Clonality of the immunoglobulin heavy chain genes in B cell non-Hodgkin lymphoma using semi-nested PCR

Identification of gene rearrangements and clonality analysis are important techniques for the diagnosis of malignant lymphoproliferative diseases. These methods have various sensitivities based on the type of primer used and method of determination of polymerase chain reaction [PCR] products. This study aimed at determining the clonality of B cell non-Hodgkin lymphoma in Iranian patients using PCR method and 2 primers of FR2 and FR3. Paraffin embedded blocks of 67 patients with B cell lymphoma and 19 cases with lymphoid hyperplasia of the lymph nodes who presented to NRITLD, Masih Daneshvari Hospital were retrospectively reviewed. After extracting the genomic DNA using phenol and chloroform, clonal analysis was performed using semi-nested PCR by using two primers: FR2 and FR3. PCR products were determined using 2 techniques of heteroduplex analysis, polyacrylamide gel and silver staining and the conventional method of agarose gel and ethidium bromide staining. Appearance of 1 or 2 bands in the desired location were considered as a sign of clonality. Monoclonal gene rearrangement was observed in 62 out of 67 patients [92.5%] as one or two discrete bands appeared within 60-120 base pairs [bp] and 200-300 bp range. Of the mentioned patients, 53 cases [79.1%] had FR2 and 51 [76.1%] had FR3 rearrangement. Heteroduplex analysis along with silver nitrate staining detected 3 out of the remaining 5 cases of lymphoma to be monoclonal. These cases had been reported negative by the conventional technique. In total, 65 out of 67 patients [97%] showed monoclonal gene rearrangement using both the abovementioned techniques. All hyperplasia cases were polyclonal by this method. Our study showed that evaluation and detection of clonality using PCR, FR2 and FR3 primers along with heteroduplex analysis is a rapid sensitive technique for the diagnosis of malignant lymphomas

Transient localized hemophagocytosis in pleural effusion

Epstein-Barr virus [EBV] is one of the most important causes of hemophagocytic syndrome. We report a 76-year-old man who presented with pneumonia like symptoms and pleural effusion following upper respiratory tract infection. He underwent thoracentesis and pleural fluid cytology revealed large number of histiocytic macrophages that had phagocytosed RBCs and other inflammatory cells like lymphocytes and neutrophils. Pleural fluid analysis showed Epstein-Barr virus Antigen [EBNA] as the causing agent and corticosteroid therapy was initiated. A few days later, pleural effusion subsided and further cytologic examinations revealed no trace of hemophagocytosis

Role of quantiFERON-TB test in detection of children infected with mycobacterium tuberculosis

Latent TB infection can persist for many years with about 10% lifetime risk of reactivation to active disease. However, in children with latent TB infection, disease develops within 2 years of infection. Recently, a new diagnostic test [QuantiFERON-TB Gold] which measures the production of interferon [IFN] gamma in whole blood upon stimulation with Mycobacterium tuberculosis has been introduced. The aim of this study is to compare the performance of the IFN-gamma assay with tuberculin skin test [TST] for the identification of latent TB infection in children in contact with active TB in the pediatric pulmonary ward. This cross-sectional study was conducted on 100 children, aged 2months-15 years admitted to the Pediatric Ward of Masih Daneshvari Hospital during 2007-2008. Whole blood was collected for measuring Interferongamma using QuantiFERON-TB Gold kit [QFT-Cellestis Comp]. In this procedure, Mycobacterium tuberculosis specific antigens [ESAT-6 and CFP-10] are used. In the present research, 100 children were studied and divided into 3 groups of case [TB], contact and control. PPD test was performed by injecting 0.1 ml of the 5 unit solution [Pasteur Institute of Iran] for all cases. Twenty-eight percent of the contacts, 60% of the cases and 10% of the controls were Afghans; the remaining were Iranians. Smear of the gastric washing [3X] was prepared in contact and case [TB] groups; 30% of the cases [TB] were AFB positive, while all of the contacts had negative smears. History of BCG vaccination during neonatal period and BCG scar were present in all cases. Positive PPD test [PPD >/= 10 mm] was observed in 90% of the cases and 24% of the contacts. PPD test was negative in the control group. Out of 50 contacts, 18 [36%] showed positive QFT test; and of 20 TB patients, 18 [90%] had positive tests. Regarding age, children with positive QFT test belonged to the older age group. To our knowledge, this is the first study to investigate the performance of the whole blood IFN-gamma assay in diagnosing latent TB infection in children in Iran. This study found a fair correlation between the TST and the whole blood IFN-gamma assay in children at high risk of latent TB infection. Our study also highlighted fair and moderate agreement in contact and TB groups respectively between the TST and QFT-TB test in children at high risk for latent TB infection. More studies are required to clarify this relationship

Idiopathic pulmonary fibrosis and mutation of TGF-beta gene, codon 10

Idiopathic pulmonary fibrosis [IPF] is associated with histological appearance of usual interstitial pneumonia. These fibrotic changes in lung interstitium are mostly attributed to cytokine production such as TGFbeta which stimulate migration and differentiation of fibroblast to myofibroblasts. The polymorphism of TGFbeta gene was found to be associated with development of IPF. We investigated whether TGFbeta1 gene polymorphism in codon 10 is associated with interstitial pulmonary fibrosis in Iranian population. The different genotypes of TGFbeta1 at [+ 870] position [in codon 10] was studied in41 cases and 83 control subjects. The allele specific PCR method was used for genotyping. In the patient group, the frequency of T allele [NO: 58] was 70.7% and C allele [NO: 24] was 29.3%. The frequency of TT genotype [NO: 20] was 48.8%, followed by T/C [NO: 18] 43.9% and CC [No. 3] 7.3% while in the control group, the frequency of T allele [N:117] was approximately 70.5% and C allele [NO: 49] was 29.5%. The frequency of TT genotype in control group [NO: 41] was 49.4%, followed by T/C [NO: 35] 42.2% and C/C [NO: 7]8.4% In comparison with the control group, there was no association between TGFbeta1 codon 10 T/C polymorphism in our cases with IPF

Solitary fibrous tumor of the pleura: a case report and review of the literature

Solitary fibrous tumor of the pleura [SFTP] is a rare mesenchymal cell tumor that can be benign or malignant. The best treatment of this tumor is a complete surgical resection. We present clinical and histopathologic characteristics of the 4 patients and their outcomes

Genetic diversity of pseudomonas aeruginosa strains isolated from hospitalized patients

Pseudomonas aeruginosa is one of the most common nosocomial pathogens often causing major problems in Intensive Care Units. This study aimed to investigate the genotypic diversity of Pseudomomas aeruginosa strains isolated from hospitalized patients in National Research Institute of Tuberculosis and Lung Disease [NRITLD] with random amplified polymorphic DNA [RAPD] method and also to determine the antibiotic resistance pattern. Seventy three P. aeruginosa isolates from different specimens were analyzed. These strains were isolated from patient admitted in Intensive Care Unit [ICU] [31], non-ICU inpatient [40], and two environmental specimens one from ventilator and one from soap specimen in ICU. All strains were identified with biochemical testing and antimicrobial susceptibility testing which carried out according to National Committee for Clinical Laboratory Standards [NCCLS]. Random Amplified Polymorphic DNA typing [RAPD] was used to study the genetic diversity of Pseudomonas aeruginosa using 2 sets of primers and electophoretic banding patterns were analyzed visually and by GelCompar ?? software. Phylogenic analysis of the RAPD pattern showed rates of genetic similarity ranging from 40-100%. Four epidemiologically and genetically related isolates [clones] each containing 2-3 isolates were identified. Most of them were from ICU. We detected high antimicrobial resistance rate to Chloramphenicol, Ceftriaxon, Cefepime, Ceftazidime [75-97%] and relatively low resistance rate to Imipenem, Amikacin, Ciprofloxacin and Gentamicin [42-53%]. Although a few epidemiologically related clones are found with RAPD method, most of the isolates are probably emanate from the host itself. There is also a high rate of antibiotic resistance especially in ICU

Assessment of angiotensin-converting enzyme gene in idiopathic pulmonary fibrosis

Interstitial pulmonary fibrosis [IPF] is a progressive fibrotic interstitial lung disease with a distinct histopathological form referred to as usual interstitial pneumonia [UIP]. Evidence has indicated that a local renin-angiotensin system is present in distal lung parenchyma. Expression of the component of this system is present in a number of fibrotic lung diseases. In this study, we assessed the association of Insertion/Deletion [I/D] polymorphism of angiotensin-converting enzyme [ACE] gene in IPF. By using semi-nested PCR, we determined the I/D polymorphism of ACE gene in 23 paraffinembedded open lung biopsy specimens from patients having clinical and imaging findings of IPF and pathologic diagnosis of UIP at National Research Institute of Tuberculosis and Lung Disease [NRITLD]. Afterwards, we compared the results with I/D polymorphism of ACE gene in a healthy control group [n= 88]. The frequency of I allele was 71.7%[33 out of 46] and the frequency of D allele was 28.3% [13 out of 46]. The frequent genotype was I/D [56.5%] which was statistically significant comparing with healthy group [27.3%]. We had no D/D genotype .There was a difference in the distribution pattern of ACE genotype between patients and controls [P < 0.05]. Our study revealed an association between carriage of I allele and I/D genotype in IPF

Fiberoptic bronchoscopy: correlation of cytology and biopsy results

Fiberoptic bronchoscopy is a diagnostic method for respiratory diseases. At present, its diagnostic yield has been increased by different cytologic and histologic procedures by convention. This study was conducted to evaluate the concordance and agreement between cytologic and histologic findings in conventional diagnostic bronchoscopic methods [washing and biopsy] for lung malignancies. This was a cross-sectional study performed on 2076 cases of bronchial biopsy and bronchial washing between 1996 and 2003. Of 2163 patients who underwent fiberoptic bronchoscopy after omitting 87[4%] cases due to unsatisfactory specimens, 2076 cases were studied including 832 [36.9%] females and 1244 [63.1%] males in the age range of 2 to 100 years, [mean age 57.7 +/- 16.3 yrs]. Male to female ratio was 1.5. Malignancy was diagnosed in 657[31.6%] biopsy and 283[13.6%] cytology specimens. Two hundred and sixty-five cases had malignant lesions according to both bronchial biopsy and bronchial washing; therefore, Kappa coefficient in both methods was 46.7% [P value = 0.000]. Concordance rate was 77.4%. Ninety-seven point three percent of malignant cases were diagnosed by biopsy and 41.9% by cytology. Cytology contributed to an additional diagnostic rate of 2.6%. Kappa agreement is classified as fair and although there is a very good concordance between the two sampling techniques, the diagnostic yield of cytology for malignancy must be improved by combination of multiple assays

Comparison of propofol and remifentanil administration on lipid profile

This study aimed to investigate the serum level of triglyceride, cholesterol, high density lipoprotein, low density lipoprotein, and very low density lipoprotein during administration of propofol and comparing it with infusion of remifentanil in patients undergoing sedation in ICU of Masih Daneshvari Hospital during 2005-2007. All patients with pulmonary disease, undergoing intubation and mechanical ventilation were enrolled in our study. The patients were randomly divided into two groups, first receiving propofol and second receiving remifentanil as the sedative agent. Lipid profile [triglyceride, cholesterol, high density lipoprotein, low density lipoprotein, and very low density lipoprotein] was checked before, immediately after, and the day after drug administration. A total of 40 patients were enrolled in this study, 20 of which took propofol and the remaining took remifentanil. The mean age of the patients was 58.67 +/- 18.57 yrs. Triglyceride and very low density lipoprotein[VLDL] were the two factors with statistically significant rise after infusion of propofol [p < 0.002]. Such a change was not detected in the remifentanil group. The other understudy factors did not show similar changes. Propofol infusion can induce dramatic rises in triglyceride and VLDL concentration even after low dose infusions and therefore special attention must be paid to patients prone to hyper-triglyceridemia and pancreatitis

TNF alpha gene polymorphism in idiopathic pulmonary fibrosis

Idiopathic pulmonary fibrosis [IPF] is characterized by a chronic inflammatory process and abnormal wound healing. Tumor Necrosis Factor alpha [TNF alpha] is considered to play a key role in fibroblast proliferation and increased collagen synthesis. It appears that there is a genetic predisposition to IPF. The genetic associations of TNF-alpha with IPF have been reported in different cohorts and revealing conflicting results. This study was conducted to evaluate the association of TNF- alpha-308 G/A polymorphism with IPF in Iranian patient by PCR-RFLP method. Materials and Methods: TNF alpha gene polymorphism at position 308 G/A was examined on DNA extract of 41 cases with IPF defined clinically, radiologically and histologically and compared with 82 unrelated healthy controls who were kidney donors. Results: The understudy population included 20 males and 21 females with the mean age of 50.4 yrs. Data showed that the frequencies of G allele [NO:72] and A allele [NO:10] were 87% and 12% respectively. The frequency of G/G genotype [NO:31] was 76%, followed by G/A [No:10] being 24%.we had no A/A genotype. Conclusion: There was no association between TNF alpha-308 G/A polymorphism and IPF in Iranian patients

Proliferative Marker in distinction between benign and malignant mesothelial proliferations

Differentiation of benign from malignant mesothelial proliferations is a major problem in the pathology of the serosal membranes, particularly in small biopsy specimen. This study was conducted for the evaluation of proliferative marker for distinction between malignant mesothelioma [MM] and mesothelial hyperplasia [MH]. Thirty six cases of malignant mesothelioma [MM] with the mean age of 62.94 years [range: 36-80 years, M/F: 3.58] and 22 cases of mesothelial hyperplasia [MH] were evaluated for proliferative status by immunohistochemical [IHC] method with monoclonal antibody, Ki-S5 [Ki-67]; the labeling indices [LI] were evaluated. Average count revealed a significant increase in MM as compared with reactive MH [p value < 0.0001]. Considering a threshold of 9% for ki-67, a sensitivity of 88% and specificity of 94% were resulted. Proliferative marker of Ki-67 can be useful in distinction between malignant mesothelioma and mesothelial hyperplasia [p-value < 0.0001]

Recovery of mycobacteria from clinical specimens and assessing drug susceptibility test of mycobacterium tuberculosis specimens by mycobacteria growth indicator tube [MGIT]

Tuberculosis is a disease of global importance. Indeed, the lack of sensitive methods for the diagnosis and inappropriate therapy may lead to increased multidrug-resistance [MDR] cases. However, early detection and identification of acid fast bacilli [AFB] in clinical specimens can lead to effective intervention. The sputum specimens from 156 clinically suspected tuberculosis patients and 40 non- tuberculosis patients were digested, examined microscopically for acid- fast bacilli, and inoculated into "Mycobacterium Growth Indicator Tube" [MGIT], BACTEC -12B vial and onto Lowenstein- Jensen slants by standard procedures. The result showed that smear was positive in 82 [52.5%] and negative in 74 [47.5%] of 156 clinically suspected tuberculosis patients. The culture positive rate with Lowenstein- Jensen, MGIT, and BACTEC-12B vial were 122 [78%], 136 [87%], and 143 [91%], respectively. Thereafter, MGIT indirect and direct susceptibility tests were performed on 15 sputum-positive specimens and the results were compared with proportional method. The results have revealed that accordance with proportional method was higher in MGIT indirect [83.5%] than direct [75%] susceptibility test, the difference was significant [p< 0.05]. In another set of experiments, the indirect MGIT drug susceptibility test in 25 mycobacterium tuberculosis isolates were performed and compared with proportional method. The results showed that MGIT could correctly detect susceptibility to streptomycin, ethambutol, rifampin and isoniazid for 77.8%, 33%, 77.2% and 80%, respectively. Also, the agreements with proportional method for resistance were 88% for streptomycin, 80% for ethambutol, 80% for rifampin and 89% for isoniazid. Furthermore, by combining MGIT technology with L.J media, the mean time required for culture to grow for identification test was reduced from 22-28 to 12-16 days [p<0.05]. MGIT is an efficient system to be used in center/ referral mycobacteriology laboratories of developing countries along with routine solid or liquid culture media