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Original Article:
Improvement of Soluble Production of Reteplase in
Escherichia Coli
by Optimization of Chemical Chaperones in Lysis Buffer
Behnaz Fazeli, Vajihe Akbari, Asgar Barkhordari, Hamid Mir Mohammad Sadeghi
Adv Biomed Res
2019, 8:65 (31 October 2019)
DOI
:10.4103/abr.abr_212_18
PMID
:31737582
Background:
Reteplase is a nonglycosylated derivative of recombinant tissue plasminogen activator, a thrombolytic agent, which can be easily expressed in
Escherichia coli
. However, overexpression of reteplase in
E. coli
usually leads to accumulation of insoluble and inactive aggregates and inclusion bodies. In the present study, we aimed to optimize chemical additives of lysis buffer to avoid the initial aggregation and formation of inclusion bodies of reteplase at cell disruption step.
Materials and Methods:
After protein expression in
E. coli
BL21 (DE3), the bacterial cells were disrupted in different lysis buffers using microsmashing. Eleven chemical additives at two concentration levels were combined based on a Plackett–Burman design to prepare 12 different lysis buffers used at cell disruption stage. Then, three additives with the most positive effect on improvement of solubility of reteplase were chosen and used for the second screening based on Box–Behnken model.
Results:
The primary screening results showed that among 11 additives, arginine, K
2
PO
4,
and cetyltrimethylammonium bromide (CTAB) had the most positive effect on solubility of reteplase. Our final results based on 14 runs of Box–Behnken design showed that the optimum buffer additive condition is 0.005 mg/ml CTAB, 0.065 mg/ml arginine, and 0.026 mg/ml K
2
PO
4
. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis analysis and Western blotting of soluble and total fraction of samples confirmed that these additives significantly improved soluble production of reteplase compared with control.
Conclusion:
Our study indicates that the application of chemical additives in cell lysis can improve the solubility of reteplase. Further studies are still required to understand the exact mechanism of chemical additives as a chemical chaperone during cell lysis.
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Original Article:
Interaction of Sex Hormones and the Renin–Angiotensin System in Ovariectomized Rats Subjected to Ischemia-Reperfusion Induction
Halimeh Lakzaei, Tahereh Safari, Gholam Reza Komeili
Adv Biomed Res
2019, 8:64 (31 October 2019)
DOI
:10.4103/abr.abr_172_19
PMID
:31737581
Backgrounds:
Ischemia-reperfusion (IR) injuries occur in a variety of clinical conditions, which lead to kidney damage. Most of the tissue damages after IR are due to the activation of the renin–angiotensin system (RAS). Hence, in this study, the interaction of sex hormones and RAS in ovariectomized (OV) rats subjected to IR induction has been studied.
Materials and Methods:
The animals were divided into different groups. Groups 1 (OV + E, OV rat + estradiol) and 2 (OV rat) each one consisted of three separate IR-induced subgroups treated with losartan, angiotensin 1–7 (Ang 1–7), and their combination, Group 3, as control and Group 4, as sham. Next, 72 h after IR, blood samples were collected, the right kidneys were homogenized, and left kidneys were fixed in 10% formalin.
Results:
Findings show that serum blood urea nitrogen, creatinine, and kidney tissue damage score levels increased significantly with induction of IR (
P
< 0.05). Mean serum levels of these factors in OV + E groups are higher than those of the OV. The presence or absence of estradiol did not affect the levels of antioxidants in the different groups receiving Los, Ang 1–7, and their combination. Los, Ang 1–7, and their combination reduced serum and kidney malondialdehyde levels in both OV and OV + E groups.
Conclusion:
Estrogen not only fails to improve renal functioning but it can also exacerbate it. While the treatments used in this study, in the absence of estradiol, it had a better effect on kidney damages and improved its functions.
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Review Article:
Histone Deacetylases and Histone Deacetylase Inhibitors: Molecular Mechanisms of Action in Various Cancers
Masumeh Sanaei, Fraidoon Kavoosi
Adv Biomed Res
2019, 8:63 (31 October 2019)
DOI
:10.4103/abr.abr_142_19
PMID
:31737580
Epigenetic modifications such as histone modification play an important role in tumorigenesis. There are several evidence that histone deacetylases (HDACs) play a key role in cancer induction and progression by histone deacetylation. Besides, histone acetylation is being accessed as a therapeutic target because of its role in regulating gene expression. HDAC inhibitors (HDACIs) are a family of synthetic and natural compounds that differ in their target specificities and activities. They affect markedly cancer cells, inducing cell differentiation, cell cycle arrest and cell death, reduction of angiogenesis, and modulation of the immune system. Here, we summarize the mechanisms of HDACs and the HDACIs in several cancers. An online search of different sources such as PubMed, ISI, and Scopus was performed to find available data on mechanisms and pathways of HDACs and HDACIs in different cancers. The result indicated that HDACs induce cancer through multiple mechanisms in various tissues. This effect can be inhibited by HDACIs which affect cancer cell by different pathways such as cell differentiation, cell cycle arrest, and cell death. In conclusion, these findings indicate that the HDACs play a major role in carcinogenesis through various pathways, and HDACIs can inhibit HDAC activity by multiple mechanisms resulting in cell cycle arrest, cell growth inhibition, and apoptosis induction.
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Original Article:
Comparison of Nutritional Behaviors and Physical Activities Between Overweight/Obese and Normal-Weight Adults
Anahita Babak, Reza Rouzbahani, Razie Khalili Nejad, Aryan Rafiee Zadeh
Adv Biomed Res
2019, 8:62 (31 October 2019)
DOI
:10.4103/abr.abr_134_19
PMID
:31737579
Background:
Overweight and obesity are major problems with increasing rates among adult populations. Nutritional behaviors and physical activity are the most important influencing factors. In this article, we conducted a survey on the nutritional behaviors and physical activities among normal-weight and overweight/obese adults.
Materials and Methods:
We conducted a survey on some of the nutritional behaviors and physical activities of 729 middle-aged adults. Data regarding desirable or undesirable use of fruits, fish, dairy products, oil, fast foods, and carbonated drinks and the intensity and duration of physical activities were collected from our data bank and analyzed using the SPSS software.
Results:
Our results indicated that overweight/obese people had an improper usage of fruits, oil, fast foods, and carbonated drinks and had lower physical activity. There was also a significant increase in intensity, duration per day, and weekly days of physical activity in healthy groups. Logistic regression analysis of nutritional behaviors, physical activities, and obesity also indicated that undesirable usage of oil, fast foods, and carbonated drinks and undesirable physical activity are associated with risks for overweight/obesity (odds ratio [OR] =10.70, OR = 7.45, OR = 7.48, and OR = 2.16, respectively) (
P
< 0.05).
Conclusion:
This article puts emphasis on the role of proper nutritional behaviors and higher physical activities in decreasing the risks of developing obesity, and we suggest that further interventions could be made based on the results of our study.
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Original Article:
Evaluation of different concentrations of imatinib on the viability of
Leishmania major
: An
In Vitro
study
Mohsen Moslehi, Fatemeh Namdar, Mahsa Esmaeilifallah, Seyed Hossein Hejazi, Fatemeh Sokhanvari, Amir Hossein Siadat, Seyed Mohsen Hosseini, Fariba Iraji
Adv Biomed Res
2019, 8:61 (31 October 2019)
DOI
:10.4103/abr.abr_58_19
PMID
:31737578
Background:
Leishmaniasis is an infectious disease caused by an intracellular parasite of
Leishmania
and is transmitted through the female sandflies bite and may lead to severe skin lesions. Although drugs such as antimony compounds are available, their side effects such as toxicity, low efficacy, and emergence of resistance have raised the importance of effective replacement. Imatinib, as an inhibitor of tyrosine kinase (TK) of
Leishmania
, stops abnormal function of TK such as Bcr-Abl through assembling into transmembrane pores in a sterol-dependent manner. Hence, the evaluation of killing effects of different concentrations of imatinib against
Leishmania major
amastigotes and
promastigotes in vitro
were the objectives of the present study.
Materials and Methods:
The killing effects of different concentrations of imatinib (25, 50, and 100 μg) and 25 μg amphotericin B (as positive control) were evaluated against RPMI 1640-cultured promastigotes and the amastigote/macrophage model by MTS cell proliferation assay kit (ab197010) and Giemsa staining method during 24, 48, and 72 h.
Results:
The results showed anti-
Leishmania
effect of imatinib in concentration and time-dependent manner. The lowest number of live promastigotes and amastigotes were obtained due to treat with 100 μ/ml imatinib at 72 h. Furthermore, 100 μg concentration of imatinib had the same effect as 25 μg amphotericin B on both
L. major
promastigotes and amastigotes (
P
< 0.001).
Conclusion:
The anti-
Leishmania
effect of imatinib was confirmed by MTS and direct microscopy. Further study is recommended for evaluating possible therapeutic effects of imatinib on leishmaniasis
in vivo.
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2022
July
[
8
]
June
[
11
]
May
[
9
]
April
[
9
]
March
[
8
]
February
[
9
]
January
[
10
]
2021
December
[
10
]
November
[
11
]
October
[
5
]
September
[
6
]
August
[
2
]
July
[
4
]
June
[
3
]
May
[
1
]
March
[
2
]
February
[
4
]
January
[
6
]
2020
December
[
8
]
November
[
7
]
October
[
13
]
September
[
10
]
August
[
10
]
July
[
7
]
June
[
4
]
May
[
5
]
April
[
5
]
March
[
3
]
February
[
4
]
January
[
5
]
2019
December
[
2
]
November
[
5
]
October
[
5
]
September
[
7
]
August
[
6
]
July
[
3
]
June
[
7
]
May
[
5
]
April
[
8
]
March
[
6
]
February
[
8
]
January
[
10
]
2018
December
[
9
]
November
[
9
]
October
[
11
]
September
[
5
]
August
[
10
]
July
[
10
]
June
[
13
]
May
[
17
]
April
[
17
]
March
[
19
]
February
[
19
]
January
[
20
]
2017
December
[
13
]
November
[
21
]
October
[
12
]
September
[
9
]
August
[
15
]
July
[
22
]
June
[
11
]
May
[
14
]
April
[
14
]
March
[
21
]
February
[
8
]
January
[
8
]
2016
December
[
17
]
November
[
20
]
October
[
12
]
September
[
9
]
August
[
19
]
July
[
16
]
June
[
17
]
May
[
17
]
April
[
16
]
March
[
36
]
February
[
14
]
January
[
14
]
2015
December
[
2
]
November
[
15
]
October
[
25
]
September
[
29
]
August
[
29
]
July
[
31
]
June
[
11
]
May
[
39
]
March
[
15
]
February
[
29
]
January
[
34
]
2014
December
[
23
]
November
[
25
]
October
[
18
]
September
[
19
]
August
[
22
]
July
[
13
]
June
[
11
]
May
[
22
]
April
[
2
]
March
[
24
]
February
[
11
]
January
[
77
]
2013
November
[
10
]
October
[
5
]
August
[
1
]
July
[
23
]
June
[
11
]
March
[
42
]
2012
December
[
4
]
October
[
17
]
August
[
34
]
July
[
15
]
May
[
10
]
March
[
7
]
1900
January
[
1
]
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