Differences in growth response of human hair follicle mesenchymal stem cells to herbal extracts and a growth factor

Background: One of the key questions in biochemistry is why cell becomes aged and what are the involved factors? Why cell growth is stopped after some divisions and cells become senescent? This occurs in a greater frame in the whole body and cells dye after a while. Androgenetic alopecia [AGA] is characterized by a loss or decrease in hair follicle size, which could be related to the loss of hair follicle stem cells. Therefore, it is of great importance to develop novel therapies to increase hair follicle stem cells viability and proliferation

Objective: In this study, we examined the effects of bFGF and aqueous Rosemary leaf and Marshmallow root extracts on human hair follicle mesenchymal stem cells [hHF-MSCs] proliferation in order to identify their potential for hair growth

Methods: hHF-MSCs were isolated from hair follicle tissues and their mesenchymal nature confirmed by detecting cell surface antigens via flow cytometry. Bromodeoxyuridine [Brdu] incorporation assay was used to study the cell proliferation effect of herbal extracts in hHF-MSCs

Results: Human hair follicle-derived mesenchymal stem cells [hHF -MSCs] were obtained by organ culture. They exhibited surface markers of mesenchymal stem cells as shown by positive staining for CD44, CD90 and CD105. Herbal extracts and bFGF were found to induce significant proliferation of human hHF-MSCs at concentrations ranging from 10 to 20 micro l/ml and 15 to 25 micro l/ml

Conclusion: These results suggest that herbal extract may produce positive effects on the hair growth promotion of hHF-MSCs and suggesting that herbal extracts may be a good candidate for helping hair growth promotion

[Behavioral factors related to musculoskeletal disorders in nurses based on theory of planned behavior]

Musculoskeletal disorders are the most common and costly occupational injuries. Musculoskeletal disorders represent a significant occupational problem among nurses. The aim of current study was to determine behavioral factors related to musculoskeletal disorders among nurses by using the theory of planned behavior. A cross-sectional study was carried out among nurses in hospitals of Qom University of Medical Sciences in 2011. A total number of 264 registered nurses with working experience at least one year, and different shifts were included in the study. A reliable and valid questionnaire based on the theory of planned behavior and Nordic was used for data gathering. Data were analyzed using statistical software SPSS for windows V. 16 Independent t-test, chi-square, Mann - Whitney and Pearson correlation tests were used for testing the correlations among variables. The study showed that 47. 5% of nurses in the past 12 months and 36. 2% during the past 7 days have suffered from musculoskeletal disorders. The most work-related disorders were in the low back and neck. Also statistically significant difference was found between the mean scores of the constructs of attitude, subjective norm, perceived behavioral control and behavioral intention in nurses with and without musculoskeletal disorders. Musculoskeletal disorder rate still is high in nurses and attention and action is needed to improve nurses' health. Theory of planned behavior is effective at identifying behavioral and attitudinal factors related to the correct ergonomic principles in the through working position

Effects of mesenchymal stem cells with injectable scaffold on cardiac function in myocardial infarction in Rabbit

Bone marrow-derived mesenchymal cells can transdifferentiate into Cardiomyocyte cells and improve heart function after transplantation. Since biomaterials can improve the cell retention in the site, cell survival and differentiation, heart tissue engineering is now being explored as an applied solution to support cell-based therapies and increase their efficacy for myocardial diseases. Chitosan in combination with Glycerol Phosphate [GP] can produce a thermo sensitive material that in body temperature can form a jellylike material. The aim of this study was to evaluate the effects of a combination of autologous undifferentiated bone marrow mesenchymal stem cells [MSCs] and injectable scaffold on cardiac function improvement in rabbits after inducing myocardial infarction. The Left Anterior Descending [LAD] coronary artery was ligated by No. 6-0 poly amide suture material, and autologous MSCs with injectable scaffold were injected into the margins of the infarcted zone at the time of surgery. At 4 weeks after transplantation, the cardiac function and structure was detected using echocardiography. There was no significant difference among the three groups [MI only, MI Scaffold, and MI+Scaffold+MSCs] in the Echocardio-graphic parameters including, heart rate [HR], Ejection Fraction [EF], Fractional Shortening [FS], Left Ventricular Diameter [LVD] and Left Ventricular Parietal Wall Diameter [LVPW]. A combination of autologous undifferentiated bone marrow MSCs and injectable scaffold made of Chitosan+ Glycerol Phosphate in echocardiographic evaluation did not have a positive influence on achieving functional improvement

[Effects of frozen storage at -18°C on the quality changes of raw brushtooth lizardfish [Saurida undosquamis] burgers without coating]

Developing minced-based products is a good opportunity for production of value-added products. Fish burgers are an example of an acceptable value-added fast food. In the present study, fish burgers were produced from deep flounder fish with no coating and their chemical, microbial, and sensory characteristics during storage at ?18°C for 5 months were determined periodically after thawing at 4°C. Fish burgers were produced and packaged in PVC/PE polymer bags. Microbial load [including total plate count, total coliform count, Escherichia coli, Staphylococcus aureus, and psychrotrophic bacteria], chemical characteristics [moisture, pH, total volatile base nitrogen [TVB-N], and TBA], and sensory characteristicswere were determined monthly. Data were analyzed using the SPSS for Windows program. The TVB-N and TBA values increased significantly by the end of the first and second month, respectively. After that no further statistically significalt changes occured in the former, while the latter decreased until the end of storage period, presumably due to reaction of the MDA produced with protein breakdown products and formation of new peoducts. The data also showed that pH increased significantly by the end of the first month; no changes were observed afterwards. All of the microbial counts and sensory parameters had decreased significantly [p<0.05] at the end of storage period. The best expiry time for lizardfish burgers [if stored at 18°C] is 3 months after production, and sensory evaluation is the most reliable way for predicting their shelf life

Carbenoxolone induces apoptosis and inhibits survivin and survivin-AEx3 genes expression in human leukemia K562 cells

Leukemia is a malignant disorder of the blood progenitor/stem cells which is characterized by abnormal proliferation of white blood cells. Although anti-cancer drugs induce apoptosis in cancerous cells, drug resistance is the significant problem mainly due to over-expression of inhibitors of apoptosis proteins [lAPs] such as survivin. In this content, it has been reported that an anti-inflammatory drug, Carbenoxolone [CBX], could induce apoptosis and growth inhibition in several types of cancerous cells. In the present study, effects of CBX on apoptosis and level of the expression of survivin gene and its deltaEx3 splicing variant have were evaluated in K562 cells. K562 cells were cultured and treated with different concentrations of CBX: [50-300 microM] at different time intervals [12-48 hrs]. Trypan blue exclusion test was used to evaluate cell viability. Fluorescent microscopy [Acridine Orange/Ethidium Bromide double staining] and DNA fragmentation assay were used to study apoptosis. The expression level of survivin and its deltaEx3 splice variant were studied by RT- PCR. It was found that both growth inhibition and apoptosis occurred in K562 cells. In addition, down-regulation of survivin and survin-deltaEx3 were observed, after 2-4 hrs treatment with 150 microM of CBX. However, the expression level of survivin and its deltaEx3 splice variant increased in subsequent time [6-12 hrs] nearly to the level of control cells. From the results of this study, it may be concluded that CBX can be considered as a candidate for further studies in CML treatment, especially in the case of drug- resistant leukemia cells

Immobilization of Pichia pastoris cells containing alcohol oxidase activity

The attempts were made to describe the development of a whole cell immobilization of P. pastoris by entrapping the cells in polyacrylamide gel beads. The alcohol oxidase activity of the whole cell Pichia pastoris was evaluated in comparison with yeast biomass production. Methylotrophic yeast P. pastoris was obtained from Collection of Standard Microorganisms, Department of Bacterial Vaccines, Pasteur Institute of Iran [CSMPI]. Stock culture was maintained on YPD agar plates. Alcohol oxidase was strongly induced by addition of 0.5% methanol as the carbon source. The cells were harvested by centrifugation then permeabilized. Finally the cells were immobilized in polyacrylamide gel beads. The activity of alcohol oxidase was determined by method of Tane et al. At the end of the logarithmic phase of cell culture, the alcohol oxidase activity of the whole cell P. Pastoris reached the highest level. In comparison, the alcohol oxidase activity was measured in an immobilized P. pastoris when entrapped in polyacrylamide gel beads. The alcohol oxidase activity of cells was induced by addition of 0.5% methanol as the carbon source. The cells were permeabilized by cetyltrimethylammonium bromide [CTAB] and immobilized. CTAB was also found to increase the gel permeability. Alcohol oxidase activity of immobilized cells was then quantitated by ABTS/POD spectrophotometric method at OD[420]. There was a 14% increase in alcohol oxidase activity in immobilized cells as compared with free cells. By addition of 2-butanol as a substrate, the relative activity of alcohol oxidase was significantly higher as compared with other substrates added to the reaction media. Immobilization of cells could eliminate lengthy and expensive procedures of enzyme separation and purification, protect and stabilize enzyme activity, and perform easy separation of the enzyme from the reaction media

Pre-hospital emergency medical services in developing countries: a case study about EMS response time in Yazd, Iran

Pre-hospital emergency medical services, a subsystem of Emergency Medical Services [EMS], provides early care to critically ill and injured patients in the field. Time is an important factor in pre-hospital care and the length of time it takes to reach care, has a significant impact on patient outcome. This study aims to calculate the response time in EMS stations of Yazd, Iran. During 2008-2009, 11961 run report forms from all 7 EMS stations in Yazd that had been submitted to central station were surveyed. By using Excell statistical software and descriptive statistics [percent, cumulative distribution and standard deviation], we calculated fractile response times for Yazd EMS. 81.15 percent of response times were within 8 minutes as a referenced standard for EMS. The majority of call services in Yazd EMS have been responded in a suitable time interval comparing with reference standards and country's approved EMS response time goal, but there are still rooms for improvement

[Apoptosis induction and S-phase cell cycle arrest by adenosine 5-triphosphate through its degradation to adenosine in human myeloid leukemia K562 cell line]

Adenosine 5-triphosphate [ATP] is not only the current energy sources for all cells but also plays a critical role in triggering signaling pathways leading to apoptosis or differentiation. During the past years many investigations have been reported the anti-cancer activity of ATP on different cell lines. Also several mechanisms have been proposed for its action and it appears that its mechanism of action depends on the cell type. In the present study effects of ATP on human leukemia K562 cell line as an experimental model of CML and it mechanism of action were studied. K562 cells were cultured and treated with different concentrations of ATP [50-1000 microM] at various time intervals [24-72 h]. Effect of ATP on cell proliferation was studied by MTT assay. Apoptosis was studied by flow cytometry and DNA fragmentation assay. Cell cycle and DNA contents were analyzed by flow cytometry. To evaluate the mechanism of action of ATP, effects of ATPgammaS [an undegradable agonist of ATP] and products of ATP degradation such as AMP, ADP and adenosine were studied. ATP with doses of 100-1000 microM inhibited growth of the cells and induced S-phase cell cycle arrest at the time intervals of 24-72 hour. These effects of ATP led to cell death by apoptosis. In addition, the results showed that these effects of ATP were through its degradation to adenosine and eventually induction of pyrimidine starvation. Because current CML therapy methods which are based on chemotherapy are not very effective and have side effects such as drug resistance, ATP can be proposed as an effective compound alone or in combination with other drugs for treatment of CML

[ protective effect of FXIII Val34Leu polymorphism against deep vein thrombosis: a report from Iranian Blood Transfusion Organization [IBTO]]

The coagulation factor XIII is a pro-transglutaminase enzyme with tetrameric structure. An exchange of G for A in exon 2 of A subunit results in replacement of valine with leucine in amino acid 34. As a result of this substitution mutation, the clots produced are fragile and loose therefore it seems that FXIII Val34Leu polymorphism acts as a factor for individual protection against thrombosis Objectives: To determine the prevalence and role of FXIIIA Val34Leu polymorphism against deep vein thrombosis. This was a retrospective case-control study performed on 116 patients with DVT who were referred to Thrombosis and Homeostasis Laboratory affiliated to Iranian Blood Transfusion Organization. Also, 100 healthy individuals [blood donors] were recruited as control. Following DNA extraction and application of PCR and RFLP techniques in presence of restriction enzyme Cfo1, the genotypes of FXIII Val34Leu polymorphism were identified. The data were analyzed using chi square test as well as calculation of OD ratio and 95% confidence interval. The prevalence of FXIII Val34Leu polymorphism among the case and control groups was 22.4% and 37.4%, respectively. While the allele frequency of leucine in case group was 14.7% it was 20.2% in control group. No significant correlation between polymorphism and sex was established. According to our data, no association between the FXIII Val34Leu polymorphism and protection against deep vein thrombosis was demonstrated. Therefore, it seems that this polymorphism occurs as a natural phenomenon and unaffected by gender

Electromagnetic fields near transmission lines -problems and solutions

Nowadays, people are highly concerned about the effects of high voltage transmission lines on their health. Probable risk for leukemia, breast cancer, neuropsychological disorders and reproductive outcomes has been reported due to this exposure. In this study, several measurements around different areas such as overhead transmission lines, GIS compartments and some appliances have been conducted and compared with the standard tolerances. The emphasis of this research is on high voltage substations and publics. Field magnitudes above l0kV/m have been measured under wires. Results show that there is no serious concern for the people living near the transmission lines but for the individuals who are beneath those lines for long. Recent achievements about electric fields' effect on human health are reviewed in the present paper. In a case study, three types of 230kV structures are analyzed and the best phase arrangement for reducing the electric and magnetic fields is determined [the circular arrangement]. It is concluded that the most effective solution is for the governments to use the best phase arrangement and replace outdoor substations with GIS to reduce the radiations and for the people to be near the high voltage overhead lines as rarely as possible

[Prevalence of overweight, obesity and underweight among high school students in Sanandaj: 2006-2007]

Childhood overweight and obesity, major public health concerns, show a drastically rising tread worldwide. Associated with an increased risk of developing cardiovascular and endocrine problems, overweight children are more likely to be overweight adults. This study was done to evaluate the prevalence of overweight, obesity, and underweight among high school students in Sanandaj, Kurdistan, Iran. This study was performed in 2007 on 694 students from Sanandaj high schools. Socio-demographic information was collected using a questionnaire, completed during interviews. Weight and height were measured. Body mass index [BMI] was calculated and overweight/obesity was defined using age-and sex-specific BMI cut-off points, proposed by the World Health Organization Expert Committee on Physical Status [World Health Organization, 1996]. Children with BMI-for-age at or above the 95th percentile, between 85 and 95th percentile and less than 5th percentile were considered as being obese, overweight and underweight respectively. Descriptive and analytical statistics were used to analyze the data. The average of BMI was 20.7 and prevalences of overweight, obesity and underweight were 11.2, 3.2 and 7.6% respectively. Levels of overweight/obesity [14.4%] differed significantly between boys [18.2%] and girls [10.7%]. The highest percentage of overweight/obesity [20%] was found among 14-years olds. Boys showed a consistent decrease in overweight/obesity with increasing age, a trend not demonstrated in girls. Results indicated that the prevalence of overweight/obesity among high school students in Sanandaj is near national averages for adolescents. Overweight/obesity were observed more in boys than in girls. As a group, girls demonstrate a lower prevalence of obesity [1.1%] when compared to national averages. Based on the results of this study, 22 percent of the high school students in Sanandaj have inappropriate weight, which calls for appropriate intervention programs

[ risk factors affecting blood transfusion in patients undergoing coronary artery bypass graft procedure related to gender]

Females were reported to have significantly lower hematocrit [Hct], which might be the cause of increased need for blood transfusion. The goal of this study is to determine risk factors affecting blood transfusion in patients undergoing coronary artery bypass graft procedure related to gender A cross sectional review of 500 [306 males and 194 females] ASA physical status I or Il patients undergoing bypass procedures with saphenous vein graft performed over a one year period were included in this study. The conducting of anesthesia was standardized. For each patient, we recorded the gender, age, body weight, height, body surface area [BSA], and duration of surgery. Hematocrit levels prior to surgery and the end of surgery were recorded. Packed red blood cell [PRBC] administration and use of fresh frozen plasma [FFP] and platelets were noted. Differences between the data for female and male patients were evaluated using Student's t-test, Chi-square test and using regression analysis. Approximately 57.2% [111] female and only 39.9% [121] male patients received PRBCs. On average, females received 1. 6 units of PRBCs intraoperatively and 2. 6 units of PRBCs during the entire hospital stay, while the males received 1.1 units and 1.8 units for similar periods [P=0.0001]. When females and males were compared within the same subgroups for age, body surface area [BSA], duration of surgery, and preoperative Hct, etc. ...had a significant correlation with age, postoperative Hct and BSA in females but had a significant correlation with preoperative Hct, preoperative PRBC and duration of surgery in males [P<0.05]. Multiple logistic regression analysis showed that the probability of a patient receiving or not receiving PRBC transfusion is significantly influenced by preoperative Hct, postoperative Hct, duration of surgery, preoperative PRBC mass, and gender. Gender is an independent essential determinant of blood transfusion in CABG patients, and may interact with BSA, preoperative Hct, duration of surgery and other factors in determining the probability of transfusion

[ case repost: vagal paraganglioma with neck mass presentation]

Paragangliomas are neoplasms that arise from paraganglionic "bodies" of the autonomic nervous system. These paraganglia are distributed symmetrically and segmentally in the para-axial regions of the trunk and are typically related to arterial vasculature and cranial nerves of the ontogenetic gill arches. We repot a case of vagal paraganglioma with all diagnostic therapeutic approach which has not been reported previously in this manner

[Fire work-related injuries in Tehran's Charshanbe Soori]

Fireworks are the leading cause of injuries in the ceremony of Chaharshanbeh Soori in Iran. The present study was designed to assess the prevalence and the severity of such injuries, the high-risk population, the causes and the accompanying morbidities in such ceremonies. This cross-sectional study was performed on individuals referred to different medical centers by Tehran Emergency Service due to fireworks- and Chaharshanbe soori-related injuries in 2007. The patients' demographic data, the cause of the accident, the type of the firework, the severity and the site of the injury, the performed prehospital and hospital care, and the patients condition at the time of discharge were extracted from the patients medical records. The information on the severity of the remaining morbidities in 8 months after the accident was also gathered. About 197 patients were enrolled in this study. The mean age of the patients was about 20.94 +/- 11.31 years. The majority of them were males. Crackers and home-made grenade were the most frequent cause of injury. The most frequently reported injuries consisted of bums, laceration and contusion, soft tissue injury and bone injuries, respectively. Fireworks should be produced and distributed based on standard regulations. Educating the families and students, and holding public ceremonies under the direct control of the families are considered as important strategies in reducing such injuries

Early outcome of concurrent mitral valve replacement and coronary artery bypass grafting

Concomitant coronary artery bypass surgery [CABG] in patients undergoing mitral valve replacement [MVR] has been shown to be an important risk factor for hospital mortality. We evaluated preoperative characteristics, postoperative complications, in-hospital mortality rate, and length of stay in hospital for patients undergoing concurrent CABG with MVR. Preoperative and postoperative clinical data from 175 patients undergoing concurrent CABG with MVR operation at Tehran Heart Center from 2002 through 2006 were collected and entered into a database. Information was obtained by clinical and case note review as well as detailed questionnaires to physicians and patients. Mean age of patients was 57.95 +/- 10.54 years and 51.4% were male. Mean New York Heart Association [NYHA] score was 2.46 +/- 0.84. Among studied patients, 18.3% and 2.9% underwent aortic and tricuspid valve replacement, respectively. In-hospital mortality was 6.9% and 96.0% of patients were hospitalized >/= 14 days. History of congestive heart failure [P = 0.027] and postoperative brain stroke [P = 0.004] were independent predictors for in-hospital mortality. Exact considering of congestive heart failure and postoperative brain stroke related to in-hospital mortality in concurrent CABG with MVR operation are necessary

[Evaluation of transfusion reactions in thalassemic patients referred to the Tehran adult thalssemia clinic]

Thalassemia is a hereditary hemolytic anemia requiring life-long transfusion treatment. The frequent blood transfusion has some side effects, including immunologic reaetions and nonimmunologics such as iron overload and transmission of some infections such as HBV, HCV and HIV. This cross-sectional study was aimed to investigate the transfusion effects in patients referred to the Adult Thalassemia Clinic, Tehran. For data collection, the patients were asked for the history of transfusion reactions, their files of medical records were surveyed with concern to transfusion status and reactions, and some laboratory data including serum ferritin and TTI were also collected. The results were analyzed using SPSS software. Foun hundreds and forty one patients comprises of 53% males and 47% females with mean age of 22.6 years were studied. They consisted of 362 [82.1%] thalassemia major and 79 [17.9%] thalassemia intermediate. The mean of ferritin level was 1856.4 ng/dl. From the studied subjects, 7 [1.6%] had history of delayed hemolytic disorder, 25 [6.1%] presented with allergy and 41 [9.9%] showed febrile transfusion reaction. The remaining 368 subjects [82.4%] showed no evidence of transfusion reactions. The records showed that 117 patients [28.4%] were HCV Ab positive, 369 [88.7%] were HBsAb positive and 80 [19.2%] were HBcAb positive, 3[0.7%] HBsAg positive. According to this results, 300 patients [70.3%] were vaccinal positive; 66 [15.5%] were immune to HBV from past infections; 40 [9.4%] were negative and 3 [0.7%] were carrier of HBV. Totally, 366 [85%] patients were immune to HBV. We had no case of HIV positive. Transfusion reaction and their documentation are very important for blood transfusion services and programming. Unfortunately, there is no statistic records of these reactions in the hospitals or there may be some unclassified data in the patients. files. Regular co-operation between hospitals and blood transfusion centers is required for collecting the information and finding total incidence of transfusion reactions. Hemovigilance or computerized network between blood bank centers in hospitals and Iran transfusion organization is the best way for collecting such data

[ efficacy of pre-hospital care in traumatic patients referred to emergency ward of "Sina Hospital" between September 2003 and September 2004]

The accuracy of the pre-hospital care has not been evaluated in Iran so that the purpose of the present study was to evaluate the efficacy of the pre-hospital care performed by Tehran Emergency Service in traumatic patients referred to "Sina Hospital". This prospective study was conducted on traumatic patients referred to emergency department of "Sina hospital" by Tehran Emergency Service from September 2003 to September 2004. The accuracy of the pre-hospital care performed by Tehran Emergency technicians, regarding the patients' general condition was evaluated. The on-call physicians in emergency department of "Sina hospital" were responsible for gathering the data and filling the questionnaires based on their observation and the EMS forms. 994 patients were enrolled in the study. Wound bandage and homeostasis were done correctly in 80% of the cases. Splint was applied correctly in 50% of the patients, while other therapeutic procedures such as collar and spinal bed were not performed among 80% of the cases. IV-line was obtained in 91.2% of the patients, while serum infusion was initiated in only 20% of patients. It is recommended to compile evidence-based protocols for the pre-hospital procedures and train the emergency technicians to follow the rules, in order to improve the efficacy and the accuracy of the pre-hospital care. According to the principal changes recently made in the quality and quantity of the care offered by Iran's Emergency Service, it is necessary to perform a new study to evaluate the present condition and compare the results of these, qualitative and quantitative changes with the current study

Accuracy of spiral CT in non-traumatic acute abdominal pain in Sina University Hospital

Acute abdominal pain is a common complaint in an emergency setting. An immediate and precise diagnosis is necessary for reducing morbidity and mortality. Several studies have reported that CT scan increases diagnostic accuracy for patients with acute abdominal pain. This study was designed to evaluate the sensitivity, specificity and accuracy of spiral CT scan compared to that of abdominal ultrasound and plain radiography. We assessed the data of 91 consecutive patients, including 45 males and 46 females, ranging in age from 8 to 84 years [mean age 52.38 years] presenting to the emergency department of Sina Hospital, Tehran, Iran, with acute non-traumatic abdominal pain during the years 2003-2005. All patients underwent spiral CT scanning; in addition, sonography and plain radiography were performed for 66 and 64 of the patients, respectively. The sensitivity, specificity and accuracy were calculated and compared, based on the final diagnosis, which was established with surgical, pathologic, and clinical follow-up. The data was analyzed by SPSS 13.0 software. Among the 91 patients examined, CT scan was reported to be normal in 15 patients [16.5%]. The most common CT findings were aortic aneurysm [12.1%], pancreatitis [9.9%], ovarian cyst [7.7%], intestinal distension [7.7%], and hepatic cyst [7.7%]. The overall sensitivity, specificity, and accuracy of unenhanced spiral CT were 92.2%, 92.86%, and 92.3%, respectively, whereas those of plain radiography were 13.2%, 72.7%, and 23.4%, respectively, and ultrasound 73.2%, 90%, and 75.7%, respectively. This study suggests that plain radiography is an insensitive technique in the evaluation of nontraumatic acute abdominal pain presenting in the emergency department. Unenhanced spiral CT is accurate for adult patients with nontraumatic acute adominal pain and should be considered as an alternative to radiography as the initial imaging modality

[Improved Diagnosis of brucella in human serum samples by double PCR assay]

Brucellae are gram-negative intracellular pathogens which cause zoonotic disease in humans. Clinical manifestations of brucellosis in human are variable and often are non-specific, and the diagnosis requires fast and accurate confirmation. Since the usc of serum instead of whole-blood samples offers several advantages for nucleic acid amplification methods, in this study we developed an improved PCR assay for the rapid and specific laboratory diagnosis of human brucellosis directly from serum specimens. DNA was extracted from 100 microL of serum from 30 patients with acute serologic brucellosis. The PCR reaction was carried out with Specific primers. Second PCR reaction for reamplification of the first reaction products was designed. A 223 bp conserved region on the sequence encoding the 31-KDa immunogenic outer membrane protein which is specific to the genus Brucella [BCSP31] and present in all its biovars was amplified in all serum samples. For confirmation and efficient amplification of the specific target, reamplification of the first PCR products had a sharper banding patterns with high sensitivity and specificity that might be considered as a new useful method for diagnosis of human brucellosis in serum specimens