[Preventive behaviors related to women's genital tract infections]

Introduction: Reproductive tract infections are recognized as one of the most serious health problems in the world. These infections can be associated with many consequences

Objective: This study aimed to determine the condition of preventive behavior related to genital tract infections and its effective factors

Methods: This is a descriptive analytical study on preventive behaviors related to reproductive tract infections and its risk factors in women admitted to health centers in Tabriz city in 2013. Twenty-two health centers and stations [including 9 centers and 13 stations] out of 27 centers and 38 stations were randomly selected. In the next stage, according to sample size, 25-30 records were randomly selected from all women's records. The sample size was measured according to the pilot study and using the ratio formula for 384 patients and considering coefficient of 1.5, the final sample size equaled 584. Our study included married 15 to 49-year-old women who had referred for health care and had health records, higher than primary education level and willingness to participate in research. Women who were unwilling to participate for complete more than 20% of the questionnaire were excluded. The data collection tool was a researcher-made questionnaire including questions related to behavioral prevention assessment of reproductive tract infections and questions about factors influencing these behaviors as well as demographic characteristics and obstetrical and fertility history, and questions on knowledge of the risk factors. In order to determine the preventive behavior, the scores of 2, 1, 0 were given to items "always", "sometimes" and "never", respectively. The sum of scores was calculated. The mean score was considered as the score of study tool. Of all the behavioral prevention questions, the ones with the lowest and highest score were reported. To determine the knowledge about non-sexually transmitted infections, "correct" and "incorrect/I don't Know" answers were given 1 and zero scores, respectively. Then the total scores were calculated. Knowledge score

Results: Most women [56.1%] were in age range of 20-29 years old. Most of them were high school graduates [48.3%], homemaker [90.2%] and had sufficient income [59.4%] in their own opinion. A statistically significant relationship was found between behavioral prevention with age, education level, occupation and income [p<0.05]. Half of the subjects had a history of RTIs. Anal, oral, and vaginal-anal sex, sex with husband only and replacement or transfusion of blood in husband were significant related to behavioral prevention score [p<0.05]. Mean score of behavioral prevention equaled 25.39 +/- 4.6 of possible scores ranging from zero to 34. Mean score of knowledge of genital non-sexually transmitted infections was 9.6 +/- 3.19 of possible scores ranging from zero to 15. Thus, research subjects had average level of knowledge. Mean score of knowledge on sexually transmitted infections was 17.08 +/- 7.42 of possible scores ranging from zero to 37. Hence, they had average to poor knowledge. A statistical significant relationship was observed between the level of knowledge and behavioral prevention score [p<0.001]. In other words, behavioral prevention had a significant increase with increased score of knowledge. After adjusting the possible confounding factors, multivariate analysis showed a statistically significant relationship between preventive behaviors and knowledge score [R2-0.06, P-0.002], family income [R2-0.023, P<0.001], sex only with husband [R2-0.015, P-0.047], anal sex, oral sex [R2-0.054, P-0.003] and blood transfusion history [R2-0.028, P-0.005]. A statistical significant association was detected between the level of knowledge and behavioral prevention score [p<0.001] so that increased knowledge significantly raised the score of behavioral prevention

Conclusion: Trainings on these infections especially in school and pre-marital ages seem necessary as well as providing related public services. Health trainers should be consistent with this strategy. Primary prevention of such infections should be prioritized in health training

Teaching emotional intelligence to intensive care unit nurses and their general health: a randomized clinical trial

Emotion and how people manage it is an important part of personality that would immensely affect their health. Investigations showed that emotional intelligence is significantly related to and can predict psychological health. To determine the effect of teaching emotional intelligence to intensive care unit nurses on their general health. This randomized clinical trial [registered as IRCT201208022812N9] was conducted on 52 of 200 in intensive care unit nurses affiliated to Shiraz University of Medical Sciences. They were recruited through purposeful convenience sampling and then randomly categorized into two groups. The intervention group members were trained in emotional intelligence. Bar-on emotional intelligence and Goldberg's general health questionnaires were administered to each participant before, immediately after, and one month after the intervention. While the mean score of general health for the intervention group decreased from 25.4 before the intervention, to 18.1 immediately after the intervention and to 14.6 one month later, for the control group, it increased from 22.0, to 24.2 and to 26.5, respectively [p<0.001]. Teaching emotional intelligence improved the general health of intensive care unit nurses

Prevalence of trichomonas vaginalis infection in Hamadan city, Western Iran

Infection with Trichomonas vaginalis is one of the most common sexually transmitted diseases [STDs] in humans. The prevalence of infection in Iran has been reported between 2 to 8%, depending on deferent socio-cultural conditions. This study aimed to determine the prevalence of T. vaginalis in women referred to gynecologic clinics in Hamadan city, West of Iran. This descriptive cross-sectional study was conducted on 750 women who referred to Gynecologic clinics in Hamadan from November 2010 to July 2011. Vaginal samples were obtained from them and examined by wet mount and culture methods for the detection of T. vaginalis. Sixteen out of 750 vaginal swab specimens [2.1%] were culture positive for T. vaginalis and 13 of these positive specimens [1.7%] were wet mount positive. Only 12 of 42 patients who were clinically diagnosed as having T. vaginalis infection, confirmed by culture method. Five hundred and fifty of the participants women [73.3%] had at least one of signs and symptoms of trichomoniasis. No statistical correlation was observed between clinical manifestations and parasitological results [p>0.05]. This study showed low prevalence of T. vaginalis infection in the study population. Since clinical signs of trichomonal vaginitis are the same of other STDs, a confirmatory laboratory diagnosis is necessary. Wet smear as well as culture are sensitive for detection of T. vaginalis

[Evaluation of the relationship between pulsatility index and prognosis of the patients with moderate head trauma]

Trauma is the third leading cause of death and is one of the most common causes for referrals to the emergencies. Assessment of prognosis in the patients with head trauma can help find high risk patients and provide more complete care to them. The evaluation includes assessment of intracranial pressure and the brain circulation with Trans Cranial Doppler [TCD]. The evaluation of prognosis in patients with moderate head trauma using pulsatility index [PI], obtained by Trans Cranial Doppler [TCD]. In this descriptive study, 52 patients with moderate head trauma [GCS= 9-13] were studied during one year [2010]. In the first 24 hours after trauma, for all the patients TCD by their middle cerebral arteries [MCA] was performed to assess PI value. After seven days, all the patients were followed up for neurological deterioration and prognosis. Finally, data were entered in SPSS software v.17 and analyzed by Independent T-test, Chi- square test with 95% Confidence Interval [CI=0.95]. In total, 47 [90.4%] were men and 5 [9.6%] were women [with mean age of 33.81 +/- 17.91 years]. In the group with low PI [<1.2], 29 [69.05%], patients were with good prognosis, and 13 [30.95%] with poor prognosis. There was a significant relationship between GCS decline >/= 2 and average of PI [p<0.021]. Average of PI in the patients who needed intubation [1.08 +/- 0.26] was higher than that in other patients [0.91 +/- 0.24, p<0.037]. Also, 32 [61.54%] patients were with good prognosis, finally. Average values of PI in good and poor prognosis groups were 0.88 +/- 0.24 and 1.08 +/- 0.24, respectively; with significant difference [p<0.005]. There was no significant relationship between average values of PI and gender, need for operating, osmol therapy and type of defect in CT scan. By using Roc curve, the best cut off point for PI was estimated at about 0.9 with sensitivity of 75% and specificity of 43%. The results demonstrated that the PI is a valuable parameter besides the other factors such as GCS decline, to determine the prognosis in the patients with moderate head injury admitted to emergency units

Iranian Azeri's Y-chromosomal diversity in the context of turkish-speaking populations of the Middle East

The main goal of this study was to conduct a comparative population genetic study of Turkish speaking Iranian Azeries as being the biggest ethno-linguistic community, based on the polymorph markers on Y chromosome. One hundred Turkish-speaking Azeri males from north-west Iran [Tabriz, 2008-2009] were selected based on living 3 generations paternally in the same region and not having any relationship with each other. Samples were collected by mouth swabs, DNA extracted and multiplex PCR done, then 12 Single Nucleotide Polymorphisms [SNPs] and 6 Microsatellites [MS] were sequenced. Obtained data were statistically analyzed by Arlequin software. SNPs and Microsatellites typing were compared with neighboring Turkish-speaking populations [from Turkey and Azerbaijan] and Turkmens representing a possible source group who imposed the Turkish language during 11-15[th] centuries AD. Azeris demonstrated high level of gene diversity compatible with patterns registered in the neighboring Turkish-speaking populations, whereas the Turkmens displayed significantly lower level of genetic variation. This rate of genetic affiliation depends primarily on the geographic proximity. The imposition of Turkish language to this region was realized predominantly by the process of elite dominance, i.e. by the limited number of invaders who left only weak patrilineal genetic trace in modern populations of the region

[ effect of variable CO[2] concentrations on HSP70 gene in Trichophyton rubrum and Microsporeum canis]

Trichophyton rubrum is considered as the most common causes of dermatophytosis in human skin and nail tissues. Microsporeum canis is a zoophile dermatophyte which can be transmited to human. HSP70 is a 70 KD heat shock protein in fungi. In this study, the effects of variable CO[2] concentrations were examined on HSP70 expression in T. rubrum and M. canis. Strains used in this study were obtained from skin scales and nails of the patients who were suffering from onychomycosis. Samples were cultured on Sabouraud dextrose broth [SDB] and incubated at 25°C for 2, 4 and 7 days under 3%, 5%, and 10% of CO[2] concentrations. Control cultures maintained for 7 days without CO[2] concentrations. Then, RNA was isolated from the harvested mycelia mass, and HSP70 gene expression was studied in T. rubrum and M. canis by RT-PCR. The obtained results were compared to the Beta actin as a house keeping gene. The results of this study revealed the maximum variations under 3%,5%, and 10% of CO[2] concentrations in maximum 7 days incubation period, and the expression of HSP70 gene showed different variations under different CO[2] concentrations. Our results showed a negative effect of CO[2] concentrations in the expression of HSP70 in T. Rubrum and a positive effect in M. canis comparing to the controls

[ comparative study of an experimental model of tendon healing process between healthy rats and streptozotocin induced-diabetic rats by a biomechanical evaluating method]

The aim of the present investigation was to compare the healing process of repaired Achilles tendons of healthy rats and streptozotocin induced diabetic [STZ-D] rats by evaluating strength and toughness of repaired tendons via tensiometrical test. 14 healthy male adult rat were divided into control [n=6] and experimental rats [n=8]. Type 1 diabetes was induced in experimental rats by an injection of 55 mg/kg STZ intraperitoneally. Control rats were received distilled water. Blood Sugar of all rats was recorded after seven days. Animals of experimental group that failed to develop average blood glucose concentration greater than 250mg/dc were excluded from the study. Right Achilles tendon of all rats was transected completely 30 days after STZ injection. Repairing Achilles tendons were extracted and were submitted to a tensiometerical examination10 days after surgery. Data were compared by student t test. Independent sample t test analysis showed that Young's modulus of elasticity [18.5 +/- 12.1MPa/mm] and stress high load [3.04 +/- 1.38, MPa/mm] of control group were significantly higher than those of experimental group [2.5 +/- 2 and 0.975 +/- 0.269 respectively], [p=0.003 and p=0.004 respectively]. It is concluded that induction of type one diabetes by STZ in rats after 30 days reduced significantly tensiometrical parameters of repairing Achilles tendon in comparison with control rats

sensitive and specific PCR based method for identification of cryptosporidium sp. using new primers from 18S ribosomal RNA

The main goal of the present study was to develop a new sensitive and specific PCR based method for Identification of Cryptosporidium sp. using novel primers from 18S ribosomal RNA. Cryptosporidiosis in high-risk host groups particularly in neonates and immuno-compromised individuals may result in death. To the best of our knowledge this is the first study regarding develop a new PCR based method to diagnose the cryptosporidiosis in Iran. A total of 850 human fecal samples from patients clinically suspected to cryptosporidiosis and 100 healthy and diarrheic cattle stool specimens were collected. The simplified formol-ether concentration method was carried out for all samples. They were then examined microscopically by modified Ziehl-Neelsen staining method. Total DNA was extracted by QIA amp DNA stool mini kit. PCR and nested-PCR was carried out by using designed primers. Twenty nine cases of cryptosporidiosis infection in human and 30 samples from cattle microscopically were positive. The described primary and nested PCR method could detect all Cryptosporidium positive samples from human and cattle. Regards to suspected negative samples in primary PCR examination, the Nested PCR could approve two more positive results. Furthermore, Nested PCR analysis was able to detect one more case which was negative in both microscopically examination and primary PCR. Specificity of the test was 100%. Sensitivity of Nested PCR in comparison to our gold standard; microscopy after Ridley concentration modified ziehl-Neelsen, was 100%. Our developed PCR based method by using new primers devised from 18S ribosomal RNA revealed the ability for identification of the Cryptosporidium species such as C. parvum and C. huminis with high specificity and sensitivity

Molecular identification and sequencing of mannose binding protein [MBP] gene of acanthamoeba palestinensis

Acanthamoeba keratitis develops by pathogenic Acanthamoeba such as A. palestinensis. Indeed this species is one of the known causative agents of amoebic keratitis in Iran. Mannose Binding Protein [MBP] is the main pathogenicity factors for developing this sight threatening disease. We aimed to characterize MBP gene in pathogenic Acanthamoeba isolates such as A. palestinensis. This experimental research was performed in the School of Public Health, Tehran University of Medical Sciences, Tehran, Iran during 2007-2008. A. palestinensis was grown on 2% non-nutrient agar overlaid with Escherichia coli. DNA extraction was performed using phenol-chloroform method. PCR reaction and amplification were done using specific primer pairs of MBP. The amplified fragment were purified and sequenced. Finally, the obtained fragment was deposited in the gene data bank. A 900 bp PCR-product was recovered after PCR reaction. Sequence analysis of the purified PCR product revealed a gene with 943 nucleotides. Homology analysis of the obtained sequence showed 81% similarity with the available MBP gene in the gene data bank. The fragment was deposited in the gene data bank under accession number EU678895 MBP is known as the most important factor in Acanthamoeba pathogenesis cascade. Therefore, characterization of this gene can aid in developing better therapeutic agents and even immunization of high-risk people

Optimization of reactive blue 19 decolorization by ganoderma SP. using response surface methodology

Synthetic dyes are extensively used in different industries. Dyes have adverse impacts such as visual effects, chemical oxygen demand, toxicity, mutagenicity and carcinogenicity characteristics. White rot fungi, due to extracellular enzyme system, are capable to degrade dyes and various xenobiotics. The aim of this study was to optimize decolorization of reactive blue 19 [RB19] dye using Ganoderma sp. fungus. Response Surface Methodology [RSM] was used to study the effect of independent variables, namely glycerol concentration [15, 20 and 25 g/L], temperature [27, 30 and 33 °C] and pH [5.5, 6.0 and 6.5] on color removal efficiency in aqueous solution. From RSM-generated model, the optimum conditions for RB19 decolorization were identified to be at temperature of 27°C, glycerol concentration of 19.14 mg/L and pH=6.3. At the optimum conditions, predicted decolorization was 95.3 percent. The confirmatory experiments were conducted and confirmed the results by 94.89% color removal. Thus, this statistical approach enabled to improve reactive blue 19 decolorization process by Ganoderma sp. up to 1.27 times higher than non-optimized conditions

Evaluation of a single PCR assays on Cp5 gene for differentiation of Entamoeba histolytica and E.dispar

We examined a molecular method with a single-PCR for amplification of a part of CP5 gene enabling us to differentiate the pathogenic species, Entamoeba histolytica, from the non-pathogenic species, E. dispar. We developed a single PCR method for this purpose. After investigation of GenBank, primer pairs were designed from highly conserved regions of cysteine proteinase [CP5] gene. The primers were utilized in PCR using isolated genomic DNA template of E. histolytica and the PCR products were then sequenced. The same primer and method for PCR was used for isolated genomic DNA template of E. dispar. A fragment of about 950 bp was isolated in PCR by using DNA from E. histolytica, however, no banding pattern was produced by using the same primers for E. dispar. We characterized CP5 gene at molecular level in E. histolytica isolates from 22 positive; including 20 non-dysentery samples isolated from both cities as well as two dysentery samples isolated only from Tabriz. Nucleotide sequence comparison in gene data banks [NCBI, NIH] revealed significant homology with CP5 gene in E. histolytica isolates. We developed a PCR method, which could detect simply and rapidly E. histolytica by amplifying a specific PCR fragment

Comparison of a PCR-based method with culture and direct examination for diagnosis of Acanthamoeba keratitis

The aim was to compare three different methods [direct examination, culture and PCR methods] for the diagnosis of Acanthamoeba keratitis [AK] in corneal scrapes. Twenty eight corneal scrapes and contact lenses were collected from keratitis patients and referred to the Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences. Corneal scrapes were divided in three parts for direct examination, culture on non-nutrient agar and PCR analysis. PCR analysis was also performed using a 18S rRNA gene primer pair [DF3 region]. DF3 [Diagnostic fragment 3] is a region of the nuclear small subunit ribosomal RNA gene which is specific for detecting Acanthamoeba strains. Acanthamoeba was the causative agent of keratitis in 50% of the patients. Direct smear of all prepared corneal scrapes in AK patients was negative and culture was positive in only 14.3% of the isolates. PCR analysis was positive in 71.4% of AK patients. These three methods were negative in corneal scrapes of non-AK patients. The sensitivity and specificity of PCR technique for the detection of Acanthamoeba sp. were calculated as 71.4% and 100%, respectively. According to high sensitivity and specificity of PCR-based method, this study confirmed that PCR using 18S rRNA gene primers [DF3 region] is more useful for detecting AK cases compare to culture and direct microscopy methods

Molecular characterization of cyclophilin protein gene in skin normal microflora: malassezia furfur

Malassezia are dimorphic, lipid-dependent yeasts, which are responsible for causing several cutaneous and systemic conditions. Although cyclophilins [CyPs] are highly conserved cytosolic proteins that catalyze the peptidyl-prolyl cis-trans isomerazation reaction before protein folding process, it has been suggestive of an allergen in a few numbers of fungi such as Aspergillus fumigatus and Malassezia species. Allergenic cyclophilins are IgE-binding components, which have been characterized in other species of Malassezia; and are considered as Mala s 6 in Malassezia sympodialis. In the present study we tried to identify the molecular characterization of cyclophilin gene in M. furfur. Pairs of oligonucleotide primers were designed from highly conserved regions of the gene counterparts in other fungi. The primers were then applied to amplify the primer-specific DNA fragment. Afterward, PCR product fragments were sequenced to be used in further analysis. About 573 nucleotides, encoding a polypeptide of 190 amino acids, have been sequenced. Sequence comparison was performed in Gene Bank, both for the nucleotides and their deduced amino acid sequence. It revealed a significant homology with cyclophilin genes and proteins of other eukaryotic cells. The amino acid sequence of the encoded protein was about 86% identical to the sequence of cyclophilin protein from other fungi. The molecular characterization of cyclophilin gene may open the way to disclosure of the functional characteristics of cyclophilin and is a fundamental step for understanding the molecular basis of its pathogenesis in AEDS disease

Prevalence of Trichomonas vaginalis using parasitological methods in Tehran

Trichomonas vaginalis is a parasitic protozoan with a predilection for human urogenital tract and causative agent for vaginitis, cervicitis and urethritis in females. T. vaginalis is known as a cofactor in transmission of human immunodeficiency virus and may lead to adverse outcomes in pregnant women. The goal of this study was to determine the prevalence of T. vaginalis infection in females attending Mirzakuchak Khan Hospital, Tehran, Iran. During May 2008 to March 2009, 500 vaginal discharges samples were obtained from women attending sexual transmitted disease [STD] clinic of Mirzakuchak Khan Hospital in Tehran, Iran. The samples were examined by Dorsse culture medium and wet-mount methods. The prevalence of T.vaginalis was determined using culture based method and wet-mount examinations. Sixteen positive [3.2%] and 484 negative [96.8%] samples for T. vaginalis were detected by culture based methods. The wet mount examination revealed 13 positive [2.6%] and 487 negative [97.4%] samples. In the above population, prevalence of trichomoniasis was estimated as 3.2% based on culturing method. Due to adverse outcomes of vaginal trichomoniasis and its correlation with HIV transmission, there is a great need for public education regarding implementation of personal hygienic measures and prevention of inappropriate sexual contacts

enzymatic activity and molecular characterization of a secreted subtilisin-like protease in Microsporum gypseum and Trichophyton vanbreuseghemii

Subtilisin -like proteases are the group of proteases including keratinases found in dermatophytes which degraded keratin. Determination of the proteases activity of Trichophyton vanbreuseghemii isolates which were obtained from soil and clinical and soil isolates of Microsporum gypseum in Iran and characterization of their genome were aim of present study. Ezymatic activity was determined by use of chromogenic substrates. The genes, which coded subtilisin-like proteases in above-mentioned dermatophytes, was identified and amplified by using specific primers in PCR. The highest yield of enzyme production was observed in only one isolate of T. vanbreuseghemii Ir-84 whereas low enzyme activity was observed in M. gypseum isolates. Homology study of obtained nucleotide as well as amino acid sequences indicated different rates of homology with other subtilisin-like proteases genes in other pathogenic dermatophytes. Intra-strain differences were observed in production of serine proteinases and molecular characterization of genes encoding such enzymes could be of great interest for studies on pathogenicity and other purposes

Study of griseofulvin effects on the expression of ATPase-subunit G gene in dermatophyte pathogen Trichophyton rubrum

Trichophyton rubrum is one of the anthropophilic dermatophytes with worldwide distribution. This fungus is a common causative agents of Tinea cruris, Tinea corporis, Tinea pedis and tinea manuum. Several properties of this fungus have been studied so far; however few investigations were carried out in the field of molecular biology of this microorganism. The main goal of this survey was the evaluation of ATPase-subunit G gene expression of this fungus due to different amounts of griseofulvin. Serial dilutions of griseofulvin with fungal growth were prepared and compared with control. In this study, the control and griseofulvin-treated samples were micro scopically investigated and the RNAs were then extracted. Consequently RT-PCR was performed simultaneously in order to evaluate griseofulvin influence on ATPase-subunit G gene expression. It was indicated that the rate of shortened and twisted mycelia in 10 micro g/ml of griseofulvin was much higher than micro g/ml. Meanwhile, in micro g/ml of this drug there is an obvious up-regulation in ATPase-subunit G gene in comparison with 10 micro g/ml. It can be concluded that in certain amounts of griseofulvin, a significant increment in ATPase-subunit G gene expression as well as mycelial abnormalities could be occurred

[Investigation of antipsychotic induced parkinsonism in patients with schizophrenia]

Antipsychotic drugs have an important role in psychiatric treatment. Their side effects such as drug induced Parkinsonism, which has been a historical challenge for patients and physicians, account a major cause of treatment rejection by the patients. Drug induced Parkinsonism is the second cause of Parkinson syndrome. The aim of this study was to evaluate antipsychotic induced Parkinsonism in patients with schizophrenia. This cross sectional descriptive study was done in the year 1999 in Noor and Shariaty Hospitals of Isfahan. 200 patients with schizophrenia, affected with Parkinsonism complication, were investigated. Variables were sex, age, dosage and group of drug, duration of treatment, Parkinsonism criteria and simultaneous anti cholinergic prescription. Data was gathered in a questionnaire and analyzed by descriptive statistics and frequency distribution tables. 122 men and 78 women were studied. 26.5% of patients had drug induced Parkinsonism, which was mostly seen in women [32% versus 22.9% in men], higher age [10-19 years: 0%, 50 years and higher: 33%], and when anticholinergic was not used simultaneously [35.7% versus 25% in anticholinergic users group]. Prevalence of Parkinsonism, in high, medium, and low drug potentials was 28.7%, 29.4% and 19.2%, respectively. Differences in all of the above groups were not significant. Prevalence increased in dosage of less than 100mg [chlorpromazine equivalent dosage] versus 101-300mg [p>0.05], and in 3-6 months after onset of treatment [p<0.05]. The most prevalence criterion was rigidity [84.9%]. Parkinsonism was diagnosed in 11.76% of patients using atypical drug [clozapin]. Anti psychotic induced Parkinsonism increased in higher ages, women and when anticholinergic was not used simultaneously. This side effect was found in all groups even with clozapin. It had a greater prevalence in the beginning of treatment but decreased with treatment continuation and anticholinergic prescription. Future studies particularly on the atypical groups are suggested

Identification of aspergillus species using morphological characteristics

Although molecular methods continue to improve and become more rapidly available, microscopy and culture remain commonly used and essential tools for identification of Aspergillus spp. In this study we emphasize on morphological methods including; macroscopic and microscopic characteristics for identification of Aspergillus species isolated from environmental and clinical specimens. We used four differential media: czapek dox agar [CZ], czapek yeast agar [CYA], malt extract agar [MEA], and czapek yeast 20% sucrose agar. Morphological features of colonies on above culture media as well as microscopically characteristics for the major strains were studied and then compared with those of standard Aspergillus strains. Our major subjects were Iranian Aspergillus strains isolated from clinical and environmental specimens. Standard Aspergillus strains for study development included; A. fumigatus, [JCM 10253], A. flavus [JCM 2061], A. niger [JCM 10254], A. nidulans [JCM 02728], A. tereus [JCM 10227]. Morphological features of Aspergillus cultures were studied, the major and remarkable macroscopic features in species identification were the colony diameter, color [conidia and reverse], exudates and colony texture. Microscopic characteristics for the identification were conidial heads, stipes, color and length vesicles shape and seriation, metula covering, conidia size, shape and roughness also colony features including diameter after 7 days, color of conidia, mycelia, exudates and reverse, colony texture and shape. Finally we compared the morphological characteristics of tested Aspergillus isolates with those of the standard species Aspergillus isolates were identified in the level of species using the differential culture media. A total of 205 Aspergillus isolates studied included: 153[75%] environmental Aspergilli and 52 [25%] clinical isolates. Within 11 Aspergillus species identified, A.flavus [55%], A.niger [31.7%] and A. fumigatus [8.7%] were the most common Aspergillus isolates from all of the specimens. In our view morphological method using the differential media is the most reliable and sensitive assay to identify more medically important Aspergillus species

Molecular characterization of 70 kDa heat shock protein [HSP70] gene in Entamoeba dispar

Amebiasis caused by Entamoeba histolytica is still mentioned as one of the major health problems in tropical and subtropical areas. E. histolytica has recently been redescribed as two distinct species; E. histolytica and E. dispar. In the present study, we characterized the 70 kDa Heat Shock Protein [HSP70] of E. dispar at molecular level and compared it with that of E. histolytica. With these findings, we were able to distinguishe E. dispar from the infectious E. histolytica. Pairs of 21 nucleotide primers were designed from highly conserved regions of the same gene in other eukaryotic cells. Mentioned primers were utilized in PCR by using isolated genomic DNA template of E. dispar and the PCR fragments were then sequenced. By the time, 1020 nucleotides have been sequenced and characterized within open reading frame of this new gene which encode a polypeptide with 337 amino acids. Nucleotide sequence comparison in gene data banks [NCBI, NIH] for both the partial DNA and its deduced amino acid sequence revealed significant homology with members of the eukaryotic 70 kDa HSP family. Small parts of the mentioned sequences from E. dispar were about 100% identical to the sequences of 70 kDa HSP from E. histolytica other eukaryotic cells. The new partial gene fragment and its encoded protein have been submitted to the gene data banks [NCBI, NIH] and registered under the accession number of AY763790

Molecular characterization of subunit G of the vacuolar ATPase in pathogen dermatophyte trichophyton rubrum

Trichophyton rubrum is an anthropophilic fungus causing up to 90% of chronic cases of dermatophytosis. Several properties of this fungus have been investigated so far. However, a few studies were carried out in the field of molecular biology of this fungus. In the present study, we tried to identify the subunit G of its vacuolar ATPase [V-ATPase]. Pairs of 21 nt primers were designed from highly conserved regions of the V-ATPase subunit G genes in other fungi. Mentioned primers were utilized in PCR using isolated genomic DNA template as well as cytoplasmic RNA of T.rubrum and the PCR and RT-PCR fragments were then sequenced. About 469 nucleotides were sequenced which encoded a polypeptide with 119 amino acids. Nucleotide sequence comparison in gene data banks [NCBI, NIH] for both the DNA and its deduced amino acid sequence revealed significant homology with V-ATPase subunit G genes and proteins of other eukaryotic cells. The amino acid sequence of the encoded protein was about 84% identical to the sequence of V-ATPase subunit G from other fungi. In summary, we have cloned the first V-ATPase subunit G of dermatophytes and characterized it as a member of this gene family in other eukaryotic cells