Understanding the Basics of Peer Review: Part 1 – Receiving a Manuscript

1. This is a the first of a two part on-line tutorial for new student reviewers for IMPULSE. IMPULSE is an on-line Journal for Undergraduate Neuroscience Research. Both undergraduate-authored neuroscience research articles and neuroscience review articles are accepted from undergraduates around the globe. Articles then are sent out to undergraduate, peer reviewers around the globe for peer review. These student reviewer teams have been set up at numerous undergraduate colleges and universities using a variety of models. Following peer review the manuscript is accepted as is, requires revision, or found unacceptable. This first tutorial will walk you through the process of peer review. Understanding the Basics of Peer Review: Part 1 – Receiving a Manuscript IMPULSE Journal for Undergraduate Neuroscience Developed by: Matthew Fisher, 2011 Advisor: Sarah M. Sweitzer, PhD

2. IMPULSE is a peer reviewed scientific journal. Below are the definitions of peer review and peer-reviewed journal. Peer Review IMPULSE IMPULSE • Definition • Peer review is the evaluation of creative work or performance by other people in the same field in order to maintain or enhance the quality of the work or performance in that field • Peer Review Journal • A peer review journal comprises literature of similar type that has been through the peer review process.

3. The Executive Editor receives and determines if the manuscript should be sent to associate editors to send out for review. Peer-Review Hierarchy IMPULSE IMPULSE Executive Editor Associate Editors answer to the Executive and are placed in charge of a review team. Associate Editor Associate Editor Associate Editor Reviewers are asked to review the scientific content of the manuscript. Reviewer Reviewer Reviewer Reviewer Reviewer Reviewer Reviewer Reviewer Reviewer

4. STEP 5: The Associate Editor sends the manuscript to reviewers. STEP 1: A manuscript is received. Manuscript Submitted Associate Editor STEP 4: If the paper is deemed appropriate, it is sent to an Associate Editor. IMPULSE STEP 2:The Executive Editor must decide if the overall paper is appropriate for the journal. STEP 6: Reviews are sent back to Associate Editor who compiles them and sends to Executive Editor. STEP 7: Based on the reviews the Executive Editor and Editor-in-Chief decide if the paper is accepted, requires revisions, or is rejected. Reviewer Reviewer Reviewer Executive Editor STEP 3:If the paper is not appropriate for the journal or if there are major problems with the format, the paper is returned to author without review. STEP 8: Manuscript is returned to authors indicating accepted for publication, requires revision, or rejected. Paper accepted for Publication Paper sent back to author

5. Manuscript Formatfor Original Research IMPULSE IMPULSE • Abstract • Introduction • Materials and Methods • Results • Discussions • Acknowledgments • Works Cited Listed to the left are the common sections of an original research manuscript. In the next few slides some common guidelines for each of the sections are provided.

6. Abstract IMPULSE IMPULSE • The abstract is a concise summary of the manuscript. • The abstract should include a brief part of each section • Introduction • Hypothesis • Methods • Results • Conclusions • The abstract should “grab” the potential reader’s attention with as few words as possible. Generally <250 words Why do we care? Why is this important? What was being tested? How was it being tested? What was found? What does it mean?

7. Introduction IMPULSE IMPULSE Usually 500-750 words • The introduction relates background information related to the manuscript. • The introduction should include: • Significance of work • Background science • Hypothesis Why is this important? Citation of the scientific literature What does the author expect? Funnel Approach Broad Narrow Begin an introduction with broad concepts and then “funneling” to more specific information relevant to the paper at-hand.

8. Materials and Methods IMPULSE Unlimited words IMPULSE • The materials and methods section describes the methodology used when gathering data. • This section should include: • Materials used • Methodology *Another investigator should be able to read this section and re-create the work Narrative format. NO LISTS! Step-by-step procedures*

9. Results IMPULSE Unlimited words IMPULSE • The results section presents all data collected throughout the project. • The results section should include: • numerical data • associated graphs, charts, tables, etc. • Statistical significance should be indicated *Only data should be presented here. The discussion is later! In narrative format Clearly labeled P<0.05 is significant P>0.05 is not significant If it is close to significance then perhaps a larger sample size is needed.

10. Discussion IMPULSE Usually <1500 words IMPULSE • The discussion draws conclusions from the project as it relates to other research in the field and potential future direction of the field. • The discussion section should include: • Restatement of hypothesis/thesis • Hypothesis refuted or accepted? • Interpretation of data • Big picture relevance • Future directions What was being tested? What was found? What does it mean? How does it relate to the literature? Why do we care? Why is this important? What questions do these results raise?

11. Acknowledgements IMPULSE IMPULSE • The acknowledgements sections allows the author to thank anyone or anything he/she deems appropriate. All financial support mustbe listed. • Often included in this section: • research mentors • fellow student investigators • grant organizations MUST HAVE PERMISSION TO ACKNOWLEDGE SOMEONE!

12. References IMPULSE IMPULSE The references section allows the author to provide a detailed description of all sources used in the manuscript. • These sources are arranged in a particular format specified by the journal.

13. IMPULSE IMPULSE This is the second half of a two part on-line tutorial for new student reviewers for IMPULSE. This second tutorial will walk you through the peer review of an original research manuscript. While some of the generalities of peer review also are true for a review article, some of the details in this module are specific for original research manuscripts. Understanding the Basics of Peer Review: Part 2 – How to Review the Manuscript IMPULSE Journal for Undergraduate Neuroscience Developed by: Matthew Fisher, 2011

14. Once You Receive A Manuscript… • Be sure to understand the correct journal format for the submitted manuscript. • Read any necessary background information to become familiar with subject area. • Identify the main points and thesis of the manuscript. • Assess methods and findings. • Associate related graphs, charts, tables, etc. to the results. http://impulse.appstate.edu/submissions.php Yes, you may have to do a little research. Are you the appropriate person for reviewing this manuscript? What was their question? Why is their question important? Do you understand what they did? Could you repeat the experiment? Are the figures/tables/charts effective in presenting the main findings?

15. Reviewer Training Article We are using a particularly good paper published by IMPULSE as a teaching tool.  With the authors’ permission we have introduced errors to show how to recognize and correct such problems; the modifications to the article to represent the original, first and second revisions are purely fictional and for educational purposes only. • Title: Oral Self-Administration Of Ethanol In Transgenic Mice Lacking β-Endorphin • Authors: Sidney B. Williams, Ashley Holloway, Kevin Karwan, Stephani Allen, Judith E. Grisel

16. How to use this tutorial • The best way to use this tutorial is to read the sections as typed and try to identify problems based on what you learned in the first tutorial. • Then click through the slide and suggested revisions will be highlighted and identified. • These suggestions are not exhaustive and in fact you may find some additional changes to suggest.

17. Abstract-Original Submission EtOH modifies the production and/or release of endogenous opioid peptides, including -endorphin (Gianoulakis, 2004; Przewlocka et al., 1994; Schulz et al., 1980). Opioids subsequently influence the reinforcing properties of EtOH and the development of alcoholism (Terenius, 1996; Van Ree, 1996). In this study, beta-endorphin deficient mutant mice were used to examine the effects of a specific opioid peptide on EtOH consumption. Mice were obtained from The Jackson Laboratory, Bar Harbor, ME, USA. Male and female, adult naïve mice were single housed in Plexiglas cages with corn cob bedding and ad lib access to food (mouse chow) and water. A two-bottle free choice EtOH oral self-administration paradigm was administered to homozygous mutant mice (void of all beta-endorphin), heterozygous mice (50% beta-endorphin expression), and sibling wildtype mice (C57BL/6J). Subjects received increasing concentrations of EtOH (0%, 3%, 6%, 12%, and 15%) each given over an eight day span, and were evaluated for preference and consumption each day. Bottles were switched every other day to avoid the development of a side preference. Overall, females drank more than males. Homozygous mutant mice (KO) showed decreased preference for EtOH at all concentrations, and self-administered significantly less than heterozygous mice (HT) and wildtype mice (C57). The HTs had a tendency to drink the most followed by the C57s, and the KOs drank the least. These data support the hypothesis that beta-endorphin influences the reinforcing effects of EtOH.

18. Abstract – Edits References belong in introduction What does this mean? Make sure to identify abbreviations. This should appear in Discussion Material and Methods knock-out Inaccurate Information EtOH modifies the production and/or release of endogenous opioid peptides, including -endorphin (Gianoulakis, 2004; Przewlocka et al., 1994; Schulz et al., 1980). Opioids subsequently influence the reinforcing properties of EtOH and the development of alcoholism (Terenius, 1996; Van Ree, 1996). In this study, beta-endorphin deficient mutant mice were used to examine the effects of a specific opioid peptide on EtOH consumption. Mice were obtained from The Jackson Laboratory, Bar Harbor, ME, USA. Male and female, adult naïve mice were single housed in Plexiglas cages with corn cob bedding and ad lib access to food (mouse chow) and water. A two-bottle free choice EtOH oral self-administration paradigm was administered to homozygous mutant mice (void of all beta-endorphin), heterozygous mice (50% beta-endorphin expression), and sibling wildtype mice (C57BL/6J). Subjects received increasing concentrations of EtOH (0%, 3%, 6%, 12%, and 15%) each given over an eight day span, and were evaluated for preference and consumption each day. Bottles were switched every other day to avoid the development of a side preference. Overall, females drank more than males. Homozygous mutant mice (KO) showed decreased preference for EtOH at all concentrations, and self-administered significantly less than heterozygous mice (HT) and wildtype mice (C57). The HTs had a tendency to drink the most followed by the C57s, and the KOs drank the least. These data support the hypothesis that beta-endorphin influences the reinforcing effects of EtOH.

19. Abstract – Revised Ethanol (EtOH) modifies the production and/or release of endogenous opioid peptides, including β-endorphin. Opioids subsequently influence the reinforcing properties of EtOH and the development of alcoholism. In this study, β-endorphin deficient mice were used to examine the effects of a specific opioid peptide on EtOH consumption. A two-bottle free choice EtOH oral self-administration paradigm was administered to homozygous β-endorphin knock out, heterozygous, and wildtype mice. Subjects received increasing concentrations of EtOH (0%, 3%, 6%, 12%, and 15%) given over an eight day span, and were evaluated for preference and consumption each day. Overall, females drank more than males. Homozygous knock out mice showed decreased preference for EtOH at all concentrations, and self-administered significantly less than heterozygous and wildtype mice. These data support the hypothesis that β-endorphin influences the reinforcing effects of EtOH.

20. Introduction-OriginalPart 1 Add the information from the human study here to enhance the background information. This would give human clinical relevance. EtOH has already been explained. There is no need to do this twice. The meaning of this is unclear. A reference should be added to clarify this statement. The structure of this sentence is confusing. Split into two sentences. This has already been discussed and should be moved up in the Introduction. This is a good example of an Intro sentence. Biological contributions to excessive ethanol (EtOH) drinking that can lead to alcoholism have been investigated for decades (Nestler and Aghajanian, 1997; Cowen et al., 2004). Animal models evaluating the chemical and genetic substrates for self-administration of alcohol (EtOH) have been used in an attempt to identify mechanisms that promote the development of alcoholism in humans. The relationship between EtOH and endogenous opioid peptides is of particular interest (For instance, EtOH modifies the synthesis and release of β-endorphin , and these changes in turn affect dopamine release in the mesolimbic pathway, a critical neural substrate for reward (Widdowson and Holman, 1992). Numerous studies in animals (Altshuler et al., 1980; Froehlich et al., 1990) and in humans (O’Malley et al., 1992; Volpicelli et al., 1992) have shown that administration of an opiate antagonist can decrease EtOH consumption. TheEtOH-mediated release of dopamine in the nucleus accumbens can also be blocked by opiate antagonists (Gonzalez and Weiss, 1998). Thus, β-endorphin is thought to play an important role in the reinforcing effects of EtOH, mediating consumption (Griseletal., 1999; Roberts et al., 2000), craving (VanRee, 1996; Marinelli et al., 2000), and relapse (Terenius, 1996). Some genotypes of rodents that prefer EtOH have lower endogenous β-endorphin when compared to non-preferring lines (Aguirre et al., 1995; del Arbol et al., 1995). Moreover, humans with a positive family history for alcoholism also tend to have lower basal levels of β-endorphin, as

21. Introduction-OriginalPart 2 It is unclear if this statement deals with a previous study. Be sure that background information is relevant to any previous study. This belongs in Discussion. Make sure to look for a thesis. well as an exaggerated EtOH-mediated release of this peptide, than those without a genetic liability for alcoholism (Gianoulakis et al., 1989). From data such as these it has been argued that those prone to alcoholism are “self-medicating” an opioid deficiency and/or especially benefiting from an opioid surge, following administration of EtOH (Oswald and Wand, 2004; Reid et al., 1991). In an earlier study, Grisel et al. (1999) found evidence to support the hypothesis that EtOH-mediated reward depends upon β-endorphin. We investigated EtOH consumption in a two-bottle, freechoice paradigm in transgenic mice with varying levels of endogenous β-endorphin. Although strain differences were small, there was a tendency for mice with low levels of β-endorphin to consume the most EtOH, supporting the idea that consumption would be especially reinforcing for subjects deficient in this peptide. Unfortunately, in this study, we only evaluated two concentrations of EtOH: 7 and 10%, and so were unable to fully elucidate the dose response relationship as it varied with respect to β-endorphin levels. In the present experiment we expanded the range of concentrations tested and also included sufficient numbers of male and female subjects to test for sex-dependent effects. Furthermore, we evaluated sucrose preference to test for differences in sweet responsivity. We predicted that mice lacking all β-endorphin (knockout; KO) would consume the least, because they would not be able to benefit from any β-endorphin release. Mice with half the normal amounts of β-endorphin (heterozygotes; HT) were predicted to drink the most EtOH and wildtype mice (B6) were expected to drink intermediate levels. Our data support our hypothesis that β-endorphin does influence the reinforcing properties of EtOH. HT mice with 50% β-endorphin expression prefer and consume the most EtOH, KO the least, and B6 in between. With this data we can further examine the neurological substrates that modulate EtOH consumption and addiction.

22. Introduction-Revision 1Part 1 This sentence can be started at “Rodents with lower…” This statement is not needed. To meet limits, be sure to only include relevant information. What changes? This section is very vague. The structure of this sentence is confusing. This concept is related to a genetic liability. This statement can be made more concise. Biological contributions to excessive ethanol (EtOH) drinking that can lead to alcoholism have been investigated for decades (Nestler and Aghajanian, 1997; Cowen et al., 2004). Animal models evaluating the chemical and genetic substrates for self-administration of EtOH have been used in an attempt to identify mechanisms that promote the development of alcoholism in humans.The relationship between EtOH and endogenous opioid peptides is of particular interest. Some genotypes of rodents with lower endogenous β-endorphin levels exhibit an increased preference for EtOH (Aguirre et al., 1995; del Arbol et al., 1995). Moreover, humans with a positive family history for alcoholism also tend to have lower basal levels of β-endorphin, as well as an exaggerated EtOH-mediated release of this peptide (Gianoulakis et al., 1989). It has been suggested that those prone to alcoholism are “self-medicating” an opioid deficiency and/or especially benefiting from an EtOH-induced opioid surge (Oswald and Wand, 2004; Reid et al., 1991). It has been shown that EtOH modifies the synthesis and release of β-endorphin (Scanlon et al., 1992 and Froehlich, 1995). These changes in turn affect dopamine release in the mesolimbic pathway, a critical neural substrate for reward (Widdowson and Holman, 1992). Numerous animal (Altshuler et al., 1980; Froehlich et al., 1990) and human (O’Malley et al., 1992; Volpicelli et al., 1992) studies have shown that administration of an opiate antagonist can decrease EtOH consumption. EtOH-mediated release of dopamine in the nucleus accumbenscan also be blocked by opiate antagonists (Gonzalez and Weiss, 1998).

23. Introduction- Revision 1Part 2 This citation appears twice. It should be removed from the beginning of the citation. This statement is not needed. Instead of saying that it is “unfortunate,” just tell what the new study is researching. It is unclear is this was actually the prior Grisel study. This entire section can be simplified. Only state the pertinent points. Thus, β-endorphin is thought to play an important role in the reinforcing effects of EtOH, mediating consumption (Griseletal., 1999; Roberts et al., 2000), craving (VanRee, 1996; Marinelli et al., 2000), and relapse (Terenius, 1996). In an earlier study, Grisel et al. (1999) found evidence to support the hypothesis that EtOH-mediated reward depends upon β-endorphin. Using a two-bottle, freechoice paradigm in transgenic mice with varying levels of endogenous β-endorphin, reported that mice with low levels of β-endorphin consumed the most EtOH, supporting the idea that consumption would be especially reinforcing for subjects deficient in this peptide (Grisel et al., 1999). Unfortunately, in this study, only two concentrations of EtOH (7 and 10%) were examined. Thus, this study was unable to fully elucidate the dose response relationship as it varied with respect to β-endorphin levels. In the present experiment we expanded the range of concentrations tested and also included sufficient numbers of male and female subjects to test for sex-dependent effects. Furthermore, we evaluated sucrose preference to test for differences in sweet responsivity. We predicted that mice lacking all β-endorphin (knockout; KO) would consume the least, because they would not be able to benefit from any β-endorphin release. Mice with half the normal amounts of β-endorphin (heterozygotes; HT) were predicted to drink the most EtOH and wildtype mice were expected to drink intermediate levels.

24. Introduction-Revision 2Part 1 Biological contributions underlying alcoholism have been investigated for decades (Nestler and Aghajanian, 1997; Cowen et al., 2004). The relationship between ethanol (EtOH) and endogenous opioid peptides is of particular interest. Rodents with lower endogenous β-endorphin levels exhibit an increased preference for EtOH (Aguirre et al., 1995; del Arbol et al., 1995). Moreover, humans with a positive family history for alcoholism have lower basal levels of β-endorphin, as well as an exaggerated EtOH-mediated release of this peptide, compared to humans without a family history of alcoholism (Gianoulakis et al., 1989). It has been suggested that those prone to alcoholism are “self-medicating” an opioid deficiency and may be especially benefiting from an EtOH-induced opioid surge (Oswald and Wand, 2004; Reid et al., 1991). It has been shown that EtOH modifies the synthesis and release of β-endorphin (Scanlon et al., 1992 and Froehlich, 1995). These changes in release of β-endorphin alter dopamine release in the mesolimbic pathway, a critical neural substrate for reward (Widdowson and Holman, 1992). Numerous animal (Altshuler et al., 1980; Froehlich et al., 1990) and human (O’Malley et al., 1992; Volpicelli et al., 1992) studies have shown that administration of opiate antagonists can decrease EtOH consumption and EtOH-mediated release of dopamine in the nucleus accumbens (Gonzalez and Weiss, 1998). Thus, β-endorphin is thought to play an important role in the reinforcing effects of EtOH, mediating consumption (Griseletal., 1999; Roberts et al., 2000), craving (VanRee, 1996; Marinelli et al., 2000), and relapse (Terenius, 1996).

25. Introduction- Revision 2Part 2 An earlier study, (Grisel et al.,1999) using a two-bottle, freechoice paradigm in transgenic mice with varying levels of endogenous β-endorphin, reported that mice with low levels of β-endorphin consumed more EtOH, supporting that consumption would be especially reinforcing for subjects deficient in this peptide (Grisel et al., 1999). The present study expands on this previous study by exploring EtOH preference across a larger range of EtOH concentrations in male and female mice and compare this to sucrose preference. We hypothesize that that mice lacking all β-endorphin (knockout; KO) would consume the least EtOH while mice with half the normal amounts of β-endorphin (heterozygotes; HT) would consume the most EtOH with wildtype mice consuming an intermediate level. Notice that the revision has a much shorter and focused Introduction. The accepted Introduction contains only the relevant information to understanding the study and is void of unwanted information.

26. The materials and methods section should let someone replicate the study without the need for a “miracle.”

27. Materials & Methods - Original Part 1 Where did the mice originate from? What sex was used? Were both sexes used and in what ratio? Was all of this approved by an approval committee? = ETHICS Homozygous (B6, KO) or heterozygous mating pairs were group housed by sex and genotype following weaning at 20- 21 days, and maintained on a reverse 12:12 light: dark cycle at 21° C ± 2° with ad libetum access to food and water. On Day 1, adult subjects were taken from the colony room to a procedural room where they were weighed and single housed in corn-cob bedding lined Plexiglas cages with wire lids. Two 25 mL graduated cylinders containing tap water were placed on each cage, food hoppers were filled with rodent block chow, and the tube volumes were recorded. Cage locations were counterbalanced so that genotype and sex were equally distributed on the rack. A sentinel cage was added to each side of the rack in order to obtain control volumes of leakage/evaporation. Approximately 24 h later, tube volumes were recorded again, and refilled as necessary. For the following 48 days, tube readings occurred every day, and after every 48 h tube positions were switched to control for development of side preferences. Following 8 days of water drinking, one of the tubes (counterbalanced Much of the missing information from this part of the Materials and Methods is crucial to understanding the study. Remember to be as specific as possible so that another researcher could recreate the study!

28. Materials& Methods – OriginalPart 2 There is no mention of statistical analysis. Although the author does mention some ratio information, the reader also needs to know any statistical programs, software programs, etc. used. for side across cages) was filled with 3% EtOH for 8 days. Remaining concentrations of EtOH and a test for sweet preference (6%, 12%, 15% EtOH, and 8% sucrose) were also tested for 8 days each. EtOH consumption was expressed as g/kg/day, and preference ratios were calculated. The experiment was conducted in three separate runs, which were procedurally identical, but separated by 12 days.

29. Materials & Methods – RevisedPart 1 The statement explains that the study was approved by an animal care committee. The section does a good job of giving detailed information about the mice and where they originated. This explains that both sexes of mice were used. Progenitor mice used to obtain subjects for this study were derived from those made by Rubinstein et al. (1996) and obtained from The Jackson Laboratories, Bar Harbor, ME. The original lines were constructed by inserting a point mutation in exon 3 of the POMC gene, causing a shortened proopiomelanocortin (POMC) prohormone. The gene has since been fully backcrossed onto the C57BL/6J inbred line. All experimental animals were born and reared at Furman University in the animal care facilities. Mating pairs were arranged and offspring produced by either homozygous (B6, KO) or heterozygous mating pairs. Mice were group housed by sex and genotype following weaning at 20- 21 days, and maintained on a reverse 12:12 light: dark cycle at 21° C ± 2° with ad libetum access to food and water. All experimental procedures were in accordance with the Furman University Institutional Animal Care and Use Committee and the principles of laboratory animal care from the National Institutes of Health guidelines. On Day 1, adult male and female subjects were taken from the colony room to a procedural room where they were weighed and single housed in corn-cob bedding lined Plexiglas cages with wire lids. Two 25 mL graduated cylinders containing tap water were placed on each cage, food hoppers were filled with rodent block chow, and the tube volumes were recorded. Cage locations were counterbalanced so that genotype and sex were equally distributed on the rack. A sentinel cage was added to each side of the rack in order to obtain control volumes of leakage/evaporation. Approximately 24 h later, tube volumes were recorded again, and refilled as necessary. For the following 48 days, tube readings occurred every day, and after every 48 h tube positions were switched to control for development of side preferences. Following 8 days of water drinking, one of the tubes (counterbalanced

30. Materials & Methods – RevisedPart 2 This statement explains more details of the mice in the study. This explains the statistical analysis techniques used. for side across cages) was filled with 3% EtOH for 8 days. Remaining concentrations of EtOH and a test for sweet preference (6%, 12%, 15% EtOH, and 8% sucrose) were also tested for 8 days each. EtOH consumption was expressed as g/kg/day, and preference ratios were calculated. The experiment was conducted in three separate runs, which were procedurally identical, but separated by 12 days. A total of 77 test subjects were used in the study. Twenty four of these subjects were wildtype (B6), 28 heterozygous (HT), and 25 knockout (KO). There were 37 females and 40 males, approximately equally divided across genotype and test run. Data were analyzed by two-way (genotype and sex) repeated measure ANOVA on average g/kg of EtOH administered and preference at each dose. Because of significant effects of sex, males and females were subsequently evaluated separately by single factor ANOVAs at each concentration. Post-hoc analysis of significant differences was evaluated using the Scheffe test. Criterion for significance was set at p ≤ .05. It is important to note that, unlike other sections of a manuscript, the Materials and Methods section often needs more information to be added by the writer.

31. Results This is a descriptive word and should be avoided. This is unclear. These are good key words to look for. There was no difference in any measure of EtOH consumption across runs, so data from all three runs were combined following two-way analysis with run and genotype. No differences in water or sucrose intake were observed among the runs or between strains (data not shown), nor were there any significant interactions with sucrose consumption. Preference β-endorphin levels influenced the preference for EtOH as evidenced by a significant effect of genotype in the two way repeated measures ANOVA on preference across the dose range: (F(2,69) = 8.361, p < .01). The KOs tended to drink the least, the HTs drank the most, and the B6s were intermediate drinkers, with this pattern becoming more evident at higher concentrations of available EtOH (Figure 1). Although there was no main effect of (F(1,69) = .103, p > .05), there was a significant sex x genotype interaction (F(2,69) = 3.496, p < .05). In addition, although there was no overall effect of EtOH concentration (F(3,207) = 1.898, p > .05), there was a significant Interaction between EtOH concentration and strain (F(6,207) = 6.306, p < .01), but not significant interactions with sex (F(3,207) = .693, p > .05). Post hoc analysis of EtOH preference was evaluated in males and females separately at each concentration. KO males drank significantly less than either B6 or HT males at 3 or 6 % EtOH, and less than B6 males at 12 or 15 % EtOH, and there was a strong tendency for them to drink less than HTs at these concentrations (p = .056 and .091, respectively). In addition, there was a tendency for HT males to prefer 6% EtOH more than B6 males (p = .056). There were only genotypic differences in females at 6% EtOH, as KO’s consumed relatively less EtOH than either other line.

32. Discussion This whole section seems to repeat the Introduction. This citation leads the reader to question the reasoning for this study. A citation should reference this information This WAS NOT tested in the study. This citation is inappropriate and unclear here. Current theories of addiction emphasize the rewarding effects produced by the drug (Wise, 1988). One mediator of these rewarding effects is β- endorphin (Herz, 1997). Many studies show that EtOH drinking in mice is correlated with the animal’s genetic ability to release β-endorphin (Gianoulakis et al., 1989) It has also been shown that people with a family history of alcoholism have an increase in β-endorphin release after EtOH administration compared to controls with no family history of alcoholism (Gianoulakis et al., 1989) The purpose of this study was to see how β-endorphin influenced EtOH drinking in an animal model. The results support our hypothesis that β-endorphin modulates the rewarding properties of EtOH. Mice completely lacking β-endorphin (KO) self-administered less EtOH in a freechoice paradigm. Overall, females drank more EtOH than the males, which is a well known result in rodents (Jones and Whitfield, 1995). Notably, the effect of β-endorphin on drinking was also sex-specific. This peptide did not influence EtOH self-administration in females, but did effect drinking in males. The male KOs, entirely lacking β-endorphin, appeared to be insensitive to the rewarding effects of EtOH as they self administered at chance levels (50% of the fluid they consumed was from the EtOH-spiked tube). On the other hand, male HTs showed a pattern of drinking consistent with enhanced EtOH reward, and tended to drink more than control mice. Both of these findings are consistent with a theory implicating β-endorphin release in EtOH reinforcement though why this effect should be sex-specific is unclear at present.

33. Discussion This is not a scientific term. Although the writer may understand the term, not all readers will. This is unclear. Never speculate without explaining. The writer should expand and explain these ideas. A new sentence about future directions would be appropriate here. This is a great example of a closing thought. This is very poorly worded. One possibility is that sex specific hormones, such as progesterone metabolites, make up for the lack of β-endorphin in females (Morrow, 2007; Morrow et al., 2006) though further research is needed to clarify the mechanisms of this sex-dependent effect. There is a main concern that should be acknowledged when interpreting data collected from induced mutant mice. It is possible that the observable phenotypic behaviors could be the cause of a “hitchhiking” gene polymorphism and not purely a consequence of the intentional genetic mutation (Gerlai, 1996; Low et al., 1998). A more likely possibility is that some compensatory reaction to the genetic mutation could cause the different phenotypic behaviors and not the mutation alone (Mogil and Grisel, 1998). The use of inducible mutant mice in future studies will lead to a better understanding of the mechanism by which β-endorphin induces behavior, including behaviors related to alcoholism. Further investigation should be done to evaluate the role of β-endorphin in EtOH sensitivity more generally . Currently, we are performing place conditioning tests with the β-endorphin KO and HT mice. This is also a test for EtOH reinforcement, but β-endorphin may affect other consequences of EtOH too. The main purpose of this study was to evaluate the neurobiological substrates that modulate EtOH consumption in an animal model where β-endorphin levels vary. We demonstrated that β-endorphin, especially in males, influences the reinforcing properties of EtOH. A more complete assessment of a possible vulnerability to alcoholism, giving us a broader understanding of the predispositions to addiction and aiding in the development of successful interventions to that would ameliorate the negative consequences of excessive alcohol use.

34. Writing your review • A review should include: • A brief summary of the main hypothesis and findings of the manuscript • Overall opinion of the manuscript. Move from positive to things requiring revision. • Detailed review • Major strengths and weaknesses • Minor weaknesses

35. Writing your review • How you write it is EVERYTHING! • Be courteous. • Remove the personal. • If you would not like to be on the receiving end of your review, or you would not sign your name to it, then you likely need to change how you phrase comments and suggestions.

36. Examples of Feedback What to avoid: This manuscript sucks. It has numerous problems with the abstract, introduction, and methods. The students did not do a very good job designing their experiments. I would not accept it! Why this is not acceptable? • Sucks is negative and not constructive! • Numerous problems is useless feedback….make lists of corrections/additions/deletions that would improve the manuscript. • “the students….” - This is a personal attack! • I would not accept it! - You as a reviewer do not get to decide if the manuscript is accepted (only the Executive Editor has that job) and your opinion does not belong in the review that is being sent out to the authors.

37. Examples of Feedback An example of part of a good review: • This manuscript explores the hypothesis that both male and female mice lacking beta-endorphin will voluntarily consume less alcohol than wild type mice. Several major and minor concerns have been identified and are listed below. • Abstract: • Minor concerns: • The abstract could be written more concisely. • References should not be included in an abstract. • All abbreviations need to be defined the first time they are used. • Details of mice and procedures need to be moved to Materials and Methods.

38. Examples of Feedback An example of part of a good review cont’d: • Introduction: • Minor concerns: • The introduction could be written more concisely. • Several references are missing. • Moving the animal and human beta-endorphin behavioral studies to the first paragraph may enhance the significance of the current study. • The paragraph on the previous study as it relates to the current study is unclear as to what was done previously and what is being currently proposed. • The explanation of the current study can be streamlined. • The final 3 sentences belong in the discussion.

39. Examples of Feedback An example of part of a good review cont’d: • Materials and Methods: • MAJOR concerns: • What is the source of the mice? • Were these methods approved by an institutional animal care and use committee? • What sex animals were used? • How many animals were used? • How was the data analyzed? These are major concerns as opposed to the previous minor concerns because these concerns impact the experimental design and interpretation of results (aka:The meat of the science!)

40. Examples of Feedback What to avoid: “This sentence is wrong.” “Put a comma here.” “Your data is wrong.” Better ways to phrase: “As written the meaning of this sentence is unclear.” “A comma would be appropriate here.” “It would appear that the written results do not match what is shown in Figure 1.”

41. Congratulations!You have completed the IMPULSE peer reviewer on-line training module.Try to remember the feelings of the author as they read your reviews….we don’t want them to feel like the scientist in the cartoon!