Specification of Bacteriophage Isolated Against Clinical Methicillin-Resistant Staphylococcus Aureus

OBJECTIVES: The emergence of resistant bacteria is being increasingly reported around the world, potentially threatening millions of lives. Amongst resistant bacteria, methicillin-resistant Staphylococcus aureus (MRSA) is the most challenging to treat. This is due to emergent MRSA strains and less effective traditional antibiotic therapies to Staphylococcal infections. The use of bacteriophages (phages) against MRSA is a new, potential alternate therapy. In this study, morphology, genetic and protein structure of lytic phages against MRSA have been analysed. METHODS: Isolation of livestock and sewage bacteriophages were performed using 0.4 μm membrane filters. Plaque assays were used to determine phage quantification by double layer agar method. Pure plaques were then amplified for further characterization. Sulfate-polyacrylamide gel electrophoresis and random amplification of polymorphic DNA were run for protein evaluation, and genotyping respectively. Transmission electron microscope was also used to detect the structure and taxonomic classification of phage visually. RESULTS: Head and tail morphology of bacteriophages against MRSA were identified by transmission electron microscopy and assigned to the Siphoviridae family and the Caudovirales order. CONCLUSION: Bacteriophages are the most abundant microorganism on Earth and coexist with the bacterial population. They can destroy bacterial cells successfully and effectively. They cannot enter mammalian cells which saves the eukaryotic cells from lytic phage activity. In conclusion, phage therapy may have many potential applications in microbiology and human medicine with no side effect on eukaryotic cells.

Mutation and Efflux System are Responsible for Ciprofloxacin Resistance in Klebsiella pneumoniae / Las mutaciones y el sistema de eflujo son responsables de la resistencia a la ciprofloxacina en la Klebsiella pneumoniae

ABSTRACT The main mechanism of quinolone resistance in Klebsiella (K) pneumoniae is caused by mutation of porin-related proteins and efflux pumps. This study aimed to investigate the prevalence of ciprofloxacin-resistant K pneumoniae in burns patients and to understand the role of the AcrAB multidrug efflux system on minimal inhibitory concentration (MIC) of ciprofloxacin. For this reason, 52 K pneumoniae samples were collected from burns patients and evaluated for the mechanism of ciprofloxacin resistance. The results demonstrated that 40 isolates of K pneumoniae were ciprofloxacin-resistant and 35 showed the mutation on gyrA locus. By inhibition of the efflux system, the MIC yield showed a significant decrease. Therefore, it could be concluded that the high rate of mutation on the gyrA locus in combination with quinolone resistance was responsible for ciprofloxacin resistance and by inhibition of AcrA, the resistance rate showed a significant decrease in K pneumoniae isolated from burns patients.

RESUMEN El principal mecanismo de resistencia a la quinolona en las Klebsiella (K) Pneumoniae tiene como causa la mutación de las porinas y las bombas de eflujo. Este estudio tuvo por objetivo investigar la prevalencia de las K pneumoniae resistentes a la ciprofloxacina en pacientes con quemaduras, así como entender el papel del sistema de eflujo multidroga AcrAB en la concentración inhibitoria mínima (CIM) de la ciprofloxacina. Por esta razón, se recogieron 52 muestras de K pneumoniae de pacientes con quemaduras, a fin de evaluar el mecanismo de resistencia a la ciprofloxacina. Los resultados mostraron que 40 aislados de K pneumoniae eran resistentes a la ciprofloxacina y 35 mostraron la mutación en el locus gyrA. Con la inhibición del sistema de eflujo, el rendimiento de CIM tuvo una disminución significativa. Por lo tanto, se pudo concluir que la alta tasa de mutación en el locus gyrA en combinación con la resistencia a la quinolona era responsable de la resistencia a la ciprofloxacina, y por la inhibición de AcrA, la tasa de resistencia mostró una disminución significativa en las K pneumoniae aisladas de los pacientes con quemaduras.

Presence of bla[PER-1] and bla[VEB-1] beta-lactamase genes among isolates of Pseudomonas aeruginosa from South West of Iran

Pseudomonas aeruginosa isolates have acquired resistance to antibiotics such as novel beta-lactams. The aim of this study was to investigate the bla[PER-1], bla[VEB-1], and bla[PSE-1] genes among isolates of P. aeruginosa among intensive care unit [ICU] patients. Sixty-five isolates were collected. The antibiotic susceptibility testing and combined disk tests were performed to detect the isolates producing extended spectrum beta-lactamases [ESBLs] among ceftazidime-resistant isolates. Polymerase chain reaction [PCR] amplification of blaPER-1, blaVEB-1, and blaPSE-1 genes was conducted. Ten [15.3%] isolates were ESBL-positive, of which 40% [n = 4] belonged to males and 60% [n = 6] were collected from females. Moreover, two and one isolates harbored blaPER-1 and blaVEB-1 genes, respectively

The rationale behind antibiotic resistance pattern in Klebsiella pneumoniae

Presently, there is an increase in antibiotic resistance in bacteria, due to relax prescription of antibiotics, especially in Iran. Undoubtedly, in toxin antitoxin (TA) system, a toxin neutralized by antitoxin, which known as a potent antimicrobial target; but there is no extensive survey on the prevalence of TA loci in large scale of Klebsiella pneumoniae. Therefore, this study aims to determine the prevalence of different TA loci in clinical and environmental K. pneumoniae isolates. For this reason, 48 K. pneumoniae clinical isolates and 49 K. pneumoniae environmental isolates were subjected for evaluation of different TA loci. The results of current study indicated that there is no association between antibiotic resistances and presence of TA loci in clinical and environmental K. pneumoniae. The role of TA loci as a potent target in antibiotic resistant K. pneumoniae has been complicated. Therefore, more studies should be performed to explain why TA loci are presented in K. pneumoniae and what is the rationale behind antibiotic resistant K. pneumoniae?

Antibacterial, anti-swarming and anti-biofilm formation activities of Chamaemelum nobile against Pseudomonas aeruginosa

Chamomile ( Chamaemelum nobile ) is widely used throughout the world, and has anti-inflammatory, deodorant, bacteriostatic, antimicrobial, carminative, sedative, antiseptic, anti-catarrhal, and spasmolytic properties. Because of the increasing incidence of drug-resistant bacteria, the development of natural antibacterial sources such as medical herbs for the treatment of infectious diseases is necessary. Extracts from different plant parts such as the leaves, flowers, fruit, and bark of Combretum albiflorum, Laurus nobilis , and Sonchus oleraceus were found to possess anti-quorum sensing (QS) activities. In this study, we evaluated the effect of C. nobile against Pseudomonas aeruginosa biofilm formation

Acinetobacter baumannii clonal lineages I and II harboring different carbapenem-hydrolyzing-beta-lactamase genes are widespread among hospitalized burn patients in Tehran

The aim of this study was to analyze antimicrobial resistance patterns and their encoding genes and genotypic diversity of Acinetobacter baumannii isolated from burn patients in Tehran, Iran. The presence of extended-spectrum beta-lactamase- and bla[OXA]-encoding genes among 37 multidrug resistant [MDR] A. baumannii strains isolated from patients hospitalized in a teaching hospital in Tehran was evaluated. Susceptibility to 7 antibiotics was tested by disk agar diffusion and to polymyxin B and colistin was tested by E-test, according to CLSI guidelines. All isolates were then analyzed by PCR for the presence of bla[IMP],bla[VIM], bla[SIM] bla[OXA-23], bla[OXA-24], and bla[OXA-58]-like carbapenemase genes, and bla[OXA-51] like, bla[TEM],bla[SHV], bla[PER], bla[VEB], and bla[GIM] genes. Genotyping of A. baumannii strains was performed by repetitive sequence-based [REP]-PCR and cluster analysis of REP-PCR profiles. A. baumannii isolates were assigned to international clones by multiplex PCR sequence group analysis. Twenty-five A. baumannii isolates were classified as MDR, and 12 were classified as extensively drug resistant. All isolates were susceptible to colistin and polymyxin B. Eighty-one percent of the isolates was resistant to imipenem or meropenem and harbored at least one or both of the bla[OXA-23]-like or bla[OXA-24]-like carbapenemase genes. Co-existence of different resistance genes was found among carbapenem-resistant isolates. Multiplex PCR sequence group analysis most commonly assigned A. baumannii isolates to international clones I [18/37; 48.6%] and II [18/37; 48.6%]. An alarming increase in resistance to carbapenems and the spread of bla[OXA-23]-like and/or bla[OXA-24]-like carbapenemase genes was observed among A. baumannii strains belonging to clonal lineages I and II, isolated from burn patients in Tehran

pidemiologic study of 80 patients with ulcerative colitis referred to Imam Hospital in Ardabil city during 2004-2011.

Background: Ulcerative colitis is one of the inflammatory bowel diseases with unknown etiology. Genetic and environmental factors are thought to be effective in this disease. The aim of this study is to assessment of demographic features and clinical symptoms of ulcerative colitis patients refereed to Emam hospital in Ardabil city. Methods: In this retrospective cross-sectional study, 80 cases of ulcerative colitis referred to Emam hospital in Ardabil city were evaluated during 2004-2011. The diagnosis was confirmed based on clinical features, colonoscopy, and pathology and resulting of other causes. Data were collected through direct interview and analyzed by statistical method in SPSS software. Results: Mean age of patients was 36.4 (SD=18.4). Duration of symptoms onset until diagnosis was 8 months. Male to female ratio was 0.8/1. 38(47.5%) of patients were male and 42 (52.5%) were female. 3 (3.75%) of patients have history of positive UC and 4 (5%) history of appendectomy. According to colonoscopy finding, 1 (1.25%) have rectum involvement, 27 (33.75%) recto sigmoid, 23 (28.75%) left side colon and 4 (5%) have pan colitis. Conclusion: Results showed that in compare with other places, clinical signs of ulcerative colitis in Ardabil province are different and so doing other d epidemiologic studies based on population to determine incidence and prevalence ulcerative colitis in Ardabil province is necessary.

The mazEF toxin-antitoxin system as a novel antibacterial target in Acinetobacter baumannii

Although analysis of toxin-antitoxin (TA) systems can be instructive, to date, there is no information on the prevalence and identity of TA systems based on a large panel of Acinetobacter baumannii clinical isolates. The aim of the current study was to screen for functional TA systems among clinical isolates of A. baumannii and to identify the systems’ locations. For this purpose, we screened 85 A. baumannii isolates collected from different clinical sources for the presence of the mazEF, relBE and higBA TA genes. The results revealed that the genes coding for the mazEF TA system were commonly present in all clinical isolates of A. baumannii. Reverse transcriptase-polymerase chain reaction analysis showed that transcripts were produced in the clinical isolates. Our findings showed that TA genes are prevalent, harboured by chromosomes and transcribed within A. baumannii. Hence, activation of the toxin proteins in the mazEF TA system should be investigated further as an effective antibacterial strategy against this bacterium.

First report of c. 1499G>C mutation in a 6-month-child with cystic fibrosis.

So far, more than 1800 mutations identified in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. In this case report, we presented first report of c. 1499G>C mutation in a 6‑month‑old girl with cystic fibrosis (CF) diagnosis. A 6‑month‑old girl with weakness and meconium Ileus referred to the pediatric clinic in Ilam, in the west of Iran. Patient’s skin was dark and suffered from bronchiectasis. The sweat test was performed, and the concentration of chloride and sodium in patient’s sweat was 130-135 mmol/L and 125-128 mmol/L, respectively. The exon 10 mutation analysis of a CF patient was performed. CFTR mutation analysis revealed the identification of 2 mutations in patient, the mutations were p.F508del (ΔF508) and c. 1499G>C (cd500), respectively. The mutation c. 1499G>C (cd500) were found for the first time in the world. Assessing this mutation in future study and genetic investigation is recommended.

Synthesis and evaluation of monoclonal antibody against Plasmodium falciparum merozoite surface antigen 2

OBJECTIVE@#To assess the quality of expressed MSP-2 and also to confirm the immune response against different domains of these proteins.@*METHODS@#Mice were immunized with a schizont extract to stimulate the immune system to make antibodies against different antigens of the late stage parasite including production of antibodies against different domains of Plasmodium falciparum (P. falciparum) MSP-2. B lymphocytes of immunized mice were extracted from the spleen and the fusion was performed using NS-1 myeloma cells and the hybridoma cells were assayed by ELISA either with a schizont extract or different domains of MSP-2 and/or by IFAT with whole schizont preparation. Fusion of NS-1 and spleen cells was performed. The positive hybrids were cloned and ELISA was applied against different dilutions. The positive clones were transferred to a small tissue culture flask and after developing they were assayed against schizont extract and the different MSP-2 domains. The positive clones were expanded to large (75 cm(2)) flask and cultured under the same conditions, checking them using both ELISA and IFAT and the positive cells were frozen as soon as possible.@*RESULTS@#A total number of 7 fusions including 26 plates (2 496 wells) were performed, of which 1 336 hybrids were produced and the overall efficiency (1 336/2496 × 100) was about 53%. ELISA was performed to detect the positive hybrids against crude schizont extract by which the highest frequency to crude schizont extract was found for the supernatant of the hybrids produced in fusion number 3 (66 out of 315 hybrids). The supernatant of both B5 and F1 hybridoma cells were more positive against domain 2 of the MSP-2 recombinant protein in Western blotting test. Western blotting results also showed that different domains of the MSP-2 recombinant protein and also the MSP-2 of the P. falciparum parasite were recognized by some of the positive clones and also immune sera.@*CONCLUSIONS@#Bringing together all the results of this study it has been confirmed that some clones have recognized both schizont extract and different domains of the MSP-2 recombinant protein and therefore confirming the quality of the MSP-2 domains.

Incidence of extended-spectrum beta-lactamase-producing Klebsiella pneumoniae in patients with urinary tract infection / Incidência de Klebsiella pneumoniae produtoras de beta-lactamases de espectro estendido em pacientes com infecção do trato urinário

CONTEXT AND OBJECTIVES: Resistant bacteria are emerging worldwide as a threat to favorable outcomes from treating common infections in community and hospital settings. The present investigation was carried out to study the incidence of extended-spectrum beta-lactamase (ESBL)-producing Klebsiella pneumoniae in patients with urinary tract infection in different seasons of the year, in order to determine the prevalence of the genes blaTEM, blaSHV and blaCTX-M, which are responsible for ESBL production among ESBL-producing K. pneumoniae, in three cities in Iran, and to investigate the antimicrobial susceptibility pattern of K. pneumoniae in different seasons. DESIGN AND SETTING: Retrospective study carried out among patients with urinary tract infections in five hospitals in Iran. METHOD: Two hundred and eighty-eight clinical isolates of K. pneumoniae were collected between March 2007 and April 2008 from five hospitals in three cities in Iran. ESBLs were identified by phenotypic and genotypic methods. ESBL-producing Klebsiella pneumoniae were evaluated against non-beta-lactam antibiotics. Genes coding for ESBLs (blaSHV, TEM and CTX-M) were screened. RESULTS: Among the 288 clinical isolates of K. pneumoniae, 37.7 percent, 46.7 percent and 15.6 percent were obtained from hospitals in Ilam, Tehran and Tabriz, respectively, of which 39.4 percent, 50.7 percent and 45.8 percent were ESBL-producing K. pneumoniae in Ilam, Milad and Emam Reza hospitals, respectively. CONCLUSION: According to the results from this study, resistance to third-generation cephalosporins is higher during the cold months than during the warm months.

CONTEXTO E OBJETIVOS: As bactérias resistentes estão surgindo em todo o mundo como uma ameaça ao resultado favorável no tratamento de infecções comuns em ambientes comunitários e hospitalares. O presente estudo foi realizado com o objetivo de avaliar a incidência de Klebsiella pneumoniae produtoras de beta-lactamases de espectro estendido (BLEEs) em pacientes com infecção do trato urinário em diferentes estações do ano, para verificar a prevalência dos genes blaTEM, blaSHV e blaCTX-M responsáveis para a produção de BLEEs em K. pneumoniae productoras de BLEEs, em três cidades do Irã, e investigar o perfil de sensibilidade antimicrobiana de K. pneumoniae em diferentes estações. DESENHO E LOCAL: Estudo retrospectivo realizado em pacientes com infecções do trato urinário em cinco hospitais no Irã. MÉTODO: Duzentos e oitenta e oito isolados clínicos de K. penumoniae foram coletados entre março de 2007 e abril de 2008 em cinco hospitais de três cidades no Irã. BLEEs foram identificados por métodos fenotípicos e genotípicos. K. pneumoniae produtoras de BLEEs foram avaliadas contra antibióticos não beta-lactâmicos. Os genes codificadores de BLEEs (blaSHV, TEM e CTX-M) foram investigados. RESULTADOS: Entre os 288 isolados clínicos de K. pneumoniae, 37,7 por cento, 46,7 por cento e 15,6 por cento eram provenientes dos hospitais em Ilam, Tehran e Tabriz, respectivamente, dos quais 39,4 por cento, 50,7 por cento e 45,8 por cento eram K. pneumoniae produtoras de BLEEs nos hospitais em Ilam, Milad e Eman Reza, respectivamente. CONCLUSÃO: De acordo com os resultados deste estudo, a resistência às cefalosporinas de terceira geração é maior nos meses frios do que nos meses quentes.

Evaluation of chronic hepatitis B infection in patients with seronegative HbsAg

It is estimated that about 370 million people are chronic carriers of HBV worldwide. Apparently 3% of Iranian populations are chronic carriers of this virus. We aimed to evaluate the viral DNA in biological fluids of chronic hepatitis patients compared to a control group. The current case-control study was designed to evaluate the viral DNA in biological fluids of 70 chronic hepatitis patients compared to a control group using ELISA, PCR and Real Time. All individuals [100%] in case group were HBsAg positive while in control group only 2 individuals [2.8%] were HBsAg positive. Three individuals, in control group were positive using PCR and Real Time PCR indicating that about 7% of those in control group were chronic carriers of HBV. The interesting point was the copy of viral DNA; [5.49 x104, 2.162x103and 7.26x106] for 3 chronic carriers using sera while it was about [5.71x103, 1.45x102 and 2.56x105] using ear cerumen confirming the necessity of investigating for the carriers of HBV in different biological fluid and by different methods. It can be concluded chronic carriers of hepatitis B are much more than what is diagnosed by routine diagnostic tests. On the other hand ELISA alone can not be relied on as a complete test for screening of chronic carriers in hepatitis B. PCR and Real Time PCR are more reliable tests for this purpose

Antimicrobial susceptibility, plasmid profiles, and RAPD-PCR typing of Acinetobacter bacteria

Background: Multiple-drug resistant Acinetobacter have widely spread in the last decades imposing a serious nosocomial source of infection. Nevertheless, little knowledge was gaimed on tracing the development of antibiotic resistance in Acinetobacter species.Objectives: Explore Acinetobacter spp. via antimicrobial susceptibility, plasmid profiles, and random amplified polymorphism DNA polymerase chain reaction (RAPD-PCR) typing.Methods: One hundred twelve Acinetobacter isolates (including 66 A. baumannii and 46 non-Acinetobacter baumannii strains) were obtained from three university hospitals. The source of infection of these isolates included blood, urine, wound, and respiratory tract. Their susceptibilities to 17 antibiotics were tested and then allAcinetobacter isolates were typed by plasmid analysis and RAPD-PCR method.Results: A. baumannii isolates revealed nine different patterns of antibiotic resistance. Of those, non-A. baumannii, were associated with plasmid and RAPD-PCR typings (p 0.05).Conclusion: There is a wide spread of multi-drug resistant Acinetobacter spp., particularly A. baumannii, in the Middle East region that can be traced efficiently by plasmid and genotyping typing of Acinetobacter. More care should be taken for tracing the development of antimicrobial resistance of Acinetobacter using precisemolecular typing techniques.Keywords: Acinetobacter baumannii, antimicrobial susceptibility, molecular typing, plasmid profiles RAPD-PCR

Qnr prevalence in extended spectrum beta-lactamases [ESBLs] and none-ESBLs producing escherichia coli isolated from urinary tract infections in central of Iran

Extensive use of quinolones has been associated with raising level of resistance, in the current, we focused on assessing the prevalence of Escherichia coli resistance to quinolones and frequency of qnrA, qnrB and qnrS in non ESBLs [extended spectrum beta-lactamases] and ESBLs producing E. coli with blaSHV and blaTEM. One hundred and fifty E. coli isolates were identified during Mar. 2007 to Apr. 2008 in Milad [Tehran] hospital. They were tested for ESBLs production as well as quinolone resistance. PCR was performed for detection of blaSHV and blaTEM as well as qnrA, B and S. Of 150 isolates, forty-two [28%] ESBLs producing and one hundred and eight [72%] non-ESBLs producing E. coli were identified. 64.2% [n= 24] of E. coli producing ESBLs and 4.62% [n= 5] of non-ESBLs E. coli were resistance to ciprofloxacin. 95.2% [n= 40] and 26.1% [n= 11] of the isolates harbored blaTEM and blaSHV, respectively. 23.8% [n= 10] had both genes. 37.5% [n= 9] and 20.8% [n= 4] of ESBLs producing E. coli were positive for qnrA and qnrB respectively. qnrS was not identified in any isolate. Our study showed high frequency of ESBLs producing E. coli as well as quinolone resistance genes [qnrA, qnrB] in Milad hospital

Cloning, expression and purification of Pwo polymerase from Pyrococcus woesei

Pyrococcus woesei is a hyperthermophilic archaea and produces a heat stable polymerase [Pwo polymerase] that has proofreading activity. In this study, this microorganism was cultured, its DNA was extracted and the pwo gene polymerase was cloned, expressed and purified. The DNA sequence of the cloned gene was verified by sequencing. The pwo polymerase gene consists of 2,328 bps [775 amino acids with about 90 kD molecular weight]. Cloning was done by GATEWAY Cloning System and for purification of recombinant protein; His6x-Tag was added to the C-terminus of the recombinant protein. We could purify Pwo polymerase enzyme by Ni-NTA resin. PCR assay showed that Pwo polymerase activity is comparable to a commercial Pfu polymerase activity

[Efficacy of acupuncture in control of primary dysmenorrhea related pain in 15-30 years old women at the gynecology clinic of Bouali Hospital between 2007 and 2008]

Dysmenorrhoea is the most common gynaecological disorder among adolescents. Acupuncture is a treatment that has been shown to be effective for pain relief in a variety of conditions. We evaluated the effect of acupuncture in control of primary dysmenorrhea related pain. In a randomized, double-blind, clinical trial study, 100 patients with primary dysmenorrhea were randomized to acupuncture [9 sessions over 3 months] or control group [placebo acupuncture]. All subjects were allowed to receive usual medical care. Pain was rated on a Numerical Pain Score [0-10] at three months. Patients' data were analyzed by t- test using SPSS software. This study showed significant effectiveness of acupuncture in control of primary dysmenorrhea pain after 3 months [p<0.0001]. No complications or side effects were reported due to acupuncture. Acupuncture was associated with relief of pain in patients with primary dysmenorrheal, and it is a safe and effective method in reduction of primary dysmenorrhea related pain

Evaluation of a PCR based approach to study the relatedness among Shigella sonnei strains

Infections caused by Shigella re a major cause of diarrheal disease in the developing and developed countries. The present study was conducted to apply and evaluate arbitrarily primed PCR [AP-PCR] for investigation of genetic relatedness among the strains of Shigella sonnei isolated from cases of acute diarrhea in Tehran. Totally, 60 S. sonnei strains isolated from children hospitalized due to enteritis at five hospitals in Tehran during 2003 and two sporadic isolates recovered in 1984 were investigated. Molecular typing was performed by AP-PCR. Depending on the number and size of amplified DNA bands, the strains were clustered into AP-PCR profiles. All strains of S. sonnei were typeable with this approach. AP-PCR generated nine indistinguishable bands ranged from 0.35 to 2.5 kbp in all studied strains. Only a single AP-PCR pattern was observed among the S. sonnei strains recovered in 2003. Two sporadic isolates recovered in 1984 showed different AP-PCR patterns compared to recent clinical isolates. Results suggest that a very homogeneous AP-PCR cluster types might be responsible for shigellosis caused by S. sonnei in Tehran in 2003. Further molecular analysis conducted on a larger selection of isolates could confirm our findings

[ study of analgesic effect of hydroalcoholic extract of Artemisia herba alba in acute and chronic models of pain in male rats]

In the present time, analgesic and anti-inflammatory drugs such as monsteroidal anti-inflammatory drugs and opioid compounds are used for the releif of pain, but due to their side effects and economical issues, the significance of research on finding analgesic drugs with less side effects and their ability to substitute these synthetic drugs and substituting newer analgesic compounds is obvious. There is very little information about physiologic effects of Artemisia herba alba, one of species of Herba alba family. In the present study the possible analgesic effects of hydroalcoholic extract of this plant were investigated in chemical and thermal models of nociception in rats. In this experimental study, percolation method was used for extraction. Specified amounts of dried extract were dissolved in twin to produce desired concentration. The extract were used orally or injected intraperitoneally and analgesic activity was assessed with two methods, Tail flick and Formalin test. In addition, anti nociceptive effect of this plant was compared with sodium salicylate as standard drug. The plant extract significantly increased tail flick latency in tail flick test and decreased significantly pain perception in both phases of formalin test. The anti-nociception effect of herba alba was dose dependent. These data confirm peripheral and central antianalgesic effect of this plant

Genotyping of Helicobacter pylori strains isolated from patients with NUD, DU, GU and GC by RAPD-PCR

Helicobacter pylori is a genetically diverse gastric pathogen that chronically infects billions of people worldwide, typically beginning in infancy and lasting for decades. It is a major cause of peptic ulcers and it is an early risk factor for gastric cancer which is the most frequently lethal malignancy globally. This project was designed to genotype H. pylori isolates isolated from patients with NUD, DU, GU and GC by the polymerase chain reaction [PCR]-based on Randomly Amplified Polymorphic DNA [RAPD] fingerprinting technique. Eighty patients admitted to the gastroenterology unit at Sharyati hospital in Iran were included in this study. Gastric biopsy specimens were inoculated onto selective medium then were cultured for 3 to 5 days at 37 °C under microaerobic conditions. Genomic DNA was extracted using a commercially available Qiagen kit. RAPD-PCR was used to genotype isolates. Six different RAPD patterns [A-F] were seen in more than one isolate which were as follow; pattern A: 9 [16.98%], B: 6 [11.33%], C: 5 [9.43%], D: 3 [5.66%], E: 2 [3.77%] and F: 2 [3.77%]. Twenty six [49.06%] of 53 isolates showed a unique RAPD pattern that were not similar to each other. A significant relationship was not seen between a single RAPD pattern and a gastric disorder [P>0.05]. The results of this study suggest a high level of DNA sequence diversity among H. pylori isolates and it is better to use sequencing method for surveying of Helicobacter pylori genome rather than RAPD-PCR

Antimicrobial susceptibility and AP-PCR typing of acinetobacter Spp. strains

Acinetobacter spp., as important opportunistic pathogens, have been found to be responsible for an increasing number of nosocomial infections. This study was undertaken to investigate the antimicrobial susceptibility and molecular typing of Iranian isolates of A. baumannii. The study was conducted over a period of 19 months in three hospitals in Tehran, Iran. Acinetobacter spp. Were isolated from different clinical specimens using standard bacteriological methods. Antimicrobial susceptibility test was performed according to the standard CLSI guideline using 17 antibiotic disks. The AP-PCR fingerprinting was carried out using ARB11 primer. The PCR product was run and visualized in 2% agarose gels and stained with ethidium bromide. The AP-PCR profiles were grouped depending on the patterns of the amplified bands. Sixty seven strains of Acinetobacter spp. [including 21 A. baumannii and 46 non- A. baumannii] were isolated. The sources of these isolates were blood, urine, wound, and respiratory tract. A. baumannii isolates were further studied. Results showed that all A. baumannii isolates were resistant to at least 11 antibiotics tested. AP-PCR analysis of A. baumannii strains resulted in 7 different patterns. The dominant AP-PCR pattern was E [57.1%]. Acinetobacter spp. are still important nosocomial pathogens in the region studied and most of isolates were multi-drug resistant. Our results also indicate that the AP-PCR technique represents a rapid and simple means for typing of A. baumannii