Comparative evaluation of bacterial microleakage of three root canal filling materials, Resilon, gutta flow and Gutta-Percha with AH26 sealer

Apical leakage of tissue fluids around root canal space was cited as the most common cause of root canal treatment failures, for which, fillings of root canal with root filling materials was suggested to achieve adequate apical seal and to prevent leakage. Due to the importance of adequate apical seal, different root canal filling materials were developed and used for this purpose. The purpose of this study was to compare bacterial apical leakage through root canal filling materials of Resilon, GuttaFlow and Gutta-Percha with AH26 sealer. In this in vitro experimental study, 55 single-rooted teeth with healthy roots meeting the study inclusion criteria were selected and randomly divided in 3 experimental groups each containing 15 specimens and 2 positive and negative control groups each with 5 samples. The teeth crown were sectioned near the CEJ, the teeth canals were instrumented with step- back technique while group 1 was obturated with Gutta-Percha, and group 2 with Resilon using a cold condensation technique. Group 3 was obturated with GuttaFlow as recommended by manufacturer. Two-chamber bacterial methods were used to assess bacterial apical leakage by means of Enterococcus Faecalis bacterium during a 60 days time interval. The data were analyzed by Kaplan-Meier test. In root canals filled with Resilon, the median of the day in which leakage was occurred was 17, in Gutta-Percha specimens it was 18 and in canals filled with GuttaFlow the median was 19. 93.3% of the roots filled with each Resilon or Gutta-Percha showed apical bacterial leakage during 60 days while in canals filled with GuttaFlow, the leakage was observed in 80%. Kaplan-Meier test showed no significant differences comparing the day in which the leakage was occurred or the overall incidence of leakage in three materials. Under the conditions of the present study, three root canal filling materials showed similar rates of apical bacterial leakage as no significant statistical differences were noted among them

effectiveness of Mtwo and step back instrumentation techniques on the elimination of enterococcus faecalis from root canal

The aim of this study was to compare the effect of Mtwo rotary system and step back hand system in reduction of enterococcus faecalis bacteria from the root canals of human extracted teeth. In this study, 62 human extracted teeth were divided into two equal experimental groups with 23 teeth each and one control group two extra control groups with 5 teeth each as negative and positive were chosen for confirmation of the canal reinfection too. All of the samples were prepared by K Flie No 20 and Gates Glidden No 2 and 3 before sterilization. Then the teeth were autoclaved and reinfected with enterococcus faecalis. The experimental groups were instrumented either with Mtwo system or step back system. Bacteriological samples were taken after instrumentation to determine the amount of remaining bacteria. In the group 1, fourteen samples and in the group 2, twelve samples represented complete reduction of bacteria. There was not statistically significant difference between two techniques [p>0.05]. Both Mtwo and step back systems are the same on reduction of the bacteria from root canal system, up to the same apical size

Gene cloning of Iranian Leishmania major mannose-1-phosphate guanyltransferase

Leishmania is an obligatory intracellular protozoan parasite, which infects human beings when infected sand fly vector takes a blood meal. Most efforts are towards designing an effective vaccine to prevent leishmaniasis. In this way, development of candidate antigen for vaccine has special important. In this study, we cloned mannose-1-phosphate guanyltransferase gene of Iranian L .major in pET32a expression vector. Primers based on L. major mannose-1-phosphate guanyltransferase sequence gene was designed and synthesized. DNA of Leishmania promastigotes was extracted and PCR reaction was done. PCR product was cloned into pTZ57R and sub cloned into pET32a expression vector. Recombinant plasmid containing 1140 bp as L. major mannose-1-phosphate guanyltransferase gene was extracted and confirmed by restriction analysis. PCR product was sequenced and deposited to GenBank. There were some differences in amino acid sequences between Iranian L. major mannose-1-phosphate guanyltransferase and others previously accepted in GenBank. We amplified and cloned Iranian L. major mannose-1-phosphate guanyltransferase successfully

[Comparative study of antibacterial activity of propolis and Ca[OH]2 against lactobacillus, entrococus feacalis, peptostreptococus and candida albicans]

Elimination of root canal microorganisms is an important step in endodontic treatment. Each one of currently used medicaments has problems such as microbial resistance and tissue toxicity. Therefore, research is still needed to identify better alternatives. Propolis, as one of the honey bee products has been the focus of attention by many investigators in order to evaluate its biological properties. The aim of present study was to compare the activity of Calcium hydroxide and Propolis against lactobacillus, Enterococcus faecalis, Pepto-streptococcus and Candida albicans. This experimental study was conducted to evaluate antimicrobial activity of ethanol extract of Propolis and calcium hydroxide powder mixed with saline solution. Agar diffusion test and dilution methods were used to compare the results. There were separate plates to control diffusion of two substances in agar and antimicrobial activity of solvents. Figures about diameter of inhibition zone and minimal inhibitor concentration [MIC] and minimal bacterial concentration [MBC] were calculated using SPSS program. Paired T-test was used to compare the MIC experimental group differences. Propolis was more effective against Lactobacillus, Enterococcus Faecalis and Pepto-streptococcus with 8.6984mm compared with 7.0833mm mean diameter of inhibitory zone for Ca[OH]2. The difference was statistically significant [P<0.001] indicating that Ca[OH]2 was less effective against experimental microorganisms. The inhibitory zone of the two drugs demonstrated stronger effect of Propolis on contaminating microorganisms. On the other hand, the Minimal Inhibitory concentration of Propolis for all studied microorganisms was at least 4 times less than Calcium hydroxide. Propolis was more effective than calcium hydroxide against lactobacillus, Enterococcus faecalis and Pepto-streptococcus

Prevalence of coxiella burnetii in human, animal hosts and hard ticks in west Mazandaran province; Iran, 2003-4

Query [Q] fever is caused by hard ticks infected by Coxiella burnetii. It belongs to a group of diseases, classified as zoonosis, that are common between human-beings and animals. This study was conducted with the objective of defining the prevalence of Coxiella burnetii in humans, animal hosts and hard ticks in the western part of the Mazandaran province. Blood samples were collected from subjects randomly selected from individuals working in professions that brought them in close contact with animals. We also obtained blood samples from randomly selected farm animals, and a limited number of samples from stray dogs in the community. Hard ticks were collected from the bodies of farm animals and also from the shrubs around the farms. The ticks were identified by genus, species and developmental stage. All blood samples were tested by PCR. With the aid of two pairs of primers especially designed 16S rRNA for Coxiella burnetii, PCR and then Nested-PCR was done on each sample. A total of 2417 hard ticks were removed from: animal bodies [1644] and from the shrubbery [773]. The hard tick species were identified as follow: - Ixodes ricinus [72%] - Hyalomma anatolicum anatolicum [15%] - Boophilus annulatus [9%] - Haemaphysalis sulcata [3%] - Dermacentor marginatus [1%] No positive case of Coxiella burnetii was observed in 1052 investigated samples in this study [120 humans, 135 sheeps, 102 cows, 60 goats, 20 dogs, 10 hedgehogs and 605 hard ticks]. This study did not find any evidence of contamination with Coxiella burnetii in the samples collected from the rural areas of Western Mazandaran. To define the prevalence of this microorganism in different parts of northern Iran further epidemiological studies are necessary

[Detection of human papilloma virus among women with cervical cancer using PCR method]

Cervical cancer is the second leading cancer among women, worldwide. It is also the second malignant cause of death, particularly in women aged 25-65. In order to progress a cancer from dysplasia to invasive carcinoma, a cascade of cellular changes should occur. Since genital HPV carries oncogenes responsible for these essential changes, today HPV is considered as the major risk factor of cervical cancer. It is believed that HPV can increase the rate of cancer progression when associating with other risk factors such as smoking, taking contraceptive drugs, immunosuppression, etc. Paraffin-embedded cervical tissues of 70 patients with cervical cancer were analyzed by PCR method for presence of HPV. In addition, high risk typing of HPV positive samples was performed using HPV high risk typing PCR kit. Among all patients 49% were positive for HPV. HPV16 was the most common type detected in HPV-positive cases. Investigation of age classification showed that a majority of HPV positive cases aged between 35 and 44 years. Considering the prevalence of HPV among young women with cervical cancer and its long premalignant period, we suggest to examine all the women above 20 years of age and also check the suspected cases for HPV

Cloning and expression of TRYP6 gene from ieishmania major [MRHO/IR/75/ER]

Leishmania, needs to detoxify the macrophage derived potent peroxides [H2O2]. Tryparedoxin pathway contains tryparedoxin peroxidase [TSA or TRYP]. The aim of the study was to detect the full-length gene sequence and its encoded protein of the LmTRYP6 gene [EU251502], and comparison the gene sequence with LmTRYP6 [LmjF15.1140], another previously reported member of this gene family. L.major [MRHO/IR/75/ER] promastigotes were cultured, DNA and RNA were extracted and the interested gene was amplified using PCR and RT-PCR methods. PCR/ RT-PCR fragments were purified and cloned first in pTZ57R/T and then in pET15b expression vector. The expressed protein was verified using western blot method. Characterization of the expressed protein was performed bioinformatically. Molecular evaluation revealed that the cloned LmTRYP6 gene [EU251502] encoded a predicted 184 amino acid long protein with a theoretical isoelectric point of 6.1101. Alignment showed a number of changes in amino acid composition including the replacement of highly conserved Trp177 by Cys in LmTRYP6 [ABX26130]. So far no study has been done on this group, i.e. TRYP6 gene, from tryparedoxin peroxidase family. The low homology with LmTRYP6 [LmjF15.1140] and vast array of differences observed in the gene under study [LmTRYP6; EU251502] could open new windows in the field of anti-Leishmania combat. Based on its important role in the viability and successful establishment of the parasite in the host organism it looks to be very good candidate for vaccine development and any other sort of novel drug development

Causes of burns and their outcomes in patients hospitalized in the burn division of Zare hospital 2006-2007

It has been reported that the cause of burn is an important factor in the determination of prognosis and outcome. Due to different results of various centers, we conducted this study to determine the relationship between cause of burn and outcomes including rate of infection, costs and mortality in Zare Hospital. This prospective study, has been preformed on 113 patients from September 2006 to April 2007. The inclusion criteria were ages more than 10 years old and partial thickness or full thickness burn. Based on the clinical situation of patients, appropriate samples, such as swab, tissue biopsy, urine, blood and sputum were obtained. The relationship between cause of burns and different variables, including demographic data, infections, costs and mortality have been assessed by Chi-square test. Flame injury was the most common type of burn [53.2% cases]. Burn percent and days stayed in the hospital were 30.6 +/- 21.2% and 21.6 +/- 11.9 days, respectively. These variables were significantly higher in the patients with flame injury. The mean antibiotics and total cost of patients were 9834102 +/- 11818945 and 24301863 +/- 18872089 Rials. Electricity was the cause of burns in 11.5% of patients. The overall mortality was 29.3%. However, it was significantly different in patients with various causes of burns. Inappropriate outcomes in patients with flame injury were much severe and more common than other causes. There was a relationship between the cause of burn and demographic and psychosocial variables such as sex, marital status, residency, educational state, suicide intent and past medical history of patients

Detection of Chlamydia trachomatis, Mycoplasma hominis and Ureaplasma urealyticum by multiplex PCR in semen sample of infertile men

The aim of this study was to detect Chlamydia trachomatis, Mycoplasma hominis and Ureaplasma urealyticum from semen samples of infertile men by Multiplex PCR and investigation of influence of bacteriospermia on semen parameters. Semen samples of 200 infertile men were evaluated by Multiplex PCR. In addition, analysis of semen parameters was performed according to the WHO guidelines. All the patients were without clinical symptoms of urogenital tract infection. Thirty three percent of cases showed at least one bacterium. We found a noticeable relation between the presence of bacteriospermia and the rate of non motile and morphologically abnormal sperms [P< 0.0001]. In addition, sperm concentration was lower in positive cases [P< 0.04]. There was no relation between leukocytospermia and bacteriospermia [P> 0.05]. Asymptomatic existence of Chlamydia and Mycoplasmas in urogenital tracts might play an important role in sperm impairment due to infertility. Bacteriospermia can influence sperm's motility, morphology and concentration

[Isolation of anaerobic, aerobic bacteria and mycobacterium from foot skin infections of diabetic patients]

Diabetic foot infections are a potentially severe complication of diabetes. Diabetic foot infections can sometimes lead to long-term debilitation and, in the most severe cases, amputation. They are the most common infections in patients with diabetes, whose weakened immune systems put them at an increased risk of acquiring antibiotic resistant infections. For this descriptive study, 120 diabetic patients [30 women and 90 men age ranged between 45-65 years and disease duration of 0.5 to 37 years] were investigated. Immediately after the hospitalization, specimens from infected foot lesions were taken using Thio and BHI as transport medium. Aerobic cultures were carried out in all cases according to conventional methods while anaerobic cultures were performed when appropriate. Finally, susceptibility tests were performed on isolated microorganism. Totally, 75% of cases were polymicrobial infections. We isolated gram positive cocci 95%, gram positive bacilli 35%, gram negative 55% and mycobacterium 10%. Meanwhile, we found that 12.5% of our bacteria were anaerobic and 87.5% were facultative aerobic bacteria. In antimicrobial susceptibility testing, Rifampin was the most effective antibiotic against S.aureus and peptostreptococcus. Surprisingly, E.coli was resistant to all tested antibiotics. Diabetic foot infections have a polymicrobial nature. Antibiotic treatment of infections should be prescribed on the results of microbiological investigation

[Nocardiosis; a case report]

Nocardia could be transmitted to lungs through dust particles; then transmitted to other organs via vascular system. We describe a 11-year old boy presenting with headache and vomiting. CT studies revealed hemorrhage in his right hemisphere as well as cerebral edema. He was hospitalized with primary diagnosis of hydrocephaly and pseudotumor cerebri. Further studies showed nocardia astroides in acid fast and blood agar culture

[Detecting legionella pneumophila with sputum culture and urinary antigen test in patients with acute pulmonary infection]

Community acquired pneumonia [CAP] is a common health problem and one of the main mortality factors worldwide. Legionella pneumophila is one of the most common responsible microorganisms for CAP and may lead to severe complications if left untreated. The present study was conducted to determine the frequency of this organism in patients with CAP.We enrolled 118 patients with CAP, COPD and asthma in Masihdaneshvari Medical center during 2004-2005. For microbiological purposes sputum culture and legionella urinary antigen measurement were achieved. The study population included 32 females and 86 males, their age range 58-77 years. The most frequently isolated respiratory microorganism were: streptococcus pneumonia [88%], candida spp. [76.2%], streptococcus beta-hemolytic [61.8%], staphylococcus [40.6%], klebsiella spp. [27.1%], fungi [16.1%], E.coli [8.4%], pseudomonas spp. [5.1%]. Dyspnea, cough, sputum production, and fever were the most common findings. Smoking was the most commonly found risk factor. Atypical pathogens are responsible for 40% of community-acquired pneumonia and several studies have ranked legionella pneumophila among the three most common microbial cause of CAP in patients admitted to the hospitals. So this is important to notice that urinary antigen test is particularly useful, simple and rapid test for legionella positive cases, because it is often easier to obtain urine in ill patients and the results can be available within hours and also reliable to commence treatment

Genital trct infection in asymptomatic infertile men and its effect on semen quality

Male urogenital tract infection plays an important role in men infertility. Asymptomatic bacteriospermia has been paid attention as a major cause of male infertility. The aim of this study was to microbiological investigation of semen sample of infertile men attending to infertility clinic and evaluation of the effects of bacteriospermia on semen quality. Eighty eight infertile men were evaluated by standard bacterial culture method. Standard semen analysis was performed according to WHO guidelines. Among total cases, 35.22% [31 cases] showed at least one pathogen: 10.22% E.coli, 9.09% Coagulase Negative Staphylococci [Saprophyticcus], 6.81% Group B Streptococci, 5.88% Entrococci, 5.68% Candida sp., 2.27% Gonococci, 2.27% Staphylococcus aureus, 1.13% Klebsiella sp. and 1.13% Providencia sp. There was a significant relation between the bacteriospermia and the rate of no motile and morphologically abnormal sperms [P<0.0001]. The quality of sperm motility was significantly decreased in contaminated semen. The percentage of morphologically normal sperm was lower. E.coli and Entrococci were the most effective agents against sperm parameters. Asymptomatic bacteriospermia has a negative effect on sperm quality. E.coli and Entrococci are the most common bacteria with negative influence on sperm motility and morphology. Moreover, presence of bacteriospermia and leukocytospermia did not correlate with each other [P>0.05]. It seems that leukocytospermia is a poor marker to predict bacteriospermia

Septic polyarthritis caused by staphylococcus aureus

Staphylococcus aureus-induced septic polyarthritis is a rare entity characterized by the involvement of multiple joints. IV drug abusers are at increased risk, and it could be fatal if prompt intervention is delayed. A 30-year old male addict, [IV drug abuser], was admitted in Loghman hospital with high fever [38.5 degree signC], pulmonary symptoms, [cough and suppurative sputum], and pain and swelling in knees, ankles and both wrist joints. Blood and synovial fluid cultures were taken and the diagnosis of staphylococcus aureus-induced septic polyarthritis was confirmed. He was treated with vancomycin, penicillin and cephazolin. Although septic polyarthritis is a rare life-threatening condition, it should be kept in mind in immunocompromised subjects and IV drug abusers. Complete recovery is feasible with prompt diagnosis and intervention

Molecular methods [16s-rRNA] compared to bacteriological diagnosis of meningitis

For treatment of patients with meningitis, rapid diagnosis of the agent is very important. Nowadays all of researches have approved qualification and efficiency of molecular tests. Detection of bacteria from CSF and blood is the major problem as a result of usage of antibiotics by patients. So we researched on CSF samples by PCR test and used DG74 and RDR80 primers for 16s rRNA sequence. Our cases were 51 children with meningitis symptoms that had referred to Mofid Hospital in Tehran. Theses samples were different from culture, cell counter and protein glucose amounts. After researching we reached to these results that 23.5% of cases were positive for bacterial culture and 41.1% of them were positive for PCR test. So sensitivity of PCR was 95.23%, specificity of PCR was 96.66% and efficiency of PCR was 96%. In first group 8 specimen were PCR positive [88.8%]. In second group, all of 12 specimens were PCR positive [100%]. In third, 8 specimens were suspected for viral meningitis, only one case was PCR positive, so it had bacterial agent. In fourth group, all of 22 specimens were PCR negative. Therefore sensitivity and specificity of PCR test with 16s rRNA gene sequence in identification of bacterial agent in CSF was 95.23% and 96.66% respectively

Purification and molecular analysis of BCG antigen 60

Tuberculosis remains as an important socioeconomical and medical problem throughout the world and especially in Iran. Early and timely diagnosis of pulmonary and extrapulmonary tuberculosis is vital to initiate prompt treatment. Current diagnostic methods are either slow or lack enough sensitivity or specificity. Several mycobacterial antigens are involved in the complex interaction with the immune system of the host. Their identification is important for both diagnosis and protection against mycobacteria. Antigen 60 [A60] is a thermostable antigen found in the cytosol of M. bovis and M. tuberculosis. An ELISA test using A60 is designed for diagnosis of tuberculosis with satisfactory results. In previous studies, A60 has also showed a protective effect against experimental infections and useful immunotherapeutic effects in promotion of cancer development. In the present work we tried to purify A60 from the cytoplasm of BCG. A60 was purified by exclusion gel chromatography using sepharose 4B. A60 was recognized by bidimensional immunoelectrophoresis with anti-BCG and anti-A60 antiserum, where it appears as the less mobile component. In agarose electrophoresis, A60 showed only one band but in immunodiffusion it showed two immunoprecipitinogen lines with anti-BCG anti-serum. In analyzing with dot blotting, both cytoplasm and cell wall of BCG showed positive reaction with anti-A60 anti-serum. When A60 was fractionated by SDS-PAGE and analyzed by western blot using anti-A60 antibody, 65,46,40, 38 and 35 KDa protein fractions were identified. It is concluded that A60 is a macromolecular antigen of BCG with a molecular weight of 10[6]-10[7] Da and is a lipoprotein-polysaccharide complex which contains several proteins. A60 is present in both cytoplasm and cell wall of BCG and can easily be purified from BCG vaccine using exclusion chromatography by sepharose 4B, to be used for designing diagnostic tests for TB

Detection of genomic DNA of chlamydia pneumonia in atherosclerotic plaques of coronary arteries

Respiratory pathogen, chlamydia pneumoniae, has been implicated in the pathogenesis of coronary artery disease. Previous studies have demonstrated an antichlamydial seroresponse to be a cardiovascular risk factor and coronary atheromata to contain chlamyial components in varing proportions. Endovascular demonstration of bacterial DNA is required to provide evidence for an infectious component in atherosclerosis. Material and Atherectomy specimens from 102 patients who under- went surgery were examined for the presence of chlamydia pneumoniae by polymerase chain reaction [PCR]. Each specimen was devided into multiple sections of 0.3 Cm2, and frozen at -20°C. Specimens were treated with glacial acetic acid and with a solution containing 0.32 Ml sucrose, 10 mM Tris - Hcl, 5 mM MgCl2 and triton x-100 [1% Vol/Vol] and incubated at room tempreture over night. There after DNA was extracted by first boiling for 20 minutes in NaOH and then by use of Tris buffer. DNA was percipitate with sodium acetate -ethanol by standard method. C. Pneumoniae DNA was detected by PCR in atherosclerotic plaques of 23 patients [22%]. Results of this study demonstrate a moderate prevalence of C. Pneumoniae DNA in atherosclerosis plaques. This study provides data on the possibility of C. Pneumoniae involvment in the pathogenesis of atheroclerosis and additional evidence for an association between this agent and atherosclerosis