. Species, total number of animals, pre-test anaesthesia, prespecies, gender, disease induction) test surgery, pre-test antibiotic and pre-test recovery time. Sound Conditioning exposure (intensity of sound exposure for each group, during of the exposure per hours and per days for each group). Post-Sound Conditioning exposure Intervention characteristics (e.g. recovery time (recovery time for each group). Traumatic intervention, timing, duration) acoustic exposure (intensity of sound exposure for each group, during of the exposure per hours and per days for each group). Permanent auditory threshold shift (time after traumatic acoustic exposure, auditory threshold at 500 Hz, 1000 Hz, Outcome measures 2000 Hz, 4000 Hz, 5600 Hz, 8000 Hz and 16000 Hz for each group). Histological findings (anaesthesia, percentage of the outer and inner hair cells losses for each group). Drop-out after pre-test exposure. Drop-out after Sound Other (e.g. drop-outs) Conditioning exposure. Drop-out after traumatic acoustic exposure. Assessment risk of bias (internal validity) or study quality The assessing of the risk of bias in each study will be Specify (a) the number of reviewers performed by two independent reviewers. Discrepancies assessing the risk of bias/study quality will be resolved by discussion. If the conflict persists, a third in each study and (b) how author will be called to assist in the final decision. Before discrepancies will be resolved any incomplete information, the authors will contact three times at maximum the authors of the included studies.

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Define criteria to assess (a) the internal validity of included studies (e.g. selection, performance, detection and attrition bias) and/or (b) other study quality measures (e.g. reporting quality, power)

[X]By use of SYRCLE's Risk of Bias tool4

□By use of SYRCLE’s Risk of Bias tool, adapted as follows: □By use of CAMARADES' study quality checklist, e.g □By use of CAMARADES' study quality checklist, adapted 22

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□Other criteria, namely:

Collection of outcome data

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The data of the permanent auditory threshold shift will be extracted as a continuous measure. The unit of measurement of this first outcome will be decibel. The data of the loss of the outer and inner hair cells will be extracted as percentage that will be as well the unit of measurement of the second outcome. The data extraction will be done, firstly, when available, Methods for data extraction/retrieval obtaining the data from the text or tables. If not available, (e.g. first extraction from graphs using the extraction will be performed using a digital ruler on a digital screen ruler, then contacting graphs. If necessary, the original authors will be contacted authors) to obtain the complete data of the study. The data extraction will be performed by two independent reviewers. Discrepancies will be resolved by Specify (a) the number of reviewers discussion. If the conflict persists, a third author will be extracting data and (b) how called to assist in the final decision. Before any incomplete discrepancies will be resolved information, the authors will contact three times at maximum the authors of the included studies. Data analysis/synthesis Specify (per outcome measure) how The data of the permanent auditory threshold shift and the you are planning to combine/compare data of the percentage of the loss of the outer and inner the data (e.g. descriptive summary, hair cells will be compared through a meta-analysis. meta-analysis) Specify (per outcome measure) how it If two or more studies are included, a meta-analysis will be will be decided whether a metaconducted. analysis will be performed If a meta-analysis seems feasible/sensible, specify (for each outcome measure): The effect measure that will be used on meta-analysis for The effect measure to be used (e.g. the permanent auditory threshold shift and for the mean difference, standardized mean percentage of the loss of the outer and inner hair cells will difference, risk ratio, odds ratio) be the mean difference. The statistical model of analysis that will be used on metaThe statistical model of analysis (e.g. analysis for the permanent auditory threshold shift and for random or fixed effects model) the percentage of the loss of the outer and inner hair cells will be a random effect model. The statistical method to assess hetererogeneity that will The statistical methods to assess be used on the meta-analysis for the permanent auditory heterogeneity (e.g. I2, Q) threshold shift and for the percentage of the loss of the outer and inner hair cells will be the I² test. Which study characteristics will be The study characteristic that will be examined as a potential examined as potential source of source of heterogeneity and will be included on a subgroup heterogeneity (subgroup analysis) analysis will be the species of the rodents. For each outcome measure, define the type of data to be extracted (e.g. continuous/dichotomous, unit of measurement)

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Any sensitivity analyses you propose to perform Other details meta-analysis (e.g. correction for multiple testing, correction for multiple use of control group) The method for assessment of publication bias

Final approval by (names, affiliations):

In case of heterogeneity, the studies will be removed, one at a time, to investigate if any particular study is the sole source of heterogeneity.

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The assessment of publication bias will be performed using a funnel plot.

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Felipe C. S. Veloso1 Kelly C. L. de Andrade1 Danielle S. da S. G. Ferreira1 Gabriella O. Peixoto1 Nassib B. Bueno2 Pedro de L. Menezes1 1.Hearing and Technology Lab, State University of Health Sciences of Alagoas, Brazil 2.Federal University of Alagoas, Brazil

Date: 05-Mar-2016

Protocol from Felipe Veloso (Final Version).pdf

permanent auditory threshold shift nor the percentage of. the outer and inner hair cells losses. X. Page 3 of 7. Protocol from Felipe Veloso (Final Version).pdf.

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