International Journal of Medical Laboratory

8

Authors: Mahmood Vakili *, Zahra Rostami, Mohsen Mirzaei

Background and Aims: Since the epidemiological, clinical and experimental characteristics of meningitis are different for various locations, this study was conducted to investigate the characteristics of meningitis patients admitted to hospitals of Yazd in the 2009 to 2013. Materials and Methods: In this cross-sectional and descriptive study, all 261 patients with meningitis were considered in Yazd hospitals in the years 2009 to 2013. The type of sampling method was census. The data were collected from patients’ files using special forms that were designed for this purpose. Results: In this study, 158 men (60.5%) and 103 women (39.5%) were considered. Among the patients, 53 people (20.30%) were under one-year-old, 63 people (24.14%) were between one to seven-year-old, and 85 people (32.57%) were adult. Among 261 patients, 126 ones (48.28%) and 135 ones (51.72%) were hospitalized with a diagnosis of non-bacterial meningitis and bacterial meningitis respectively. Generally, the cultivations of just 4 people (1.5%) were positive. The percentage of neutrophils and lymphocytes, the amount of glucose and the protein of cerebrospinal fluid in bacterial and aseptic meningitis was significantly different (p<0.001), but the total number of white blood cells, red blood cells and lactate dehydrogenase levels were not significantly different between the two groups. Conclusions: Since there is no significant difference between epidemiological characteristics and clinical symptoms in bacterial and no-bacterial meningitis, it seems that doing laboratory tests, despite aggressive, is the best practice for differentiating between types of meningitis and the fast performance of it could somewhat prevent the improper antibiotic therapy.

8

Authors: Hamed Hajipour, Hamed Hamishehkar, Sina Raeisi, Siamak Barghi, Akbar Hasani *

Background and Aims: Epigallocatechin-3-gallate (EGCG) is a polyphenolic compound from green tea, which its anticancer effects on many types of cancers have been confirmed, but the molecular mechanism by which EGCG induces apoptosis remains unknown. The aim of the present study was to investigate anti-proliferative properties and apoptotic signaling pathway of EGCG on PC3 human prostate cancer cells. Materials and Methods: Cytotoxic effect of EGCG on prostate cancer cell line (PC3) was evaluated by MTT assay. DAPI staining was carried out to determine the morphological appearance of cells. Finally, the expression of Bax and Bcl-2 (apoptosis-regulating genes) were evaluated by quantitative Real-time polymerase chain reaction (PCR). Results: Cytotoxicity evaluations demonstrated that EGCG prevented prostate cancer cells growth in a time and dose depended manner, but the effect of treatment duration is more significant than effect of concentration. Cell growth inhibition was found to be accompanied by nucleus condensation or chromatin fragmentations which are signs of apoptosis, as assessed by DAPI staining. Quantitative Real-time PCR results demonstrated that EGCG causes up-regulation of Bax as a pro-apoptotic protein, and down-regulation of Bcl-2 as an anti-apoptotic protein, thus shifting the Bax/Bcl-2 ratio in favor of apoptosis. Conclusions: It is tempting to suggest that consumption of EGCG could be an effective strategy to inhibit prostate cancer. Our results demonstrated that increase in the ratio of Bax/Bcl-2, is the probable molecular mechanisms through which EGCG induces apoptosis in PC3 cells.

12

Authors: Bahareh Rahmatizadeh, Farzaneh Tafvizi *, Masoud Salehipoure

Background and Aims: Prostate cancer is the second most common cancer in men worldwide in men. Matrix metalloproteinase 2 (MMP2) has a role in the invasion and destruction of the basement membrane and the extra-cellular matrix and facilitating the process of tumor cell invasion. The present study was conducted to compare the expression of MMP2 gene in prostate cancer (PCa) and benign prostatic hyperplasia (BPH). Materials and Methods: 48 samples of PCa and 48 samples of BPH (as controls) were examined. RNA was extracted from paraffin tissue blocks and then used for cDNA synthesize. The real time polymerase chain reaction method was used to quantify the expression of MMP2 gene. The level of gene expression was assessed using participants’ clinicopathological characteristics. Results: The expression of MMP2 gene increased in the PCa samples compared with the BPH samples (p=0.003). MMP2 gene increased in all the grades of this group compared with the controls. The highest expression, however, was observed in the moderately differentiated grade. No significant correlations or relationships were observed in the PCa samples between prostate specific antigen and the other study variables such as age, MMP2 expression, disease grade and the Gleason score; however a positive correlation was observed between prostate specific antigen and the samples' pathological stage. Conclusions: The expression of MMP2 increased in the PCa samples compared with the BPH samples, with the highest expression occurring in the samples with a Gleason score of 6. Further tests and studies conducted with larger sample sizes may help to use this marker in differentiating malignant from benign samples.

13

Authors: Hamidreza Famitafreshi, Morteza Karimian *

Background and Aims: Alcohol abuse is an important concern of many societies. Hippocampal neurogenesis regulates abusing drugs in a positive manner. The aim of this study was to identify factors that regulate neurogenesis in isolation period that increase preference for alcohol and salt. Materials and Methods: In this study sixteen rats were randomly divided into two groups: Pair (social) and isolation. Rats in the isolation group were isolated for 14 days plus one week for acclimatization. Rats in pair group also were kept in the same condition for 14 days. In this period BrdU (50 mg/kg/day/i.p.) was injected. At the end of the experiment, rats examined for copper, zinc, malondialdehyde (MDA), neurogenesis, salt consumption and alcohol preference. Results: Zinc in serum reduced in isolated rats, but copper in serum paradoxically increased in isolated rats. Neurogenesis reduced in isolated rats. Also, MDA in serum, salt consumption, and alcohol consumption increased in the isolated group. Conclusions: Social isolation with reduction of neurogenesis predisposes rats to consume more alcohol and also salt. The reduction in neurogenesis is associated with paradoxical balance of zinc and copper and increase in MDA in serum. So, regulation of copper and zinc may have beneficial effects on neurogenesis, sensitization and alcohol preference.

4

Authors: Zakieh Rostamzadeh *, Nasim Valizadeh, Mahshid Mohammadian

Background and Aims: Human T-lymphotropic virus (HTLV) is a human retrovirus which has been known to cause adult T-cell leukemia/lymphoma and some other inflammatory disorders. Patients with hereditary bleeding diseases are at high risk for these viruses. In this study, we evaluated serological HTLV-I/II infection among these patients in west Azerbaijan of Iran. Material and Methods: We studied 50 hemophiliacs including 43 males and 7 females. Serum samples were screened for Anti-HTLV-I and II using enzyme-linked immunosorbent assay. Western blot method was applied on borderline results for confirmation. Results and Conclusions: All studied patients were seronegative for HTLV-I/II and serologic evidence of HTLV I/II infection was not found in any of patients. Based on our results, blood donor population of Urmia must be increased to achieve more accurate results.

7

Authors: Amir Cheraghi, Shahla Mohammad Ganji *, Bita Najmi

Background and Aims: Recently it has been proved that some of Enterobacteriacea like Klebsiella pneumoniae, which carry PKS islands, damage dsDNA by encoding Colibactin genotoxin; and finally they induce some apoptosis in damaged mucosal cells. This study aimed to isolating the clbN and clbB genes, which are the markers of the PKS genomic island, from PKS cluster in these bacteria Materials and Methods: In this study, 110 biopsies were obtained from colorectal cancer patients referred to the clinic of Imam Khomeini Hospital. Then, all samples were cultured in LB medium for bacterial isolation. In the next step, Klebsiella peunomoniae was detected by biochemical and microbiological tests. Finally, the genomes were extracted by boiling method and then were amplified by PCR with specific clbN and clbB gene primers. Results: Our findings have showed 30 Klebsiella pneumonia isolated out of 110 biopsies (27.3%) from patients with colorectal cancer; in which all of biochemical and microbial experiments including Gram-negative staining confirmed this bacteria. Also, 23.23% frequency of clbN were positive, 20% frequency of clbB were positive genes in the isolated Klebsiella pneumonia, and 13.33% frequency were positive for both clbB and clbN genes as simultaneous. Conclusions: It is expected to find a significant correlation between Klebsiella bacteria which carries PKS genes and colorectal cancer by increased number of samples. The lower frequencies for Klebsiella with clbB and clbN positive strain in patients with colorectal cancer were shown in European countries based on the results of this study rather than the similar studies.

7

Authors: Reza Dolatyari Eslami, Asghar Tanomad *, Jamal Hallajzadeh, Mehdi Ariana

Background and Aims: Probiotics are defined as live microorganisms that are beneficial to human health. Probiotics are commonly found in some foods such as dairy products or complementary tablets. A recent research shows that probiotics have a significant role in diabetes type 1 and 2 treatment. Among the variety of lactic acid bacteria, Lactobacillus strains are well known in probiotics for their different properties. The aim of this study is to investigate the effect of Lactobacillus delbrueckii PTCC1057 on diabetic mice blood sugar. Materials and Methods: sample mice were selected from the age range of 6 to 8 weeks. Then for converting the normal mouse to diabetic type, 180 mg/kg streptozotocin was injected intraperitoneally. 10 days later, the blood sugar of the mice was measured and the mice with higher than 200 mg/dl of blood sugar were considered as diabetic mice. Finally, the diabetic mice were divided into two groups of experimental and control for next investigation. For a survey of probiotics effect on blood sugar, an amount of 3×106 CFU of L. delbrueckii was fed by gavage feeding approach daily during 4 weeks and the blood sugar level was determined at the end of the week after streptozotocin injection. Results: the result showed that the diabetic mice blood sugar level significantly decreased conspicuously by feeding L. delbrueckii in comparison to diabetic control group. Conclusion: The result of this study suggests that L. delbrueckii can be impressive in diabetes treatment. However, another clinical study is needed for proving the exact effect of this probiotic on diabetes.

9

Authors: Niko Rostaei Rad, Mahmood Vakili, Javad Zavar-reza *, Sona Rezaie, Ahmad Reza Shirvani

Backgrounds and Aims: Thyroid hormones have a crucial physiological role to maintain balance of metabolism of body. Thyroid hormones have an important role in metabolism and proliferation of blood cells. Thyroid gland also has a vital influence on erythropoiesis by induction of erythropoietin secretion and proliferation of erythroid progenitors. In the present study, it was tried to evaluate the effect of hypothyroidism on hematological parameters and also body iron store. Materials and Methods: The research population consisted of 90 subjects divided into two groups: 45 people with hypothyroidism and 45 age-matched healthy individuals. This study checked the amounts of total triiodothyronine (T3), total thyroxine (T4), thyroid stimulating hormone (TSH), and some iron status and hematological parameters. Results: The mean TSH levels were significantly increased in hypothyroid patients compared to control group. On the other hand, the levels of T4 and total iron binding capacity, serum iron, ferritin, Hemoglobin, and Hematocrit were significantly lower in hypothyroid patients compared to control group (p<0.05). However, there was no significant difference in the mean of T3 between hypothyroidism and control. Conclusion: Our data suggest that alterations in thyroid status change serum iron metabolism and hematological index. Hypothyroidism is often associated with anemia,depleted body iron store. As there is no significant clinical manifestation of hypothyroidism at earlier stages with anemia, it is advisable to routinely investigate it for early detection allowing its early management.

6

Authors: Shokouh Rajabi Firoozabadi, Seyed Mostafa Shiryazdi, Fateme Keshavarz, Tahere Nazari, Nasrin Ghasemi *

Backgrounds and Aims: P53 gene is regarded important in pathogenesis of different cancers. Therefore, this study aimed to investigate the frequency of p53 gene codon 72 Arg/Pro polymorphism in women suffering from breast cancer. Materials and Methods: A total of 90 patients with breast cancer and 83 matched healthy control women participated in this case-control study. Genomic DNA was extracted from peripheral blood circulation, and codon 72 polymorphism of the p53 gene was examined using the amplification refractory mutation system polymerase chain reaction (ARMS-PCR) method with 4 different primers contemporaneously. Results: A significant difference was observed between patients and controls (p˂0.05) in regard with allele frequency of the p53 gene codon 72 Arg/Pro. The distribution of genotypes frequencies was also proved to be significantly different between the cases and controls (p˂0.05). Conclusions: The results of the present study revealed that Pro/Pro homozygous and its Pro/Arg heterozygous genotypes of p53 gene codon 72 constitute breast cancer in our community. However, the complex instinct of the breast cancer and polymorphisms needs to be further studied in regard with different polymorphisms along p53 and various genes in this population in order to clarify the role of codon 72 in susceptibility of the breast cancer.

7

Authors: Hojatollah Moemeni, Durdi Qujeq, Alijan Ahmadi Ahangar *, Karimollah Hajian, Hadi Parsian

Backgrounds and Aims: Stroke incidence rate has increased during the past 2 decades. The question of whether calcium, chloride, potassium, sodium levels are associated with stroke has remained controversial. The present study aimed to evaluate the serum levels of potassium, sodium, calcium, and chloride within stroke patients. Materials and Methods: The study participants consisted of 53 cconsecutive stroke patients as well as 53 healthy subjects.The stroke was defined by focal neurological signs. After collection of blood samples, serum levels of calcium, chloride, potassium and sodium concentration were measured by standard laboratory method. Results: Serum calcium level in the patient group was significantly increased as compared to the control group [11.0±1.3) vs. 9.4±0.7, mg/dl], whereas serum potassium level in the patient group was decreased as compared to the control group [3.5±0.6 vs. 3.9±0.4, mmol/L]. Serum sodium level in the patient group was slightly low as compared to the control group [136.3 ±0.6 vs. 136.7 ±5.5, mmol/L]. Moreover, serum chloride level in the patient group was slightly high as compared to the control group [102.4±7.9 vs. 100.7±6.2, mmol/L]. Conclusions: The study results demonstrated that a statistically significant positive correlation was observed between serum electrolytes or ionic levels and stroke status. Changes in calcium, chloride, potassium and sodium levels during stroke may be a good biochemical marker of diagnosis for this disease. Therefore, these biochemical factors could play a useful role in the stroke. However, further studies are demanded to confirm the validity of study.

9

Authors: Hassan Momtaz *, Amir Ghafari, Mostafa Sheikh-Samani, Ali Jhazayeri

Backgrounds and Aims: Mastitis is a mammalian disease which is considered important due to its potential economic damages. Trueperella pyogenesis is one of the important opportunistic pathogens of the mammary glands of cattle. This bacterium can produce acute mastitis infection in dairy cattle. In fact, this bacterium has several virulence genes which contribute to its pathogenicity. Therefore, this study aimed to determine the frequency of this bacterium in the sick cattle in addition to identifying the effective virulence gene in this disease and finally investigating the antibiotic resistance of T. pyogenes. Materials and Methods: In total, 126 milk samples from mastitic cows, were collected and immediately transferred to the laboratory. Isolation and identification of T. pyogenes strains were performed using bacteriological methods. Moreover, antimicrobial susceptibility testing was performed using the disk diffusion technique polymerase chain reaction was used to detect plo, fimA, fimC, cbpA, nanH, and nanP virulence genes. Results: T. pyogenes was detected in 46 milk samples. The expression of plo and fimA virulence genes was observed in all samples while fimC expression was detected in 84.7% of cases. The most resistance was observed against tetracycline (97.8%) and gentamicin (86.9%), whereasthe least resistance was reported for chloramphenicol (4.3%) and nitrofurantoin (10.9%). Conclusions: T. pyogenes has been regarded as one of the bacteria causing mastitis disease in the cattle. Plo, fimA, and fimC virulence genes seem to be the main virulence factors of this bacterium.

12

Authors: Fahima Danesh Pouya, Javad Zavar-reza *, Beman Ali Jalali, Naser Danesh Pouya

Backgrounds and Aims: Methylglyoxal (MGO) is an –α, β dicarbonyl aldehyde inevitably produced from triose-phosphate intermediates of glycolysis, and amino acid. Increased MGO in blood leads to alterations in coagulation, clot permeability and thus, atherosclerosis in children with diabetes; however, the precise mechanism is not clear. The present study aimed to compare different concentrations of MGO and aspirin on coagulation and clot permeability in the plasma of healthy individuals in vitro. Materials and Methods: Different concentrations of MGO (5, 50, 100, 500 µM) and aspirin (1, 10, 100 mg/l) were added to the plasma citrate. They were incubated at 37°C for 24 h. Then, coagulation parameters were analyzed by the turbidimetric procedure and then clot permeability was investigated. Results: MGO at 500 µM with aspirin 100 mgl/ made significant changes in the coagulation maximum velocity (0.253±0.006), total coagulation time (803±8.88s) and permeation coefficient (0.778×10-6±0.099) compared to MGO at 500 µM (0.271±0.007), and (500±10.00), (0.446×10-6±0.017), respectively (P< 0.05). MGO at 500 µM with aspirin 1 mg/l did not significantly change in either parameter (p>0.05). MGO at 100 µM with aspirin 1 mg/l did not significantly change in either parameter (p> 0.05), compared to MGO at 100 µM. MGO at 5 µM with aspirin (1, 10, 100 mg/l) changed in all coagulation and clot permeability parameters (p<0.05), compared to MGO at 5 µM. Conclusions: Our findings revealed that aspirin (≥1 mg/l) was held to have more effects on higher concentrations of MGO. Moreover, it decreased the velocity of coagulation and increased permeability of clot.

7

Authors: Ali Jebali *, Bahram Kazemi, Seyedhossein Hekmatimoghaddam

Background and Aims: Some nanoparticles can be used in immunoassays to increase sensitivity. This study aimed to evaluate a novel nano-immunoassay based on bovine serum albumin nanoparticles (BSA NPs). Materials and methods: At first, the nanostructure was synthesized, and then applied as a tag in the nano-immunoassay. Then the concentration of β-subunit of human chorionic gonadotropin (βHCG) in the clinical samples was quantified by traditional enzyme-linked immunosorbent assay (ELISA), and then checked by the nano-immunoassay. Results: The Pearson’s correlation coefficient between ELISA and nano-immunoassay was high, i.e., 0.80. Relative sensitivity and specificity of this nano-immunoassay were reported 97.4% and 96.6%, respectively. Conclusions: BSA NPs can be applied in nano-immunoassays as a new structure, and as an example, βHCG can be detected by this novel assay.

7

Authors: Ali Aliabadi, Mahin Izadi *, Mohammad Ebrahim Rezvani, Mansour Esmaeili-dehaj

Background and Aims: Kindling is regarded as an experimental model of temporal lobe epilepsy that reflects a process of progressive and persistent intensification of seizure. Experimental and clinical evidence suggest that there is an interrelationship among epileptogenesis, local inflammation and antioxidant activity. In this study, the possible antioxidant and anti-inflammatory effects of thymol was investigated in epileptic rats. Materials and Methods: Kindling was induced by repeated injections of intraperitoneal pentylenetetrazole (PTZ) every other day. Then, the rat hippocampi were isolated, weighed and prepared as a 5% tissue homogenate in ice-cold 0.9% saline solution. Serum malondialdehyde (MDA) and superoxide dismutase (SOD) levels were assessed by thiobarbituric acid reacting substance and nitroblue tetrazolium methods respectively. Hyppocampal tumor necrosis factor alpha (TNF-α) and interleukin-1 beta (IL-1β) were evaluated using enzyme-linked immunosorbent assay method. Results: As the study findings revealed, epileptic seizures increase the serum level of MDA, hyppocampal levels of TNF-α as well as IL-1β and decrease the SOD activity. Conclusions: thymol exerts anticonvulsant activity through the antioxidant and anti-inflammatory mechanisms.

10

Authors: Fahimeh Nourbakhsh *, Amirmorteza Ebrahimzadeh Namvar, Hassan Momtaz

Background and Aims: Staphylococcus aureus is one of the important pathogens around the world. The present investigation was carried out to study the distribution of Staphylococcal Cassette Chromosome Mec (SCCmec) types and antibiotic resistance properties in methicillin-resistant Staphylococcus aureus isolated from Isfahan hospitals. Materials and Methods: A total of 250 clinical specimens were collected from three major Isfahan hospitals. The samples were cultured, and biofilm producer isolates were subjected to several polymerase chain reaction methods. The patterns of antibiotic resistance were studied using the disk diffusion method. Results: In the present study, 110 out of 250 samples (44%) were found to be positive for Staphylococcus aureus, and all the isolates produced the biofilm in different levels. The most commonly infected samples were collected from wounds (44.5%). The incidence of mecA, tetK, ermA, ermC, tetM, aacA-D, linA, msrA, vatA, vatC and vatB antibiotic resistance genes were 93.6%, 34.84%, 28.20%, 29.30%, 21.87%, 18.71%, 9.48%, 8.65%, 7.18%, 4.43% and 3.71%, respectively. The distribution of SCCmec III (42) was found to be the most type out of 103 mec positive strains. Conclusions: In the prsent study, the highest resistance belonged to methicillin (90.2%), erythromycin (89.7%), ciprofloxacin (89.5%) and penicillin (88%) and the lowest resistance was reported for vancomycin (10%) and nitrofurantoin (8%). These infections with these strains require more advanced hospital care with an emerging demand for the novel antibiotics.

14

Authors: Samere Kiani, Hassan Momtaz *, Amir Arsalan Serajian, Elahe Tajbakhsh

Background and Aims: Acinetobacter baumannii is regarded as an important nosocomial pathogen around the world, especially in the intensive care unit that today seems to be resistant to the most antibiotics. Therefore, this study aimed to trace classes 1, 2, and 3 integrin in the isolates resistant to Acinetobacter baumannii. Materials and Methods: In this descriptive study, Acinetobacter baumannii of 67 patients in Ahvaz hospitals were all isolated and their antibiotic resistance pattern was determined by the disk diffusion. The presence of genes coding for antibiotic resistance as well as integrons (class 1, 2 and 3) were analyzed using polymerase chain reaction method. Results: Out of 67 isolates, the most resistance was observed for the antibiotic tetracycline (89.5%) and the most sensitivity to antibiotic was reported for chloramphenicol, meropenem and nitrofurantoin (2.9%). The distribution of dfrA1, sul1, aac(3)-IV, tet(B), tet(A), aadA1, blaCITM, vim, qnr, blaSHV, sim, Oxa-24-like, Oxa-51-like, Oxa-58-like, Oxa-23-like, imp, cmlA and cat1 were 42 (62.6%), 40 (59.7%), 36 (53.7%), 34 (50.7%), 31 (46.2%), 17 (25.3%), 17 (25.3%), 16 (23.8%), 11 (16.4%), 11 (16.4%), 8(11.9%), 5(7.4%), 5(7.4%), 4 (5.9%),4 (5.9%), 4 (5.9%), 2 (2.9%), 1 (1.4%) respectively. Moreover, frequency of class 1, II and III integrons was 67 (100%), 22 (32.8%), and 3 (4.4%) respectively. Conclusions: High prevalence of integrons among Acinetobater baumannii isolated strains in Ahvaz hospitals indicate the importance role of integrons in multidrug resistance in this bacteria. Therefore, unnecessary use of antibiotics are recommended to be avoided.