مجله دانشکده پزشکی اصفهان
پیاپی 471 (هفته سوم اردیبهشت 1397)

In spite of the wide use, low proteome coverage and fuzzy patterns are the most important deterrents for the clinical applications of gel-based proteomic studies. So herein, we tried to increase the 2-dimentional proteome coverage of Hs578T breast cancer cells via investigating the efficacy of the three common techniques, usually used for interfering removal.
Hs578T cells were incubated in a lysis solution to obtain raw cell extracts. Cellular soups of each extraction were then pooled, homogenized, and aliquoted to be further treated by three different protein-specific purification methods including acetone, acetone-methanol, and trichloroacetic acid (TCA)-acetone, each in triplicates. All the purified protein pellets were then dissolved in a standard rehydration buffer solution, loaded into the 17-cm immobilized pH gradient (IPG) strips, and separated according to their isoelectric points. Proteins were then separated once more according to their molecular weights in an OFarrell separation system. Finally, by the visualization of the protein spots on the 2-dimentional profiles, quality and quantity of these 2-dimentional proteome patterns were then analyzed using the ImageMaster software.
Findings: The obtained proteome recovery yields and total protein counts for acetone, acetone-methanol, and trichloroacetic acid-acetone methods were 0.100 ± 0.001, 0.070 ± 0.002, and 0.120 ± 0.005 ng/cell, and 1299 ± 9, 1698 ± 14 and 1973 ± 17, respectively. The results represent data obtained from three independent experiments.
Trichloroacetic acid-acetone purification not only represented the highest recovery yield, suitable for expensive assays, but also showed the most suitable proteome coverage. So, the method is recommended for the comparative proteomic studies. However, the acetone-methanol procedure is more recommended for serological proteome analysis (SERPA); since it represents stronger protein spots which are more fitted to the immunoblotting procedure.

Stress detection is essential and important in order to control, manage, and reduce it. On the other hand, there are many theories about the stress-related back pain. The most important point in all of these theories, is the psychological and emotional factors that cause some kinds of physiological changes, and as a result, back pain. Therefore, it seems that the electromyogram (EMG) signal of the lower back muscles can be used as a marker for stress detection.
In this research, stress was detected using the electromyogram signal of erector spinae muscles. The signals of 15 persons were recorded from left and right erector spinae muscles. After extracting seven time and frequency domain features, stress was detected in two-level (stress and no stress) and four-level (no stress, low stress, moderate stress, and high stress) modes applying an efficient support vector machine (SVM) classifier. It was also attempted to improve the performance of the proposed approach by using feature selection methods.
Findings: In two-level mode, stress was detected with 100% of accuracy. In the four-level mode, the efficiency of the right erector spinae muscle was higher and reached 100% of accuracy.
The results denote that the electromyogram of the erector spinae muscles is an appropriate indicator of stress. The right erector spinae muscle is more effective than the left one. Furthermore, feature selection reduces the computation amount, and improves the efficiency of stress detection. The findings of this study can be used to detect stress for controlling and managing it.

Ventilator-associated pneumonia (VAP) is a prevalent infection occurs in intensive care unit (ICU) accompanied with increased complications and mortality. In this study, effects of chlorhexidine mouthwash versus herbal stop-snoring mouthwash on VAP among ICU admitted patients have been compared.
86 patients admitted at ICU were randomly divided to two equal groups including; group C treated with chlorhexidine mouthwash and group S treated with stop-snoring mouthwash. Patients of both groups were similarlytreated for three times a day (including amount, technique and times of mouth washing). Time of incidence and frequency of VAP were compared in two groups.
Findings: 74.41% of group S and 62.79%of group C presented VAP (P-value = 0.353). 22.22% of patients presented VAP in first-5-days and 77.77% after 5 days and in group S while 6.25% of patients of group C presented VAP in first-5-days and 93.75% after 5 days(P-value = 0.126).
Time of VAP incidence or frequency of VAP was not different using chlorhexidine mouthwash versus herbal stop-snoring mouthwash for prevention of VAP.

Clinical diagnostic kits for detecting hepatitis B are based on antibodies, and have inefficient sensitivity, stability, and cost. Therefore, researches about aptamers and using them instead of antibodies may make these defects up. In this study, a biotinylated anti-hepatitis B virus surface antigen (anti-HBsAg) aptamer sequence was used to evaluate the possibility of specific detection of HBsAg via enzyme-linked aptamer assay (ELAA) method using biotin-streptavidin system.
HBsAg was immobilized in Maxibinding plate (SPL, Korea) by using carbonate-bicarbonate buffer (pH: 9.4), and by considering different variables. Immobilization of HBsAg was evaluated by commercial kit. Aptamer sequence was cloned and imbalance polymerase chain reaction (PCR) was improved and performed using plasmid as DNA template. Then, biotinylated aptamer was utilized in enzyme-linked aptamer assay method via taking advantage of streptavidin-biotin signal amplification system.
Findings: Antigen immobilization was set up using conjugates number 1 and 2 of commercial kit. Colony polymerase chain reaction confirmed aptamer cloning in Escherichia coli (E. coli) Top10 host. The best gradient polymerase chain reaction result was achieved at 64 ºC annealing temperature. Control group in assays showed accuracy of immobilization; but no repetitive specific signal was obtained that could prove joining of aptamer to HBsAg, and detection of that.
Three-dimensional structure of an immobilized antigen on a solid surface may vary from that which exists on the surface of viruses or cells. Negative results might be due to the ability of aptamers to attach into antigens with totally intact shape. Conjugation of aptamer with biotin may interfere in aptamer conformation; so, a connective arm between aptamer and biotin could be a suggestion, which needs more assessments.

Endometriosis is an estrogen-dependent disorder in which endometrial cells that are in the normal state of the uterus are seen outside the uterus. The disease is one of the most common and costly diseases in women with an estimated 176 million female infections in the world and a financial burden of about 11 billion dollars per year. In addition to existing therapies, the use of epigenetic factors, and in particular, assessment of the expression of microRNAs is effective in the diagnosis and treatment of the disease. The aim of this study was to evaluate the expression of miR-135b in endometriosis, so that through identifying changes in these microRNAs, it could be used in diagnosis and treatment.
From 25 cases with endometriosis and 25 healthy subjects (control) tissue and serum samples were taken. First, RNA extraction was performed and then, using the polyA reverse transcription-polymerase chain reaction (polyA RT-PCR) and real-time polymerase chain reaction, miR-135b expression was investigated.
Findings: The expression of mir-135b significantly increased in endometrial tissue of patients with endometriosis in both ectopic and uterine tissues compared to healthy women; while the expression of this microRNA in the serum samples of patients decreased compared to control group.
Considering significant changes in both eutopic/ectopic samples and serum samples in patients with endometriosis compared with controls, miR-135b may be considered as a diagnostic and therapeutic biomarker for endometriosis.