International Journal of Medical Laboratory

10-16

Authors: Niayesh Mohebbi, Sara Pakzad Karamad, Keyvan Gohari Moghadam, Mohamadreza Jafary, Poorya Rajabi, Afsaneh Nejat, Amir Kasaeian, Mona Talaschian

Background and Aims: In 2019, a novel coronavirus called SARS-COV-2 spread throughout the world, causing a pandemic a few months later. As the virus is not well-known and highly contagious, it is important to observe patients for all possible symptoms and comorbidities. This study aimed to demonstrate some demographic, clinical, and paraclinical features of the deceased patients with COVID-19 in Iran. Materials and Methods: A retrospective observational study was designed to demonstrate the clinical and paraclinical features of deceased patients who had a positive real time-polymerase chain reaction test result for COVID-19. The study included all COVID-19 patients who visited Shariati Hospital, Tehran, Iran, from February 20th until May 20th, 2020. All data were analyzed using IBM SPSS version 21 (Mann-Whitney or Fisher’s test). Results: The patients had a mean age of 63.86 ± 16.428, and most were males (59.65%). Diabetes and hypertension were the most frequent comorbidities. The most common symptoms among the patients were dyspnea (64.91%), weakness (61.40%), fever (60.71%), and having ground-glass opacities in the CT scans (85.96%). There was also a rise in lactate dehydrogenase, serum ferritin, D-dimer, and procalcitonin. Conclusions: Older COVID-19 patients are more likely to get more severe forms of the disease. Most of the deceased COVID-19 patients had ground-glass opacities in their CT scans. Dyspnea, fatigue, and fever were the most frequent symptoms. Hypertension and diabetes were the most important comorbidities. There was almost no significant difference between the two genders regarding clinical and paraclinical symptoms.

27-34

Authors: Vahid Nasiri

Background and Aims: In most of the studies, Toxoplasma gondii is maintained in laboratory mice or studied in vitro using non-lymphoid cell lines or primary mouse macrophages. The target of our research was to design a new axenic culture of Toxoplasma gondii tachyzoites to providing a sufficient quantity of them. Material and Methods: Theileria annulata-infected lymphoblastoids, which had been maintained up to 260 sub-cultures to attenuate the Theileria annulata, were evaluated for their suitability to the cultivation of Toxoplasma gondii tachyzoites. This cultivation process was carried out continuously for up to 10 passages, and after each 5 sub-culture, 0.1 ml of culture suspension (1×106 tachyzoites) was inoculated to each BALB/c mouse. Results: It was observed that the tachyzoites have attacked the lympho-blastoids, multiplied inside them, and many fresh tachyzoites with typical shape and gliding movement were present in the culture suspension. In all processes of cultivation, the pathogenesis of parasites remained stable, and they were able to proliferate in mice and eventually lead to the death of the animals. Conclusions: We describe here a new protocol for prolonged maintenance of tachyzoites of Toxoplasma gondii, which is more efficient (both in terms of yield and cost (it does not need fetal calf serum)) than other traditional methods for maintenance of the parasite.

44-54

Authors: Hamid Reza Jamshidi , Sina Negintaji

Background and Aims: Hepatotoxicity induced by Co-amoxiclav has been indicated in multiple studies. Thymol is the main constituent of the Thymus vulgaris essential oil that has antioxidant properties. Even though thymol can exhibit antioxidant activity in vivo models, there is a lack of evidence about the thymol’s effectiveness in drug-induced liver injury. Thus, the present study was conducted to explore the thymol anti-hepatotoxic effects. Materials and Methods: Thirty male rats were randomly divided into five groups of six. The control group received corn oil (0.25 ml/100 g body weight). CoA group was given only co-amoxiclav in doses of 10 mg/kg daily by gastric tube. CoA+T50, CoA+T150, and CoA+T300 groups orally received Co-amoxiclav at the same dose as the second group along with thymol at a daily dose of 50, 150, and 300 mg/kg for 7 consecutive days. At the termination of the treatment, all animals fasted overnight, and then blood samples were collected to determine alanine transaminase, aspartate transaminase, alkaline phosphatase, glutathione S-transferases, and bilirubin. Results: Administration of thymol at the dose of 300 mg/kg with co-amoxiclav resulted in a significant decrease in direct and total bilirubin levels. Findings also revealed that the concomitant administration of thymol at the 150 mg/kg dose caused a significant reduction in the total bilirubin level. Additionally, the concomitant administration of thymol at the doses of 150 mg/kg and 300 mg/kg resulted in a significant reduction of alanine transaminase, aspartate transaminase, and alkaline phosphatase serum activities along with increased plasma Glutathione S-transferase activity compared to co-amoxiclav group. Conclusion: Administration of thymol can cause a significant ameliorative effect against co-amoxiclav-induced hepatotoxicity in rats.

62-69

Authors: Atefeh Sharifi, Ata Ghadiri, Anayatallah Salimi, Pegah Ghandil, Seyed-Alireza Esmaeili

Background and Aims: Asthma is a complicated chronic inflammatory disease associated with pulmonary inflammation, severe immune responses of the respiratory tract, and change or destruction in the respiratory tract structure. Several factors, especially genetic and environmental factors, contribute to the pathogenesis of asthma. Interlukin (IL)-13 is considered the most important inflammatory mediator of asthma that play an effective role in the different stages of B cell maturation and differentiation. It also increases the expression of major histocompatibility complex II and CD23 and is effective in IgE isotypes switching. Single nucleotide polymorphisms (SNPs) cause the wide genetic diversity in the genome, and SNP analysis helps evaluate and diagnose disease-related genes. The present study was conducted to investigate the association of -1112C / T and + 2044 G /A polymorphisms in the IL-13 gene with asthma. Material and Methods: Blood samples (5 ml) from asthma patients (167) and controls (172) after spirometry test were collected into tubes, and then DNA was extracted to determine the genotype of asthma patients. Real-time polymerase chain reaction was performed by the Taq Man method, and the results were analyzed using SPSS software version 22. Results: There was a significant difference between the genotype of IL-13 -1112C / T polymorphism in the patient group and the control group (p = 0.028). The IL13 + 2044G / A polymorphism results showed no significant difference between the two groups (p = 0.319). Conclusions: The present study showed that IL-13 -1112C / T polymorphism positively correlates with the induction of asthma, but there was no significant association between the polymorphism + 2044G / A IL-13 and the risk of asthma.

247-255

Authors: Zivar Zangeneh, Alireza Andalib, Gholamreza Khamisipour *, Hamid Saadabadimotlagh, Sareh Zangeneh, Niloofar Motamed

Background and Aims: Cells and secreted molecules by the innate immune system are the essential factors in the pathogenesis and determining the severity of inflammation in COVID-19 patients. Severe inflammation results from increased activity of neutrophils, macrophages, and other cells with their products. Inflammatory cytokines such as tumor necrosis factor-α (TNF-α) increases the severity and pathogenesis of the disease caused by the virus. Phagocytes are armed with inducible nitric oxide synthase (iNOS), that upon stimulation by proinflammatory cytokines augment an immune response against pathogens. Materials and Methods: Two groups of patients were included with COVID-19 infection from the intensive care unit (ICU, n=52) and (non-ICU-care, n=54). Blood samples were collected to measure cells and serum parameters, including lymphocytes, neutrophils, platelet counts, accompanied with C-reactive protein, lactate dehydrogenase, TNF-α and iNOS levels. Results: In the ICU group, increased white blood cells (p=0.048), decreased lymphocytes (p=0.0007), increased neutrophils (p=0.001), decreased platelets, increase serum levels for lactate dehydrogenase (p =0.0001), c-reactive protein (p=0.003), TNF-α (p=0.018), and iNOS (p=0.008) were statistically obtained. Positive correlations were calculated between TNF-α and iNOS (r=0.65, p=0.0002) and with c-reactive protein (r=0.52, p=0.003) and with lactate dehydrogenase (r=0.68, p=0.0001). Conclusion: Inflammation due to macrophages and neutrophils activity in COVID-19 patients and increased mediators correlate with disease progression. It seems that control of the cell activity and their inflammatory cytokines would be considered for therapeutic goals. Changing the polarization of inflammatory macrophages to anti-inflammatory macrophages with therapeutic applications could prevent the severity of the provocative course of the disease.

264-274

Authors: Abbas Dabbaghzadeh, Javad Ghaffari *, Jamshid Yazdani Charati, Marziyeh Mohammadi kordkheyli, Fatemeh Pouresmaeil

Background and Aims: Allergic disorders such as asthma, urticaria, eczema, and allergic rhinitis are common worldwide, and allergens are the most common etiology and exacerbating factors. So, This study aimed to find the allergens in these patients with skin prick tests. Materials and Methods: All the allergic cases referred to an allergic clinic in the north of Iran were visited by an allergist and clinical immunologist. Based on patient history, physical examination, and diagnosis, we select food allergens and aeroallergens. A standard skin prick test was performed on all patients, and all data was then analyzed SPSS 20. Results: Two thousand one hundred and twenty-eight cases entered the study with a mean age of 27.65 ± 15.52 years old. 1235 (58.04%) females and 893 males (41.96%) participated in this study. Prevalence of allergic rhinitis, asthma, urticaria, and eczema were 717 (33.7%), 611 (28.8%), 550 (25.8%), and 250 (11.7%), respectively. The most common aeroallergens were Dermatophagoides farinae (75.9%), Dermatophagoides pteronyssinus (65.4%), feather (56.1%), and Candida (51.1%), respectively, in all patients. The most common food allergens were egg white (31.1%), cacao (29.7%), and egg yolk (28.9%), respectively, in all cases. Conclusion: Both aeroallergens and food allergens were the most common in all allergens by skin prick test. Therefore, it is important to find the relationship between sensitization and allergy.

284-290

Authors: Samira Hajimaghsoodi , Abbas Ali Jafari Nodoushan , Mohamad Hassan Akhavan Karbassi, Yasaman Yazdanparast *

Background and Aims: Candida albicans is the most prevalent opportunistic fungal species in the oral cavity. To date, several studies have been investigated the various factors associated with oral candidiasis. On the other hand, it has been proven that blood types antigens lead to some infectious factors. This study aimed to evaluate Candida albicans colonies in the saliva of dentistry students based on their blood type to detect a relationship between blood group and incidence of oral candidiasis. Materials and Methods: In this descriptive cross-sectional study, 200 dentistry students were selected by a simple sampling method, including 100 individuals with blood type O and 100 with other blood types. The unstimulated salivary samples of all the participants were collected by spitting, cultured on Sabouraud medium, and then the isolated Candida albicans colonies were enumerated and recorded. Results: In the present study, samples comprised 77 males and 123 females, of whom 15.5% (31 individuals) carried colony-forming units > 40. The mean of Candida albicans colonies in the individuals' saliva with blood type O was 21.55, and it was 10.68 in the other groups. Besides, the differences were statistically significant (p = 0.024). There was no significant difference in Candida albicans colony count between O positive and O negative blood groups. Conclusions: The result of this study showed a significant relationship between the number of Candida albicans colonies of saliva and the individual’s blood type.

304-315

Authors: Vijayalakshmi Kuttath * , Shyamily Mohan

Background and Aims: This study aims to identify the level of motivation among blood donors and their intention to donate again and suggest remedial measures to rectify de-motivating influences. Materials and Methods: A self-administered questionnaire was distributed among donors to collect data on variables related to the objective. Results: Most donors were men aged 18-24 years, graduates, and employed. The statistical analysis indicated a significant correlation between socio-demographic variables and intention to donate. Fear is the major de-motivating factor, and waiting hours at the blood bank were rated as inconvenient. Conclusion: Recognition of the positive and negative motivation factors and measures to remove misconceptions of donors will go on a long way in the recruitment and retention of blood donors.

168-179

Authors: Samira Soltanmoradi M.Sc., Fatemeh Kouhkan Ph.D., Iman Rad * Ph.D.

Multiple Sclerosis (MS) is the most prevalent neurological disability in young adults. The pathogenesis of MS is characterized by demyelination and neurodegeneration in the central nervous system (CNS) as the ruinous result of chronic activation of the immune system. All clinical forms of MS, including relapsing-remitting multiple sclerosis (RRMS), secondary progressive multiple sclerosis (SPMS), and the primary progressive MS (PPMS), demonstrate inflammation as a common symptom. In various autoimmune diseases like MS, the ability of the immune system to set a balance between pro-inflammatory and anti-inflammatory responses is lost. In this review, the imbalance between proinflammatory and anti-inflammatory responses of immune cells and their role in MS progression is discussed. Disturbing the balance of Th1/Th2 and Th17/Treg cells and M1/M2 phenotypes of macrophages and microglial plays a key role in the development and progression of MS. In this review, we first depict an outline of regulatory immune cells involved in inflammation. Second, we discuss shreds of evidence that confirm how B cells play both pathogenic and protective roles in MS disease. Third, we point out the pros and cons of B cell/T cell-targeted therapies in clinical trials.

188-195

Authors: Naser Nazari 1 Ph.D., Tooran Nayeri 2,3* Ph.D., Farkhondeh Hazrati 4 M.Sc.

Background and Aims: Echinococcus granulosus (E. granulosus) is a cestode parasite that causes cystic hydatid disease in humans worldwide. Iran is one of the endemic regions for infection that indicate the importance and presence of infection in this country. Therefore, the current research aimed to characterize the seroprevalence of human cystic echinococcosis in Sanandaj city, Kurdistan province, western Iran. Materials and methods: Totally, 500 serum samples were collected from patients referred to different health centers in Sanandaj city using cluster sampling in 2018-2019. All the sera were examined using the enzyme-linked immunosorbent assay test. Results: The seroprevalence of human hydatidosis was reported at 2.2% by ELISA test in Sanandaj city. This rate was 9 (1.9%) in women and 2 (0.4) in men. The age group of 20-30 years old had the highest positivity rate (1.0%). Also, the subjects that consumed home slaughtered meat had the highest infection rate at 4 (0.8%). There was no significant difference regarding factors studied such as sex, education, residence, consumed water, keeping a dog, and the seropositivity. Conclusions: Seroprevalence of human cystic echinococcosis in Sanandaj city is lower than the general prevalence in Iran. Our research team hopes to provide accurate data on the prevalence of hydatidosis in Sanandaj encourage more extensive research to help prevent this parasite in Iran and worldwide.

206-214

Authors: Fatemeh Safari 1 Ph.D., Reza Atarodi 2, Pharm.D., Hamid Reza Jamshidi 2,3* Ph.D.

Background and Aims: Long-term surge of heart loads causes cell hypertrophy. Left ventricular hypertrophy is an adaptive response of the heart to pathological stimuli such as hypertension. B-cell lymphoma 2 (Bcl-2) family members play an essential role in this process regulation. This study aimed to evaluate the effect of thymol on the transcription level of Bcl-2 family factors in the rat model of left ventricular hypertrophy. Materials and Methods: Male Wistar rats were divided into four groups: 1- Control 2-Untreated hypertrophy (H), 3 and 4 groups which received 25 and 50 mg/kg/day of thymol (H + Tym25 and H + Tym50 groups, respectively). Hypertrophy was induced by abdominal aortic banding, and the real time polymerase chain reaction technique was used for gene expression. Results: Data showed that in the H group, the mRNA level of the BAD was increased significantly (p ˂ 0.001). However, the transcription level of BAX was increased in the H and H+Tym25 compared with the control group. In the H + Tym50 group, BAX mRNA level decreased significantly compared to the H group (p ˂ 0.05). Conclusions: Our findings demonstrated that the expression rates of the antiapoptotic factor, Bcl-2, was significantly increased in the H group (p < 0.01) and thymol-treated hypertrophy groups (p < 0.001). Interestingly, the upregulation of Bcl-2 mRNA was statistically significant in the H+Tym50 group compared with H and H + Tym25 groups (p < 0.01). The results showed that thymol could protect heart hypertrophied by increasing the expression of anti-apoptotic factors.

223-232

Authors: Hamid Reza Jamshidi 1,2* Ph.D., Faezeh Taheri 1 Pharm. D.

Background and Aims: Mercuric chloride is highly toxic once absorbed into the bloodstream, especially the kidneys in which it is collected. Mercury chloride increases hydrogen peroxide and enhances the destruction of protective enzymes such as superoxide dismutase (SOD) and glutathione peroxidase (GPX), leading to oxidative stress. Besides, thymol has anti-oxidant effects and can increase the activity of SOD and GPX. This study aims to evaluate the efficacy of thymol on mercury chloride-induced toxicity. Materials and Methods: In this study, 30 rats, consisting of 6 groups of 5, were used. Control group receiving a single dose of 0.5 mg/kg mercuric chloride for 15 days, third, fourth, and fifth group received intraperitoneal injection of mercuric chloride at a dose of 0.5 mg/kg for 15 days plus thymol at a dose of 10, 30, 50 mg/kg. The sixth group received mercuric chloride at a dose of 0.5 mg/kg for 15 days plus thymol at 30 mg/kg per day for ten days. Results: Results showed a significant difference in the activity of catalase enzyme in kidney tissue samples test. According to the results of SOD, there is a significant difference between the group of corn oil and the group of mercury chloride and between the group of mercury chloride and the group that receives thymol at a dose of 10, 30, 50 mg/kg (p ≤ 0.05). Conclusions: It can be concluded that mercury chloride-induced kidney toxicity and thymol have anti-oxidant protective effects for SOD and GPX.

96-102

Authors: Alireza Tabibzadeh 1 Ph.D., Masood Naseripour 2 M.D., Mohammad Hadi Karbalaie Niya 3 Ph.D., Davod Javanmard 1 Ph.D., Alireza Sadeghipour 4 M.D., Maryam Esghaei 1* Ph.D.

Background and Aims: Retinoblastoma tumors are the most common intraocular malignancy in childhood, leading to death after two years. The Human Adenovirus (HAdV) infection could be critical in the retinoblastoma pathogenesis due to the virus and retinoblastoma 1 interactions. The objective of the current study was to investigate the possible presence of the HAdV genome in the retinoblastoma patient's tumors. Materials and Methods: In this study, we evaluated the HAdV infection in 96 pathological confirmed retinoblastoma samples. The DNA was extracted from formalin-fixed paraffin-embedded blocks, and the virus infection was assessed using polymerase chain reaction. SPSS version 22 was used for statistical analysis. Results: The mean age ± SD of the retinoblastoma patients was 28.89 ± 17 months. In addition, the demographic evaluation indicated that 43 (46.7%) of patients were female. The retinoblastoma laterality assessment indicates 87 (90.4%) unilateral and 9 (9.4%) bilateral tumors. Growth pattern analysis indicates endophytic 58 (77.3%), exophytic 8 (10.7%), and 9 (12%) of tumors with mix endophytic and exophytic patterns. The polymerase chain reaction results could not found any evidence of HAdV infection in all 96 formalin-fixed paraffin-embedded samples. Conclusions: The study results suggest that there is not any association between HAdV infection and retinoblastoma tumors in studied samples. The HAdV infection may not a concern in retinoblastoma pathogenesis. Further investigations are recommended in this field of study.

114-121

Authors: Jamile Salmanzade 1 M.Sc., Zahra Tahmasebi Fard 2* Ph.D., Zahra Deilami khiabani 1 Ph.D. 1

Background and Aims: The genes involved in detoxification and the elimination of toxic metabolites have a vital role in cancer pathogenesis. Also, there is evidence that higher amounts of body fat are associated with increased risks of several cancers. The current study aims to identify the relationship of age, body mass index (BMI), smoking, family history, and polymorphism rs1056827 of CYP1B1 with prostate cancer. Materials and Methods: A total of 103 patients and 103 healthy men as control groups were enrolled in the current study. The DNAs were extracted using the salting-out method after venesection, and the genotypes were determined by polymerase chain reaction-restriction fragment length polymorphism. The obtained data were analyzed by SPSS version 23 using X2 statistical tests and logistic regression. Results: The results showed a significant association between the study groups regarding age, BMI, and family history of the disease (p = 0.024, p = 4.08⨯10-4, and p = 3.58⨯10-19, respectively). Investigation of genetic models except to additive model showed a significant relationship between TT (Ser/Ser) genotype and prostate cancer. It also showed a strong association between this genotype with BMI and family history. Conclusion: The current study results showed that the carriers of the TT genotype with a high BMI have a higher chance of developing prostate cancer. Further studies in this area will provide stronger results.

131-137

Authors: Mohadeseh Kamali 1 M.D., Mohammad Shariati 2 M.D. Mehdi Taheri Sarvtin 3* Ph.D.

Background and Aims: Vitamin D plays an important role in bone health, cardiovascular health, brain development, immune system regulation, mood regulation, and cancer prevention. This study aimed to evaluate the vitamin D status and the prevalence of vitamin D deficiency in Jiroft city, Kerman Province, south of Iran. Materials and Methods: In this cross-sectional study, 5243 people were selected by random sampling during 2015-2019. Serum 25 (OH) D level was measured by VIDAS 25-OH vitamin D total testing kits. Levels used for definitions were deficient, D3 < 20 ng/ml; insufficient, D3 20-30 ng/ml; adequate, D3 30-100 ng/ml, and potential toxicity, >100 ng/ml. Results: The mean of vitamin D was 26.03 ng/ml and 24.19 ng/ml in men and women, respectively (p=0.003). The prevalence of vitamin D deficiency was 41.55% and 51.7% in men and women, respectively. A significant relationship was seen between age and level of vitamin D (p=0.001). Conclusions: The results of our study showed that vitamin D deficiency is common among the general population, especially in women in Jiroft city, Kerman Province, south of Iran. Serious plans are needed to improve the status of vitamin D in the people living in this region.

147-153

Authors: Mohadeseh Kamali 1 M.D., Seyyedeh Sedigheh Seyyedi 2 M.D., Mehdi Taheri Sarvtin 3* Ph.D.

Background and Aims: Cow's milk is a daily staple food for many individuals that can be contaminated with many toxins such as aflatoxin M1 (AFM1). AFM1 is a chemical form of the aflatoxin B1 produced by some species of Aspergillus genus like A. ochraceus, A. flavus, A. nomius, and A. parasiticus that can contaminate feed and forage cattle. This toxin enters into the milk after eating contaminated feed by cows. AFM1 can cause various dangerous diseases such as cancer and immune deficiency in humans. The present study is aimed to investigate the level of AFM1 in cow's milk in Jiroft, Kerman Province, Iran. Materials and Methods: A total of 90 cow’s milk samples were collected in spring and summer 2019 from available stores in Jiroft city. Enzyme-linked immunosorbent assay was used to measure AFM1 in all cow’s milk samples. Results: In the present study, AFM1 was found in 88 (97.8%) milk samples with a range of 0.2-90.62 ppt (mean, 20.07±24.46 ppt). AFM1 concentrations exceeded 50 ppt (maximum tolerance level of AFM1 in the European Union) was seen in 12 (13.3%) samples. Conclusions: The results of this study showed the presence of AFM1 in cow's milk in Jiroft city. So, in this region, many people are exposed to dangerous diseases such as cancer due to the consumption of milk contaminated with AFM1.