Iranian Journal Of Pathology
Volume:3 Issue: 4, Autumn 2008

Formalin-fixed paraffin-embedded tissues are a valuable source of DNA for molecular studies. We designed and optimized an efficient procedure for DNA extraction from formalin-fixed paraffin embedded tissues. Seventy three blocks of cervical paraffin-embedded tissues were investigated. DNA was extracted using 45 minutes boiling in alkaline solution together with 10 beads of Chelex-20, followed by phenol-chloroform extraction and alcohol precipitation. This method produced DNA suitable for amplification using primers specific for human SMN and β globin genes in 98.63% and 82.2% of samples respectively. We also detected human papillomavirus DNA in 58.33% of appropriate samples. This procedure provides simple and efficient method for recovery of amplifiable genomic and viral DNA from paraffin wax embedded tissues.

Vascular endothelial growth factor (VEGF) and its receptors (VEGF-R1 and R2) are major regulators of angiogenesis. This study was designed to assess serum levels of VEGF and VEGF-R1 and their prognostic significance in newly diagnosed childhood acute leukemia. For this purpose, VEGF and VEGF-R1 were determined using enzyme linked immuno-sorbant assay (ELISA) in samples obtained before treatment. Demographic data were recorded. Bone marrow blast percentage was counted on diagnosis and 2 weeks after induction therapy. A p value less than 0.05 considered significant. Fifty-three children (22 boys and 31 girls) with newly diagnosed acute leukemia were enrolled in the study. Most cases (56.6%) were pre B cell ALL. Mean value of VEFG-A in good responders was 55.13 ± 24.96 pg/ml and in poor responders it was 94.46 ± 15.75 (p<0.0001). Mean valve of VEGF-R1 in good and poor responders was 0.132 ± 0.0653 and 0.1665 ± 0.0857 pg/ml respectively (p>0.05). Using ROC curve, we found out a cut-off point of 76 pg/ml to discriminate poor response to chemotherapy. Soluble VEGF-A is an independent factor for response to therapy in childhood leukemia and leukemic patients with sVEGF-A level over 76 pg/ml will have poor response to treatment.

To estimate the risk of human papilloma virus (HPV) infection for cervical malignancies, we conducted a case-control study in southern Iran (Hormozgan province). For this purpose, 52 paraffin embedded blocks with exact diagnosis of cervical carcinoma(50 carcinomas and 2 carcinomas in situ) from 2001 to 2006 and 52 praffin embedded blocks of cervical tissue specimens with normal histopathology as the control group were tested for the presence of HPV DNA using PCR based assay. HPV DNA was found out in 16 out of 52 patients (30.7%), while it was not detected in any of the control group samples. Considering the fact that unrestrained sexual behavior increases risk of becoming infected with HPV, our finding is in favor of the concept of low frequency of HPV infection and thus its less important role in women with cervical cancer in islamic countries.

There have been concerns over possible association between mercury and neurodevelopmental outcomes in infants. In this study we aimed to determine whether blood levels of mercury are above safe values in Iranian infants or not. A total of 85 infants (0, 2, 4 and 6 months old) were enrolled in this study. All of them received vaccines according to Iranian immunization schedules. We measured total mercury in all blood samples by cold vapor atomic absorption. The mean concentration of blood mercury in our subjects were as follows: newborns as 33.95 ± 11.86 nmol/l (with a range of 23.93-52.84), 2 months as 32.94 ± 11.76 nmol/l (with a range of 23.92-52.84), 4 months as 30.44 ± 10.44 nmol/l (with a range of 23.92-50.85) and 6 months as 37.93 ± 12.97 nmol/l (with a range of 21.43-52.34). There was not any significant difference for the mean concentration of blood mercury in those age groups. The lowest level of blood mercury detected was 21.43 nmol/l and the highest level was 52.84 nmol/l. The finding of this study showed that approximately 33% of the infants had blood mercury levels above the U.S. Environmental Protection Agency recommended reference dose of 5.8 μg/l (29 nmol/l). Therefore, it is needed to reduce exposure of infants to mercury from all sources including thimerosal containing vaccines (TCVs) in Iran.

Urinary tract infection (UTI) is one of the most common diagnosed infectious diseases. In order to determine the spectrum of bacterial etiology and antibiotic resistance pattern of the uropathogens that cause community acquired UTI, a retrospective study was performed in Research Center of Reference laboratories of Iran on urine samples of referred patients during a 4 years period (from 2002 to 2006). During the study period, 4207 urine specimens were cultured. Cultures with a significant bacterial growth were selected and susceptibility testing was performed using disk diffusion method. Out of 4207 urine specimens, 476 (11%) yielded positive culture. Escherichia coli (44.5 %) and Klebsiella pneumoniae (8.5%) were the most predominant isolated bacteria respectively. Escherichia coli revealed the highest susceptibility among oral antibiotics to nitrofurantoin (76%) and the highest rate of resistance to carbenicillin (94%) and ampicillin (89%) respectively. In total isolates, the highest level of susceptibility was for imipenem (99%) and among oral antibiotics for nitrofurantoin (70 %), and the highest level of resistance was for carbenicillin (89%) and ampicillin (84%). The results of this study revealed a considerable resistance to fluoroquinolones and cotrimoxazole and a high sensitivity to nitrofurantoin among isolated bacteria. We strongly recommend nitrofurantoin as the drug of choice for empiric therapy of UTI in our country.

Pneumocystis pneumonia (PCP) has been historically the most prevalent opportunistic infection in patients infected with the human immunodeficiency virus. Culture of the organism has not been faced with suitable success in artificial media, while various results have been reported for cell culture media. The aim of this study was proliferation of Pneumocystis carinii on the Razi Bovine Kidney (RBK) cell line and to compare growth rate with ‘Vero’ and ‘MRC-5’ cell lines. We used 6 rats (Sprague-Dawley) provided from Razi Institute to infect with Pneumocystis carinii after suppressing the immune system with methylprednisolone acetate (40 mg/kg). Methylprednisolone acetate was used subcutaneously once a week for 8 weeks. Samples were homogenized after separation of the lung tissue. Microscopic examination was applied for prepared smears to confirm the presence of Pneumocystis carinii. Purified trophozoites were then inoculated into the cell line flasks. Growth rate was estimated by counting the trophozoite in each day. Number of cultivated organisms was increased after 5 days incubation in all applied cell lines. Growth rate of Vero, MRC-5 and RBK were 3, 3, and 3.75 times more respectively in comparison with number of the calculated cells in first day. Hence the difference between RBK and two other cell lines was significant (p = 0.023). RBK cell line is suitable to proliferate Pneumocystis carinii.

According to the fact that neonatal infection is a challenging diagnosis field, several studies have tried to test sensitivity and specificity of diagnostic tests. This study was conducted to evaluate the sensitivity and specificity of procalcitonin (PCT) as a single early marker of neonatal sepsis. In this study, 150 neonates admitted to NICU and neonatal ward in Shahrekord Hajar hospital were enrolled. A full workup including blood cultureand other tests and PCT was conductede. Sensitivity, specificity, positive and negative predictive values for PCT was determined. It was found out that 8 patients had definite infection, 15 patients had possible infection, and 127 patients had no infection. Although PCT was not able to significantly differentiate between those with definite and possible infections (p>0.05), but there was a significant difference for frequency of abnormal PCT between non-infectious patients and other patients. Sensitivity, specificity, positive and negative predictive values of PCT was 87.5%, 87.4%, 30.4%, 99.1%, and 87.41% respectively. Beside the limitation of the sample size, the satisfactory diagnostic characteristics of PCT highlight it as a good measure for diagnosis of neonatal sepsis. Further studies are essential to be carried out.

Non-specific granulomatous prostatitis is an uncommon diffuse inflammatory condition of the prostate. It is important because it may be mistaken for prostatic carcinoma. The aim of the study was to determine the prevalence of non-specific granulomatous prostatitis (NSGP) and comparing the results of transrectal ultrasonography, serum prostate specific antigen (PSA) and free prostate specific antigen levels, digital rectal exam (DRE) in NSGP with the prostate carcinoma and benign prostate hyperplasia. During a cross-sectional study, the prostate needle biopsy cases with the granulomatosts prostatitis diagnosis that had been referred to one of the largest pathology centers(1 year: 2006) were found and their clinical files were revised from the point of DRE, transrectal ultrasonography (TRUS), fPSA, and PSA. Some clinical and pathology findings such as age, microscopic findings, sonography information and experimental findings that had been necessary for the study were gathered and analyzed using SPSS software. Out of 783 needle biopsies of prostate, 8 (1.02%) cases were non-specific granulomatous prostatitis. The age range of patients was 55-76 years (with a mean of 66.1 years). Mean of PSA level was 19.45 ng/ml and fPSA level was 0.7 ng/ml. In 2 patients, TRUS showed focal hypoechoic areas and in other 2 of these DRE revealed asymmetry and mild nodularity. There is no pattern of clinical, biochemical or ultrasound findings that allows a specific diagnosis of granulomatous prostatitis to be made or differentiate it from prostatic carcinoma and the biopsy is still necessary for the certain disease diagnosis.

Celiac disease is an autoimmune disorder, characterized by inflammation, villous atrophy, and crypt hyperplasia of the small bowel mucosa. In this study we considered and compared sensitivity and specificity of serological tests in patients with celiac disease. In this cross-sectional study we prospectively recruited children with suspected celiac disease. An intestinal biopsy specimen was obtained from all patients. Celiac disease diagnosed on the basis of histologic findings of Marsh classification. A serum sample was taken at the time of biopsy for serologic tests. Findings were analyzed using SPSS program, t-test, and chisquare tests. Out of a total of 134 children in this study, seventy (52.21%) patients were boy and sixty four (47.8%) patients were girl. Celiac disease was diagnosed in 14 (10.4%) of the patients. In serologic tests, 11 patients (78.6%) were positive for antigliadin-Ab, 4 (28.6%) for anti tissuetransglutaminase Ab, and 9 (64.3%) for antiendomysial antibody. Sensitivity of antigliadin-Ab was 78.6% and its specificity was 95.9%. Sensitivity of anti tissue-transglutaminase Ab was 28% and its specificity was 95%. Sensitivity of antiendomysial Ab was 64% and its specificity was 96%. Positive serologic tests are supportive of the diagnosis in those with characteristic histopathologic changes on small intestinal biopsy. The best tests for this purpose are the IgA antiendomysial antibody or IgA anti tissue-transglutaminase, both of which are highly sensitive and specific.

Inflammatory pseudotumor is a lesion composed of proliferating spindle cells with mixed inflammatory infiltrates. Some authors have proposed the name inflammatory myofibroblastic tumor as a proper descriptive term rather than the vague inflammatory pseudotumor. The aim of this study was to verify the myofibroblastic origin of spindle cells in idiopathic orbital inflammatory pseudotumor by immunohistochemical staining. We reviewed a series of 32 inflammatory pseudotumors arising in orbit for expression of smooth muscle actin, vimentin, desmin and anaplastic lymphoma kinase using immunohistochemical staining. There were 21 females and 11 males aged 3 to 64 years with a mean age of 31. Immu-nohistochemically, spindle cells of 51.75%, 79.3%, and 17.2% of lesions expressed smooth muscle actin (15/29), vimentin (23/29) and desmin (5/29). All lesions (32/32) were negative for anaplastic lymphoma kinase In this study, reactivity for smooth muscle actin in spindle cells can be demonstrated as myofibroblastic differentiation. The absence of anaplastic lymphoma expressionin all cases of orbital inflammatory pseudotumor in this study strongly suggests that these lesions, albeit histologically similar, are biologically distinct from their soft tissue counterparts or those inflammatory myofibroblastic tumor that negative for anaplastic lymphoma immunoreactivity may be characterized by one or more chromosomal aberration involving regions other than 2p23 is as yet unknown.

Endometriosis is the presence of benign endometrial glands and stroma outside the uterus. Urinary system involvement by endometriosis is a rare occurrence accounting for only 1.5 % of all endometriosis cases.Case 1. The patient was a 41 years old woman admitted for right flank pain. The physical and gynaecologic examination was unyielding. Intravenous urography (IVU) revealed stenosis in distal part of right ureter, unfortunately associated with hydronephrosis. Case 2. A 26 years old woman who suffered from colic pain in low abdomen and pelvis. The only positive finding was microscopic hematuria. Abdominal sonography showed hydronephrosis of right kidney and proximal part of ureter. Ureterolysis by an open surgical procedure performed for both patients and histologic examination revealed endometriosis of ureter.Endometriosis of ureter is mainly asymptomatic and unfortunately ends in functionless kidney due to prolonged hydronephrosis. Early diagnosis needs high index of suspicion and intended use of paraclinic aids to save patients normal renal function. An individualized therapy plan depending on the patient’s age and the extent of the endometriosis should be attempted.