Treatment modalities for granulomatous mastitis, seeking the most appropriate treatment with the least recurrence rate: a systematic review and meta-analysis.

BACKGROUND: Granulomatous mastitis (GM) is a rare, benign, inflammatory breast disease with an unknown etiology that predominantly affects women of reproductive age. The definitive treatment of GM is currently controversial; an appropriate therapeutic strategy has yet to be identified, and the disease's high recurrence rate remains. This study aims to determine the recurrence rate for each GM treatment strategy to identify the most appropriate treatment modality. METHODS: The search for relevant articles was undertaken using three international databases, including Medline, Scopus, and Web of Science. Articles published in English until the end of 2021 evaluating the recurrence rate of GM were included. Using Stata 13.0, the pooled incidence and 95% confidence interval (CI) for the recurrence rate were determined. RESULTS: Sixty-five eligible studies were included in our study. The recurrence rates of systemic steroid use, topical steroid use, antibiotic use, methotrexate use, observation, drainage, excision, antibiotic use and surgery, steroid use and surgery, antibiotic and steroid use, methotrexate and steroid use were 24% (95% CI: 21-27%), 11% (95% CI: 6-21%), 18% (95% CI: 14-22%), 13% (95% CI: 7-22%), 11% (95% CI: 7-17%), 65% (95% CI: 50-78%), 13% (95% CI: 10-16%), 23% (95% CI: 14-36%), 7% (95% CI: 5-11%), 11% (95% CI: 6-18%), and 4% (95% CI: 2-8%), respectively. Drainage had the highest recurrence rate, while combined methotrexate and steroid treatment had the lowest rate. CONCLUSION: The optimal treatment strategy for GM depends on the disease's severity, consequences, and the patient's features. The study results indicate that combination therapy is preferable for minimizing the risk of relapse and reducing treatment complications.

An HIV-positive woman with massive brain lesion due to toxoplasmosis: A case report.

Key Clinical Message: Toxoplasmosis-related huge brain lesions may require decompressive craniectomy and lesion excision to avoid brain damage. In this situation, injectable cotrimoxazole is a better choice for treatment. Abstract: Toxoplasma gondii is a worldly distributed obligate intracellular protozoa. Toxoplasmosis is a prevalent opportunistic infection in HIV-infected people, but it was rarely recorded prior to the identification of HIV infection. Here, we report a toxoplasmosis brain lesion in an Iranian HIV-positive patient. A 45-year-old woman with a complaint of malaise was referred to the Valiasr Hospital in Arak city. In her past clinical history, the patient had a history of anemia, deep vein thrombosis (DVT), and positive HIV. The patient was informed of the diagnosis of massive brain toxoplasmosis as a definite diagnosis. The patient was then taken to the operating room for a left decompressive craniectomy, during which the ensuing brain lesion was excised. After a few days, she was discharged from the hospital in good condition and without any complications.

Evaluation of the effect of favipiravir on patients with COVID-19.

Background: The novel coronavirus disease 2019 caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV2) started in December 2019 in Wuhan, China. No specific drug has been accepted for COVID-19 treatment up to now. Favipiravir as an antiviral drug affects RNA viruses like influenza and Ebola. Accordingly, the aim of this study is the evaluation of favipiravir's effect on COVID-19 outcomes. Method: This is a randomized controlled study including 97 patients with COVID-19 randomly allocated into favipiravir or control groups. The primary outcomes were improvement of clinical manifestations and atrial oxygen saturation (SpO2), and the secondary outcome was the length of hospitalization. Results: Clinical manifestations recovery of COVID-19 patients was better in the favipiravir group, and the mortality rates were less than in the control group (P = 0.0001 for both). The level of blood oxygen saturation (SpO2) was significantly higher in the favipiravir group (P = 0.0001). The mean lymphocyte count was lower in the control group (P = 0.004). In addition, levels of blood urine nitrogen (BUN) were higher in the favipiravir group (P = 0.033). The length of hospitalization was similar in both groups (P = 0.586). Conclusion: Favipiravir can be effective for clinical and laboratory improvement of COVID-19 patients, and it is a promising drug for decreasing of mortality rate in these patients.

Post-COVID-19 splenic infarction in a patient with chronic atrial fibrillation: A case report.

We describe splenic infarction (SI), an infrequent condition, in an 82-year-old COVID-19 patient with chronic atrial fibrillation (AF). COVID-19 may cause thrombosis, and AF is a predisposing factor for splenic infarction. Suspicion of SI may be warranted in COVID-19 patients with abdominal pain, especially if a predisposing factor exists.

Pap Smear miR-92a-5p and miR-155-5p as potential diagnostic biomarkers of squamous intraepithelial cervical cancer.

BACKGROUND: one of the female-specific diseases with a high incidence and mortality is cervical cancer. The main cause of cervical cancer is infection with Human papilloma virus (HPV). Low-grade squamous intraepithelial lesions (LSIL) and High-grade squamous intraepithelial lesions (HSIL) usually is caused by an HPV infection. Considering the role of microRNAs (miRNAs) as diagnostic biomarkers for a variety of cancers, the aim of this study was to determine miR-92a-5p and miR-155-5p expression levels in LSIL and HSIL Pap Smear samples. METHODS: After initial bioinformatic studies, A total of 75 samples (25 samples of patients with LSIL, 25 patients with HSIL and 25 healthy individuals) were subjected to RNA extraction and cDNA synthesis. The expressions levels of confirmed miRNAs in samples of patients with LSIL, HSIL and healthy individuals were evaluated by Real time PCR analysis. To demonstration the role of predicted miRNAs as novel biomarkers in diagnosis of LSIL and HSIL, ROC curve analysis was done. RESULTS: Bioinformatics results showed that miR-92a-5p and miR-155-5p target the HPV E6 and E7 genes. The expression levels of these miRNAs were strikingly higher in Pap smear of patients with LSIL than in the healthy individuals (35.36, P = 0.001) (62.23, P = 0.001). Similarity, expression levels of miR-92a-5p and miR-155-5p were amazingly higher in patients with HSIL than in the healthy individuals (33.62, P= 0.001) (69.07, P= 0.001). Although, the levels of miR-92a-5p (0.95, P = 0. 85) and miR-155-5p (1.11, P = 0.84) exhibited no statistical differences between patients with LSIL and HSIL. Also, ROC curve analyses verified that miR-92a-5p and miR-155-5p are specific and sensitive and may serve as new biomarkers for the early detection of cervical cancer. CONCLUSION: These data suggest miR-92a-5p and miR-155-5p, which are upregulated in LSIL and HSIL, can be consider as predictive biomarkers for the prognosis of cervical cancer patients.
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Clinical manifestation and epidemiological findings of human linguatula serrate infection in Iran: systematic review.

The aim of this study was to collect data and provide information on clinical diagnosis of disease, methods of dissemination, prevention and treatment of human linguatuliasis in order to increase people's awareness and health care. In this study case reports of individuals in Iran those infected with the Linguatula serrata were taken from the international databases (e.g., Medline (PubMed), Scopus, Iranmedex, Scientific Information Database, and Irandoc) and reviewed. The review covers the case reports of human tongue worm infection published either in Persian or in English between the years 1980-2018. An individual researcher undertook the bibliographic search and evaluation of articles based on inclusion/exclusion of specific criteria. Cases of infection have been reported in eight provinces of Iran. Dietary habits play the important roles in the transmission of the parasite. Clinical diagnosis was achieved by obtaining a patient's history, then completing a clinical examination of the mentioned patients. Symptoms of infection were mainly observed in the upper respiratory tract in most reports, the patient's status was improved by removing the parasite from the infected organ. Given that viscera of slaughtered livestock plays a significant role in the transmission of this infection to humans, Careful inspection of slaughtered livestock in slaughterhouses and Preventing the slaughter of livestock illegally and unsanitary, outside the slaughterhouse, is important. Accordingly, it is very important to train the employees working in the slaughterhouses and the people who consume these organs.

Cryptosporidium animal species in Iran: a systematic review and meta-analysis.

BACKGROUND: Cryptosporidiosis is an acute and short-term infection which can lead to severe diarrhea (intestinal cryptosporidiosis) associated with a persistent cough in the host with immune system defect. This systematic review and meta-analysis was conducted to estimate the prevalence of animal Cryptosporidium species and the corresponding epidemiological aspects in Iran. METHODS: In this study, all original research articles relating to the animal cryptosporidiosis in Iran were collected from reliable databases using keywords. A meta-analysis was conducted separately for each subgroup, and heterogeneity among the studies was performed using the Q and I2 tests. Furthermore, it should be noticed that the significance level in the statistical analysis with the Comprehensive Meta-analysis software was considered to be less than 0.05. Finally, meta-analysis results were shown in forest plot with a 95% CI. RESULTS: In total, 4795 studies were included in the initial screening. Duplicated or non-original studies and the ones which did not meet our considered criteria were excluded from the list. Out of the 100 articles included in our first list for the meta-analysis, 40, 16, 13, 10, 9, 7, and 5 were done on cattle and calves, birds, dogs, sheep, rodents, camels, and horses, respectively. The prevalence rate of cryptosporidiosis among the birds, horses, rodents, camels, dogs, cattle, and sheep in Iran was estimated to be 7.5%, 19.5%, 20.8%, 8.4%, 4.9%, 14.4%, and 9.1%, respectively. CONCLUSION: The different Cryptosporidium species have been found in different regions of Iran. Geographical region, climate, and domestic animals are considered as factors responsible for animal cryptosporidiosis prevalence in the area. Moreover, this parasite is zoonotic which causes disease in animals as well as humans which can result in economic loss.

Fungal and parasitic contamination of indoor public swimming pools in Arak, Iran.

BACKGROUND: Swimming is a popular exercise for different types of people at different ages. Public swimming pools are places where fungal infections can be easily transferred. The purpose of this study is to evaluate the quality of mycological, parasitological, and physicochemical parameters of swimming pools of Arak city. METHODS: This cross-sectional study was done for 12 months from April 2013 to March 2014 in six indoor active swimming pools of Arak city (A, B, C, D, E, and F). Samples were collected in four seasons, two times/season; each time, two samples were obtained from six specified locations (shallow level pool, deep level pool, dressing rooms, showers, margin of pool walls, and foot-washing sink) from each pool with a total of 576 samples. Physicochemical parameters including water temperature, pH, turbidity, and the residual chlorine were measured on-site. In order to isolate and detect the fungal agents, special filters and culture Sabouraud's dextrose agar, chloramphenicol, and mycosel agar media were applied. Furthermore, non-nutrient agar medium enriched with Escherichia coli was used to detect and to separate the eggs of the worms, cysts, and parasitic protozoa from centrifuges of samples. In order to investigate their sediment, optical microscope and culture media were used. RESULTS: We found that 456 (79.1%) samples were positive regarding the fungal elements, and 516 fungal species were isolated. The most common isolates were saprophytic species (8.74%), yeast species (25%), and dermatophyte species (2.5%). The most contaminated surfaces were foot-washing sinks and showers. In this study, Acanthamoeba parasites were isolated from one pool only. All the investigated physicochemical parameters of pool water except for the temperature were found to be in the standard range. CONCLUSIONS: Existence of saprophytic fungi and yeast in pools' water is plausible to be considered as an indicator of water resistance to the detergent agents. This high degree of contamination is due to the huge number of visitors, the complexity of construction, the choice of materials, and the long opening hours. Isolation of dermatophytes and Acanthamoeba parasite from the pools' area and foot-washing sink reveals the important role of the public swimming pools in disease transmission.

Evaluation of Toxoplasma gondii infection in western Iran: seroepidemiology and risk factors analysis.

BACKGROUND: Toxoplasmosis is a parasitic infectious disease, and Toxoplasma gondii is the causative factor of this intracellular protozoan disease. Due to the lack of information about the rate of T. gondii in general papulation of Markazi Province in Iran, the current study was conducted to determine the prevalence of toxoplasmosis and the related risk factor analysis in the general population of Markazi Province. METHODS: This cross-sectional study was performed within 6 months on individuals who were referred to diagnostic laboratories in Markazi Province. The demographic and background information of the subjects were collected using a questionnaire. Three milliliters of blood samples was collected from the participants under sterile conditions. The sera were separated and evaluated for levels of anti-Toxoplasma IgG antibody using a commercial enzyme-linked immunosorbent assay (ELISA) method. The collected data were analyzed by the SPSS software using descriptive statistics and chi-square test. RESULTS: Out of 824 people from the general population of Markazi Province who were investigated in this study, 276 (33.5%) had anti-Toxoplasma antibodies in their blood. According to the logistic regression model, gender variables, location, marital status, and having a cat at home do not affect the chances of contracting the parasite. Furthermore, the chance of contracting the parasite in 41- to 50-year-olds is 0.85 times the one in the 20- to 30-year-olds. The prevalence of toxoplasmosis in men and women in Markazi Province was 33% and 34.5%, respectively. CONCLUSION: The mean prevalence of T. gondii infection in the age groups of 20-40, and ≥ 40 years was estimated to be 24.7%, and 40.8%, respectively. These rates were significantly lower than the national results (44%, and 55%, respectively). Therefore, regarding to the health authorities, it is necessary to raise the level of awareness of people of the region, especially at-risk groups about the transmittance and prevention methods, and infection risk factors in order to prevent the occurrence of T. gondii infection and reduce the prevalence and incidence of the disease.

Elimination of Staphylococcus aureus nasal carriage in intensive care patients lowers infection rates.

This study surveys the clinical relevance of the nasal Staphylococcus aureus colonization status on intensive care unit (ICU)-acquired S. aureus infections and compares molecular characteristics of isolates from the nose and infectious sites. The 390 patients included comprised 278 non-carriers and 112 carriers. Among the carriers, 56 were decolonized with mupirocin. Decolonization was verified through a second (negative) culture. Spa typing and virulence gene profiling were performed for all isolates. Twenty six S. aureus infections were detected in the carriage group and 20 in the non-carriage group. Eighteen of these 26 (69.2%) infections were among carriers, and 8 of these 26 (30.8%) infections occurred among decolonized carriers (p = 0.02). Overall, 31/112 (27.7%) of the colonized patients and 25/46 (60.1%) of infection were due to methicillin-resistant S. aureus (MRSA). The highest frequency virulence genes were sea and hlg (both 100%) in nasal isolates and sea, hlg, fnb, and clf (100%) for infectious isolates. t030 was the most abundant spa type overall. S. aureus carriers were more likely to develop S. aureus infection compared with decolonized and non-carrying patients. The sources of ICU S. aureus infection appear to be exogenous mostly, and a predominant clone (spa type 030) plays an important role. We confirm that nasal mupirocin treatment prevents ICU infections even when there is an increased prevalence of nosocomial MRSA.

Plasma Level of MicroRNAs, MiR-107, MiR-194 and MiR-210 as Potential Biomarkers for Diagnosis Intestinal-Type Gastric Cancer in Human

Background: Timely and sensitive diagnosis of gastric cancer is crucial for efficient treatment and survival of the patients. microRNAs have been considered as diagnostic biomarkers in different type of cancers including gastric cancer. In the present study, the expression profile of four microRNAs, miR-103, miR-107, miR-194 and miR-210 were evaluated in patients with intestinal-type of gastric cancer (IGC) in order to assess their diagnosis utility as noninvasive biomarkers. Methods: A total number of 100 plasma samples from patients with gastric cancer and healthy controls were obtained and total RNA was extracted using a commercial monophasic solution of phenol and guanidium thiocyanate. Reverse transcription (RT) reactions were performed by specific stem-loop RT primers and M-MuLV RT-enzyme. The expression patterns of microRNAs were assessed using reverse transcription quantitative real-time PCR (RT-qPCR) method and the expression of SNORD47 RNA was used as the reference for normalization. Results: The results indicate that the plasma levels of miR-107, miR-194, and miR-210 were significantly lower in patients. Receiver operating characteristic (ROC) curve analysis showed that the patients could be distinguished from healthy individuals at the cutoff levels of 0.504, 0.266, and 0.394 of miR-107, miR-194, and miR-210, respectively. On the other hand, the expression levels of these miRNAs were not significantly different in different clinicopathological stages of the disease. Conclusion: These findings suggest that the plasma levels of miR-107, miR-194 and miR-210 were downregulated in patients with ICG and propose these molecules as potential non-invasive biomarkers for detection of IGC.

High prevalence of methicillin resistant and enterotoxin gene-positive Staphylococcus aureus among nasally colonized food handlers in central Iran.

This study defined the prevalence of enterotoxin gene-positive Staphylococcus aureus strains among food handlers and non-food processing healthy nasal S. aureus carriers in central Iran. Meticillin-resistant S. aureus (MRSA) strains were diagnosed by cefoxitin disk diffusion. PCR was used to detect the mecA, Sa442, and enterotoxin genes. Out of the 1113 food handlers, 224 (20.1%) were nasal carriers of S. aureus and 157 (70.1%) of these isolates were positive for one or more enterotoxin genes. The most prevalent enterotoxin gene was sei (40.2%), followed by seg (35.3%), sea (23.5%), seb (15.2%), sec (5.5%), and seh (2.7%). See and sed genes were not found. Sixty seven (42.7%) of enterotoxin gene-positive isolates possessed a single enterotoxin gene, and 64 (40.8%), 23 (14.7%), and 3 (1.9%) contained two, three, or four enterotoxin genes, respectively. The most frequently detected gene combination was sei/seg (n = 35, 22.3%). Thirty seven (16.5%) isolates were diagnosed as MRSA, and 27 (73%) of these strains were positive for at least one enterotoxin gene. Out of 546 healthy controls, 100 individuals were identified as S. aureus nasal carriers; among the strains, 39 (39%) were positive for at least one enterotoxin gene. Only one (1%) CA-MRSA was identified among the strains from the volunteers. A high prevalence of meticillin resistant and enterotoxin-positive S. aureus were documented in food handlers. We suggest that this may be due to the frequent handling of contaminated foodstuffs and that this is possibly related to the elevated frequencies of acquired staphylococcal food poisoning in this population.

Novel Mutations in pncA Gene of Pyrazinamide Resistant Clinical Isolates of Mycobacterium tuberculosis.

In clinical isolates of Mycobacterium tuberculosis (MTB), resistance to pyrazinamide occurs by mutations in any positions of the pncA gene (NC_000962.3) especially in nucleotides 359 and 374. In this study we examined the pncA gene sequence in clinical isolates of MTB. Genomic DNA of 33 clinical isolates of MTB was extracted by the Chelex100 method. The polymerase chain reactions (PCR) were performed using specific primers for amplification of 744 bp amplicon comprising the coding sequences (CDS) of the pncA gene. PCR products were sequenced by an automated sequencing Bioscience system. Additionally, semi Nested-allele specific (sNASP) and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) methods were carried out for verification of probable mutations in nucleotides 359 and 374. Sequencing results showed that from 33 MTB clinical isolates, nine pyrazinamide-resistant isolates have mutations. Furthermore, no mutation was detected in 24 susceptible strains in the entire 561 bp of the pncA gene. Moreover, new mutations of G→A at position 3 of the pncA gene were identified in some of the resistant isolates. Results showed that the sNASP method could detect mutations in nucleotide 359 and 374 of the pncA gene, but the PCR-RFLP method by the SacII enzyme could not detect these mutations. In conclusion, the identification of new mutations in the pncA gene confirmed the probable occurrence of mutations in any nucleotides of the pncA gene sequence in resistant isolates of MTB.

Comparison of antibacterial effects of a carrier produced in microemulsion system from aqueous extract of Aloe vera with selected antibiotics on Enterobacteriacea.

BACKGROUND AND OBJECTIVES: Antibiotics resistance has recently increased. The aim of this study was the evaluation of antibacterial efficacy of Aloe vera carrier produced in microemulsion system in comparison with ordinary antibiotics against some Enterobacteriacea. MATERIALS AND METHODS: The aquatic extract of Aleo vera was produced by the Soxhlet method and a nonocarrier in the microemulsion system was prepared by two emulsifiers. The clinical isolates of Escherichia coli, Klebsiella pneumoniae, Salmonella enterica, Shigella dysenteriae, Salmonella Typhimurium, Salmonella Paratyphi, Serratia marcescens, Proteus mirabilis, Enterobacter aerogenes, Citrobacter freundii and Morganella morganii were obtained from patients and were identified by microbiological methods. Diffusion disk was used for evaluation of antibacterial properties in comparison with selected ordinary antibiotics. Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) for tested materials were determined using MTT in the Micro Broth dilution method. RESULTS: The results proved that effect of carrier on studied isolates is dependent on concentration level. The inhibitory effect of carrier in concentration of 15 µg/ml by 18 mm zone of inhibition for Klebsiella pneumoniae was comparable to Ceftazidime and Cefalothin. The lowest MIC and MBC determined by the Microbroth dilution method with MTT belonged to Klebsiella pneumoniae as 0.1 and 3 µg/ml and higher concentrations belonged to Enterobacter aerogenes at 7 and 15 µg/ml. The greatest effect of carrier of Aleo vera aquatic extract was observed for Klebsiella pneumoniae and the lowest effect belonged to Enterobacter aerogenes, Citrobacter freundii and Morganella morganii. CONCLUSION: It was concluded that the carrier of Aloe vera produced in microemulsion system was most effective and had equal effects in comparison with ordinary antibiotics against Enterobacteriacea.

Transmission Electron Microscopy of XDR Mycobacterium tuberculosis Isolates Grown on High Dose of Ofloxacin.

The aim of the study was to investigate behavior of resistant Mycobacterium tuberculosis (MTB) isolates under a high dose of ofloxacin and its morphological changes. 19 extensively drug resistant (XDR) clinical isolates of MTB were grown on Löwenstein-Jensen medium containing progressively increasing concentrations of ofloxacin (2, 4, 8, 16, 32 mg/L). Ultra-structure analyses of resistant isolates grown on ofloxacin were conducted by transmission electron microscopy (TEM). Fixation was carried out by 4% glutaraldehyde in 0.1 M sodium cacodylate buffer on 300 mesh carbon formvar copper grid. The samples were negatively stained with uranium acetate suspension. All19XDRMTBisolatesweregrownandformedcoloniessuccessfullyon2,4,8mg/L,sevenisolates on16mg/L,andfourisolateson32mg/Lofloxacin. Morphologicalchangesandunusualformswere detected in 8, 16 and 32 mg/L ofloxacin at 43%, 76.5% and 81% of cells, respectively. Swollen form (protoplast like), ghost-like cell, degraded forms, and in a few cases, detached cytoplasm from cell wall were clearly detected in high drug concentrations in comparison to control. Changes in morphology were increased with increasing ofloxacin concentrations (p < 0.05). Some XDR isolates could be successfully grown on high doses of ofloxacin (32 mg/L), but with changes in morphology. It was concluded that several magnitudes of the drug doses could not prevent growth of drug resistant forms.

An In Vitro Evaluation of Ozonized Organic Extra-Virgin Olive Oil on Giardia Lamblia Cysts.

BACKGROUND: Giardia lamblia is a common intestinal parasite that has been reported all over the world. OBJECTIVES: This study was conducted to evaluate the effect of ozonized organic extra-virgin olive oil on the cyst of G. lamblia. METHODS: The olive oil was ozonized based on international standards and confirmed by the world health organization (WHO) at various times in a generator. The ozone concentration of olive oil was adjusted at 32, 64, 96, 128, 160 mg/g based on ozone absorption. Giardia lamblia cysts were isolated from heavily infected stool samples and the sucrose gradient flotation technique. Five groups of triple tubes containing Giardia cysts were exposed to olive oil with 32, 64, 96, 128, 160 ozone concentrations, and the sixth and seventh groups were exposed to non-ozonized olive oil and normal saline, respectively. The tubes were placed at room temperature, and every four hours, the mortality of the Giardia cysts was assessed. RESULTS: The results showed that the first five groups' mortality rate of Giardia cysts reached 100% in 100 hours. An increasing concentration of ozone in olive oil leads to an increase in the mortality rate of Giardia cysts. The results showed a significant difference in the mean time of the mortality in all the groups (P ≤ 0.05). Furthermore, the higher fatality effect of ozonized organic extra-virgin olive oil (Ozonized Olive Oil = OZO) was proved in comparison with metronidazole in vitro. CONCLUSIONS: We concluded that ozonized organic extra-virgin olive oil was a growth inhibitor of Giardia cysts, and concerning its compatibility with a biological system, it is recommended for further clinical trials.

Antimycobacterial activity assessment of three ethnobotanical plants against Mycobacterium Tuberculosis: An In Vitro study.

OBJECTIVE/BACKGROUND: Resistances to herbal medicines are still not defined and finding natural remedies against drug resistant Mycobacterium tuberculosis (MTB) has research priority. The antimycobacterial susceptibility method for herbal extracts is unclearly defined and there is no standard method for assessment of the materials against bacteria. In the present study, time kill of three medicinal plants was determined against MTB. METHODS: The clinical isolate of MTB from a patient who harbored confirmed tuberculosis was used in the study. Aqueous extracts of Aloe vera leaves, mint, and Hypericum perforatum were prepared using reflux distillation. Disk diffusion methods were conducted in Petri dishes and McCartney bottles containing Löwenstein-Jensen medium to measure the sensitivity of plant extracts in serial concentrations of 0.25-8mg/mL. A pour plate method was performed by mixing 0.7mL of each concentration of extract in 5mL Löwenstein-Jensen medium followed by surface culturing of MTB fresh cells. The time kill method was conducted by bacterial suspension in equal amounts of the extract and viable evaluation in fresh culture at the beginning, and at 24-h, 48-h, 72-h, and 1-week intervals. All cultures were incubated at 37°C for 4weeks. Inoculum concentrations were considered as a variable. RESULTS: The zones of inhibition of A. vera, H. perforatum, and mint extracts in the disk diffusion method in McCartney bottles were 60mm, 41mm, and zero, respectively, but Petri dishes did not have repeatable results. In the pour plate method, an extract concentration up to 1mg/mL could inhibit cell growth. In mint extract, colony forming was four times more than the others at 0.5mg/mL. Time kill of 95% of cells occurred when exposed to extracts of A. vera and H. perforatum separately, but was 50% in 24 h and 20% in 10 min. The time kill for mint was 95% in 1week. CONCLUSION: The results give some scientific basis to the use of plant extracts for growth control of MTB cells. Clinical trials are recommended for assessment of the extract as complementary medicine, as well as for antisepsis.

Study of carD gene sequence in clinical isolates of Mycobacterium tuberculosis.

Mycobacterium tuberculosis growth rate is closely coupled to rRNA transcription which is regulated through carD gene. The aim of this study was to determine the sequence of carD gene in drug susceptible and resistant clinical isolates of M. tuberculosis and designing of a PCR assay based on carD sequence for rapid detection of this bacterium.Specific primers for amplification of carD gene were carefully designed, so that whole sequence of gene could be amplified; therefore primers were positioned at the upstream (promoter of this gene and ispD gene) and downstream (in ispD gene). DNA from 41 clinical isolates of M. tuberculosis with different pattern of drug resistance was used in the study. PCR conditions and annealing temperature were designed by means of online programs. PCR products were sequenced by ABI system.PCR product of carD gene was a 524 bp fragment. This method could detect all resistant and susceptible strains of M. tuberculosis. The size of amplified fragment was similar in all investigated samples. Sequence analysis showed that there was similar sequence in all of our isolates therefore probably this gene is considered to be conservative. Translation of nucleotide mode to amino acids was showed that TRCF domain in N-terminal of protein CarD was found to be fully conservative.This is the first study on the sequence of carD gene in clinical isolates of M. tuberculosis. This conservative gene is recommended for use as a target for designing of suitable inhibitors as anti-tuberculosis drug because its importance for life of MTB. In the other hand, a PCR detection method based on detection of carD gene was recommended for rapid detection in routine test.

Rapid detection of coliforms in drinking water of Arak city using multiplex PCR method in comparison with the standard method of culture (Most Probably Number)

<p><b>OBJECTIVE</b>To analyse molecular detection of coliforms and shorten the time of PCR.</p><p><b>METHODS</b>Rapid detection of coliforms by amplification of lacZ and uidA genes in a multiplex PCR reaction was designed and performed in comparison with most probably number (MPN) method for 16 artificial and 101 field samples. The molecular method was also conducted on isolated coliforms from positive MPN samples; standard sample for verification of microbial method certificated reference material; isolated strains from certificated reference material and standard bacteria. The PCR and electrophoresis parameters were changed for reducing the operation time.</p><p><b>RESULTS</b>Results of PCR for lacZ and uidA genes were similar in all of standard, operational and artificial samples and showed the 876 bp and 147 bp bands of lacZ and uidA genes by multiplex PCR. PCR results were confirmed by MPN culture method by sensitivity 86% (95% CI: 0.71-0.93). Also the total execution time, with a successful change of factors, was reduced to less than two and a half hour.</p><p><b>CONCLUSIONS</b>Multiplex PCR method with shortened operation time was used for the simultaneous detection of total coliforms and Escherichia coli in distribution system of Arak city. It's recommended to be used at least as an initial screening test, and then the positive samples could be randomly tested by MPN.</p>