Medical Laboratory Journal

Applicability of Saliva for Evaluation of Some Biochemical Parameters of Kidney and Liver Function in Healthy Individuals

Nasir Idkaidek

ABSTRACT
          Background and Objectives: Proper diagnosis of clinical conditions is a major goal of clinical and biochemical analyses. Recently, increasing efforts have been put on the use of less invasive sampling techniques with optimal sensitivity and specificity. The aim of this study was to investigate the applicability of saliva instead of blood for measuring biochemical parameters of liver and kidney function in healthy individuals.
          Methods: Plasma and saliva samples were collected from 100 healthy volunteers to measure level of alkaline phosphatase (ALP), alanine transaminase (ALT), aspartate aminotransferase (AST), total bilirubin, gamma-glutamyl transpeptidase (GGT), urea and creatinine using a fully automated chemistry analyzer (ACE Alera) with ready to use validated kits. Receiver operating characteristic (ROC) analysis was carried out using MediCal program to calculate sensitivity and specificity and area under ROC (AUC).
          Results: The mean (standard deviation) salivary level of ALP, AST, ALT, GGT, total bilirubin, creatinine and urea was 20.9 (20.7) U/L, 25.8 (17.9) U/L, 10.6 (11.8) U/L, 9.6 (4.37) U/L, 0.16 (0.13) mg/dL, 0.09 (0.05) mg/dL and 35.6 (15.2) mg/dL, respectively. Saliva to blood ratios of ALP, AST, ALT, GGT, total bilirubin, creatinine and urea was 14%, 113%, 65%, 45%, 19%, 12% and 130%, respectively. The suggested normal saliva ranges of ALP, AST, ALT, GGT, total bilirubin, creatinine and urea were 7-98 (U/L), 31-104 (U/L), 6-31 (U/L), 15-24 (U/L), 0-0.13 (mg/ dL), 0.14-0.31 (mg/ dL) and 45-74 (mg/ dL), respectively. The calculated sensitivity and specificity values were 38% and 85% for ALP), 80% and 76% for AST, 75% and 45% for ALT, 60% 91% for GGT, 49% and 38% for total bilirubin, 20% and 91% for creatinine and 100% and 75% for urea. The AUC was higher than 0.7 for urea, GGT and AST, indicating good sensitivity and specificity of saliva testing for evaluation of these enzymes.
          Conclusion: Based on the results, saliva could be as a noninvasive method of assessing kidney and liver function. Saliva may be a favorable alternative to plasma for measuring level of urea, GGT and AST in humans.

Presence of Silver Resistance Genes is not associated with Minimum Inhibitory Concentration of Silver Nanoparticles against Staphylococcus aureus and Escherichia coli

ezzat allah ghaemi

Background and objectives: Drug resistance in Staphylococcus aureus and Escherichia coli, as severe pathogenic bacteria, has become a health challenge. However, nanoparticles have been introduced as effective candidates for their eradication. In this study, we investigated presence of genes involved in conferring resistance to silver nanoparticles in S. aureus and E. coli isolates and evaluated its association with minimal inhibitory concentration (MIC) of the nanoparticles against these isolates.
Methods: The MIC of silver nanoparticles against 121 clinical isolates of E. coli and 183 S. aureus isolates was assessed by broth microdilution assay. Presence and expression of the silver resistance genes (silE, silR/S) in the isolates were investigated by PCR and real-time PCR, respectively.
Results: The silE gene was found in three (1.6%) S. aureus and four (3%) E. coli isolates. MIC of silver nanoparticles against S. aureus isolates with the silE gene was 1, 2 and 8 µg/ml. Moreover, the MIC of the nanoparticles against silE-positive E. coli isolates was 16 μg/ml in three cases and 8 μg/ml in one case. None of the S. aureus isolates contained the silR/S gene, but presence of both silE and silR/S was confirmed in two E. coli isolates. Real-time PCR showed no sil expression in the isolates containing the resistance genes.
Conclusion: The frequency of the silver resistance genes among S. aureus and E. coli isolates is very low. There is no relationship between presence of the resistance genes and the MIC value of silver nanoparticles.

Effect of Clove Essential Oil (Syzygium aromaticum) on Some Virulence Factors of Staphylococcus aureus

Azizollah Ebrahimi

ABSTRACT
          Background and objectives: Clove (Syzygium aromaticum) essential oil is a food additive with proven antimicrobial and antioxidant properties. Thus, it may be a good candidate for controlling foodborne pathogens, such as Staphylococcus aureus. The aim of the present study was to evaluate effects of sub–minimum inhibitory concentrations (MICs) of clove oil on some virulence factors of S. aureus.
          Methods: The standard strain and 12 field isolates of S. aureus were obtained from our microbial collections. The broth tube dilution method was used to determine the MIC of clove oil against the isolates. Sterile 96-well flat bottom poly styrene microtiter plates were used for planktonic growth and biofilm formation assays. Slide coagulase test was used for assaying effect of clove oil on clumping factor production. Production of α- and β-hemolysins was assessed by culture on 5% bovine blood agar.
          Results: The results showed that sub-MIC concentrations of clove oil inhibited α- and β-hemolysins and biofilm production and planktonic growth of the examined isolates. However, clumping factor was not affected by sub-MIC concentrations of clove oil.
          Conclusion: Our results indicate the favorable inhibitory effects of sub-MIC concentrations of clove oil against growth and biofilm and hemolysins production of S. aureus isolates.

Phylogenic Relationship of Salmonella Serovars Isolated from Different Foodstuffs in Isfahan

Majid Bouzari

   Background and objectives: are divided into two species: Salmonella enterica and Salmonella Salmonella bongori. S. enterica has more than 2,500 serotypes. Serovars of S. enterica such as Typhimurium, Enteritidis, Paratyphi B, Paratyphi A and Newport are associated with human infections. Approximately 75% of human Salmonella infections have been associated with contaminated food such as eggs, chicken, beef, pork, dairy products, fruits and vegetables. The aim of this study was to determine the frequency of Salmonella strains isolated from various food sources in Isfahan, Iran.
    Methods: Forty Salmonella strains were isolated from 450 suspected cases referred to the veterinary reference laboratory of Isfahan Province. The isolates were identified by differential and serotyping tests and then confirmed by PCR. A phylogenic tree was constructed with 34 sequences by neighbor-joining method using the MEGA7 software (version 7.1).
    Results: Overall, 10 Salmonella serovars were isolated from 32 chicken meat, three beef and five egg shell samples. S. enterica serovar Ouakum (20%), S. Enteritidis (17.5%) and S. Typhimurium (17.5%) were the most common serovars, while S. enterica serovar Nitra (2.5%) was found as the least prevalent isolate.
    Conclusion: In this study, S. Typhimurium species is placed in different clusters along with sequences reported from different parts of the world, indicating that the serovars are circulating all over the world.

Screening for Azole Resistance among Aspergillus spp isolated from Soil of Hospitals and a University Campus in Gorgan, Iran

Farhad Niknejad

Background and objectives: Aspergillosis is a widely distributed infectious disease, which is difficult to manage. According to recent studies, the prevalence of resistant Aspergillus fumigatus has increased from 3.3% to 6.6%. Acquired triazole resistance in Aspergillus species is an evolving global health challenge, which has made the control of diseases caused by Aspergillus a concern. This study was performed to investigate prevalence of azole resistance in Aspergillus isolates from environmental samples.
      Methods: In this study, 316 soil samples were collected from three hospitals and a university campus in Gorgan (Iran) from July to September 2017. Two grams of each sample were suspended in 5 ml of 0.2M NaCl with 1% Tween 20. Then, 100 µl of the suspension was plated on sabouraud dextrose agar (SDA) supplemented with chloramphenicol, SDA supplemented with chloramphenicol and voriconazole (VOR, 1 mg/L) and SDA supplemented with chloramphenicol and itraconazole (ITC, 4 mg/L). The plates were incubated at 37 °C and examined for growth after 24, 48 and 72 hours.
      Results: We detected Aspergillus fumigatus, Aspergillus flavus, Aspergillus niger and Aspergillus nidulans isolates in 187(59.2%), 84(26.6%), 147(46.5%) and 65(20.6%) samples, respectively. We found no VOR resistant isolate. However, 21 (25%) A. flavus and 16 (8.6%) A. fumigatus isolates were intermediate for VOR. In addition, seven (8.3%) A. flavus, 68 (36.4%) A. fumigatus, 41 (27.9%) A. niger and three (4.5%) A. nidulans isolates were resistant to ITC.
      Conclusion: We were able to detect A.fumigatus, A. flavus, A. niger from all four sampling sites in Gorgan, North of Iran. A. fumigatus is the most prevalent and most resistant isolate in the studied area. History of previous agriculture activity and use of pesticides in the proximity of sampling sites may have affected the rate of ITC resistance.

Effect of Eight Weeks of Aerobic Exercise and Vitamin D Supplementation on 8-hydroxy-2'-deoxyguanosine and O6-methylguanine DNA methyltransferase in Lung of Rats Poisoned with Hydrogen Peroxide

mohammad ali azarbayjani

Background and Objectives: Prolonged exercise can reduce physiological capacities and cause DNA damage by inducing oxidative stress and inflammatory responses. Aerobic exercise reduces the risk of cancer by activating DNA repair enzymes and reducing oxidative stress. The aim of the present study was to investigate effects of eight weeks of aerobic exercise with and without vitamin D supplementation on DNA damage.

   Methods: Forty-eight adult male rats were randomly divided into six groups: control (C), H₂O₂ (H), H₂O₂and vitamin D (HD), H₂O₂ and exercise (HE), H₂O_2,, vitamin D and exercise (HDE), and dimethyl sulfoxide. Cancer was stimulated through intraperitoneal injection of H₂O₂(2 mmol/kg). Animals in groups HE and HDE ran on treadmill for eight weeks. Concentration of 8-hydroxy-2'-deoxyguanosine (8-OHdG) and O⁶-methylguanine DNA methyltransferase (MGMT) was measured by enzyme-linked immunosorbent assay. Statistical analysis of data was carried out using SPSS 22 at significance level of 0.05.

   Results: Vitamin D supplementation significantly lowered the level of 8-OHdG expression compared to the control group (P=0.0001). The 8-OHdG expression in the exercise group was slightly lower than control group (P=0.063). Combination of exercise and vitamin D supplementation had no significant effect on expression of 8-OHdG (P=0.281). Both exercise and vitamin D supplementation significantly increased MGMT expression compared to the control group (P=0.0001 and P=0.040). However, combination of exercise and vitamin D supplementation had no significant effect on MGMT expression (P=0.326).

Conclusion: The results showed that aerobic exercise and vitamin D supplementation can have protective effects against DNA damage, possibly by increasing antioxidant capacity and DNA repair.

Synergistic Effect of Docetaxel Combined with Quinacrine on Induction of Apoptosis and Reduction of Cell proliferation in a Lung Cancer Cell Line

Esmaeil khorshid Sofyani

      Background and objectives: Combination chemotherapy with new adjuvants has been introduced as an innovative method of treating various types of cancer. The aim of this study was to investigate potential synergistic effect of quinacrine on the anti-proliferative and anti-apoptotic activity of docetaxel in A549 lung cancer cells.
     Methods: Cell viability and apoptosis percentage were evaluated with MTT assay and annexin V staining. To understand the mechanisms through which quinacrine modulates expression of pro-apoptotic and anti-apoptotic genes, expression of Bcl-2, Bcl-xl and Bax genes were investigated using real-time RT-PCR.
     Results: The half maximal inhibitory concentration values for docetaxel and quinacrine was 3.16±1.5 nM and 4.4±0.58 μM, respectively. The combination index value of docetaxel and quinacrine was 0.66 against A549 cells, indicating strong synergism. The expression of anti-apoptotic genes Bcl-2 and Bcl-xl reduced significantly, while the expression of the pro-apoptotic gene Bax increased significantly after co-treatment with docetaxel and quinacrine (P<0.05). Treatment of cells with a combination of quinacrine and docetaxel significantly increased the inhibitory effect of docetaxel (reduced proliferation by 50%) and the percentage of apoptotic cells.
      Conclusion: Our findings suggest that the combination of quinacrine and docetaxel can be considered as a promising strategy for the treatment of patients with lung cancer.

In Vitro Effect of Some Medicinal Plants on Leishmania major Strain MRHO/IR/75/ER

Oghlniaz Jorjani

      Background and objectives: Leishmaniasis is a tropical disease caused by protozoan parasites from the genus Leishmania. In this study, we aimed at investigating the in vitro anti-leishmanial effect of essential oils of Rosmarinus officinalis, Mentha pulegium, Foeniculum vulgare, Lippia citriodora and Pelargonium graveolens.
       Methods: The essential oils were prepared from freshly dried and powdered plants with steam-distilled water. Iranian strain of Leishmania promastigotes was cultured in RPMI medium and the inhibitory effects of different concentrations (25, 32, 62.5, 125, 250, 500 and 1000 μg/ml) of the essential oils were investigated at 24, 48 and 72 hours. The number of live parasites before and after treatment with the essential oils was counted by trypan blue 10% staining and using neobar lam.
      Results: The essential oils significantly decreased the number of promastigotes in a dose-dependent manner (P<0.05). However, the inhibitory effects of F. vulgare and R. officinalis essential oils were more profound compared to other essential oils. Moreover, concentrations of 500 and 1000 μg/ml of these two essential oils exerted equal and more anti-leishmanial potency compared to glucantime, the first-line drug used for treatment of leishmaniasis.
       Conclusion: Based on the results, it is recommended to evaluate the in vivo anti-leishmanial effects of the tested essential oils, particularly F. vulgare and R. officinalis.