IRANIAN JOURNAL OF PATHOLOGY (IJP)
Year:2016 | Volume:11 | Issue:2|Papers Count:15

It is estimated that the number of HIV infected children globally has increased from 1.6 million in 2001 to 3.3 million in 2012. The number of children below 15 years of age living with HIV has increased worldwide. Published data from recent studies confirmed dramatic survival benefit for infants started anti-retroviral therapy (ART) as early as possible after diagnosis of HI. Early confirmation of HIV diagnosis is required in order to identify infants who need immediate ART. WHO has designed recommendations to improve programs for both early diagnoses of HIV infection and considering ART whenever indicated? It is strongly recommended that HIV virologocal assays for diagnosis of HIV have sensitivity of at least 95% and ideally greater than 98% and specificity of 98% or more under standardized and validated conditions.Timing of virological testing is also important. Infants infected at or around delivery may take short time to have detectable virus. Therefore, sensitivity of virological tests is lower at birth. In utero HIV infection, HIV DNA or RNA can be detected within 48 h of birth and in infants with peripartum acquisition it needs one to two weeks. Finally it is emphasized that all laboratories performing HIV tests should follow available services provided by WHO or CDC for quality assurance programs. Both clinicians and staffs providing laboratory services need regular communications, well-defined SOPs and nationally validated algorithms for optimal use of laboratory tests. Every country should use assays that have been validated by national reference laboratory.

Background: Automatic Cell Counter devises make the CBC differential very easy and delivering the results in few second. However, the problem with this device is facing a flag requires a time-consuming microscopic review of the specimen which causes unacceptable wait times for patient as well as costs for laboratories. In this study, we calculated the validity of WBC diff flags in Sysmex XT-1800i. In addition, we verified the correlation between manual and automated samples.Methods: Overall, 1095 flagged samples were selected in the period of 6 weeks (Imam Hospital complex, Tehran Iran, 2014). The results of both automated and manual counting of the samples were carefully studied and compared. Totally, 624 NRBC flags, 450 Blast flags, 155 abnormal WBC Scatter gram flags, 140 Eosinophilia flags and 468 Monocytosis flags were identified.Results: Considering NRBC and blast flags there was a significant difference between our manual counted and automated counted NRBCs and blasts (P<0.05).There was no significant difference between automated and manual counting of flags for WBC Scatter gram. A significant difference between automated and manual counting data in flags, eosinophilia and monocytosis was foun (P<0.05).Conclusion: Maspin expression was reduced in samples with grade II& III of invasive ductal carcinoma. Based on expression of Maspin Inc-erb-B2, it seems that more expression happened in normal group comparing with different scores of it.We could suggest that there was a reverse relationship between tumor formation and Maspin gene expression. These results showed possible role of Maspin as prognostic factor

Background: The breast cancer is the most prevalent cancer among women, on the other hand absence of myoepithelial cells play a pivotal role in pathogenesis of this cancer. Thus we aimed to investigate the possible abilities of the molecular assay technique to find a relationship between mammary serine protease inhibitor (Maspin) gene expression possibly secreted by myoepithelial cells, grade of breast cancer and other prognostics factors (ER, PR, and c-erb-B2).Methods: Paraffin embedded blocks of 31 breast cancer patients together with two normal breast tissues were used for IHC staining and Maspin gene RNA detection uses the real-time PCR method. Applying QIAGEN kit, we were able to measure Maspin RNA and Extract the cDNA of different samples for evaluating the Maspin RNA level.Results: We found that the RNA level was considerably lowerin these cancer samples compared with normal samples. In addition, different grades of breast cancer in the obtained results adopt some distinguishable values. The Maspin expression in samples with grades II and III is much lower than the ones in normal group (P<0.05) which could be considered as a promising way in diagnosing of this disease. The results showed no considerable differences in Maspin gene expression of the c-erb-B2 scores in the tumor group except the samples having score 0. The other observation of this research study confirmed that Maspin gene expression couldn' t show any differences between the values of both ER and PR in different scores of the tumor group. On the other hand, the cDNA of these patients showed lower values compared with normal samples.Conclusion: Maspin expression was reduced in samples with grade II& III of invasive ductal carcinoma. Based on expression of Maspin Inc-erb-B2, it seems that more expression happened in normal group comparing with different scores of it.We could suggest that there was a reverse relationship between tumor formation and Maspin gene expression. These results showed possible role of Maspin as prognostic factor.

Background: Tuberculosis (TB) remains as a major cause of death. Construction of a new vaccine against tuberculosis is an effective way to control it. Several vaccines against this disease have been developed. The aim of the present study was to cloning oftb10.4 gene in pcDNA3.1+plasmid and evaluation of its expression in eukaryotic cells.Methods: Firstly, tb10.4 fragment was amplified by PCR and the PCR product was digested with restriction enzymes. Next, it was cloned into pcDNA3.1+plasmid.Following that, pcDNA3.1+/tb10.4 recombinant plasmid was transfected into eukaryotic cells.Results: 5700 bp band for pcDNA3.1+/ tb10.4 recombinant plasmid and 297 bp fragment fortb10.4 were observed. Cloning and transfection were successful and designed recombinant vector was confirmed by sequencing.Conclusion: Successful cloning provides a basis for the development of new DNA vaccines against tuberculosis.

Background: Presence of discordance between the Gleason score on needle biopsy and the score of radical prostatectomy specimen is common and universal. In this study, we determined the accuracy of Gleason grading of biopsies in predicting histological grading of radical prostatectomy specimens and the degree of overgrading and undergrading of prostatic adenocarcinoma in our center, which is one of the referral centers in Tehran.Methods: In this retrospective study, we analyzed the results of prostate needle biopsies and subsequent prostatectomies diagnosed at the Pathobiology Laboratory Center, Tehran, Iran in 45 patients between 2002 and 2013. Preoperative clinical data and the information from biopsy and prostatectomy specimens were collected. The accuracy, sensitivity, specificity, and positive and negative predictive values of different grades and groups were assessed. Pearson and Spearman correlation coefficient were used to determine the relation of different variables.Results: The biopsy Gleason score was identical to the scores in prostatectomy specimens in 68.2% cases, while 31.8% were discrepant by 1 or 2 Gleason score. We had 9.1% downgrading and 22.7% cases upgraded after prostatectomy. The sensitivity and positive predictive value was 86% and 79% for low grade, 67% and 75% for moderate grade, and 80% and 80% for high-grade tumors, respectively.Conclusion: Overall, the reliability of Gleason grading of needle biopsies in predicting final pathology was satisfavory. Moderate grade group was the most difficult to diagnose in needle biopsy.

Background: Hepatitis B virus potentially accelerates graft rejection and mortality in renal transplantation population. Vaccination of graft candidates without prior immunization against HBV seems essential before transplantation but some candidates of transplantation have not received HBV vaccine at the time of receiving graft. We aimed to evaluate immunogenicity of an enhanced regimen (4 doses of double-strength intramuscular shots) after kidney transplantation in candidates without history of prior HBV vaccination.Methods: This quasi-experimental study was conducted, 49 renal graft recipients in Sina Hospital (Tehran University of Medical Sciences, Tehran, Iran) of age>18, receiving graft within past 6 months and negative history of hepatitis B vaccination from 2010-2011. Participants received 40 μg intramuscular (IM) shots of a recombinant vaccine in the months 0, 1, 2 and 6. The titer of HBsAb was measured 8 weeks after the 3rd and 4th injections. Cases with HBsAb titers less than 10 mIu/ml were considered as non-responder while antiHBs≥10 mIu/ml was considered protective.Results: The overall response rate was 57.14% (28/49 patients). Protective HBsAb titers were detected in 44.89% patients following 3rd dose and reached to 57.14% after injecting the 4th shots. The mean HBsAb titers were 50.00 (±88.35) mIu/ml and 229.45 (±356.56) mIu/ml after the 3rd and 4th shots respectively. Responders showed significantly younger age in comparison to non-responders (P=0.013). The vaccine was well tolerated in all patients with no side effects.Conclusion: Regarding the relative good response rate following HBV vaccination in graft recipients, we suggest a post-transplantation enhanced regimen of 4-dose double-strength IM shots against HBV in patients without prior immunization.

Background: Until now, trimodal distribution of HbS has been seen by six different studies in the world when associated with alpha-thalassemia with confirmation by corresponding alpha-genotyping studies. The RBC indices reduce as alpha-globin genes reduce in sickle cell trait (SCT) patients, which decreases the extent of intravascular sickling and thus betters the clinical course of the patients. This is a pioneer study conducted on Central Indian poor population to use the already proven six studies to screen associated alpha-thalassemia in SCT patients thus, circumventing the much costlier alpha-genotyping studies. Moreover, it aimed to study the haematological parameters in such cases.Methods: The study was performed at RHDMC, IGGMC, Nagpur, India from 2003 to 2012. The sample population was suspected cases of haemolytic anaemia. CBC and RBC indices were obtained by a cell analyzer. The sickle solubility test positively screened cases were confirmed by agar-gel haemoglobin electrophoresis at pH 8.6.Finally, quantitative assessment of haemoglobin variants was performed by HPLC.Results: Out of total 5819 cases over ten years, 933 cases were sickle heterozygotes.Overall, 180/933 subjects were predicted to be homozygous alpha-thalassemia and 338/933 were heterozygous alpha-thalassemia, based on trimodal distribution of HbS.Conclusion: Genotyping is costlier for majority of the poor non-affording patients in Indian government set-ups, so this study is suitable to screen for associated alphathalassemia in SCT patients.

Background: In most cases, prostatitis can be caused by a bacterial agent such as Ureaplasma urealyticum. Considering to the cumbersome of the culture method for the detection of Ureaplasma species in clinical samples such as prostate; PCR method that is faster and more appropriate than the cultivation methods, can be utilized for the detection of U. urealyticum and U. parvum. PCR-RFLP method can differentiate both biovars and assist in studies of the clinical diagnosis, epidemiology and pathology of this species in human. The aim of this study was to molecular detection of U. urealyticuminin prostate tissue samples based on PCR- RFLP.Methods: Two hundred prostate tissue samples were collected from patient suffering from prostatitis. The PCR assay was used to amplify a 559 bp fragment of 16S-23SRNA interspace region of Ureaplasma. After sequencing, PCR products from positive samples were digested withTaqI restriction enzyme.Results: Seven cases (3.5%) out of 200 prostate tissue samples were positive for U. urealyticum. Results of PCR products sequencing demonstrated that all isolates were U. parvumbiovar. PCR-RFLP results shown that there was not any differentiation in pattern of enzymatic digestion, in addition, all isolates were U. parvum, serovar 3.Conclusion: U. urealyticum can be one of the causing agents of prostatitis. Using PCR-RFLP with specific primer and restriction enzyme is a rapid and cost-effect method for detection and differentiation of Ureaplasma from clinical samples.

Background: Rapid diagnosis and differentiation of Brucella is of high importance due to the side effects of antibiotics for the treatment of brucellosis. This study aimed to identify and compare PCR-ELISA as a more accurate diagnositc test with other common molecular and serological tests.Methods: In this experimental and sectional study, during March 2014 to Sep 2015, 52 blood samples of suspected patients with clinical symptoms of brucellosis were evaluated in medical centers all over Iran with serum titers higher than 1: 80. Using two pairs of specific primers ofBrucella abortus, B. melitensis and DIG-dUTP, Fragment IS711 (The common gene fragment in B. melitensis and B. abortus) was amplified.DIG-ELISA was performed using specific probes of these 2 species of Brucella and patterns were subsequently analyzed, then positive responses were compared by detecting gel electrophoresis.Results: PCR-ELISA method detected all 28 samples from 52 positive samples. Its sensitivity was 6.0 pg concentration of genomic DNA of Brucella. In gel electrophoresis method, 22 samples of all positive samples were detected. PCR-ELISA was more efficient than PCR and bacterial culture method atP -value<0.05.Conclusion: PCR-ELISA molecular method is more sensitive than other molecular methods, lack of mutagenic color and also a semi-quantitative ability. This method is more effective and more accurate compared to PCR, serology and culture of bacteria.PCR-ELISA does not have false responses. The limitation of this method is detection of bacteria in the genus compared to Multiplex PCR and Gel Electrophoresis.

Background: Quality assurance in the hematology laboratory is a must to ensure laboratory users of reliable test results with high degree of precision and accuracy.Even after so many advances in hematology laboratory practice, pre-analytical errors remain a challenge for practicing pathologists. This study was undertaken with an objective to evaluate the types and frequency of preanalytical errors in hematology laboratory of our center.Methods: All the samples received in the Hematology Laboratory of Dayanand Medical College and Hospital, Ludhiana, India over a period of one year (July 2013-July 2014) were included in the study and preanalytical variables like clotted samples, quantity not sufficient, wrong sample, without label, wrong label were studied.Results: Of 4, 71, 006 samples received in the laboratory, preanalytical errors, as per the above mentioned categories was found in 1802 samples. The most common error was clotted samples (1332 samples, 0.28% of the total samples) followed by quantity not sufficient (328 sample, 0.06%), wrong sample (96 samples, 0.02%), without label (24 samples, 0.005%) and wrong label (22 samples, 0.005%) Conclusion: Preanalytical errors are frequent in laboratories and can be corrected by regular analysis of the variables involved. Rectification can be done by regular education of the staff.

Background: Overexpression of Rad51 protein in many tumor cells has been proven to increase radioresistance and can be related to the resistance of chemosensitivity of tumor cells. This preliminary study was conducted to determine the relationship between the Rad51 expression level in nasopharyngeal carcinoma and the response of the treatment based on the measurement of the tumor reduction.Methods: Thirteen cases of the NPCs were analyzed. The expression levels of the Rad51 were examined from the pretreatment biopsies. Furthermore, tumor reductions were determined based on the change in sum longest diameter of the nasopharyngeal CT-scan before and after therapy.Results: The expression level of the Rad51 was associated with the reduction of tumor mass. The P value was 0.049 and the correlation coefficient was 0.479.Conclusion: The tumor cells Rad51 expression levels may affect the tumor reduction of NPC after the therapy.

Chondrosarcoma is a mesenchymal tumor composed of tumor cells producing cartilage. It is more common in older age and often affects the axial skeleton. We report a rare case of chondrosarcoma mimicking a sellar and suprasellar mass with parasellar extension. A 40 yr woman presented with decreasing visual acuity and headache. Magnetic resonance (MR) image revealed a cystic sellar and suprasellar mass with parasellar extension showing mild enhancing solid component. It favored the diagnosis of craniopharyngioma. The patient underwent trans-sphenoidal partial resection of the tumor resulting in removal of the sellar mass. However, the suprasellar mass could not be excised completely due to limited surgical field. The pathologic diagnosis was chondrosarcoma. Eight months after the operation, pterional approach was performed to remove the remaining mass. Intraoperative findings confirmed that the mass originated from dorsum sellae.

Malignant papillary neoplasms of the breast comprise a number of microscopically distinct lesions, where apocrine metaplasia is commonly found in papillomas compared to other papillary lesions including papillary carcinomas. However, association of apocrine metaplasia in papilloma with Ductal Carcinoma in Situ (DCIS) is not very well defined. The lesions with apocrine metaplasia are not only difficult to categories, but also there is controversy regarding their relative risk of subsequent carcinoma development. A case of extensive apocrine differentiation in duct papilloma with DCIS developing in the background of papillomatosis, posing a diagnostic dilemma for the pathologist and a therapeutic challenge for the surgeon, is hereby reported for its uniqueness and rarity. Awareness of this association should be kept in mind by both the pathologist as well as clinician for optimal therapeutic intervention.

Granulomatous response in association with breast cancer and within the cancer draining lymph nodes is an extremely rare phenomenon. Granulomatous inflammation is an immune response commonly seen against infectious agents and certain non-neoplastic conditions. The etiopathogenesis of granulomas associated with malignancies is not clear but it may be because of an immunologic reaction to tumour antigens. We hereby report a 50-yr-old postmenopausal female presented to Surgical Outpatient Department, Aligarh Muslim University, India, with complaints of lump and pain in her left breast for 6 months. We have also discussed about its etiopathogenesis, final diagnosis, treatment & patient outcome.

Neurilemmomas are benign tumors of peripheral nerve sheath Schwann cells. One of the variants of neurilemmoma is the ancient type of neurilemmoma characterized by degenerative features such as cystic degeneration, calcification, hemorrhage and hyalinization which could be easily misdiagnosed. Their occurrence in oral cavity is extremely rare and intraosseous type occurring in maxilla is exceedingly rare with very few cases being published in literature. A 38 year old male patient reported with a chief complaint of swelling over the left cheek and left upper back region since 10 months. The case is of recurrent intraosseous ancient neurilemmoma in the maxilla in the patient which is distinctive for the lesion. This unique case presented with distinct histologic architectural pattern of ancient neurilemmoma showing degenerative changes such as cystic degeneration and recurred within a short duration of time.