Journal of Advances in Medical and Biomedical Research
Volume:17 Issue: 67, 2009

Many genetic studies on predisposing factors for active tuberculosis have been conducted. Study on human leukocyte antigens (HLA), vitamin D receptor (VDR), NRAMP1, mannose binding lectin (MBL), and tumor necrotizing factor (TNF) are the most studies in this field. This study was planned to identify any relationship between VDR polymorphisms (Apa I, Bsm I, Fok I and Taq I) and susceptibility to TB. This case-control study was performed on blood samples from tuberculosis cases (n=164) and controls (n=50). DNA was extracted from white blood cells and the sequences were amplified by PCR followed by restriction digestion (PCR-RFLP technique) using specific primers and enzymes for each polymorphism. VDR polymorphisms were evaluated for two mentioned groups. Two genotypes of AbfT and AabbFfTT were the only statistically significant genotypes which had a dfferent frequency between the study and control groups. Results of this study showed that genotypes of AbfT and AabbFfTT are protective factors against TB in our patients. We could not find any genotype as a predisposing factor for TB in our study group. However, other studies with larger group of samples are needed to find such a relationship.

JWH133 is known to have cannabinoid-2 (CB2) receptor agonist properties. Celecoxib, a selective cyclooxygenase-2 (COX-2) inhibitor, is also known to have antinociceptive properties. Endocannabinoids produce analgesia possibly through cyclooxygenase (COX) pathway. The aim of the present work was: to study the effect of celecoxib on JWH133 induced antinociception and to compare the effects of two different dose ranges of celecoxib (mg/kg and nano g/kg) on the JWH133 antiniciceptive effect. We have studied the possible interaction of administration of mg/kg (50-200 mg/kg) and Ultra-Low Dose (ULD) (25 and 50 ng/kg) of celecoxib on the antinociceptive effect of intraperitoneal (i.p.) injection of JWH133 using formalin test in mice. JWH133 (0.01, 0.1 and 1 mg/kg) induced antinociceptive effect just in phase I of the formalin test. Celecoxib (50-200 mg/kg) and its ULD (25 and 50 ng/kg) attenuated and potentiated, JWH133 induced antinociception, respectively. It is concluded that JWH-133 induced antinociception is modulated by celecoxib and mg/kg doses of celecoxib showed opposite effects compare to its ultra-low doses.

Spoligotyping is a method based on 36bp Direct Repeat (DR) chromosomal loci polymorphism which is connected to one or two 35-41 bp spacer sequences. There are 94 different intra DR spacer sequences which are identified so far and only 43 of them are used as usual. Mycobacterium tuberculosis complex strains can be identified based on lacking or having these sequences. Spoligotyping test was carried out on 238 TB smear positive patients. Primary separation of mycobacterium strains was done through Petrof 4% method and Lowenstein Jensen (LJ) media. Biochemical tests such as Niacin test/Catalase activity/Nitrate reduction were done in order to identify the strains. Drug sensitivity to INH (0.2Mg/ml)/ RIF (40Mg/ml)/ STM (10Mg/ml) and ETBl (2Mg/ml) identified by proportional method and according to that, the strains were divided into three groups: sensitive, multi drug resistance (MDR) and non MDR. Then DNA was extracted by CTAB method from the positive colonies. Sequences were amplified by PCR and after denaturizing, hybridization with Streptavidine peroxidase enzyme was performed by Line reverse blot method. Radiography was done after adding the Luminoscense and membrane onto the X-ray films. Serotypes were divided into 9 groups (Beijing/ CAS1/ Haarlem / U/ T2/ T1/ EAI3/ EAI2 and CAS2). Most of the strains were from Haarlem (27%) and CAS1 (25%) groups. Two strains were also identified in this method that belonged to Mycobacterium bovis. Spoligotyping method is an easy, rapid and sensitive test in order to identify Mycobacterium tuberculosis complex strains.

Identification of atypical mycobacterium (Non tuberculosis Mycobacterium NTM) is important because of the worldwide propagation of these organisms. Recently, molecular studies have identified the specific loci for mycobacterium species by DNA - finger printing methods, but these methods are time-consuming and expensive. In this study, in addition to hsp65 PCR-RFLP method, QUB3232 locus was evaluated for differentiation of atypical mycobacterium from mycobacterium tuberculosis complex. This study was performed on 371 pulmonary and non pulmonary specimens separated from patients with the symptoms of pulmonary tuberculosis (PTB). After the isolation and culturing of mycobacterium strains using the Lowenstein Jensen media, biochemical tests including production of Niacin, Catalase activity, Nitrate reduction, pigment production and growth rate were performed. Drug susceptibility testing was performed by proportional method. DNA extraction was performed by phenol-chloroform method. hsp65 gene was amplified by PCR. Subsequently the amplicons were digested with three restriction enzymes namely AvaII, HphI and HpaII and electrophoresed on 3% agarose gel. QUB3232 locus was also evaluated for differentiation of atypical mycobacterium and mycobacterium tuberculosis complex. Out of 371 isolates, 32 (8.6%) were multi-drug resistant TB (MDR-TB), 184 (49.5%) were susceptible and 155 (42.5%) were non MDR (combined resistance) that 15% of MDR cases and 25% of non MDR cases were non tuberculosis mycobacterium. Out of 31 slow growing isolates, 58% were M. simiae and 19% were M. kansasii. The sensitivity of QUB3232 locus for differentiation of the atypical mycobacterium from mycobacterium tuberculosis complex was 80%. From the total of 43 NTM samples, 12 (27.9%) were rapid growing and 72% were slow growing. QUB3232 locus has the high discriminative power for differentiation of atypical mycobacterium from the mycobacterium tuberculosis complex, therefore, it can be used as a substitute for PCR-RFLP method.

Tuberculosis (TB) caused by Mycobacterium tuberculosis, is an infectious disease in human which kills nearly three millions of people annually. Approximately, one - third of the world populations are infected with this bacteria and 5 - 10 % of them develop the active form of the disease. Individuals are different in susceptibility to TB infection. These differences might be due to the host characteristics especially genetic factors. TNF-α as a pro-inflammatory cytokine, play a key role in host defense against tuberculosis. Presence of mutation in this gene can influence the effectiveness, performance and capability of immune responses against TB infection. The Aim of this study was to investigate the frequency of TNF-α gene polymorphisms and its relation with susceptibility to the pulmonary TB. Sixty healthy controls and 60 TB patients were enrolled. Genotype of TNF-238, TNF -244, TNF-308, TNF -857 and TNF-863 were determined using PCR-RFLP method. The results were analyzed by Fisher Exact and χ 2 tests using SPSS v.14 and evaluated with Hardy–Weinberg equilibrium. The results of this study showed a significant difference in TNF-308 and TNF -857 regions between the control and study groups (P < 0.05). Presence of mutation in TNF-308 and TNF -857 regions may increase the host susceptibility to mycobacterium tuberculosis and genotyping of these regions can be used for screening of the high risk individuals.

Due to fears of postoperative complications following upper gastrointestinal surgeries (UGI), fasting before bowl function recovery is a traditional practice, but fasting following elective surgery is controversial. The aim of this study was to compare early oral feeding versus traditional oral feeding in patients who underwent UGI surgeries. Fifty two patients who underwent UGI anastomosis or surgery for various reasons were randomly divided into early oral feeding (EOF) group and traditional oral feeding (TOF) group. The nasogastric tube (NGT) removal time, tolerance of oral feeding, ileuses, nausea and vomiting, vital sign before and after surgery, postoperative stay, patients’ satisfaction and complications were recorded. The mean time of NGT removal was 1.62 ±0.49 and 4.61±1.99 days in EOF group and TOF group respectively (p=0.0005). The mean start time of oral feeding was 2.04 ± 0.19 and 5.87 ± 1.32 days in the EOF group and TOF group respectively (p=0.0005). Tolerance of oral feeding was seen in 24 (92.3%) patients and 21 (91.3%) patients in the EOF and TOF groups respectively. Duration of hospital stay following surgery was 5.62 days in the EOF group and 8.04 days in the TOF group. 24(92.3%) out of 26 patients in the EOF group were satisfied with oral feeding that started in the second postoperative day. 13 patients (56.5%) complained of delay feeding in the TOF group. The results of the present study suggest that early oral feeding following upper gastrointestinal anastomosis or surgery is safe and can result in a shorter hospital stay and less cost.

There are several techniques for the diagnosing of salmonella infectious. Several molecular methods such as PCR and hybridization assay have recently been used for the detection of this bacterium. However, these methods require precision instruments for amplification and complex procedures, which are the major obstacles to the widespread use of these methods in relatively small scale clinical laboratories, clinics and the filed laboratories. Recently, a new, rapid and sensitive technique called loop-mediated isothermal amplification (LAMP) was developed. In this study we used 7 different strains of salmonella to compare the PCR with LAMP method. For PCR test we used thermocycler, but The LAMP reaction can be conducted under isothermal conditions by using only one type of enzyme and four primers recognizing six distinct regions. The most important merit of this method is that no denaturation of the DNA template is required, so, technique is simple and no need to thermocycler machine and several temperatures cycles. Conventional PCR method for the detection of Salmonella with standard thermocylcer takes 3 hrs but, with LAMP method we were able to amplify and detect the salmonella in very simple thermal block made in IRAN. After Optimization of the process it was possible to rapidly detect and identify Salmonella typhi bacteria within 90 minutes. This method was also 100 times more sensitive comparing to the PCR method. According to the results, comparing LAMP isothermal amplification method for detection and identification of Salmonella with conventional PCR we have been able to determine the simplicity, speed (3 times) and the superior sensitivity(100 times) of the LAMP to PCR method. This Method is more simple, faster and cheaper (10 times). Another advantage is independence to cycle's temperature and thermo-cycling and replacement with one thermo block which is very simple, inexpensive and made in inside the country.

Polycystic ovarian syndrome (PCOS) is the most common endocrine disorder among reproductive-age women. There is very little information about the prevalence of PCOS in Iran. With regard to the symptoms of PCOS which begin after menarche and regarding to its side effects on women's health, we aimed to determine the prevalence of PCOS in adolescents in Zanjan, Iran. In this descriptive community based study, 1882, 14-18 year old adolescents were randomly selected from Zanjan schools. The presence of PCOS was determined by the presence of olygomenorea, hirsutism, acne and androgenic alopecia. For correlation between PCOS and obesity, BMI and central obesity was evaluated. Statistical analysis was performed using K2 test. PCOS was present in 54(2.9%), hirsutism was present in 161 (8.6%), acne was present in 220 (11.7%), androgenic alopsia was present in 130(6.9%) and menstrual irregularity was present in 281 (16.9%) of the cases. The prevalence of central obesity and over weight did not differ among the studied groups. The prevalence of PCOS in our study was similar to the results reported from other societies. With regard to PCOS side effects, we suggest that the diagnosis and treatment of PCOS is better to be started from adolescence.

Urinary tract infection (UTI) is one of the most common bacterial infections in the childhood which could result in chronic renal failure and hypertension. Antibiotic resistance is increasing due to widely using of antibiotics. The aim of this study was to determine the MIC of antibiotics which are using in the treatment of UTI in children by E-test. In this descriptive study, 87 E.coli strains were isolated from the urine samples of the patients with UTI. E.coli antimicrobial susceptibility was determined using E-test. The MIC for each antibiotic was determined and classified using NCCLS criteria. Eighty seven urine samples were collected from 57 girls (65.5%) and 30 boys (34.5%). The mean age for girls was 61 months and for boys was 41 months (p=0.015). The MIC (50 and 90 percentiles) for each antibiotic was as fallow: Ampicillin (256, 256), Amikacin (1/5, 4/8), Gentamycin (0.38, 32), Nalidixic Acid (1/5, 256), Ceftriaxon (0.023, 32), Cefixim (0.19, 256) and Trimethoprim-Sulfamethoxazole (32, 32). The antibiotic susceptibility rate for each antibiotic was as fallow: Ampicillin (21.8%), Amikacin (92%), Gentamycin (75.5%), Nalidixic Acid (64/4%), Ceftriaxon (72/4%), Cefixim (65/5%) and Trimethoprim-Sulfamethoxazole (41/4%). This study showed that the antibiotic resistance of E.coli was very high. Due to increasing rate of E.coli resistance to Ampicillin and Cotrimoxasole in children, it is better to reconsider the empirical therapy with these antibiotics. Since in this study a lower resistance rate of E.coli was observed for Amikacin and Gentamycin therefore, we could suggest these antibiotics as alternatives in the treatment of children with urinary tract infection.

Meningitis is one of the serious complications of extra pulmonary tuberculosis (TB). TB meningitis is the most common chronic meningitis in the world. Early diagnosis and treatment of TB meningitis will decrease the probability of focal neurological defects in these patients. AFB smear and culture are the methods that routinely are used for the diagnosis of tuberculosis. Polymerase Chain Reaction (PCR) of CSF samples is a specific diagnostic technique to approve the TB meningitis infectious. A 26 years old woman with a 10 days history of continuous global headache, nausea and restlessness referred to a neurologist. A brain CT scan without contrast was normal. Then a lumbar puncture (LP) was carried out and CSF sample was analyzed. An aseptic meningitis pattern in the CSF test was observed. All of the other tests were showed to be normal. There was no change in the result of the secondary LP test and CSF analysis that was done less than 24 hours after the first LP. Regarding to the seasonal prevalence of entroviruses, causes of lymphocytic viral meningitis was ruled out. Tuberculin Skin Test (TST) was positive in this patient. The PCR of CSF sample for TB was positive. So the diagnosis of TB meningitis was approved and treatment was started. The patient had a good condition after one year of treatment. Since the TB infectious has a variety of presentations that could be unusual, we should think about TB in patients with similar manifestations of meningitis and pneumonia. Due to its specificity, PCR in these conditions is a very helpful method for TB diagnosis as it can detect few copies of bacilli in biological samples.