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To: Tatas H.P Brotosudarmo, Ph.d <[email protected]>
08-Jun-2020
Dear Dr THP Brotosudarmo:
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MANUSCRIPT DETAILS
AUTHORS: Gholami, Asma; Minai-Tehrani, Dariush; Eftekhar, Fereshteh
ABSTRACT: Lipase hydrolyzes the ester bonds in triglyceride. It is an important enzyme in medicine and industry. Some pathogen bacteria use this exoenzyme to disrupt the extracellular matrix of host organisms. Pseudomonas uses various extracellular enzymes such as lipase to invade its host. In this report, for the first time, bromhexine was
introduced as an inhibitor of lipase. Bromhexine is a mucolytic drug which is used in the treatment of respiratory tract disorders. The results showed that bromhexine inhibited the enzyme by competitive inhibition. IC50 and Ki values of the drug were 0.049 mM and 0.02 mM, respectively. Arrhenius plot showed that the drug reduced activation energy.
The enzyme was purified and SDS-PAGE showed that its molecular weight is 13 kDa.
Fluorescence measurement revealed that binding of the drug to lipase could make structural changes in the enzyme. Inhibition of lipase by bromhexine could be applicable in medicine.
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To: Tatas H.P Brotosudarmo, Ph.d <[email protected]>
08-Jun-2020
Dear Dr THP Brotosudarmo:
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Agreeing to review an article for this Journal implies that you as the reviewer will adhere to the accepted ethical standards of scientific, medical and academic publishing. Material submitted for peer review is a privileged communication that should be treated in
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Thank you for submitting your review of Manuscript ID NAPB-2020-0475 for Archives Of Physiology And Biochemistry
Archives Of Physiology And Biochemistry <[email protected]>
Tue 09/06/2020 10E05 AM
To: Tatas H.P Brotosudarmo, Ph.d <[email protected]>
08-Jun-2020
Dear Dr THP Brotosudarmo:
Thank you for reviewing the above manuscript, entitled "Bromehoxine and its inhibitory effect on lipase; kinetics and structural study" for Archives Of Physiology And
Biochemistry.
We greatly appreciate the voluntary contribution that each reviewer gives to the Journal.
We hope that we may continue to seek your assistance with the refereeing process for Archives Of Physiology And Biochemistry, and hope also to receive your own research papers that are appropriate to our aims and scope.
Sincerely, Dr Müller
Editor in Chief, Archives Of Physiology And Biochemistry [email protected]
For Peer Review Only
Bromehoxine and its inhibitory effect on lipase; kinetics and structural study
Journal: Archives Of Physiology And Biochemistry Manuscript ID NAPB-2020-0475
Manuscript Type: Original Paper Date Submitted by the
Author: 06-Jun-2020
Complete List of Authors: Gholami, Asma; Shahid Beheshti University
Minai-Tehrani, Dariush; Shahid Beheshti University, Life Science
&Biotechnology
Eftekhar, Fereshteh; Shahid Beheshti University Keywords: Enzyme, lipase, Inhibition, drug
For Peer Review Only
Bromehoxine and its inhibitory effect on lipase; kinetics and structural study
Asma Gholami1, Dariush Minai-Tehrani1*, Fereshteh Eftekhar1
1- Faculty of Life Sciences & Biotechnology, Shahid Beheshti University, GC, Tehran, Iran
*Corresponding author: Dariush Minai-Tehrani (PhD),
Email: [email protected] ORCID: 0000-0003-3589-7324 Tel: +98-21-29903144
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Abstract:
Lipase hydrolyzes the ester bonds in triglyceride. It is an important enzyme in medicine and industry. Some pathogen bacteria use this exoenzyme to disrupt the extracellular matrix of host organisms. Pseudomonas uses various extracellular enzymes such as lipase to invade its host. In this report, for the first time, bromhexine was introduced as an inhibitor of lipase. Bromhexine is a mucolytic drug which is used in the treatment of respiratory tract disorders. The results showed that bromhexine inhibited the enzyme by competitive inhibition. IC50 and Ki values of the drug were 0.049 mM and 0.02 mM, respectively. Arrhenius plot showed that the drug reduced activation energy. The enzyme was purified and SDS-PAGE showed that its molecular weight is 13 kDa. Fluorescence measurement revealed that binding of the drug to lipase could make structural changes in the enzyme. Inhibition of lipase by bromhexine could be applicable in medicine.
Keyword: Enzyme, lipase, inhibition, drug.
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1- Introduction:
Lipase is an important enzyme in industry and medicine. It widely uses in industries such as processing of fats and oils, detergents, food processing, leather industry and production of cosmetics (Rubin and Dennis 1997). In medicine, lipase plays a key role in production of specialty lipids and digestive aids (Vulfson 1994). For example S. marcescens lipase was used for the asymmetric hydrolysis of 3-phenylglycidic acid ester which is a key intermediate in the synthesis of diltiazem hydrochloride (Matsumae et al 1993. The inhibition of lipase may be useful for the persons suffering from obesity. Orlistat is an inhibitor of pancreatic lipase which is used for decreasing lipid absorption by obese people (Zhi et al 1995).
Bacterial lipases are valuable enzymes in biotechnology, because of the versatility of their application and simplicity of high production. Among bacteria, Pseudomonas aeruginosa lipase is highly active and applicable in industries and medicine (Buisman et al 1998).
Pseudomonas aeruginosa is a Gram-negative and opportunistic pathogenic bacterium which is the important cause of infection, particularly in the patients with cystic fibrosis, burn patients, or hospitalized in intensive care units (Lyczak et al 2002, de Bentzmann and Plsiat 2011, Pollack 2000). In this regard, P. aeruginosa uses some extracellular enzymes to degrade the extracellular matrix of host patients. The important extracellular enzymes produced by P. aeruginosa are protease (Frimmersdorf et al 2011), elastase A and B (Cryz and Iglewski 1980), phospholipase C (Bever and Iglewski 1988) and lipases (Ostroff and Vasil 1987, Stuer et al 1986). Some lipases are expressed and secreted by pathogenic organisms during the infection which causes biofilm in skin surface of patients. As a result, any agents that inhibit lipase activity may be important for medicine. Drugs are made to interact with certain receptors or enzymes, but irregularly; some drugs may bind to other enzymes or molecules in the body, which causes some side effect. Bromhexine is a synthetic substance obtained as a synthetic analogue of vasicine, a substance found in a plant called Adhatoda vasica (Fig 1). It is a mucolytic agent which is used in the treatment of respiratory disorders associated with viscid or excessive mucus (Ellis and West 1970). In addition, bromhexine has antioxidant properties (Felix et al 1996) and disrupts the structure of acid mucopolysaccharide fibers in mucoid sputum and produces less viscous mucus. Bromhexine hydrochloride is rapidly absorbed from the gastrointestinal tract and undergoes extensive first-pass metabolism in the liver.
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The aim of this study is to introduce bromhexine as a new inhibitor for lipase. Kinetic parameters of inhibition and the type of inhibitor binding to the enzyme are also investigated. The object of this research may be useful for inhibition of fat absorption, and also refusing biofilm formation by pathogenic bacteria in patients.
2- MATERIALS AND METHODS 2-1- Materials
The chemicals used for culture media and buffer preparation were of reagent grade and obtained from Merck Company. Para-nitrophenyl palmitate (pNPP) was obtained from Sigma Chemical Company. Bromhexine was of pharmaceutical grade and obtained from Chemidaru Company.
Culture medium and cell harvesting
The salt medium was used by adding 2.5 g KH2PO4, 2.5 g Na2HPO4, 1 g NH4NO3, 0.2 g MgSO4, and 0.01 g CaCO3 to 1 litre of distilled water and pH was adjusted to 7.0. Olive oil was used as the carbon source with the final concentration of 1%. The culture medium was aerated in a rotary shaker at 30°C for 72 h. The cells were precipitated by centrifugation and the supernatant was used for enzyme assay.
2-2- Enzyme assay
The buffer for enzyme assay was prepared by adding different concentrations (0.06 to 0.5mM) of para-nitrophenyl palmitate (soluble in isopropanol) as the substrate to 0.1 M Tris buffer pH 8 containing 0.1% (v/v) Tween 80. The reaction was started by adding 20 µl of the supernatant to the working buffer in the test tube. Final volume in the test tube was always 2 ml. The lipase catalytic activity was continuously checked by following the increased yellow color of the product (p-nitrophenol) with absorption at 410 nm (Talebi et al 2018). The enzyme assay was detected in the absence and presence of bromhexine with concentrations 0.048 to 0.32 mM. The assay was monitored for 10 min; this time was enough for the enzyme to reach the plateau in the progress curve (substrate depletion). The extinction coefficient of the product, p-nitrophenol (18.8 × 103 mol-1.cm-1), was used to determine its concentration and enzyme activity.
Lineweaver-Burk plot was applied to obtain the kinetic parameters of the enzyme and type of inhibition. All the assays were carried out at room temperature (25-28oC) using a Perkin-Elmer
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visible spectrophotometer. Lowry method was used for protein determination. All the assays were repeated as three separated experiments.
The enzyme activity was also measured at different pH (pH 5 -10) and temperatures (0-80oC).
Arrhenius plot was used to compare the activation energy of the enzyme in the presence and absence of the drug by calculating the Vmax of the enzyme at different temperatures.
2-3- Enzyme purification
The enzyme purification was performed by adding 60% ammonium sulphate to medium culture supernatant which had lipase. The solution was centrifuged and the precipitates were dissolved in 1 ml of 0.1 M Tris-base buffer (pH 8) and dialyzed against the same buffer. The dialyzed solution was loaded onto the DEAE cellulose column equilibrated with 0.05M Tris- base buffer pH 8. A gradient of NaCl was used for elution of the enzyme from the column. The fractions were checked for lipase activity and protein concentration. SDS-PAGE electrophoresis was done for the active fractions to verify lipase purification.
2-4- Fluorescence measurements
The pure enzyme was used for structural study by using fluorescence spectrophotometer (Perkin Elmer fluorometer). The enzyme concentration was always 0.1 mg/ml. Structural study was performed with pure enzyme either in presence or absence of bromhexine. The excitation wavelength was at 285 nm and the emission spectra were recorded between 300 and 450 nm.
3- Results:
3-1- Kinetic parameters
Double reciprocal plot was used to resolve the kinetic parameters of the enzyme and type of inhibition. Fig. 2 shows that, in the presence of bromhexine, the Vmax of the enzyme was constant (Vmax = 2 mmol/min/mg protein), while Km was decreased by increasing the concentration of the drug. A competitive type of inhibition was observed in the enzyme in the presence of the drug. IC50 of the drug is an important factor which shows the concentration of the drug that reduces 50% of the affinity of the enzyme for its substrate (Fig 3). The results demonstrated that IC50 of bromhexine was about 0.049 mM, which determined that the drug could inhibit the enzyme with high affinity.
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To resolve Ki value (inhibition constant) of the drug, a secondary plot from the primary plot (Lineweaver–Burk, Fig. 2) was used. The slopes of line of primary plot were utilized for secondary plot (Fig. 4). The Ki value was determined to be about 0.02 mM
3-2- Effect of temperature
The enzyme activity was determined at different temperatures, both in the absence and presence of the drug. The maximum temperature was observed at 60oC in the absence of the drug, while in the presence of the drug, a plateau from 40 to 60oC was observed (Fig. 5). The enzyme was inactivated at 80oC. Arrhenius plot was used to compare the activation energy of the reaction.
Vmax of the enzyme was calculated at different temperatures for drawing the Arrhenius plot (Fig. 6). The results demonstrated that the activation energy was increased in the presence of the drug.
3-3- Effect of pH
Maximum activity was observed at pH 8 both in the presence and absence of the drug (Fig. 7).
The enzyme was inactivated at acidic pH, as at pH 5, the enzyme had minimum activity.
Conversely, the enzyme preserved its activity at alkaline pH.
3-4- Structural changes
To determine structural changes caused by interaction between the enzyme and the drug, fluorescence spectra were considered. Pure enzyme was prepared and purification was confirmed by SDS-PAGE (Fig 8). The pure lipase was excited at 285 nm to stimulate tryptophan residues.
The emission was monitored between 310 to 430 nm (Fig 9). Excitation of the pure enzyme gave an emission with a peak at 348 nm (Fig 9, graph A), while binding of bromhexine to the enzyme induced hypochromicity. This quenching was accompanied by peak blue shift to 342 nm (Fig 9, graph C).
4- Discussion:
Lipase is an important enzyme with applications in industry and medicine. Although its inhibition may not useful for industry, it is important in medicine. Many microorganisms which induce skin sores use lipase for degrading tissue layers, so inhibition of lipase in these microorganisms may reduce the invasive manner of microorganisms. It has been shown that bromhexine can prevent bacterial adherence to mammalian cells (Hafez et al 2009). On the other hand, in obese persons, inhibition of intestinal lipase helps reduce weight. These individuals use
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drugs such as Orlstat to inhibit the intestinal lipase. However, the drugs which inhibit lipase activity are not numerous. Recently, dicyclomine was also determined as potent inhibitor of lipase (Talebi et al 2018).
In this research, for the first time, bromhexine was introduced as a lipase inhibitor. bromhexine belongs to a group of medications called mucolytics, which mucus less sticky and also facilitates its removal (Olivieri et al 1991).
The results of this research showed that bromhexine inhibited lipase in a competitive manner.
Estimation of LC50 and KI values of the drug indicated that the enzyme interacted with the drug with high affinity.
Some drugs have been shown to interact with enzymes as their non-specific ligands in recent years, among which cimetidine with its imidazol ring has been demonstrated to bind to catalase and alkaline phosphatase with high affinity (Yazdi et al 2015, Minai-Tehrani et al 2011).
Cimetidine also uses hydrogen bond and hydrophobic interaction to bind catalase. It has also showed that dicyclomine attached to lipase with high affinity which induced a competitive inhibition (Talebi et al 2018).
This research showed that maximum activity of the lipase was observed at pH 8 in the presence and absence of the drug. Increasing pH decreased the activity of the enzyme. The pH profile obeyed the same pattern in the presence and absence of the drug, suggesting that pH change could not prevent the drug from binding to the enzyme and the drug does not use ionic interaction for binding to the enzyme.
The effect of temperature on the lipase activity showed that the enzyme had higher activity at 60oC both in the presence and absence of the drug. The temperature profiles in both cases were nearly the same, but the drug reduced the activity of the enzyme, suggesting that change of temperature could not refuse the binding of the drug to the enzyme. Arrhenius plot depicted two lines with different slope, which indicated that, in the presence of bromhexine, more energy was needed to proceed the reaction. In other reports, Arrhenius plot has been used to determine the effect of drugs on the progression reactions of enzymes, which showed that, in the presence of cimetidine, the activation energy of the reaction for catalysis is increased (Masoud et al 2014).
Fluorescence measurement is used to determine the structural change between ligand and enzymes. For example, Yazdi et al. (2015) demonstrated that binding of cimetidine to the enzyme caused conformational changes in the enzyme which was accompanied by
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hyperchromicity and displacement of tryptophan (Trp) residues to less polar medium. Talebi et al. (2018) implied that binding of dicyclomine to lipase not only induced hypochromicity but also revealed a red-shift in fluorescence spectrum proposed that the tryptophan residues have been transferred to more polar medium. In this research, fluorescence results revealed that binding of bromhexine to the lipase induced hypochromicity and blue shift which suggested that tryptophan residues have moved to more polar regions which confirm structural changes after attachment of the drug to the enzyme.
Conclusion: For the first time, bromhexine was shown to be a potent inhibitor of lipase. It could competitively inhibit the enzyme by attachment to the amino acids near the active site of the enzyme and could increase the activation energy of the reaction. Further experiments are needed to reveal the effect of bromhexine on pancreatic lipase, which may be a useful study for lipid absorption in the gastrointestinal tract and also more studies, must be done to reveal inhibition of biofilm formation of pathogen bacteria in patients by bromhexine.
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References:
1- Bever RA, Iglewski BH. (1988) Molecular characterization and nucleotide sequence of the Pseudomonas aeruginosa elastase structural gene. J. Bacteriol. 170, 4309-4314.
2- Buisman GJH, van Helteren CTW, Kramer GFH, Veldsink JW, Derksen JTP, Cuperus FP.
(1998) Enzymatic esterifications of functionalized phenols for the synthesis of lipophilic antioxidants. Biotechnol Lett. 20, 131-136.
3- Cryz SJ, Iglewski BH. (1980) Production of alkaline protease by Pseudomonas aeruginosa. J.
Clin. Microbiol. 12, 131-133.
4- de Bentzmann S, Plesiat P.(2011) The Pseudomonas aeruginosa opportunistic pathogen and human infections. Environ Microbiol. 13, 1655-65.
5- Ellis GP, West GB. (1970) Progress in medicinal chemistry 7. Butterworth & Co Pub.
London,; Vol7. : 44-45
6-Felix K, Pairet M, Zimmermann R. (1996) The antioxidative activity of the mucoregulatory agents: ambroxol, bromhexine and N-acetyl-L-cysteine. A pulse radiolysis study. Life. Sci.
59, 1141-1147
7- Frimmersdorf E, Horatzek S, Pelnikevich A, Wiehlmann L, Schomburg D. (2010) How Pseudomonas aeruginosa adapts to various environments: a metabolomic approach.
Environ. Microbiol. 12,1734-1747.
8- Hafez MM, Aboulwafa MM, Yassien MA, Hassouna NA, (2009) Activity of some mucolytics against bacterial adherence to mammalian cells. Appl. Biochem. Biotechnol.
158, 97-112
9- Lyczak J.B, Cannon CL, Pier GB. (2002) Lung infections associated with cystic fibrosis, Clin. Microbiol Rev. 15, 194-222
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10- Masoud M, Ebrahimi F, Minai-Tehrani D. (2014) Effect of cimetidine on catalase activity of Pseudomonas aeruginosa: A suggested mechanism of action. J Mol Microbiol. Biotechnol.
24, 196–201.
11- Matsumae H, Furui M, Shibatani T, (1993) Lipase catalysed asymmetric hydrolysis of 3- phenylglycidic acid ester, the key intermediate in the synthesis of Ditiazem hydrochoride. J Ferment Bioeng. 75, 93-98.
12- Minai-Tehrani D, Khodai S, Aminnaseri S, Minoui S, Sobhani-Damavadifar Z, Alavi S, Osmani R, Ahmadi S. 2011 Inhibition of renal alkaline phosphatase by cimetidine. Drug Metab Lett. 5, 197-201.
13- Olivieri D, Ciaccia A, Marangio E, Marsico S, Todisco T, Del Vita M, (1991) Role of bromhexine in exacerbations of bronchiectasis. Double-blind randomized multicenter study versus placebo. Respiration. 58, 117-121.
14- Ostroff RM, Vasil M.L. (1987) Identification of a new phospholipase C activity by analysis of an insertional mutation in the hemolytic phospholipase C structural gene of Pseudomonas aeruginosa. J. Bacteriol. 169, 4597-460.
15- Pollack M, (2000) In Principles and practice of infectious diseases, eds. Mandell G L, Bennet J E, & Dolin, R. Churchill Livingstone, Philadelphia,; Vol. 2, pp. 2310-2335
16- Rubin B, Dennis EA. (1997) Lipases: Part A. Biotechnology Methods in Enzymology.
Academic Press, New York, 284, 1-408.
17- Stuer W, Jaeger KE, Winkler UK, (1986) Purification of extracellular lipase from Pseudomonas aeruginosa. J. Bacteriol. 168, 1070-1074.
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18- Talebi M, Minai-Tehrani D Fazilati M, Minai-Tehrani A, (2018) Inhibitory action of dicyclomine on lipase activity, kinetics andmolecular study. Int J Biol Macromol 107, 2422–2428
19- Vulfson EN, (1994) Industrial Applications of Lipases. In: Lipases-Their Structure, Biochemistry and Application, Woolley, P. and S.B. Peterson (Eds.)., Cambridge University Press, UK. 271-288.
20- Yazdi F, Minai-Tehrani D, Jahngirvand M, Almasirad A, Mousavi Z, Masoud M, Mollasalehi H, (2015) Functional and structural changes of human erythrocyte catalase induced by cimetidine: proposed model of binding. Mol. Cell. Biochem. 404, 97–102.
21- Zhi J, Melia AT, Eggers H, Joly R, Patel IH, (1995) Review of limited systemic absorption of orlistat, a lipase inhibitor, in healthy human volunteers. J Clin Pharmacol. 35, 1103–1108.
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Figure 1: Chemical structure of bromhexine.
Figure 2: Lineweaver-Burk plot showed a competitive inhibition of lipase when bromhexine as an inhibitor present in the enzyme reaction medium. Bromhexine concentration was from 0.048 to 0.32 mM.
Figure 3: Change of Km and the affinity of the enzyme in the presence of different concentration of the drug. The affinity of the enzyme for its substrate decreased with increasing the drug
concentration.
Figure 4: Secondary plot was drawn using the slopes of lines in figure 2. The x-intercept gives the Ki value.
Figure 5: The effect of different temperature on the activity of the enzyme. The optimum temperature was about 60oC in presence and absence of the drug.
Figure 6: Arrhenius plot gives the activation energy of reaction. The slope of the line was used for determination of activation energy. In the presence of the drug, the activation energy was higher than the control (absence of the drug).
Figure 7: The effect of different pH on the activity of lipase. The enzyme activity was preserved in alkaline pH.
Figure 8: SDS-PAGE of the pure enzyme with silver nitrate staining. Lane A: protein ladder.
Lane B: pure enzyme with molecular weight of about13 kDa.
Figure 9: Fluorescence spectra of the pure lipase. The excitation was applied at 285 and the emission was traced from 310 to 430 nm. The pure lipase (A) showed a peak with maximum intensity at 348 nm. The enzyme and substrate (para-nitrophenyl palmitate) complex induces hypochromicity without any shift in peak (B). The complex of enzyme and bromhexine induces hypochromicity and peak blue shift with maximum intensity at 342 nm (C). Bromhexine in buffer had no peak around 340 nm (D).
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Figure 1
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Figure 2:
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Figure 3
0 0.05 0.1 0.15 0.2 0.25 0.3 0.35
0 10 20 30 40 50 60 70 80 90 100
0 0.2 0.4 0.6 0.8 1 1.2 1.4
Affinity Km
Affinity % Km (mM)
Bromhexine (mM)
IC
50 34 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55
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Figure 4
-0.04 0.01 0.06 0.11 0.16 0.21 0.26 0.31 0
0.05 0.1 0.15 0.2 0.25 0.3 0.35 0.4 0.45
Sl op e
Bromhexine (mM)
-Ki
3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55
For Peer Review Only
Figure 5
0 10 20 30 40 50 60 70 80
0 20 40 60 80 100 120
No Drug With Drug
Relative activity %
Temperature (oC)
3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55
For Peer Review Only
Figure 6:
2.9 3.1 3.3 3.5 3.7 3.9
1 1.2 1.4 1.6 1.8 2
2.2 No Drug
With Drug
Log Vmax
10-3K/T
3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55
For Peer Review Only
Figure 7
5 6 7 8 9 10
0 20 40 60 80 100 120
No Drug With Drug
Relative activity%
pH
3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55
For Peer Review Only
Figure 8
3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55
For Peer Review Only
Figure 9:
310 330 350 370 390 410 430
0 100 200 300 400 500 600 700 800 900 1000
Wavelength (nm)
Intensity
34 34
B
C
D A
3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55
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Archives of Physiology and Biochemistry
COUNTRY United Kingdom
SUBJECT AREA AND CATEGORY
Biochemistry, Genetics and Molecular Biology
Medicine
PUBLISHER Informa Healthcare
H-INDEX
53
PUBLICATION TYPE
Journals
ISSN
13813455, 17444160
COVERAGE
1921-1927, 1929-1931, 1933-1943, 1946- 2004, 2006-2021
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SCOPE
Archives of Physiology and Biochemistry: The Journal of Metabolic Diseases is an international peer-reviewed journal which has been relaunched to meet the increasing demand for integrated publication on molecular, biochemical and cellular aspects of metabolic diseases, as well as clinical and therapeutic strategies for their treatment. It publishes full-length original articles, rapid papers, reviews and mini-reviews on selected topics. It is the overall goal of the journal to disseminate novel approaches to an improved understanding of major metabolic disorders. The scope encompasses all topics related to the molecular and cellular pathophysiology of metabolic diseases like obesity, type 2 diabetes and the metabolic syndrome, and their associated complications. Clinical studies are considered as an integral part of the Journal and should be related to one of the following topics: -Dysregulation of hormone receptors and signal transduction -Contribution of gene variants and gene regulatory processes -Impairment of intermediary metabolism at the cellular level -Secretion and metabolism of peptides and other factors that mediate cellular crosstalk -Therapeutic strategies for managing metabolic diseases Special issues dedicated to topics in the eld will be published regularly.
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SJR
The SJR is a size-independent prestige indicator that ranks journals by their 'average prestige per article'. It is based on the idea that 'all citations are not created equal'. SJR is a measure of scienti c in uence of journals that accounts for both the number of citations received by a journal and the importance or prestige of the journals where such citations come from It measures the scienti c in uence of the average article in a journal, it expresses how central to the global scienti c discussion an average article of the
Total Documents
Evolution of the number of published documents. All types of documents are considered, including citable and non citable documents.
Year Documents
1999 41
2000 251 2001 171 2002 151
Citations per document
This indicator counts the number of citations received by documents from a journal and divides them by the total number of documents published in that journal. The chart shows the evolution of the average number of times documents published in a journal in the past two, three and four years have been cited in the current year. The two years line is equivalent to journal impact factor ™ (Thomson Reuters) metric.
Cites per document Year Value Cites / Doc. (4 years) 1999 0.234 Cites / Doc. (4 years) 2000 0.319 Cites / Doc. (4 years) 2001 0.232 Cites / Doc. (4 years) 2002 0.191 Cites / Doc. (4 years) 2003 0.277 Cites / Doc. (4 years) 2004 0.227 Cites / Doc. (4 years) 2005 0.318 Cites / Doc. (4 years) 2006 0.222 Cites / Doc. (4 years) 2007 0.285 Cites / Doc. (4 years) 2008 0.569 Total Cites Self-Cites
Evolution of the total number of citations and journal's self- citations received by a journal's published documents during the three previous years.
Journal Self-citation is de ned as the number of citation from a journal citing article to articles published by the same journal.
Cites Year Value lf
External Cites per Doc Cites per Doc
Evolution of the number of total citation per document and external citation per document (i.e. journal self-citations removed) received by a journal's published documents during the three previous years. External citations are calculated by subtracting the number of self-citations from the total number of citations received by the journal’s documents.
l
% International Collaboration Citable documents Non-citable documents Cited documents Uncited documents
1999 2002 2005 2008 2011 2014 2017 2020 0
0.9 1.8 2.7
1999 2002 2005 2008 2011 2014 2017 2020 0
200 400
Cites / Doc. (4 years) Cites / Doc. (3 years) Cites / Doc. (2 years)
1999 2002 2005 2008 2011 2014 2017 2020 0
1 2 3 4 5
1999 2002 2005 2008 2011 2014 2017 2020 0
600 1.2k
1999 2002 2005 2008 2011 2014 2017 2020 0
2.5 5
Metrics based on Scopus® data as of April 2022
Ko 3 years ago
Please I would like to know if you publish a paper with the cardiovascular effects of plant extracts reply
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