International Journal of Medical Laboratory

6

Authors: Kave Tari, Amir Atashi *, Farshid Karami, Fatemeh Kiani nodeh, Mohammad Shahjahani

Background and Aims: Iron deficiency anemia (IDA) and beta thalassemia minor (BTM) are the most common hypochromic microcytic anemias, and it is regarded important to differentiate between them. several formulae can be taken into consideration based on red blood cell (RBC) parameters in order to distinguish between these two disorders. Hence, the present study intended to evaluate the sensitivity as well as specificity of some of these formulae. Materials and Methods: In this cross-sectional study, IDA was diagnosed in 200 patients based on hypochromic and microcytic RBC appearance, reduced mean cell volume, mean cell hemoglobin and ferritin as well as increased total iron binding capacity. Furthermore, BTM was diagnosed via hypochromic microcytic appearance of RBC and increased hemoglobin A2 level. Then, IDA and BTM diagnosis were confirmed using the formulae of King-Green, Mentzer index, England and Fraser, Shine and Lal, Srivastava and Sirdah. The sensitivity and specificity of these formulae were calculated as well. Results: The study findings demonstrated that based on the study criteria, out of 200 patients with hypochromic microcytic RBC appearance, 120 were afflicted with IDA and 80 suffered from BTM. The formulae-based diagnosis demonstrated that King-Green formula was the most reliable one. Conclusions: Although King-Green formula had the highest sensitivity and specificity and was the most reliable formula, none of the formulae revealed 100% sensitivity and specificity. As a result, making definitive distinction between IDA and BTM is not possible using these formulae.

7

Authors: Donya Nikaein, Alireza Khosravi, Seyyed Shamsadin Athari, Abdolghaffar Ownagh, Mehdi Taghavi *

Background and Aims: Aspergillus fumigatus is a sporadic fungus that causes different infections and allergies in immunocompromised patients. The allergic disease caused by this fungus is called allergic bronchopulmonary aspergillosis (ABPA). ABPA is considred important in atopic and immunocompromised individuals, which can result in inflammation and epithelial damage. Therefore, the aim of this study was to evaluate the T helper (Th)2 responses in a ABPA murine model by measuring the main cytokines involved in Th2. Materials and Methods: Twenty male BALB/c mice were divided into two groups of 10 mice each: control and ABPA group. ABPA was induced by inhalation of A. fumigatus conidia intranasally. Total and specific IgE were measured in the mice sera. Levels of cytokines in broncho alveolar lavage (BAL) of under studied groups were measured by Enzyme-linked immunosorbent assay three weeks after the treatment. Results: The obtained results indicated that total and specific IgE increased in the ABPA group (p<0.05). The levels of Interleukin (IL)-4, IL-5 and IL-13 in brocho alveolar lavage of ABPA group was significantly higher than the control group (p<0.05), whereas interferon-gamma levels did not reveal any significant differences between the studied groups. Conclusions: The findings of the present study confirmed the role of Th2 cytokines in the ABPA reactions. However, more comprehensive studies are necessitated to determine the exact mechanisms of immune responses to ABPA as well as the role of Th1/Th2 responses in control of ABPA reactions. Regulation of Th2 responses could be regarded as a potential therapy for ABPA as well.

9

Authors: Parvin Hosseini Pour, Hassan Momtaz *, Amir Arsalan Serajyan, Elahe Tajbakhsh

Background and Aims: The indiscriminate use of antibiotics can lead to antibiotic resistance in the treatment of infections caused by bacteria such as Pseudomonas aeruginosa. The presence of integrons in Pseudomonas is clearly associated with multidrug resistances. Therefore, this study aimed at tracking class I, II and III integrons of antibiotic-resistant isolates of Pseudomonas aeruginosa that were isolated from nosocomial infection. Materials and Methods: In this study, 51 isolates of Pseudomonas aeruginosa were collected from different wards of Imam Khomeini hospital of Ahvaz since October of 2014 until March of 2015. After identification test and antibigram, coding genes of antibiotic resistance and class I, II and III integrons were detected by polymerase chain reaction (PCR) method. Results: Tetracycline revealed the most resistance with 84% frequency in discreted isolates. In the encoding antibiotic resistance genes with a frequency of 94% was the most common blaTEM. Class I integron had 92% prevalence, class II Integron showed 52% prevalence and class III Integron demonstrated 17% prevalence. Conclusions: In Pseudomonas aeruginosa, class I integron was more prevalent than other integrons and the integrase gene was probably one of the causes of multiple antibiotic resistance in this bacteria. Moreover, frequency of integron III was reported 17%.

10

Authors: Fereshteh Safari, Mehdi Fazeli, Meysam Dehghani, Arian Naghedi, Fatemeh Jahanifar, Fatemeh Safari *

Background and Aims: There has been scant information concerning antihypertrophic effects of vitamin D specifically on its cellular and molecular mechanisms. Sirtuin 1 (SIRT1) is regarded as a key deacetylase enzyme in cardiomyocytes which applies potential cardioprotective effects by functional regulation of different proteins. This study aimed to evaluate the effects of 1, 25-dihydroxyvitamin D on the hypertrophic markers and cardiac level of SIRT1 mRNA in rats following the aortic banding. Material and Methods: In this study, male Wistar rats (170-220g) were used, which were divided into 4 groups: rats subjected to hypertrophy without treatment (H), rats pretreated with 1,25 dihydroxyvitamin D3 (H+VD), rats received propyleneglycol as a vitamin solvent (H+P), and intact animals which were elected as the control group. Arterial blood pressure was directly measured by the carotid cannulation. Transcription level of target genes was measured by real time polymerase chain reaction technique. Results: In H+VD group, systolic blood pressure as well as heart weight-to-body weight ratio decreased significantly compared to the group H (P<0.01). Moreover, regarding hypertrophy marker genes in H+VD group, both atrial natriuretic peptide mRNA (H+VD:64.8±14% vs. H:127±26% P<0.05) and brain natriuretic peptide mRNA (H+VD:25.6±6% vs. H:84.2±12% P<0.01) levels decreased significantly. SIRT1 mRNA level was increased by 56.8±14% in group H and by 42.6±12% in group H+VD which were significant in comparison to the control group (P<0.01 and P<0.05, respectively). No significant difference was noted between H+VD and H groups. Conclusions: The results of the present study revealed that administration of 1, 25- dihydroxyvitamin D decreases myocardial hypertrophy markers in rats following the abdominal aortic banding. The pressure overload-induced hypertrophy accompanies with SIRT1 mRNA upregulation, though antihypertrophic effects of vitamin didnot participate in SIRT1 transcription level.

6

Authors: Hossein Hadinedoushan *, Mahdi Dehghan-Manshadi, Fateme Zare

Background and Aims: Hepatitis B virus (HBV) infects the liver and causes acute and chronic hepatitis, hepatocellular carcinoma and cirrhosis. HBV has been divided to eight genotypes (A–H) and subgenotypes of A, B, C and F. For the first time, we determined HBV genotypes in infected samples by INNO-LiPA method in Yazd, central province of Iran. Materials and Methods: This study was performed on samples suspected of HBV infection. The sera of fifteen out of ninety-five samples that had shown positive results by RT-PCR were used for HBV genotyping by using INNO-LiPA HBV genotyping assay. Results and Conclusions: Seven (46.7%) out of fifteen samples were female. The mean age of the patients was 37.8±14.3 years. The average number copy of HBV in infected samples was 1.04×106±4.74×105 Copies/ml. All fifteen infected samples had genotype D. Our results showed that HBV genotype D was the only detectable genotype in Yazd, central province of Iran. A further study with a larger sample size in different provinces of Iran is needed to identify HBV genotypes in Iran.

7

Authors: Seyed Hassan Hejazian, Sara Sadegh Zade, Mahila Lotfi, Fatemeh Safari *

Background and Aims: Our previous study has demonstrated that essence of Carum copticum reduces contraction activity of ileum in rat. The present study was designed to find out the effects of nonpolar fraction of Carum copticum essence (NFCCE) on mechanical activity of the isolated ileum in rat. Materials and Methods: For evaluation of spasmolythic property of fraction, different doses of the solution were added to organ bath after acetylcholine with concentration of 10-4 molar (M), and for assessment of antispasmodic effect of fraction, different doses of the solution were added to the organ bath before acetylcholine with concentration of 10-9 up to 10-2 M. Then isotonic contractions of ileum were recorded through an isolated tissue chamber in an organ bath using oscillographic device. Results: Our findings showed that the spasmolytic effect of NFCCE in concentrations of 25, 50, and 100 ng/ml significantly reduces acetylcholine (10-4M)-induced contractions (p< 0.05). Also antispasmodic effect of NFCCE on logarithmic concentrations of acetylcholine (10-9 up to 10-2M) indicates that in the presence of 10-3 M acetylcholine, the maximum (100%) and minimum (84.9%), inhibition of contraction is induced by concentrations of 100 ng/ml and 25 ng/ml, respectively (p< 0.05). Conclusions: The spasmolytic effect of this study is probably resulted by anti-cholinergic response of NFCCE on isolated ileum of rats.

7

Authors: Shirin Taleifard *, Rasoul Roghanian, Majid Buzari, Hasan Salehi

Background and Aims: Infectious mononucleosis (IM)is the clinical manifestation of primary infection with Epstein-Barr virus (EBV). Humans are the only known reservoir of EBV. Regarding the problems in diagnosis of the disease, the purpose of this study was to assess Enzyme-linked immunosorbent assay (ELISA) and Nested polymerase chain reaction (PCR) as a diagnostic tool for this disease. Materials and Methods: 50 samples were collected from the suspicious patients with EBV and 50 samples from the healthy individuals as the control and both techniques were applied for them. Results: The results showed that 76% of the patients and 14% of the control samples had EBV DNA in serum with PCR. Statistical analysis showed significant difference between the patient and the control samples for infection with EBV (P < 0.0001). Samples were classified into three groups according to the ELISA that were acute phase (20%), recent infection or convalescence phase (14%) and past infection (66%), respectively. Conclusions: Comparing the two methods, the results of the ELISA test indicated that ELISA would be the best method to be used for the diagnosis of IM. Our results suggest that serology may be more sensitive and could be performed as the initial screening test for acute EBV infection. Although, the PCR test is routinely used as an accurate method for detection of the pathogens with a higher specificity and sensitivity comparing the immunoassay, in IM, ELISA seems to be the best method for detecting antibodies against EBV.

9

Authors: Saeide Nezami Saridar, Seyedhossein Hekmatimoghaddam *, Mona Yadegar

Background and Aims: In this lab trial, the effect of scaffold based on human serum albumin (HSA) and hydroxyapatite nanoparticles (HA NPs) on mouse spermatogonial cell line (SCL) was investigated. Materials and Methods: To synthesize HA NPs, calcium nitrate and diammonium phosphate at pH 13 were gently added and heated at 100 ºC for 24 hours. Then serial concentrations of HA NPs was separately added to 500 mg/mL of HSA, and immediately placed in the 100 ºC water bath. Then, all scaffolds were cut, and incubated with SCL for 6h, 12h, and 24h at 37 ºC. Finally, the cell count was read, and homing of the cells was examined by optical microscopy. Results: It was found that the quantity of cells did not change by increase in concentration of HA NPs. On the other hand, increased incubation time led to decrease in cell count. Light microscopic observation of scaffold cavities after incubation showed the homing of spermatogonial cells. Conclusions: This promising scaffold must be more investigated in vitro and in vivo, and may be suitable for making artificial testis.

4

Authors: Afsaneh Sarabandi, Rima Manafi Shabestari, Yadolah Farshi, Shadi Tabibian, Akbar Dorgalaleh *, Samira Esmaeili Reykande, Seyyed Hossein Kia, Bijan Varmaghani, Jamal Rashidpanah

Introduction: It is believed that uremia causes destruction of white blood cells (WBC) and thus causes leukopenia. Therefore this study had an attempt to assess the effect of uremia on WBC count. Materials and Methods: This case control study was conducted on 120 uremic patients and 100 samples as control group. All samples were examined for determination of urea and creatinine in their serum and complete blood counts were determined. Results and Conclusions: In healthy individuals, the mean value of urea was 14.5±1.9 mg/dL and the mean value of creatinine was 0.9±0.2 mg/dL (male) and 0.66±3.2 mg/dL (female). In the patient group, the mean value of urea was 83±2.4 mg/dL. The mean value of creatinin in male and female were 2.4±1.3 mg/dL and 2.1±1.7 mg/dL respectively. The mean of WBC count in case and control groups were 6.08± 2.24 and 6.17± 2.43x109/L respectively (p=0.71). Our results indicate that uremia cannot change leukocyte count.

7

Authors: Mohammad Mehdi Kooshyar, Mohammad Hadi Sadeghian, Mohammad Reza Keramati, Hossein Rahimi, Seyyede Fatemeh Shams, Sepideh Shakeri, Hossein Ayatollahi *

Background and Aims: Multiple myeloma is a malignant proliferation of plasma cells derived from a single clone. The tumor, its products and the host response lead to organ damages. Some factors that are responsible in its pathogenesis are recognized. As FMS like Tyrosine Kinase 3 receptor (FLT3) mutation has been proved as a determining factor in leukemic patients the goal of this study was to find association of FLT3 internal tandem duplication (ITD) and FLT3 tyrosine kinase domain (TKD), mutations with multiple myeloma. Materials and Methods: This case-control study was conducted on 60 paraffin-embedded bone marrow biopsies (30 multiple myeloma and 30 normal bone marrow specimens) in the pathology departments of Ghaem and Imam Reza hospitals in Mashhad. After sections preparation, DNA was extracted and two PCR reactions were set up for detection of FLT3/ ITD and FLT3/TKD mutations. Results: The Mean age of samples was 64±10 years. No FLT3 mutations were detected in multiple myeloma patients. Conclusions: Our findings showed that FLT3 mutations occurrence seem unusual in multiple myeloma.

8

Authors: Arman Jalili, Hossein Ali Khazae *, Zari anchuli

Background and Aims: The immune system protects the body against many invasive foreign materials. The aim of this study was to investigate the effect of an intense anaerobic exercise session on serum IgG, IgM and IgA levels in handballists, volleybalists and climbers. Materials and Methods: In this study, 45 professional athletes with the average age of 20-30 years who had participated in the major leagues and the first batch of Zahedan city were enrolled. To assess anaerobic power the 30 second Wingate test was used. Blood samples before, immediately after, and 2h after exercises were collected, and serum levels of immunoglobulins IgG, IgM and IgA were measured by nephelometry method. Results: The results of this study indicated that the level of serum immunoglobulins IgG, IgM ˛ IgA concentration in all three study groups before and after and two hours past exercise did not show significant change (P>0.05). Conclusions: Our findings showed that short anaerobic exercise does not have any effect on the level of immunoglobulins in athletes.

9

Authors: Masouma Mowahedi, Mohammad Samet, Fateme Zare, Morteza Samadi *

Background and Aims: Recent evidence suggests that T helper (Th) 17 cells are involved in the emergence of asthma. Th17 cells have a key role in inducing inflammation in asthmatic airways thus Interleukin (IL)-17A, the main cytokine of Th17, contributes to airways inflammation. Materials and Methods: We evaluated the level of IL-17A and total immunoglobulin E (IgE) in sera of 100 asthmatic patients and 81 healthy controls by ELISA to determine how serum concentration of IL-17A is associated with asthma severity. We classified patients into three groups mild (n=28), moderate (n=33) and severe cases (n=39). Results: Respectively, serum IL-17A and IgE concentrations were significantly higher in the asthmatic patients than the control group (p=0.026 and p<0.01). Mean of serum IL-17A and IgE values were 37.73 pg/ml and 39.02 IU/ml in the control group and 68.55 pg/ml and 295.87 IU/ml in the patients group. Nevertheless there were non-significant differences between the three groups of asthmatic patients. Respectively, mean of serum IL-17A and IgE values were 94.17 pg/ml and 255.07 IU/ml in the mild group, 71.29 pg/ml and 271.27 IU/ml in the moderate group, and 47.85 pg/ml and 345.97 IU/ml in the severe group. Moreover, there was no correlation between serum levels of IL-17A and IgE. Conclusions: In this study we found that IL-17A, like IgE, rises in sera of asthmatic patients though in a different manner. IgE increases in serum consistent with disease severity though the increases of IL-17A in serum has an inverse relationship with IgE rising.

15

Authors: Mohammad Razi, Ali Dehghani, Fahimeh Beigi, Hamid Najminejad, Kazem Vatankhahyazdi, Mohammad Agha Ayatollahi, Ali Jebali *

Nanotechnology has opened a new field in medicine as well as in other sciences. The aim of this study was to seek the capability of nanotechnology for the treatment of various reproductive diseases. In this study, we analyzed all articles about “nanotechnology and reproductive medicine” published in 2000-2015, indexed in Google Scholar, PubMed and Science Direct. This study indicated that nanotechnology has been extensively used for different biomedical applications, e.g. detection, drug delivery, diagnostic imaging, etc. particularly in cancer diagnostics and treatment. Here, the emerging uses of nanotechnology in reproductive medicine and reproductive biology were found. The available evidence regarding the use of nanomaterials as experimental tools for the detection and treatment of reproductive diseases was summarized. Nanoparticles have potential applications in reproductive biology. Treatment and imaging of reproductive system-related cancers can be performed by engineered nanoparticles. Also, some non-cancerous diseases can be treated by nanotechnology, e.g. endometriosis. The benefits and concerns associated with their use in a highly delicate system of reproductive tissues and gametes have been investigated. Nano-based methods are innovative and potentially controversial approaches in the clinical settings and give us the mechanisms underlying reproductive diseases.