Therapeutic Effects of Single and Combined Anti-Disseminated Intravascular Coagulation (DIC) Drugs in a Rat Venom-Induced Consumption Coagulopathy (VICC) Model Using Yamakagashi (Rhabdophis tigrinus) Venom
Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsReference: “Therapeutic effects of single and combined anti-disseminated intravascular coagulation (DIC) drugs in a rat DIC model using yamakagashi (Rhabdophis tigrinus) venom ” Submittted to TOXINS, February, 2026.
General comments: In the manuscript submitted for this review, the authors describe their results regarding the use of a new treatment for accidents involving the snake Yamakagashi snake (Rhabdophis tigrinus), a species distributed in Japan. One of the effects of a bite from this snake is disseminated intravascular coagulation (DIC). The use of antivenom for Yamakagashi is not an approved treatment and, according to the authors, the serum is also difficult to obtain. Thus, the authors investigated the application of commercially available drugs, such as tranexamic acid and antithrombin III, in the treatment of DIC caused by Yamakagashi venom, using a rat model, and also the combination of each drug with recombinant thrombomodulin α. The data show that administration of either tranexamic acid or antithrombin III alone, did not prolong the survival of rats, nor did it alter blood coagulation markers such as prothrombin time, fibrinogen concentrations, and D-dimer levels in rats treated with yamakagashi venom. However, the combined administration of recombinant thrombomodulin α and tranexamic acid prolonged the survival of the rats and partially restored blood coagulation markers. Based on their experimental data, the authors conclude that the combination of recombinant thrombomodulin α and tranexamic acid may represent a useful therapeutic regimen for yamakagashi venom exposure. After reading the text, it is my opinion that this manuscript falls within the scope of TOXINS; the text is well-written, clear, and understandable. However, in my opinion the authors need to conduct analyses where rTM was used alone for the treatment of DIC so that they can perform comparative analyses with treatments in combination with other drugs. Additionaly, I suggest some changes that could make the text more complete and appealing to the readership.
Specific Comments:
1- Between lines 22 to 24 Key Words. I would change the order in which the words appear in Key words: Yamakagashi (Rhabdophis tigrinus) venom; rat disseminated intravascular coagulation (DIC) model; recombinant thrombomodulin alpha; Tranexamic acids; antithrombin III; Combined therapy.
2- Between lines 33 and 34 … Yamakagashi is a venomous snake belonging to the Colubridae family, and it is typified by short posterior fangs. Please include one or more references.
3- Line 39 … Severe cases can be fatal without treatment, and deaths have been previously reported. Please include one or more references.
4- Between lines 40-41 …. the high-molecular weight portion of the venom …Please change molecular weight to molecular mass throughout of the text, as proteins have masses instead weight. Although this term is accepted in the literature, it is wrong.
5- Between lines 40 to 43 … Regarding the chemical characteristics of yamakagashi venom, the high-molecular weight portion of the venom contains prothrombin activators. Exposure to its venom shortened both the prothrombin time (PT) and activated partial thromboplastin time of human plasma. Please include one or more references.
6- In lines 66 in vitro and in line 68 in vivo, These are words of Latin origin and should be cited in the text in italics as in vitro and in vivo throughout of the text.
7- Between lines 71 and 72 … Meanwhile, antithrombin III (ATIII) inhibits over activated blood coagulation and prevents thrombus formation in microvessels. Please include one or more references. I will review Materials and Methods first, before moving on to the other items.
8- Line 219. Authors wrote … Male Sprague-Dawley rats (JAPAN SLC, Inc., Shizuoka, Japan) aged 12 weeks… Is there any particular reason why the authors used only male rats? And also the age of 12 weeks? Some explanation could be given throughout the text.
9- Between lines 235 and 236 … Toxic glands were collected from 100 male and female snakes. The sentence is confusing because it doesn't make it clear whether 100 snakes were used in total or 100 male and 100 female animals? Please rewrite in the revised version.
10- Between lines 236 to 238. … The glands were excised, cut into small pieces, and centrifuged with distilled water, after which the supernatant was lyophilized [25]. This description of how the venom was obtained seems somewhat vague to me. There was no maceration of the gland to release the venom. Just mixing it with distilled water? If so, the authors could refine this methodology so that other researchers can reproduce the method if interested.
11- Between lines 245 and 246… In the model group, 300 μg of Yamakagashi venom were administered intramuscularly. It would be interesting if the authors detailed why this particular mass of venom was chosen and why the intramuscular injection route was used instead of intradermal or subcutaneous routes, which are more pathophysiologically similar to natural envenomation.
12- Between lines 246 to 249 … In the TXA group, 5 mg of TXA were administered intravenously 30 min after venom administration. In the ATIII group, 20 U of ATIII were administered intravenously 30 min after venom exposure. Each group included six rats. Blood samples were collected at 0, 2, 4, 8, 24, 48, 72, 96, and 120 h after the start of the experiment. In my opinion, the authors could refine the information by indicating some explanation for why these parameters were used.
13- The same applies to text written between the lines 253 to 263. A more refined explanation of the parameters could be provided. Results
14- About figure 1, line 88. In my opinion, the authors could include the meanings of the colored spheres in the figure, so that readers wouldn't need to read the legend to understand the results. This approach has been used more frequently in more recent publications and complements the information in the legend.
15- Still figure 1… Generally, experiments with pharmacological treatments in live animals or cells follow a concentration-dependent curve. Is there any particular reason why the authors didn't test other concentrations of TXA (5 mg/rat) or ATIII (20 U/rat)?
16- The same question also applies to the time dependency of the treatments. Only one time frame was used.
17- About figure 2, line 104. The same as comment for figure 1. The authors could include the meanings of the colored spheres in the figure.
18- I'm simply asking the authors if these results weren't expected, given that treatment with the tested drugs didn't alter animal mortality after treatment with the venom.
19- 2.3, lines 110 and 111. In my opinion authors should rewrite the sentence to … Effects of rTM, ATIII or TXA alone, and TXA or ATIII in combination with rTM on survival in a rat model of DIC induced by yamakagashi venom .
20- About figure 3. The same as comment for figure 1. The authors could include the meanings of the colored spheres in the figure.
21- About figure 3. The data shown in Figure 3 are very interesting, especially when analyzing the treatments of animals with venom and rTM plus TXA. I'm asking if analyses were performed over periods longer than 96 hours? Were the animals still protected?
22- About figure 3. Although the effects with the combination of rTM and ATIII were smaller, than combination of rTM and TXA, they also showed some protection. Did the authors not perform variations in these combinations seeking better results? Once again, tests studying combinations at a single concentration of the drugs studied may be insufficient for more detailed conclusions as previously commented.
23- About figure 3. I would like to know the authors' opinion on whether there was a lack of control over the use of rTM alone, because if there is treatment in combination with other drugs, shouldn't there also be treatment alone? This is an essential control for drawing conclusions about additive or synergistic activities.
24- Lines 116 and 117…authors wrote…Conversely, TXA (5 mg/rat) or ATIII (20 U/rat) alone did not improve rat survival. But rTM alone? For comparative analyses using two drugs authors must test each one separate and in combination.
25- About figure 4. The same comment made for Figure 3 can be applied to Figure 4. The authors failed to test rTM alone to verify if there is any additive or synergistic activity with the other drugs tested. Discussion
26- Lines 150 to 152…The dose of each anti DIC drug was calculated by converting the maximum clinical dose in humans to the weight-based equivalent in rats. This sentence should be written in the Materials and Methods section, not in the Discussion section. The Discussion section is for interpreting the results and mechanistic hypotheses, but not for methodology.
27- Lines 153 to 159 …TXA binds strongly to the lysine-binding site, which is the fibrin affinity site of plasmin and its precursor plasminogen, which participate in fibrinolysis occurring after blood coagulation. Through this binding, TXA prevents plasmin and plasminogen from binding to fibrin, thereby strongly inhibiting fibrin deg radation by plasmin. Furthermore, the antifibrinolytic effect of TXA is further enhanced in the presence of plasma antiplasmins such as α2-macroglobulin. Plasmin inhibits platelet aggregation and degrades coagulation factors, but it specifically induces fibrin degradation. Please include one or more references to sentence written.
28- Lines 167 to 169 …phrase is confuse … It is speculated that accelerated coagulation occurs rats administered venom, resulting in the secondary phenomenon of deficiency of FIB coagulation factors [31]. Better change to …. It is speculated that accelerated coagulation that occurs in rats administered with venom, resulting in the secondary phenomenon of deficiency of FIB….
29- Lines 178 to 181… phrase … Based on a previous study [26], rTM was administered intravenously at a dose of 1 mg/kg 10 min after yamakagashi venom administration. ATIII was administered intravenously at 20 U/rat, calculated from the clinical dose, 30 min after venom administration. This sentence should be written in the Materials and Methods section, not in the Discussion section.
30- Lines 190 to 192…. Considering these reports, ATIII might function as a secondary complement factor, albeit with limited efficacy, even if it does not directly affect yamakagashi venom. If the authors do not use rTM alone in the same experiment, they cannot compare the synergistic or additive activities of another drug used in combination with rTM. This is because rTM alone may produce the observed effect, and it may not be a case of additive or synergistic action.
31- Lines 193 and 194 … Although TXA did not exert therapeutic benefits alone, its combined use with rTM led to improvements in survival and blood coagulation markers in the rat DIC model. The same commentary of item 30 could be applied here. Conclusions32- Lines 213 and 214 … We investigated the effect of rTM and TXA in a rat DIC model induced by yamakagashi venom. Authors also studied ATIII and this can be pointed.
Author Response
To Reviewer 1
Thank you for the 32 insightful, detailed, and helpful comments you provided. I have responded to each comment as follows.
Specific Comments:
1- Between lines 22 to 24 Key Words. I would change the order in which the words appear in Key words: Yamakagashi (Rhabdophis tigrinus) venom; rat disseminated intravascular coagulation (DIC) model; recombinant thrombomodulin alpha; Tranexamic acids; antithrombin III; Combined therapy.
A: Change the order of key Words according to Reviewers comment.
2- Between lines 33 and 34 … Yamakagashi is a venomous snake belonging to the Colubridae family, and it is typified by short posterior fangs. Please include one or more references.
A: Added references.
3- Line 39 … Severe cases can be fatal without treatment, and deaths have been previously reported. Please include one or more references.
A: Added references.
4- Between lines 40-41 …. the high-molecular weight portion of the venom …Please change molecular weight to molecular mass throughout of the text, as proteins have masses instead weight. Although this term is accepted in the literature, it is wrong.
A: Changed “molecular-weight” to “molecular-mass”.
5- Between lines 40 to 43 … Regarding the chemical characteristics of yamakagashi venom, the high-molecular weight portion of the venom contains prothrombin activators. Exposure to its venom shortened both the prothrombin time (PT) and activated partial thromboplastin time of human plasma. Please include one or more references.
A: Added references.
6- In lines 66 in vitro and in line 68 in vivo, These are words of Latin origin and should be cited in the text in italics as in vitro and in vivo throughout of the text.
A: Change all “in vivo” and “in vitro” to italics.
7- Between lines 71 and 72 … Meanwhile, antithrombin III (ATIII) inhibits over activated blood coagulation and prevents thrombus formation in microvessels. Please include one or more references. I will review Materials and Methods first, before moving on to the other items.
A: Added references.
8- Line 219. Authors wrote … Male Sprague-Dawley rats (JAPAN SLC, Inc., Shizuoka, Japan) aged 12 weeks… Is there any particular reason why the authors used only male rats? And also the age of 12 weeks? Some explanation could be given throughout the text.
A:　 I added an explanation to the "Animal preparation" section regarding the reason for selecting the rat strain and age used.
9- Between lines 235 and 236 … Toxic glands were collected from 100 male and female snakes. The sentence is confusing because it doesn't make it clear whether 100 snakes were used in total or 100 male and 100 female animals? Please rewrite in the revised version.
A: Rewrite details of the method for extracting crude venom from Yamakagashi.
10- Between lines 236 to 238. … The glands were excised, cut into small pieces, and centrifuged with distilled water, after which the supernatant was lyophilized [25]. This description of how the venom was obtained seems somewhat vague to me. There was no maceration of the gland to release the venom. Just mixing it with distilled water? If so, the authors could refine this methodology so that other researchers can reproduce the method if interested.
A: Rewrite details of the method for extracting crude venom from Yamakagashi.
11- Between lines 245 and 246… In the model group, 300 μg of Yamakagashi venom were administered intramuscularly. It would be interesting if the authors detailed why this particular mass of venom was chosen and why the intramuscular injection route was used instead of intradermal or subcutaneous routes, which are more pathophysiologically similar to natural envenomation.
A: The details of the intramuscular administration of 300 μg of yamakagashi venom have been added.
12- Between lines 246 to 249 … In the TXA group, 5 mg of TXA were administered intravenously 30 min after venom administration. In the ATIII group, 20 U of ATIII were administered intravenously 30 min after venom exposure. Each group included six rats. Blood samples were collected at 0, 2, 4, 8, 24, 48, 72, 96, and 120 h after the start of the experiment. In my opinion, the authors could refine the information by indicating some explanation for why these parameters were used.
A: Regarding drug dosage, I have moved the section previously described in the discussion here and added information about the procedure for determining the time and amount of blood sample collection.
13- The same applies to text written between the lines 253 to 263. A more refined explanation of the parameters could be provided.
A: The drug dosages listed here have been moved from the discussion section to this page. A more refined explanation of the parameters was provided.
Results
14- About figure 1, line 88. In my opinion, the authors could include the meanings of the colored spheres in the figure, so that readers wouldn't need to read the legend to understand the results. This approach has been used more frequently in more recent publications and complements the information in the legend.
A: Following your suggestion, I have added an explanation of the meaning of the colored spheres in Figure 1.
15- Still figure 1… Generally, experiments with pharmacological treatments in live animals or cells follow a concentration-dependent curve. Is there any particular reason why the authors didn't test other concentrations of TXA (5 mg/rat) or ATIII (20 U/rat)?
A: The dosage for each anti-DIC drug was calculated by converting the maximum clinical dose in humans to a body weight equivalent in rats. If this dosage was ineffective, it would mean the anti-DIC drug could not be used in actual clinical practice, rendering this experiment meaningless.
16- The same question also applies to the time dependency of the treatments. Only one time frame was used.
A: Regarding the timing of administration for each anti-DIC drug, Reviewer 1's point is correct, but administering the drug 30 minutes after venom administration seems quite early from a clinical standpoint.
17- About figure 2, line 104. The same as comment for figure 1. The authors could include the meanings of the colored spheres in the figure.
A: Following your suggestion, I have added an explanation of the meaning of the colored spheres in Figure 2.
18- I'm simply asking the authors if these results weren't expected, given that treatment with the tested drugs didn't alter animal mortality after treatment with the venom.
A: The result that the anti-DIC drug shown in Figure 1 did not alter the mortality rate of the animals was unexpected.
19- 2.3, lines 110 and 111. In my opinion authors should rewrite the sentence to … Effects of rTM, ATIII or TXA alone, and TXA or ATIII in combination with rTM on survival in a rat model of DIC induced by yamakagashi venom.
A: I have changed the title as suggested.
20- About figure 3. The same as comment for figure 1. The authors could include the meanings of the colored spheres in the figure.
A: Following your suggestion, I have added an explanation of the meaning of the colored spheres in Figure 3.
21- About figure 3. The data shown in Figure 3 are very interesting, especially when analyzing the treatments of animals with venom and rTM plus TXA. I'm asking if analyses were performed over periods longer than 96 hours? Were the animals still protected?
A: As shown in Figure 3, the group receiving rTM and TXA in combination remained alive for more than 96 hours after venom administration, up to 144 hours after euthanasia.
22- About figure 3. Although the effects with the combination of rTM and ATIII were smaller, than combination of rTM and TXA, they also showed some protection. Did the authors not perform variations in these combinations seeking better results? Once again, tests studying combinations at a single concentration of the drugs studied may be insufficient for more detailed conclusions as previously commented.
A: The combination therapy group of rTM and ATIII shown in Figure 3 demonstrated extended survival in approximately half of the animals. While increasing the ATIII dosage is an option, as pointed out, we did not do so based on the meaning of comment #15 below:
The dosage for each anti-DIC drug was calculated by converting the maximum clinical dose in humans to a body weight equivalent to rats. If this dosage was ineffective, it would mean the anti-DIC drug could not be used in actual clinical practice, rendering this experiment meaningless.
23- About figure 3. I would like to know the authors' opinion on whether there was a lack of control over the use of rTM alone, because if there is treatment in combination with other drugs, shouldn't there also be treatment alone? This is an essential control for drawing conclusions about additive or synergistic activities.
A: Your point is correct. We did not include a group receiving rTM alone as a concurrent experimental group in this study. However, we did not achieve complete therapeutic efficacy with rTM alone in another experiment. This is currently being documented in a separate paper.
24- Lines 116 and 117…authors wrote…Conversely, TXA (5 mg/rat) or ATIII (20 U/rat) alone did not improve rat survival. But rTM alone? For comparative analyses using two　drugs authors must test each one separate and in combination.
A: Your point is correct. We did not include a group receiving rTM alone as a concurrent experimental group in this study. However, we did not achieve complete therapeutic efficacy with rTM alone in another experiment. This is currently being documented in a separate paper.
25- About figure 4. The same comment made for Figure 3 can be applied to Figure 4. The authors failed to test rTM alone to verify if there is any additive or synergistic activity with the other drugs tested.
A: Your point is correct. We did not include a group receiving rTM alone as a concurrent experimental group in this study. However, we did not achieve complete therapeutic efficacy with rTM alone in another experiment. This is currently being documented in a separate paper.
Discussion
26- Lines 150 to 152…The dose of each anti DIC drug was calculated by converting the maximum clinical dose in humans to the weight-based equivalent in rats. This sentence should be written in the Materials and Methods section, not in the Discussion section. The Discussion section is for interpreting the results and mechanistic hypotheses, but not for methodology.
A: Following the comments, I moved my personal writing to "Materials and Methods".
27- Lines 153 to 159 …TXA binds strongly to the lysine-binding site, which is the fibrin affinity site of plasmin and its precursor plasminogen, which participate in fibrinolysis occurring after blood coagulation. Through this binding, TXA prevents plasmin and plasminogen from binding to fibrin, thereby strongly inhibiting fibrin deg radation by plasmin. Furthermore, the antifibrinolytic effect of TXA is further enhanced in the presence of plasma antiplasmins such as α2-macroglobulin. Plasmin inhibits platelet aggregation and degrades coagulation factors, but it specifically induces fibrin degradation. Please include one or more references to sentence written.
A: Based on the points you raised, I have added two references.
28- Lines 167 to 169 …phrase is confuse … It is speculated that accelerated coagulation occurs rats administered venom, resulting in the secondary phenomenon of deficiency of FIB coagulation factors [31]. Better change to …. It is speculated that accelerated coagulation that occurs in rats administered with venom, resulting in the secondary phenomenon of deficiency of FIB….
A: I have replaced the text with the suggested version, taking into account the points you raised.
29- Lines 178 to 181… phrase … Based on a previous study [26], rTM was administered intravenously at a dose of 1 mg/kg 10 min after yamakagashi venom administration. ATIII was administered intravenously at 20 U/rat, calculated from the clinical dose, 30 min after venom administration. This sentence should be written in the Materials and Methods section, not in the Discussion section.
A: Following the comments, I moved my personal writing to "Materials and Methods".
30- Lines 190 to 192…. Considering these reports, ATIII might function as a secondary complement factor, albeit with limited efficacy, even if it does not directly affect yamakagashi venom. If the authors do not use rTM alone in the same experiment, they cannot compare the synergistic or additive activities of another drug used in combination with rTM. This is because rTM alone may produce the observed effect, and it may not be a case of additive or synergistic action.
A: Your point is correct. We did not include a group receiving rTM alone as a concurrent experimental group in this study. However, we did not achieve complete therapeutic efficacy with rTM alone in another experiment. This is currently being documented in a separate paper.
31- Lines 193 and 194 … Although TXA did not exert therapeutic benefits alone, its combined use with rTM led to improvements in survival and blood coagulation markers in the rat DIC model. The same commentary of item 30 could be applied here.
A: Your point is correct. We did not include a group receiving rTM alone as a concurrent experimental group in this study. However, we did not achieve complete therapeutic efficacy with rTM alone in another experiment. This is currently being documented in a separate paper.
Conclusions
32- Lines 213 and 214 … We investigated the effect of rTM and TXA in a rat DIC model induced by yamakagashi venom. Authors also studied ATIII and this can be pointed.
A: A description regarding the combined administration of ATIII and rTM was added to the conclusion.
Author Response File: 
Author Response.pdf
Reviewer 2 Report
Comments and Suggestions for AuthorsThis study examines the effectiveness of combining recombinant thrombomodulin (rTM) with standard treatments for disseminated intravascular coagulation in reversing the coagulopathy caused by a colubrid snake venom in a murine model, supporting further investigation of this combination therapy. This manuscript seems to be a follow up from in vitro studies, where rTM neutralized the pro-coagulant mechanism of the venom, and provides an important alternative route to treat these envenomings, considering the lack of an approved antivenom.
Despite the relevance of the study, the authors must improve the manuscript and the data presented to make this paper suitable for publication. First, the term disseminated intravascular coagulation (DIC) for the pathophysiology caused by the Yamakagashi venom is inaccurate. It has been well established and documented that snake venom toxins caused a coagulopathy known as venom-induced consumption coagulopathy (VICC), which is a distinct process that lacks the initiation mechanism and outcomes seen in DIC. Please revise this information and modify the content accordingly.
In methodology and discussion, the authors must explain the experimental design beyond referring solely to a previous publication. Why do the authors mention the lethal dose 50 (LD50) after i.v. injection, if the route employed was i.m.? The LD50 is usually expressed in mg/kg.
How were the blood samples collected?
Given the study designs, ANOVA followed by appropriate multiple comparisons should be used. Similarly, please consider plotting individual data points.
The data presentation should be improved. Specify in the legends against what the statistical analysis was performed and include in the graphs themselves a chart explaining to which experimental group each color belongs to. Moreover, the results could be summarized into two figures to avoid repetitive data.
In discussion, there is a lack of information about the pathophysiology of this envenoming model regarding the venom composition (which toxins besides metalloproteinases could be involved in the hemostatic dysregulation) and the pro-fibrinolytic components involved, to support the drugs chosen as potential treatment.
Author Response
To Reviewer ï¼’
Thank you for your five valuable and helpful comments. I will respond to each comment as follows:
- Despite the relevance of the study, the authors must improve the manuscript and the data presented to make this paper suitable for publication. First, the term disseminated intravascular coagulation (DIC) for the pathophysiology caused by the Yamakagashi venom is inaccurate. It has been well established and documented that snake venom toxins caused a coagulopathy known as venom-induced consumption coagulopathy (VICC), which is a distinct process that lacks the initiation mechanism and outcomes seen in DIC. Please revise this information and modify the content accordingly.
A: Following your feedback, I have corrected the expression in the text from DIC model to VICC model. I have also rewritten the blood coagulation disorder caused by yamakagashi bites as VICC. Furthermore, since there are no anti-VICC drugs other than antitoxin preparations, I have added a statement to the objectives and discussion to indicate that the administration of anti-DIC drugs, which is the purpose of this study, will be applied to the treatment of yamakagashi bites.
- In methodology and discussion, the authors must explain the experimental design beyond referring solely to a previous publication. Why do the authors mention the lethal dose 50 (LD50) after i.v. injection, if the route employed was i.m.? The LD50 is usually expressed in mg/kg.
A: I have made revisions to the methodology and considerations you pointed out. I have also corrected the notation for LD50.
３) How were the blood samples collected?
A: Blood samples were collected by inserting a cannula into the femoral artery of rats.
4) Given the study designs, ANOVA followed by appropriate multiple comparisons should be used. Similarly, please consider plotting individual data points.
The data presentation should be improved. Specify in the legends against what the statistical analysis was performed and include in the graphs themselves a chart explaining to which experimental group each color belongs to. Moreover, the results could be summarized into two figures to avoid repetitive data.
A: We have made some corrections to the way the figures are displayed, as pointed out, and added statistical methods such as significance testing to the legend.
5) In discussion, there is a lack of information about the pathophysiology of this envenoming model regarding the venom composition (which toxins besides metalloproteinases could be involved in the hemostatic dysregulation) and the pro-fibrinolytic components involved, to support the drugs chosen as potential treatment.
A: Following your suggestion, I have added a description and discussion regarding toxic components other than metalloproteinases contained in yamakagashi venom
Author Response File: Author Response.pdf
Round 2
Reviewer 1 Report
Comments and Suggestions for AuthorsAnswer to Authors
Dear authors, after carefully reading the reply letter sent, it is my opinion that the responses were technically coherent and extremely polite, making the revised text more complete from a scientific point of view. My concern was regarding the absence of an experimental group where rTM had been tested alone, and regarding this, the response you provided indicates that you have already studied this question, or are preparing a new manuscript in this regard. I understand that this gap needs to be addressed in the literature to truly affirm the synergistic or additive potential of the new therapy indicated. In my opinion, the new text has the potential to be approved for publication by TOXINS.
Author Response
Dear authors, after carefully reading the reply letter sent, it is my opinion that the responses were technically coherent and extremely polite, making the revised text more complete from a scientific point of view. My concern was regarding the absence of an experimental group where rTM had been tested alone, and regarding this, the response you provided indicates that you have already studied this question, or are preparing a new manuscript in this regard. I understand that this gap needs to be addressed in the literature to truly affirm the synergistic or additive potential of the new therapy indicated. In my opinion, the new text has the potential to be approved for publication by TOXINS.
Thank you so much for your insightful comments in your initial feedback, and for your continued appreciation of our approach in this latest comment. Thank you also for your understanding of our explanation regarding rTM monotherapy, which you had concerns about. Your numerous suggestions for improvement have greatly contributed to the development of our paper. Thank you so much.
Reviewer 2 Report
Comments and Suggestions for AuthorsThe authors have addressed the majority of the comments from the previous version. However, it still needs to improve the following:
Figures have been improved; however, authors need to remove the redundancy in the legends. It is enough to indicate the samples one time. The legend in Figure 3 is incomplete, only showing 3 groups out of 5, also, the blue color does not allow to distinguish well between the venom control group and the rTM/ATIII treated group. Please consider plotting individual data points for each group since it will allow visualizing better the trend of the results.
In the experimental protocol 5.3, revise the following sentence “The model groups used were those previously reported to have been used to investigate the routes and doses of toxin administration for the creation of a rat DIC model using yamakagashi venom”, by modifying DIC with VICC. Furthermore, revise line 277 where it says “300 μg of Japanese keelback snake venom” instead of “300 μg of yamakagashi venom”
Author Response
The authors have addressed the majority of the comments from the previous version. However, it still needs to improve the following:
A: Thank you for your specific comments, Professor. I have addressed the following two points you raised.
- Figures have been improved; however, authors need to remove the redundancy in the legends. It is enough to indicate the samples one time. The legend in Figure 3 is incomplete, only showing 3 groups out of 5, also, the blue color does not allow to distinguish well between the venom control group and the rTM/ATIII treated group. Please consider plotting individual data points for each group since it will allow visualizing better the trend of the results.
A: Thank you for your comments regarding the legend in Figure 3 and the data points for each group.
First, I have revised the legend in Figure 3 so that all five groups are visible. Also, to improve clarity, I have highlighted the data points for the rTM/ATIII administration group in yellow. The explanations for the data points in Figures 3 and 4 for the rTM/ATIII administration group have also been corrected from light blue to yellow.
Furthermore, I have standardized the descriptions of all data points and legends in Figures 1 through 4. Thank you for pointing this out.
- In the experimental protocol 5.3, revise the following sentence “The model groups used were those previously reported to have been used to investigate the routes and doses of toxin administration for the creation of a rat DIC model using yamakagashi venom”, by modifying DIC with VICC. Furthermore, revise line 277 where it says “300 μg of Japanese keelback snake venom” instead of “300 μg of yamakagashi venom”
A: Thank you for pointing out the errors in the text.
The corrected parts have been replaced with blue text.
