Avicenna Journal of Clinical Medicine
Year:2004 | Volume:10 | Issue:SUPPLEMENT 4|Papers Count:12

Inventing PCR technique made a great progress in the field of medical diagnostic, legal affairs, archeology and so on. Early detection of some infections like toxoplasmosis, rubella and etc are important in view of preventing undesired outcomes. Formalin-fixed archival samples are one of the popular source of  diagnostic materials but, however are known to be poor materials for molecular biology applications. These are the easiest to store and transport. They are often used as the source of nucleic acids for retrospective molecular analyses based on DNA amplification by polymerase chain reaction. We conducted a series of experiments to understand possibility of doing polymerase chain reactions on materials fixed for a long time. Two human stool samples and two cattle samples contained Cryptosporidium oocysts, that had been stored for a long time, used for experiment. DNA extraction were done by serial freeze and thaw and phenol chloroform method. Some modifications were done in usual PCR technique, for example altering the time of anealing step, to repeat this step and other modifications. Employing some modifications in the anealing step, and repeat of this  step one or two times, showed that it can be possible doing PCR with long time formalin-fixed materials. Extraction of DNA by such method was not impossible but difficult. Also our results showed that two isolates of Cryptosporidium from human and cattle genetically were the same. Formalin-fixed clinical materials can use for diagnostic purposes.

Indium is a toxic element belonging to group IIIa. This element and it's radioisotopes (lllIn, 113In)often are used in medical sciences, including monitoring of health in workers of atomic industries, treatment and imaging of tumor and infections and also imaging of genes expression. Therefore in this study we tried to measure the concentration of indium in different parts of the hepatocytes. After measurements of LD50, 0.35 mg/Kg indium as InCl3 was injected (I.P) daily to male rat for 60 days. Then the animals were scarified and the concentration of indium was determined in serum and homogenate tissue of liver> kidney> brain and also in nucleus, mitochondrial, lysosomal and post-lysosomal fractions of hepatocytes using flamless atomic absorption. Results of this study showed that LD50 for indium ion was approximately 4.2 mg per Kg of rat weight. Chronic injection of such metal ion showed that indium was distributed in these tissues as kidney>liver> brain. It's intracellular distributionin hepatocytes was in  mitochondrial> nucleus> post- lysosomal> lysosomal respectively. Results obtained from this study indicated that indium mainly stored in the kidney, liver and mitochondrial, nucleus of hepatocytes.

The goal of present study was to determine the frequencies of antigens and their alleles of ABO and Rhesus blood systems in a group of women with recurrent abortion, and comparison with the general population in Hamadan. All women referred to "Fattemieh" hospital because of their recurrent abortion during three years (1998-2000) were our cases (no=265) and all women that were voluntary blood donner in Hamadan Blood Bank during one year (1996) were the control group (no=1217). The study was a cross sectional descriptive study. Primary information (ABO& Rh blood types) were obtained from their files in "Fattemieh" hospital (for cases) and from official records of Hamadan Blood Bank (for controls). By using suitable formula, Frequencies of genes A, B, O of ABO system and D and d genes of Rhesus system were calculated. In final step, the results of two groups were compared with each other. In ABO system, the most frequent blood types were O, A, B and AB respectively in case group. The frequent alleles were O, A, B, respectively. The frequencies of alleles were not statistically different in two groups. Also, the frequent allele in Rhesus system was "D". The frequency of D allele in two groups was statistically different (p=0.05). It seemed that Rh negative was a risk factor for recurrent abortion in our case population.

Our previous studies have shown a severe iodine deficiency in lagoon (A village in north-east of Tehran).Whole inhabitants of this village received iodized oil injection (containing 480 mg iodine) in 1989. In addition iodized salt was distributed after three years in this village. This study was carried out in order to monitor of the changes of height and weight of schoolchildren aged 6-14years of lagoon ten years after iodine supplementation. Ninety-nine students including 46 female and 53 mates in eight age groups were studied. Anthropometric measurement for weight and height were performed in all subjects. Weight was measured with minimum clothing using electronic scales that was calibrated for accuracy. Height was measured bare foot. The weight was measured to the nearest 0.1 kg and height to the nearest 0.1 cm. Height and weight determined in every age group. These data were compared with similar data in 1989 (before intervention). Anthropometric data were presented as indices of l score of height for age and weight for age. These indices were compared with reference data of the National Center of Health Statistics(NCHS). Deviation of height in subjects in 1999 were 51, 13, 36 % in below 5th percentile, between 5th and 50th percentile and upper 50th percentile (median)respectively. Deviation of weight were 5,13 and 28%.Ascompared to data in 1989 (before supplementation) median of height and weight is near to NCHS cure. These changes were not statistically significant. Weight and height in 13%of schoolchildren were under 5th percentile of NCHS. However height in 51%and weight in 59%of subjects were between 5th and 50th percentile of NCHS. This study shows that administration of iodine supplementation in this village ten years after intervention has caused improvement of physical growth specially in children below 10 years old because of the mean of height and weight were nearer to NCHS. In the future studies the socio-economical indices must be considered.

Visceral leishmaniasis or kala-azar, is a worldwide disseminated intracellular infection of the liver, spleen and bone marrow. Mortality and morbidity of visceral leishmaniasis is about 15% even in case of treatment. Determination of anti-leishimania antibody in serum of patients using indirect fluorescent technique (IFA) and detection of leishman bodies in bone marrow smear are presently used for definitive diagnosis of visceral leishmaniasis in Iran. Bone marrow puncture is an invasive method and inconvenient for patients. Moreover, because of some technical problems, the sensitivity of the test is quit low. The ability to diagnosis visceral leishmaniasis using easily obtained material, such as patients' blood, would be a major advance. The aim of this study was to determine the ability of PCR method for detection of Leishmania DNA in peripheral blood of patients with visceral leishmaniasis. The results were then compared with IFA method. A total of 67 whole blood and sera were collected from patients suspected of having visceral leishmaniasis. Polymerase chain reaction on whole blood and indirect fluorescent antibody on sera were applied for these patients. DNA extraction was carried out on whole blood by using lysis buffer and boiling method. Leishmania-specific primers (13A and 13B) were used to amplify 120 base pair fragment of Leishmania kinetoplast DNA (kDNA) minicircles. Microscopically examination of smears of bone marrow was carried out for the patients during hospitalization. In 27 patients, the diagnosis of visceral leishmaniasis was made by clinical symptoms, laboratory data, response to treatments and final clinical diagnosis. 23 of 27 patients with confirmed visceral leishmaniasis had positive PCR (85.2% sensitivity). None of 40 cases with others infections had PCR- positive (100% specificity). Sensitivity and specificity of IFA was 63% and 82.5%, respectively. There was statistically difference between specificity of these methods (P value < 0.01). These results demonstrate the high specificity and sensitivity of the non-invasive PCR. The patients with early stage of visceral leishmaniasis were detected by this method as well. Therefore, PCR is a valuable method for the diagnosis of visceral leishmaniasis in compare of IFA.

Staphylococcus aureus (S.aureus) is the most common cause of cutaneous pyogenic infection. The main source of the bacteria is human body that is commonly colonized in the anterior nares of nose. Some groups of individuals seem to be particularly prone to colonization with S.aureus. For example, physicians, nurses, patients undergoing chronic hemodialysis & users of illicit intravenous drugs, etc. The purpose of this study was determination of nasal carriage rate of S.aureus in hemodialysis patients & communication with shunt site infection & antibiogram of all isolates S.aureus from nasal carriers. This survey was carried out in a descriptive analytic cross-sectional research in 2002. 77 patients of chronic renal failure were selected who had undergoing hemodialysis in dialysis ward of Ekbatan hospital. Specimens with swabs were performed from anterior nares nose of the patients & culture on blood agar broth then antibiogram were performed for all the isolated of S.aureus to 9 types of antibiotics. The patients were 41 females (53.2%) & 36 males (46.8%). The mean of age in patients was 48.43 years & the mean of duration of dialysis was about 23.54months. 31 cases were nasal carriers of S.aureus (40.3%) versus 46 cases of noncarriers (59.7%). There was no significant statistical correlation between duration of hemodialysis in nasal carriers S.aureus & noncarrier patients (P-value=0.78). 12 patients (7 carriers & 5 noncarriers) died (15.6%) showed no significant statistical correlation between the death in nasal carriers & noncarrier patients (P-value=1.93). Follow-up of patients in 3 months showed 2 cases (6.45%) with shunt site infection in carrier cases. There was no shunt infection in noncarrier patients. Antibiogram was done for all isolated of S.aureus from individuals who were sensitive to vancomycin, rifampin, trimethoprim, sulfamethoxazole, gentamycin & cephalotin 100%.96.8%. 93.6%, 90.4%&80.6%respectively. Resistance to penicillin, cloxacillin & oxacillin has been respectively 100%, 100%.90.3% that indicate higher rate of resistant species of S.aureus to methicillin. In this survey, the nasal carriage rate of S.aureus in dialysispatients were more than the other similar studies. We recommend all the hemodialysis patients to be considered as nasal carriers of S. aureus and if possible. All the nasal carrier patients should be treated with effective antibiotics such as rifampin. trimethoprim/ sulfamethoxazole or the other similar drugs.

Anaerobic bacteria produce volatile and non-volatile fatty acids in enriched media containing glucose (FAB) in anaerobic conditions. The production of these acids by mentioned organisms could be used for identification purposes. On the other hands, the extraction of whole-cell proteins of strains have been introduced as a reliable technique for identification and typing of microorganisms. The aim of this study was, the comparison of efficacy of GLC method with extraction of structural proteins techniques (SDS-PAGE) in identification and typing of reference strains and clinical isolates. In this study 62 clinical strains isolated from patients and 4 reference strains obtained from collection including bacteroides fragilis, mobiloncus crutisii, prevotella bivia and porphyromonas gingivalis were tested by two methods. Clinical strains isolated from patients with periodontal disease and bacterial vaginosis. Volatile and non-volatile fatty acids of species were detected by gas liquid- chromatography (method of Holdman, et al). Whole-cell proteins of strains were also detected by Taylor, et al. method. Most reference strains and clinical isolates produced volatile and non-volatile fatty acids in enriched media that differentiated them on the genus level, however, this method could not discriminated them on the sub-species level. The most fatty acids produced by strains were: propionic, iso-valeric, butyric, acetic, lactic, succinic and phenylacetic acids. In this study, many protein bands from 205 KDa to 37 KDa were also detected by SDS-PAGE that differentiated them well. Protein profiles of reference strains and clinical strains isolated from patients could discriminated them on the species and sub-species levels. The species of bacteroides divided to 5 sub-species, mobiloncus to 2 sub-species, prevotella and porphyromon as each to 3 sub-species by SDS-PAGE. Our results showed that method of gas-liquid-chromatography (GLC) can be used for identification and typing of organisms on the genus level and extraction of structural proteins(SDS-PAGE)is a useful method for typing them on the species and sub-species levels, also can be use in molecular studies.

The purpose of this study was to evaluate of salivary estriol level in hospitalized women with diagnosis preterm labor. This study was prospective study, in which salivary estriol specimens were obtained from 43 women at 24-34week 6 days gestational age with symptoms of preterm labor (between 9AMand 8 PM).Samples were kept frozen at-20°C. Salivary estriol was assayed by a specific enzyme linked immunoassay. We compared maternal salivary estriol levels between two groups, case group with preterm labor and delivery (22 individuals) and control group with preterm labor and term delivery (21individuals). The results have showed that the mean ± standard error salivary estriol levels in 31, 32, 33 and 34 weeks gestational in case group were higher than control group. The results showed that the mean ± standard error salivary estriol level in case group (0.31±0.21) was higher than control group (0.25±0.21). Receiver-operator curve analyses showed that exceeding a 18.75ng/dl saliva estriol level was associated with occurrence of preterm delivery (sensitivity = 68.2%, 57.1%specificity, 62.5%positivepredictive value, 63.2% negative predictive value).

One of the most challengeable issues in reproductive medicine is management of patient who poorly response to ovarian stimulating drugs. IVF/ICSI one of the methods that has been used in infertility with different ethiology. Because of limited success, high expense and stress in treatment, multiple endocrine tests and sonography have been done by infertility  center to prediction of ovarian function. It would be clinically and economically helpful if there were better methods for prior assessment of likelihood of an inadequate ovarian responses. In prospective case control study, Data include age, duration of infertility, ethiology , ovarian volume and FSH, Estradiol in 3rd of menstrual collected from 54 woman <42 y without historyof surgery on ovaries and FSH<20who candidated for IVF/ICSI. Results included number of follicle, duration of stimulation and rate of pregnancy analyzed with SPSS. In point of response to COH patients divided two group, 36 case with good responses and 18case with poor responses(lesser than 4 follicle). Age, duration, and ethiology of infertility, FSH>12 , Estradiol >80 and ovarian volume < 3 cm3 was not significant different in two groups.

Phospholipases are the major component of all eukariotic bilayer cell membranes and serve as a scaffold for membrane proteins. Phospholipases are a class of ubiquitous enzymes that have in common both their substrate (phospholipid) and the fact that they are all estrase and due to their ability to lyses of cell membrane can play an important role for pathogenicity of microrganisims. During this study using degenerate primers based on homologous amino acid sequences of phospholipase B (PLB) search for detection of some fungal PLB was carried out. DNA extraction of Aspergillus fumigatus, A. niger, Fusarium venonatum and Aurobasidium pullulans was carried out and then using degenerate primers based on nocleotide sequences of Phospholipase B were degenerated. Predicted 550 bp product from A. fumigatus was cloned in pGEMT-Easy vector and then transformed into E. coli Top 10 F' competent cell for extraction of cloned DNA fragment. Sequence analysis of 550 bp fragments revealed a sequence for the PLB gene with a high homology to published PLB sequences. Negative result for detection of PLB gene in Fusarium venonatum can be due to unsuitable primer or the lack of PLB gene in the genome of these  microrganisim.

One of the most important etiology for obstetrical hemorrhage is plecenta previa, risk factors for this are most variation range. In this point thisstudy evaluated risk factors. In this study 105 patients have been calculated in duration of 4 years, rate of prevalence placenta previa in years 74-77was 0.31%present. More of them have 0-2 parity 25%of patients had a history of abortion and 15.2% had a historyof repeat cls, 10.5%had historyof curetage due to abortion or molar pregnancy. Between 32947NVD, 4 case had status placenta acreta or increta Sonography detected placenta previa in  6.2%cases before labor, the most chief complaint of patients was bleeding without pain the most of patient symptomized in gestational age 31-40 (64.7%). The most of patients had 21-25 years 10%of patients had symptoms of severe  abruption placenta, the patients haven't history of Tobacco using.

Congenital cutis laxa (CCL) is a very rare disorder with hanging skin, giving the appearance of premature aging. The genetic, physiologic and clinical aspects of a 13-year-old girl affected with CCL were examined. Pedigree pattern study, cardiovascular and respiratory parameters assessment were performed. Cell culture of blood cells and skin fibroblast also were done. In addition karyotyping and detection of mitochondrial deletions (mt-DNA)with standard methods were done. The results indicated that this case was affected by autosomal recessive congenital cutis laxa and pathological examinations confirm it. Biochemical and hormonal parameters were normal compared with results of normal populations at the same age. Her karyotype was normal and there was no any mt-DNAmutation. Our study showed that in CCL despite sever skin, and cardiopulmonary involvement normal conditions in other physiological aspects may appear. Also it seems that the best way for early detection of the disease is skin biopsy and study of elastic fibers employing histopathological methods.