Clinical Pathology

Introduction: Endometriosis affects up to 1 every 5 women at their reproductive age, with variable and complex symptomatology. Patients may be asymptomatic but may have pain episodes or subfertility. Its negative impact is on patients' health and quality of life. Objective: it was to investigate the serum and peritoneal fluid (PF) concentrations of Angiopoietin-2, Interleukin-1β, and Vascular Endothelial Growth Factor, aiming to evaluate their diagnostic performance in endometriosis. Methods: Serum and peritoneal fluid samples were taken from 112 women undergoing laparoscopy for infertility, pelvic pain or adnexal masses. 61 diagnosed with endometriosis and 51 controlled. Primary outcome was to estimate serum and PF concentrations of Angio-2, IL-1β and VEGF and secondarily correlate these concentrations to disease stages thus assuming their diagnostic potential. Results: Significant differences were found between patients and control as regards serum and PF concentration of all studied markers except serum IL-1β. Serum Angio-2 and PF VEGF showed a significantly higher level in more advanced stages of endometriosis. PF VEGF showed a positively significant correlation with the stage of the disease, spearman coefficient t = 0.442 p = 0.014. PF concentrations of Angio-2 and Serum VEGF did not show significant pattern changes with stage-related levels. Diagnostic potential of serum and PF concentrations of the 3 markers were assessed by the ROC curve. Angio-2 proved an excellent diagnostic ability for endometriosis. PF and serum VEGF proved an equal diagnostic performance, whereas, PF IL-1β was the least efficient. Based on the results, we suggested preliminary serum threshold values for these markers to be used as diagnostic or follow-up landmarks with relatively acceptable sensitivity, specificity, positive and negative predictive values. Conclusion: Non-invasive predictive biomarkers for endometriosis were Serum Angio-2, IL-1β, and VEGF independently or in combination with the estimated threshold values. Serum Angio-2 merit is considered as a novel marker for endometriosis due to its diagnostic power.

Hepatocellular carcinoma is one of the most prevalent life-threatening human cancers with etiological factors chronic viral hepatitis B and C infections. Tumor cell dispersion relies on the loss of homotypic cell-cell adhesion. Invasion through basement membrane and interstitial extracellular matrix is another key event for metastatic progression, which requires the action of a series of proteolytic enzymes named matrix metalloproteinases, secreted by tumor cells that enhance tumor invasiveness and metastasis. TIMPs are dominant inhibitors of MMPs and able to control MMP-mediated ECM breakdown by binding active forms of MMPs. Lipocalin-2 is known as neutrophil gelatinase associated lipocalin promotematrix degradation and tumor progression. Aim: To evaluate the importance of lipocalin for diagnosis HCC in Egyptian chronic liver disease patients. Subjects and Methods: 50 patients and 25 controls. (G-1) 25 HCC on top of hepatitis C. (G-2) 25 hepatitis C. The following done: schistosoma antibodies, ASAM, LKM-1, ANA AKA and CBC. Hepatitis B surface antigen, Hepatitis C antibodies AFP, Cupper and zinc, Matrix metaloprotinase-9, TIMP-1 and Neutrophil gelatinase-associated lipocalin. Results: Median value of MMP-9 level in G-I (206 µg/L) is significantly higher than G-2 (100 µg/L) and G 3 (49 µg/L) p < 0.001. TIMP-9 median value G-1 (48 µg/L) is significantly lower than G-2 (54 µg/L) and G-3 (113 µg/L) p < 0.001. Lipocalin-2 median levels are significantly higher in G-I (389 ng/mL) than G-2 (166 ng/ml) versus G-3 (60 ng/mL) p < 0.001. Lipocalin-2 associated with increasing lobular inflammation, ballooning & fibrosis with MMP-9 has an important role in pathogenesis of liver cirrhosis and HCC. Conclusion: We elucidated predictive value for MMPs, TIMPs, and progression metastasis of hepatocellular carcinoma. Liver cell impairment alters metabolism of trace metals as zinc and copper, with possible relationship of these changes to pathogenesis of chronic liver disease. Lipocalin-2 can be used as a future diagnostic marker with better sensitivity and specificity than MMP-9 for the progression of hepatocelluar carcinoma.

Analytical quality is an essential requirement for best practice in any medical laboratory. Lack of a harmonized approach for sigma calculation is considered an obstacle in the objective comparability of analytical performance among laboratories adopting sigma metrics. It is urgently needed that all laboratory professionals interested in the analytical quality to work hard towards harmonization protocol for sigma calculation in order to properly select their analytical goals. This study aims at harmonization of Sigma metrics calculation in four accredited Egyptian laboratories. This observational cross sectional study compared the sigma levels for certain biochemical parameters in the four participating laboratories. Coefficient of variation (CV) and bias were determined for some biochemical analytes, data assayed by different automated analysers in the four different accredited laboratories. The sigma level for the four medical laboratories was calculated for each biomedical parame...

Background: Tuberculosis (TB), is one of the major common air born infectious bacterial diseases which remains a major worldwide health problem with global mortality. Objective: To evaluate the efficiency of serum samples compared to sputum for the early diagnosis of TB, and to evaluate the levels of superoxide dismutase (SOD), catalase (CAT), total antioxidant status (TAS) and tumor necrosis factor- (TNF-) in patients with pulmonary tuberculosis (PTB). Methods: One hundred patients with clinically suspected PTB and 25 healthy individuals were enrolled in the study. According to the bacteriological results, 78 patients were diagnosed as having PTB infection. These cases were categorized into 69 culture positive cases [sputum and serum PCR positive patients (n=42), sputum PCR positive and serum PCR negative patients (n=16) and sputum and serum PCR negative patients (n=11)] and 9 culture negative and sputum PCR positive cases with radiological lung abnormalities suggestive for PTB. For these 78 cases, erythrocyte SOD, CAT, serum TAS and TNF- were determined. Twenty two patients were culture negative and negative for both sputum and serum PCR. They had no PTB and were not involved in biochemical studies. Results: In all 78 PTB patients, erythrocyte SOD, CAT and serum TAS levels were statistically lower than controls (p<0.05), while TNF- was highly significantly increased (p=0.001). There was a significant direct linear correlation between SOD and CAT and TAS (p<0.0001, r= 0.78; p<0.0001, r= 0.88; p<0.0001, r=0.80 respectively) and a significant reverse linear correlation between TNF- and SOD, CAT and TAS level (p<0.0001, r=-0.55; p<0.0001, r=-0.51; p<0.0001, r=-0.65 respectively). Conclusion: Although the sputum culture is still the gold standard for the diagnosis of patients with PTB, sputum PCR is an efficient method that could be used as an alternative to the culture for the rapid identification of PTB cases. The lower levels of SOD, CAT and TAS may be improved by the antioxidant therapy which may help in better prognosis. Anti TNF-α therapy may help in decreasing the elevated level of TNF-α shown in all PTB patients.

Objectives: Diabetic retinopathy (DR) is one of the most common microvascular complications of type 2 diabetes mellitus (T2DM). It is multifactorial with the contribution of multiple genetic factors. We questioned the association of polymorphisms in the peroxisome proliferator-activated receptor ɣ2 (PPARɣ2) gene (Pro12Ala and C161T) with DR in an Egyptian population. Methods: This case control study included one hundred healthy individuals and 252 T2DM among them 122 with DR and 130 without DR. Genotyping was done by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP). Results: The Pro12Ala Ala allele was associated with decreased risk of DR with an odds ratio (OR) of 0.484, 95% confidence interval (CI) (0.254-0.920), and a p value = 0.024. The C161T T allele was associated with increased risk of DR with OR = 2.593, 95% CI (1.672-4.020), p < 0.001. However, when considering other covariates such as glycosylated hemoglobin (HbA1c) in multivariate regression analysis only C161T was associated with increased risk of DR with OR = 3.479, 95% CI (1.907-6.346), p < 0.001, while the significant association with Pro12Ala was lost. HbA 1c was higher in Pro/Pro genotype when compared to those with

Background Hepatitis C virus infection is a global health challenge with Egypt being one of the highly affected countries. IL-10 has been suggested as a suitable marker to assess necroinflammation and to monitor the progression of liver damage. Vascular endothelial growth factor (VEGF) is a potent angiogenic factor playing a central role in many physiological as well as pathological processes. Several factors can be predictive of the response to treatment and achievement of SVR; some of which are host-related, and others are virus-related. The gene expression of IL-10 and VEGF have multiple effects for treatment response. The aim of the present work was to study the effect of treatment with directly acting agents (DAA) on the expression of VEGF and IL-10 genes in chronic hepatitis C virus-infected Egyptian genotype-4a patients. Twenty-five HCV subjects where evaluated for IL-10 and VEGF gene expression before and after treatment with DAA. Results IL-10 expression was downregulated i...

Objective: Health-care waste has not attended much attention in developing countries. Staff is involved in cleaning and collect waste may often be at greater risk due to their less education and training. The current intervention study was conducted to improve knowledge and practice skills for medical waste handlers in some selected hospitals of Alexandria, Egypt. Methods: An intervention study was conducted on medical waste handlers from some selected hospitals who accepted to participate in the study in the period of May 2015 to June 2016. A predesigned questionnaire was developed to measure knowledge, skills, and practice on medical waste management pre- and post-intervention. Results: Analysis of pre-intervention data revealed that 9.6%, 80.8%, and 9.6% of participants had high, moderate, and low knowledge levels, respectively. Whereas post-intervention, data revealed that 97.3%, 2.2%, and 0.5% of workers had high, moderate, and low knowledge levels, respectively. A significant ...

Point of Care Testing is increasingly being used for diagnosis and management of various disease states. Management of different Point of Care instruments at multiple sites can be challenging, particularly when such instruments are operated by non technical staff. Connectivity is critical for optimal management of these services which are intimately linked to operator training and competency and are important in minimising harm to the patient by reducing analytical errors. Furthermore, connectivity improves turn around time leading to faster decision making by physicians. Recent advances in technology such as 5G and artificial intelligence are likely to lead to a greater focus on personalized care as a result of big data analysis and development of algorithms.

With the objective of identifying candidate tumor suppressor genes, we used fluorescence in situ hybridization to map leukemia-related deletions of the long arm of chromosome 6 (6q). Twenty of 24 deletions overlapped to define a 4.8-Mb region of minimal deletion between markers D6S1510 and D6S1692 within chromosome 6 band q16. Using reverse transcription-PCR, we found evidence of expression in hematopoietic cells for 3 of 15 genes in the region (GRIK2, C6orf111, and CCNC). Comparison between our own and published deletion data singled out GRIK2 as the gene most frequently affected by deletions of 6q in acute lymphocytic leukemia (ALL). Sequence analysis of GRIK2 in 14 ALL cases carrying heterozygous 6q deletions revealed a constitutional and paternally inherited C to G substitution in exon 6 encoding for an amino acid change in one patient. The substitution was absent among 232 normal alleles tested, leaving open the possibility that heterozygous carriers of such mutations may be susceptible to ALL. Although low in all normal hematopoietic tissues, quantitative reverse transcription-PCR showed higher baseline GRIK2 expression in thymus and T cells than other lineages. Among T-cell ALL patients, 6q deletion was associated with a statistically significant reduction in GRIK2 expression (P ‫؍‬ 0.0001). By contrast, elevated GRIK2 expression was measured in the myelomonocytic line THP-1 and in one patient with common ALL. Finally, we detected significant levels of GRIK2 expression in prostate, kidney, trachea, and lung, raising the possibility that this gene may be protective against multiple tumor types.

Expression of BCR/ABL, a constitutively active tyrosine kinase, is a primary event in the pathogenesis of chronic myeloid leukemia (CML) and Ph-positive acute lymphoblastic leukemia (Ph+ALL). Inhibition of the BCR/ABL kinase activity in the BV173 CML cell line with STI571 resulted in a significant overexpression of a 10-kb novel mRNA, found to be the human ortholog of the murine Bach2, a B-cell-specific transcription factor. The human BACH2 cDNA is >9,120 bp long and includes an open reading frame of 2,526 bp encoding a protein with a basic leucine zipper (bZip) and a BTB/POZ domain, mediating DNA-binding and heterodimerization. BACH2 was consistently upregulated (2–10-fold) in all 10 Ph+ lymphoid lines tested following BCR/ABL inhibition. In CML myeloid cell lines (n = 8) and BCR/ABL-negative lines (n = 6), BACH2 was either undetectable by Northern blotting or did not change in response to STI571, suggesting that BACH2 repression by BCR/ABL may be specifically relevant to lymphoid transformation. Quantitative RT/PCR revealed a significantly lower level of BACH2 expression in leukocytes from patients with CML (n = 24) as compared to normal individuals (n = 23) (P < 0.0005). Moreover, CD34+ cells treated in vitro with STI571 exhibited a consistent upregulation of BACH2 in 8 of 10 CMLs but in none of the 9 normal individuals tested. Transcription regulation of BACH2 in BCR/ABL-positive cells was exerted via the MEK pathways, as shown by their responses to the U0126-specific inhibitor. Radiation hybrid mapping and FISH revealed that BACH2 is located on chromosome 6, band q15, a region frequently associated with deletions in ALL and non-Hodgkin's lymphoma, suggesting its possible role as a tumor suppressor gene. However, no rearrangement or loss of signal was observed by Southern blotting in 34 lymphomas, 10 B-cell ALLs, or seven reactive lymph nodes. The pattern of BACH2 expression in BCR/ABL-positive cells suggests that transcriptional repression by this regulator is impaired in CML and may contribute to the emergence of lymphoid blast crisis. © 2001 Wiley-Liss, Inc.

Chromosome 8 abnormalities are seen in 80% of patients with T-cell prolymphocytic leukemia (T-PLL). The abnormalities described are idic(8)(p11),t(8;8)(p11;q12), ϩ 8, and 8p ϩ with the involvement of 8p. To localize 8p11-p12 breakpoints in T-PLL, metaphases from seven cases were karyotyped. Those with idic(8)(p11) and add were probed with a panel of contiguous YACs derived from 8p11-p12 using fluorescence in situ hybridization (FISH). Analysis of FISH results showed that 8p11-p12 breakpoints cluster into two regions. The first region is telomeric to YAC 899e2, which contains the fibroblast growth factor receptor-1 gene ( FGFR1 ) and appears to cluster within a 1.5-MB YAC 807a2. The second region is more centromeric with breakpoints on either side of YAC 806e9, flanked by YAC 940f10 distally and YAC 910d7 proximally, the latter containing the MOZ gene. These findings showed that a segment of 8p was still present in the isodicentric, but the pattern of clustering does not seem to correspond to a breakpoint affecting a single gene. The clustering regions are likely to be hot spots for recombination and result in idic(8)(p11) and 8p ϩ . These changes point to the pathogenesis of T-PLL involving deletion of a gene sequence on 8p and/or gain of a copy of 8q.

Chronic lymphocytic leukemia (CLL) follows an extremely variable clinical course with overall survival times ranging from months to decades. The clinical staging systems do not allow one to predict if and at what rate there will be disease progression in an individual patient diagnosed with early stage disease. There has been intensive work on clinical and biological factors of potential prognostic relevance that may add to the classic assessment provided by the staging systems. Among these are: Laboratory parameters reflecting the tumor burden or disease activity such as lymphocyte count, lactate dehydrogenase (LDH) elevation, bone marrow infiltration pattern or lymphocyte doubling time (LDT), serum markers such as soluble CD23, β2microglobulin (β2-MG) or thymidine kinase (TK), and genetic markers of tumor cells such as genomic aberrations, the mutation status of the variable segments of immunoglobulin heavy chain genes (VH), or surrogate markers for these factors (CD38, ZAP-70, LPL). Thirty patients were included in our study, the investigation included CD38 expression, ZAP-70 expression, and interphase Fluorescence In Situ Hybridization (FISH) for the detection of trisomy 12, del 13q14.3, del 17p13, and del 11q22.3. Our results showed positive statistical significant correlation between ZAP-70 expression and CD38 expression with some of the chromosomal aberrations encompassing bad prognosis as ATM and p53. Also CD38 expression was positively correlated with trisomy 12 and p53 deletions. Chromosomal aberrations were found to be present in 76.6% of our patients with 13q deletion as the most frequent abnormality in our patients (46.7%), followed by trisomy 12 (36.7%), then ATM and p53 deletion (26.7%) each. Another interesting finding in our study is the fact that 100% of the ZAP-70 positive patients were of bad prognosis, 58.3% of the CD38 positive cases, 81.8% of the positive trisomy 12 cases, 100% of the ATM deletion, 62.5% of the p53 deletion, and 64.3% of the 13q-cases were also of bad prognosis, which indicates that ZAP-70, trisomy 12, and ATM deletion are powerful indicators of prognosis. We conclude that FISH for the detection of the most important chromosomal aberrations in CLL is an important laboratory parameter that is recommended for assessment and correlation with simultaneous evaluation of ZAP-70 and CD38 expression which could help in the prediction of outcome of CLL patients.

CXC ligand 12 (CXCL12) or stromal cell-derived factor1 (SDF-1), a chemokine abundantly produced by the bone marrow microenvironment, and its receptor CXC chemokine receptor 4(CXCR4) have crucial roles in malignant cell trafficking. In acute myeloid leukemia (AML), blasts invade the blood stream and may localize in extramedullar sites, with variations from one patient to another. It has been hypothesized that a polymorphism (CXCL12 G801A) could influence blast dissemination and tissue infiltration in AML. In the present study CXCL12 G801A polymorphism was determined by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) assay in 64 patients with de novo AML admitted to the hematology department of Alexandria Main University Hospitals as well as 50 healthy volunteers of matched age and sex as control group. The frequency of the CXCL12 genotypes among AML patients were 50 % had a GG alleles genotype while 34.

Aim: To investigate if there is an association between the different DNA repair gene polymorphisms and the risk of development of acute myeloid leukemia (AML) in a sample of the Egyptian population and to find out if there is any interaction between these polymorphisms and the NQO1 gene that acts to protect the cells from oxidative damage. Results: Our study was conducted on 90 patients with de novo AML and 60 healthy subjects with matched age and sex. We studied polymorphisms in three DNA repair genes; XRCC1, XRCC3, and XPD. We also assessed the incidence of the NQO1 gene polymorphism. We genotyped the polymorphisms by polymerase chain reaction-restriction fragment length polymorphism. Conclusion: The distribution of XRCC1Arg 339Gln genotypes showed a significant difference between patients and controls ( p = 0.025). The presence of at least one XRCC1 399Gln allele indicated an increased risk of AML and the proportion of AML patients homozygous for the Gln/Gln allele was significantly higher than in the control group ( p = 0.025). However, distributions of the XRCC3 Thr241Met, XPD Lys751Gln, and NQO1Pro 187Ser genotypes were not significantly different between patients and controls. Combined analysis of the studied DNA repair gene polymorphisms did not show an interaction with the detoxification NQO1 Pro187Ser polymorphism.

Prostate cancer is a common disease with a multifactorial and complex etiology. It is the most common male malignancy and the second leading cause of death in many countries. The widespread use of PSA testing has increased the detection of this cancer at earlier stages, although this diagnostic method has proved to be insufficient to identify the disease. DNA in most cells is regularly damaged by endogenous and exogenous mutagens. At least four main partially overlapping damage repair pathways operate in mammals. Common polymorphisms in DNA repair genes may alter protein function and an individual's capacity to repair damaged DNA; deficits in repair capacity may lead to genetic instability and carcinogenesis. In the present study, we investigated the genotypic distribution of XRCC1 and XPC polymorphisms and its association with prostate cancer risk, pathological staging and Gleason's scoring. The present study was conducted in the departments of Clinical Pathology, Pathology, and Urology Faculty of Medicine, Alexandria University-Egypt. A total number of 50 patients with pathologically confirmed prostate cancer and 50 age-matched control subjects were enrolled in this study. The diagnosis was made on the basis of histopathological findings, following radical prostatectomy or transurethral resection of the prostate (TURP). Genomic DNA was extracted from peripheral blood using QIAamp blood DNA isolation kits. PCR followed by enzymatic digestion of the PCR products for (XRCC1, XPC) was used for the genotyping of these polymorphisms. Statistical analyses were performed using SPSS statistics version 20. The genotype frequencies of the studied polymorphisms in all the samples (n = 100), PC patients (n = 50) and healthy controls (n = 50) were consistent with the Hardy-Weinberg equilibrium distribution (p-value > 0.05). There was no statistical difference in the genotypes of the XRCC1 Arg399Gln and XPC Lys939Gln between cases and controls. The "Gln" allele frequency of XPC Lys939Gln as well as the "Gln" allele frequency of XRCC1 Arg399Gln tended to be lower in controls than in PC patients. Yet, these decreases were not statistically significant. We also examined the combined effect of XPC and XRCC1 and we found a decreased PC risk when XPC 939 Lys/Lys + Lys/Gln and XRCC1 399 Arg/Arg + Arg/Gln are combined (OR = 0.370, 95% CI = 0.142-0.962).

Background: MicroRNAs (miRNAs) are short noncoding RNAs of ~21 to 23 nucleotides in length that post-transcriptionally regulate mRNA expression. Highthroughput methodologies have shown deregulated miRNA expression in an increasing number of human cancers. MiRNA expression patterns have been found to distinguish tumors of different developmental origin, even better than traditional mRNA expression profiling. Aim: To assess the plasma level of micro-RNA-92a in adult acute myeloid leukemia and to correlate it with prognostic factors and therapeutic response. Patients and Methods: This study was carried out on fifty AML patients as well as fifty healthy subjects as control. Conventional cytogenetics was performed on patients group only while measurement of the plasma level of miRNA-92a using TaqMan quantitative RT-PCR with miRNA-638 as endogenous reference for standardization and FLT3/ITD mutation was performed on patients and controls. Results: The differences in the ratio or relative quantitation (RQ) of plasma miRNa-92a to miRNA-638 in patients group to the control group have confirmed statistical significance. Also there was significant negative correlation between RQ of miRNA-92a and white blood count in patient group. Patients who achieved a response after induction chemotherapy had a mean RQ of miRNA-92a higher than non-responder with statistical significance. With regard to cytogenetics, favorable risk cytogenetics had meant RQ of miRNA-92a that was comparable to intermediate risk cytogenetics. While poor risk cytogenetics had a mean RQ which is significantly lower than both favorable and intermediate risk cytogenetics. Summary/Conclusions: Our data suggest the potential importance of the microRNA-92a as noninvasive cancer biomarkers helping in diagnosis, clinical prediction and therapeutic response.

Background: Circulating microRNAs are potential biomarkers of diagnostic and prognostic impact in various inflammatory and malignant diseases. Aim: Linking inflammation with malignancy, we studied miRNA-21 in sera of hepatitis-C-virus (HCV) and none hepatitis diffuse large B-cell lymphoma (DLBCL) patients, aiming to identify its differential expression and prognosis in DLBCL with its subtypes; germinal center B-cell (GCB) and activated B-cell-like (ABC) and to evaluate its relation with HCV. Subjects and Methods: MiRNA-21 expression was measured using Taq-Man quantitative RT-PCR in sera of 30 newly diagnosed DLBCL patients (HCV positive (n = 10), HCV negative (n = 20)) and 20 controls (HCV positive (n = 10), HCV negative (n = 10)). Results: MiRNA-21 expression was significantly higher in DLBCL patients than in control (p = 0.00). Significant positive correlations between miRNA-21 and LDH, IPI and disease stage were detected (p < 0.05). Significantly higher miRNA-21 was detected in ABC sub-type compared to GCB sub-type (p = 0.00). Higher miRNA-21 was associated with worse response (p = 0.016), 2 years overall (p = 0.017) and progression free survival with statistical significance (p= 0.003). Significantly higher miRNA-21 levels were detected in HCV positive DLBCL patients compared to HCV negative patients (p = 0.00). Higher miRNA-21 levels were detected in HCV positive ABC subtype than GCB subtype (p = 0.05). Significantly higher levels were also detected in HCV positive controls compared to HCV negative controls. Conclusion: Our study shows that miRNA-21 is over expressed in our patients with DLBCL, displaying higher levels in ABC than in GCB subtypes. MiR-NA-21 is associated with poor response to treatment and survival in DLBCL. MiR-NA-21 is a potential marker of necro-inflammation independent of its role in tumorogenesis, showing higher expression in HCV positive DLBCL patients compared to none hepatitis patients.