ATOM INDONESIA
Author's Responses
Article : #486
Name of All Authors :
Article Title : Technetium-99m Labeled Diethyl Carbamazine Citrate (99mTc-DEC) As a New Diagnostic Agent for Lymphatic Filariasis Detection
E-mail :
Line # Referee’s Comments Author's Responses
12-13
….bedside the ….values.
….detection is therefore needed.
….99mTc-DEC has been successfully prepared.
…requirement have to be evaluated.
The result of this innovation is expected to give a real contribution in improving of public health,
particularly in early detection of filariasis infection.
In addition this result is expected to be a concrete
….
It has been revised in accordance suggestions
22
Fig.1.A become Fig. 1 has to be placed a close as possible to the text which discussed the related matter. It is suggested to place Fig.1 just after 1st paragraph.
Editor’s report suggest that Fig 1 is not necessary since almost everybody knows what lymphatic filariasis looks like. Therefore the image of filariasis patient was removed.
22-23
It resulted in a significant impact…
It has been revised in accordance suggestions
39-40
This is why the World Health Organisation…
It has been revised in accordance suggestions
49-50
…between 10 pm and 2 a.m (nocturnal), therefore…
It has been revised in accordance suggestions
53 ± 4 years….it has to be written as …..around 4 years…..or… ~ 4 years….
It has been revised from ±4 years to around 4 years
56 Exx Lxx Ixx Sxx Axx (ELISA)…
It Has been added to Enzyme-Linked Immunosorbent Assay
56-57
Rapid Immuno Chromatography Card
It has been revised in accordance suggestions
63-64
…..gave an alternative expectation to solve this problem.
Or could be
…..gave an alternative expectation in solving this
It has been revised to….. gave an alternative expectation to solve this problem.
problem.
65 ….that is commonly used It has been revised in accordance
suggestions
66
…in humans and animals (Fig. 2). Fig.2. is a chemical structure of DEC. Please place Fig.2 just after 2nd paragraph (line 81)
Fig of chemical structure of DEC has been added (Fig 1)
69
…….W. bancrofti, xxxxxxxxis, do not apper…..
…it has to be explained what is W. bancrofti between two commas… as it is not clear what is its relation with lymphatic filariasis
It has been added …. W. bancrofti, human parasitic roundworm that cause lymphatic filariasis, do not appear…
72 ….to form the organo-metal… It has been revised in accordance suggestions
74 …. 99mTc-DEC is expected to be uptaken by microfilariae.. It has been revised in accordance suggestions
78 ….could be detected… It has been revised in accordance
suggestions 102 …in the previous research [Reference].
Reference has been added (ref. 17)
103 This is why…. It has been revised in accordance
suggestions
112
This paragraph should be moved up/cooporated to previous paragraph. ….to be studied. Uptake of 99mTc- DEC by filaria is also important to be studied. The verification could be done not only through in-vitro studies, but also by in-vivo testing in animals infected with filaria.
In-vitro and in-vivo have to be written in italic as they are not English but Latin
It has been revised in accordance suggestions
119 (end user) It has been revised in accordance
suggestions
121- 127
….of filariasis. The use of this new radiopharmaceutical is expected to give a real contribution in solving the
present problems in term of filariasis infection of the Indonesian community. This in turn will be supporting
"The Global Goal of Elimination of Lymphatic Filariasis as a Public Health Problem by the Year 2020 "program in particular and the Indonesian health program in general.
It has been revised in accordance suggestions
145 ….in two forms which are in fresh solution and from a
freeze dried kit It has been revised in accordance
suggestions
133- 156
The labeling of DEC with 99mTc was conducted using a direct method. Two forms consisting 4 mg DEC-citrate, 100 mg of SnCl2.2H2O, pH 4 were prepared. The first formula was freshly prepared and the second one was freeze dried and re-constituted just prior addition of
99mTc. Labeling of DEC with 99mTc- was performed by adding aliquot of 99mTc-pertechnetate into the above- mentioned forms which was followed by incubation at
It has been revised in accordance suggestions
room temperature for 10-20 minutes. The results of radiolabeling which is stated as radiochemical purity was determined using two-system chromatography. First system was chromatography using Whatman 3MM paper (1x10 cm) as stationary phase and dry acetone as mobile phase. This system was for separating of 99mTc-DEC from free 99mTc in form of 99mTc-pertechnetat (99mTcO4-). The second system was chromatography using ITLC-SG (??) (1x10 cm) as stationary phase and water as mobile phase. This system was for separating 99mTc-DEC from free 99mTc in its reduced form (TcO2). The degree of radiochemical purity labeling of 99mTc-DEC could be known by:
135- 139
This statement should be moved to the next paragraph.
(Line 161)
The stability of 99m Tc-DEC (indicated as radiochemical purity) after storage at a certain period of time was tested with the above-mentioned chromatographic methods. Stability test of ……
It has been revised in accordance suggestions
161
Stability test of freeze dried DEC/ DEC kit over certain period of time during 7 months of storage was also performed. The good stability is expressed when both labeling efficiency and purity level is higher than 90 %, and there is no changes in term of physico-chemical and biological performance during storage. The test was performed in similar manner to the radiolabeling procedure as previously mentioned which was then followed by radiochemical purity determination.
In addition, the decrease of radiochemical purity of Tc- DEC was also observed in interval specified period of time after reconstitution with the addition 99mTc–
pertechnetate. Not quite clear the meaning of this sentence I think it needs to be removed.
It has been revised in accordance suggestions
173 Biological tests on animal were conducted….. It has been revised in accordance suggestions
175 …..(KEPPHP-BATAN) No.????? Number of KEPPHP-BATAN has
been added
177 Sterility test of DEC kits. It has been revised in accordance suggestions
178
Sterility test was performed according to the procedure in Indonesian
Pharmacopoeia and published journal using two types of media which are Fluid Thioglycolate Media (FTM) for aerobic and anaerobic bacteria and Soybean Casein Digest Media (SCDM) for fungus/mold [References].
It has been revised in accordance Suggestions and reference has been added (ref 18)
183- 187
Two vial (??) DEC kits were re-constitute with one mL of saline solution. Each solution was then added into a glass tubes consisting 5 mL of FTM and 5 mL of SCDM
respectively.
It has been revised in accordance Suggestions
2 vial from 1 batch (consist of 25 DEC kits) using for sterility test
How many vial DEC kit were prepared? How many samples were taken for sterility test? Is the number of samples represent the number of kits??
According to Indonesia
Pharmacopeia 2 pieces of sample from 25 pieces were stated quite representative. If from the first test the result is not sterile then the test is repeated with the number of samples doubled.
187 FTM tubes were incubated….… It has been revised in accordance Suggestions
190- 193
Media tubes were observed up to 14 days, if there were no growth of bacteria or mold colony, then DEC kit was sterile. However, if there were a growth bacteria or mold which was indicated…
It has been revised in accordance Suggestions
197- 199
…vials of DEC kits; and if after 14 days of observation ther It has been revised in accordance Suggestions
205 …with a slight modification [17]…. It has been revised in accordance Suggestions
212 ….for up to 14 days, It has been revised in accordance
Suggestions
218- 226
I think Figure 1-A to 1-F should be labeled as Figure 2A- 2E as Fig.1A would be place just after paragraph 1(see Line 22)
Fig. 2-A to 2-E, show the sequence of events of induction activities through the injection of the blood patients infected by microfilariae to the test animals in the middle of the night (after 22:00 pm).
Blood sample from five filarial volunteers (Figure 2B) was collected and 1 mL was injected to 10 experimental Wistar rats (250-300 g) on the same day (Figure 2F).
The remain blood sample was analyzed in laboratory with microscope (Nicon) (Fig.2-C & 2-D)
It has been revised in accordance Suggestions but Fig 2A was removed
231
….every day and the blood sample was taken and
analyzed ……… It has been revised in accordance
Suggestions
237- 246
Note: Be consistent!! In British English one digit number always written as one, two…etc.
However in American English one digit number could be written as 1, 2, 3 …etc. Therefore, if one style has been chosen…be consistent with that choice. Please have a look the whole article for this matter!!
Three months post infected, three experimental rats were injected intravenously with
1 mL of 1 mCi/ ml of 99mTc-DEC, and three others were injected intra-dermally with 0.1 mL of the same
radioactive concentration of 99mTc-DEC. After 15 minutes post injection, the rats were anesthetized by Ketamine injection solution to death which was followed
It has been revised in accordance Suggestions using an American English
by dissection. Several critical organs were taken, and radioactivity accumulation of 99mTc-DEC determined by using Single Channel Analyzer.
253
After intradermal and intravenous administration of
99mTc-DEC, blood sample was taken from the tail vein of each rat at the periodic time of 5; 15; 30; 45; 60; 120 and 180 minutes post injection. Blood samples were weighed and their radioactivities measured with a Single Channel Analyzer. The biological half-life was then calculated from the kinetic profile of 99mTc-DEC by using of computerized Multivit program.
It has been revised in accordance Suggestions
264- 266
Was performed on …..…0.1 mCi (0.1 mL) of… It has been revised in accordance Suggestions
268- 271
Should be removed
It has been removed
277- 281
Fig.2 Fig 3.
Fig.3 is a picture of DEC kits that have been prepared and used throughout this research. These kits are easily distributed to the Nuclear Medicine Department in hospitals for preparing of 99mTc-DEC radiopharmaceutical
It has been revised in accordance Suggestions
294 Fig. 3. DEC Kits It has been revised in accordance
Suggestions
296 The stability of DEC kits indicated by ... It has been revised in accordance Suggestions
304
Table 1. Labeling efficiency and radiochemical purity levels of DEC kits
(in storage) labeled with 99mTc It has been revised in accordance
Suggestions 305-
306
Kolom title :
Efficiency Labeling and Radiochemical Purity ( % ) should be change to
Labeling Efficiency and Radiochemical Purity ( %)
It has been revised in accordance Suggestions
308- 311
Table 2 shows that labeling efficiency and radiochemical purity of DEC kits (stored for up 7 month) labeled with
99mTc were higher than 90%. This indicated that DEC kits were stable during this period of storage.
These results need to be compared with other Tc-99m based kits. More stable or less stable???.
It has been revised in accordance Suggestions and has been compared with 99mTc-ciprofloxacin
312- 322
Stability of DEC kit labeled with 99mTc (99mTc-DEC) when stored for up to 4 hours is shown in Table 2. This results show that 99mTc-DEC stored for up to 2 hours still have acceptable radiochemical purity (>90%). The decrease of radiochemical purity levels of 99mTc-DEC during storage might be caused by radiolysis effect or conformation of the TC-DEC complex itself. However when the stability of
99mTc-DEC compared with other Tc-99m based radiopharmaceuticals this complex was……..?????
These results need to be compared with other Tc-99m based radiopharmaceuticals. More stable or less
It has been revised in accordance Suggestions and has been compared with 99mTc-ciprofloxacin
stable???.
336- 343
Please check!!! There is procedure for pyroginity test in Experimental method….need to be added.
Pyrogenity and sterility test of 99mTc-DEC of fresh (recentus paratus) or kit form that have been stored for up to 8 months were found to be sterile and pyrogen free.
Line 339- 343 Started from ……..on the other hand…….as possible. Need to be moved to the next paragraph.
It has been revised in accordance Suggestions and procedure for pyroginity test has been added Line 339- 343 has been moved
344
There has been worry that radiolabeling of DEC with large volume of 99mTc-pertechnetate might hinder the radiolabeling process. Therefore radiolabeling of EDC with 99mTc-pertechnetate with volume ranged between 1 – 4 mL was carried out. Table 3 shows the influence of volume of 99mTc-pertechnetate slightly influenced the radiobeling efficiency of DEC. It can been seen that radiobeling with volume of 99mTc-pertechnetate larger than 1 mL decreased the radiobeling efficiency to 92.23 – 94.58%. However, these results …..
It has been revised in accordance Suggestions
351- 352
Effect of volume of 99m Tc-pertechnetate on….. It has been revised in accordance Suggestions
367 …up to 14 days in order to notice if there were… It has been revised in accordance Suggestions
370
This paragraph need to be moved to line 369.
Data shown that there was no abnormalities or death on test mice although…..
…of acute toxicity it can be stated that …….is safe to be used…
It has been revised in accordance Suggestions
390 Figure. Biodistribution of 99mTc - DEC in rats after intradermal injection It has been revised in accordance Suggestions
393
…..such as as lumbar and mesenteric. Significant
radioactivity was also found accumulating in the kidneys.
This can be understood as kidney is a main excretory organ.
I think this paragraph needs more explanation regarding the biodistribution of 99mTc – DEC. Is this patern due to the nature of 99mTc – DEC or something else.
It has been revised in accordance Suggestions, more explaination has been added
410 …arised in similar pattern for all… It has been revised in accordance
Suggestions 421
…of 99mTc. Radioactivity concentration in blood from
intradermal route… It has been revised in accordance
Suggestions 428-
426
These figures need to be labelled as Fig 5A and 5B It has been revised in accordance Suggestions, figure has been labelled as 6A and 6B
429 …at 44.32 minutes (Fig 5A)…..by 37.83 minutes (Fig 5B). It has been revised in accordance Suggestions
435- 444
99mTc-DEC which was administered via intradermal injection seem having relatively long biological half-life.
This indicated that 99mTc-DEC was retained in the body for some time. On one hand this value…. management.
However, on the other hand administration via intradermal injection would cause a difficulty in distinguishing….
It has been revised in accordance Suggestions
448
Figure 6. shows that……minutes. ….to discover the uptake of 99mTc-DEC
in the body when an imaging was performed a using a gamma camera.
It has been revised in accordance Suggestions
483- 498
I am not sure if this paragraph is really necessary in this
article. Please consider to remove it!! It has been removed 501-
510
Given that…cases, it is hoped….. It has been revised in accordance Suggestions
514- 518
…of DEC in freeze dried and sterile form ….with 99mTc to
produce 99mTc-DEC for diagnosis filariasis infection. It has been revised in accordance Suggestions
518
In-vitro and in-vivo evaluation such as testing stability of
99mTc-DEC, biodistribution, pharmacokinetic and toxicological studies which have been…..
It has been revised in accordance Suggestions
