Urinalysis: The Automated Versus Manual Techniques; Is It Time To Change?.

BACKGROUND: Urinalysis is the third major test in clinical laboratory. Manual technique imprecision urges the need for a rapid reliable automated test. We evaluated the H800-FUSIOO automatic urine sediment analyzer and compared it to the manual urinalysis technique to determine if it may be a competitive substitute in laboratories of central hospitals. METHODS: 1000 urine samples were examined by the two methods in parallel. Agreement, precision, carryover, drift, sensitivity, specificity, and practicability criteria were tested. RESULTS: Agreement ranged from excellent to good for all urine semi-quantitative components (K > 0.4, p = 0.000), except for granular casts (K = 0.317, p = 0.000). Specific gravity results correlated well between the two methods (r = 0.884, p = 0.000). RBCS and WBCs showed moderate correlation (r = 0.42, p = 0.000) and (r = 0.44, p = 0.000), respectively. The auto-analyzer's within-run precision was > 75% for all semi-quantitative components except for proteins (50% precision). This finding in addition to the granular casts poor agreement indicate the necessity of operator interference at the critical cutoff values. As regards quantitative contents, RBCs showed a mean of 69.8 +/- 3.95, C.V. = 5.7, WBCs showed a mean of 38.9 +/- 1.9, C.V. = 4.9). Specific gravity, pH, microalbumin, and creatinine also showed good precision results with C.Vs of 0.000, 2.6, 9.1, and 0.00 respectively. In the between run precision, positive control showed good precision (C.V. = 2.9), while negative control's C.V. was strikingly high (C.V. = 127). Carryover and drift studies were satisfactory. Manual examination of inter-observer results showed major discrepancies (< 60% similar readings), while intra-observer's results correlated well with each other (r = 0.99, p = 0.000). CONCLUSIONS: Automation of urinalysis decreases observer-associated variation and offers prompt competitive results when standardized for screening away from the borderline cutoffs.

Association of MicroRNA 196a and 499 Polymorphisms with Development of Cirrhosis and Hepatocellular Carcinoma Post-HCV Infection in Egyptian Patients.

Hepatocellular carcinoma (HCC) is the commonest primary tumor of the liver. Chronic HCV infection is the leading cause of end-stage liver disease, HCC and liver-related death in Egypt. Single nucleotide polymorphisms (SNPs) in microRNAs were reported to increase susceptibility to tumorigenesis; affect prognosis and as promising biomarkers in virus-host interactions. This study was conducted to investigate the role of genetic variants of miR-196a2 (rs 11614913) C>T and miR-499 (rs 3746444) A>G in the development of cirrhosis and HCC in Egyptian HCV infected patients. Genotyping of the candidate SNPs was performed by Real Time PCR in 75 HCV-related HCC patients, 75 cirrhotic patients on top of HCV and 75 healthy controls. There was significant difference in miR-499 (rs3746444) genotypes frequency between the three studied groups as the GG genotype was significantly lower in HCC cases than other groups (P = 0.009) while the combined miR-499 (AA+AG) genotypes were significantly higher in HCC cases than other groups (P = 0.005). Also a significant difference was found in miR-499 genotypes frequency when compared between HCC and cirrhosis groups as the GG genotype was significantly lower in HCC cases than cirrhosis group (P = 0.006) while the combined miR-499 (AA+AG) genotypes were significantly higher in HCC cases than in cirrhosis group (P = 0.003) [OR (95% CI) = 0.131 (0.028-0.601)]. The frequency of the G allele was significantly lower in HCC than other groups (P = 0.024) and significantly lower in HCC than normal group (P = 0.006) [OR (95%CI) = 0.501 (0.304-0.825)]. For miR-196a2 (rs11614913) C>T polymorphisms, no significant association was found with HCC risk. Our study concluded that the G allele of miR-499 is associated with lower risk of HCV related HCC development. No significant association of miR-196a2 (rs 11614913), genotypes or alleles with risk for HCC development, could be detected.
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RECK Gene Promoter rs10814325 Polymorphism in Egyptian Patients with Hepatocellular Carcinoma on Top of Chronic Hepatitis C Viral Infection.

BACKGROUND: The reversion-inducing-cysteine-rich protein with Kazal motifs (RECK) gene is a novel transformation suppressor gene that was linked to several malignancies. OBJECTIVE: To analyze any association between RECK gene rs10814325 single-nucleotide polymorphism (SNP) and HCC susceptibility along with it is association wiht various clinico-pathological and laboratory data. MATERIALS AND METHODS: RECK gene rs10814325 SNP was estimated, using real-time PCR technique, in 30 HCC patients on top of chronic HCV infection, 30 HCV related cirrhotic patients and 30 healthy controls. RESULTS: No special pattern of association could be detected on comparing the RECK gene rs10814325 genotypes(P=0.5), or alleles(P=0.49) among the studied groups. HCC patients with TT genotype had younger age (mean of 54.1±6.0 years vs 60.6±10.2 years for TC/CC genotypes, P=0.035). Abdominal distension was significantly greater in TT genotype patients (75% vs 30% of TC/CC genotypes, P=0.045). TT genotype was present in 75% of patients with lymph node metastasis. Serum GGT levels were higher in TT genotype patients [80 of (48.5-134.8) vs 40 IU/l (33-87.5) for TC/CCgenotypes], and lower limb edema was observed in 60% of TT vs 20% of TC/CCgenotypes, however, both just failed to reach significance (P=0.05 and P=0.06, respectively). CONCLUSIONS: RECK gene rs10814325 T>C could not be considered a risk factor for HCC development on top of HCV, but may be related to the disease progression and metastasis.

Value of optical coherence tomography in the detection of macular pathology before the removal of silicone oil.

PURPOSE: To assess the pathological macular changes with optical coherence tomography (OCT) before the removal of silicone oil (SiO) in eyes that had undergone pars plana vitrectomy for complicated forms of retinal detachment (RD). PATIENTS AND METHODS: Subjects included 48 patients (51 eyes) with complicated RD including proliferative vitreoretinopathy, proliferative diabetic retinopathy, recurrent RD, penetrating trauma, uveitis, giant retinal tears, and macular holes. All the eyes had undergone SiO injection. Furthermore, all eyes had been planned for the removal of SiO 6-12 months after the primary surgery. Finally, all eyes had a fundus examination and OCT examination before the silicone oil removal. RESULTS: OCT findings indicated epiretinal membrane in 41% of the eyes, macular edema in 17%, macular detachment in 13.5%, macular thinning in 13.5%, macular holes in 10%, and subretinal membranes in 2%. Preoperative OCT was normal in only 12% of the eyes, while a clinical fundus examination was normal in 43% (P<0.001). Eyes with normal OCT had significantly better mean logMAR (0.35) than eyes with pathological changes detected through OCT (1.28; P<0.001). Surgical modifications were made during the removal of SiO in 74.5% of the eyes. CONCLUSION: OCT detected significantly more pathological changes than a clinical fundus examination. This had an impact on both surgical step modification during the removal of SiO and predictability of visual outcome after the removal of SiO.

The role of PON1 and CYP2D6 genes in susceptibility to organophosphorus chronic intoxication in Egyptian patients.

BACKGROUND: Paraoxonase-1 (PON1) activity toward organophosphorus(OP) compounds shows inter-individual variations, rendering the identification of individuals' PON1 allozymes valuable in treating patients suffering from organophosphorus intoxication. One of the most important cytochrome P450 monooxygenases (CYPs) is CYP2D6. The CYP2D6 G1934A polymorphism leads to good, poor or no enzyme activity. Genetic testing helps identification of high risk individuals as well as management of chronic intoxicated patients. OBJECTIVE: to investigate a possible association between genetic polymorphisms of PON1 Q192R, and CYP2D6 G1934A as well as PON1 and pseudo-cholinesterase (PChE) enzyme activity levels and chronic organophosphate exposed patients, and hence, susceptibility for organophosphorus chronic poisoning. DESIGN AND METHODS: Thirty chronic organophosphate exposed farm workers were compared to 29 healthy controls as regards PON1 Q192R and CYP2D6 G1934A polymorphisms using PCR-RFLP technique. Also serum PON1 and PChE activities were determined spectrophotometrically. RESULTS: Serum PChE was significantly reduced in chronic intoxicated patients compared to the control group (p=0.02), while PON1 activity was increased, but just failed to reach significance (p=0.06). PON1 192 RR genotype and R allele were significantly increased in chronic OP intoxicated patients (p=0.005 &p=0.002 respectively). CYP2D6 1934A allele was significantly increased in chronic OP patients (p=0.045). combining the two SNPs showed a significant statistical difference between the two groups with PON1QQ and CYP2D6 GG genotypes being more represented in the healthy controls (p=0.001). Fatigue and motor weakness were the most prevalent neurological symptoms seen in chronic cases (56.7%), followed by headache and lacrimation (30% each), depression (23%), tingling and sensory symptoms (20%), sleep disorders and limb pain (13%). The mean duration of environmental exposure to organophosphates was 7.7±5.2years and no association was found between chronic symptoms of intoxication and duration of exposure, provided that all workers were exposed for at least 3 years. CONCLUSION: PON1 192RR genotype and CYP2D6 1934A allele were found to be related to the susceptibility to organophosphate chronic toxicity in Egyptians. Larger scale gene-environmental interaction studies are recommended to confirm results and Genotyping is recommended during selection of agricultural pesticide workers to exclude high risk group.

Paraoxonase 1 and cytochrome P450 polymorphisms in susceptibility to acute organophosphorus poisoning in Egyptians.

BACKGROUND: Organophosphates are the basis of many insecticides, herbicides, and nerve agents. They were listed as highly acutely toxic agents. Findings in knockout mice suggest that paraoxonase 1 may modulate the toxicity resulting from exposure to organophosphorus compounds. In human, there is no enough data about genetic modulation of acute organophosphorus intoxication. CYP2D6 is involved in the metabolism of about 30% of xenobiotics. Prompt accurate management of OP acute intoxication can promote patient's survival. DESIGN AND METHODS: Forty acute organophosphorus intoxicated patients were divided according to presence of clinical toxicity manifestations and serum level of pseudo-cholinesterase into two groups of acute symptomatic and acute asymptomatic patients. A third group of 29 healthy volunteers served as control. Paraoxonase 1 Q192R and CYP2D6 G1934A polymorphisms, (QQ, QR, and RR for PON1) and (GG, GA, and AA for CYP2D6), were studied using polymerase chain reaction-restriction fragment length polymorphism technique. Serum paraoxonase 1 and pseudo-cholinesterase activities were measured spectrophotometrically. RESULTS: Serum pseudo-cholinesterase was significantly reduced in both acute intoxication groups compared to the controls (p=0.000). Paraoxonase 1 was significantly reduced in the symptomatic acute intoxication patients in comparison to the asymptomatic group (p=0.002). There was a significant increase in paraoxonase 1 192 RR genotype and R allele in the symptomatic patients in comparison to the controls and asymptomatic patients (p=0.006 and p=0.01, respectively). For CYP2D6 G1934A genotypes and alleles, no significant difference was found between groups (p=0.3 and p=0.18, respectively). However, one case of the two recorded fatalities was for a symptomatic female patient with the only traced AA genotype. The combination of both single nucleotide polymorphisms revealed a significant distribution difference between groups, with QQ+GG genotypes being more represented in the controls, while RR+GA genotypes were exclusively present in the group of symptomatic patients (p=0.04), none of the participants was found to have RR+AA genotypes. Some nicotinic (fasciculation and weakness), and muscarinic symptoms (bronchospasm, salivation, lacrimation, and diarrhea), increased with high significance in the symptomatic group compared to the asymptomatic one (p<0.001 for all). Convulsions also showed significant increase (p=0.02). CONCLUSION: Paraoxonase 1 Q192R modulates patient's response, and CYP2D6 may be related to the acute organophosphorus intoxication in the context of other genetic-environmental factors. Paraoxonase 1 enzyme level is related to symptom severity in acute OP poisoning, while pseudo-cholinesterase level indicates exposure to OP rather than severity of clinical manifestations.

Soluble TWEAK and cardiovascular morbidity and mortality in chronic kidney disease patients.

INTRODUCTION: Cardiovascular disease (CVD) is a major cause of morbidity and mortality in chronic kidney patients (CKD). The aim of this study was to demonstrate the role of soluble tumor necrosis factor (TNF) weak inducer of apoptosis (sTWEAK) as a marker of cardiovascular morbidity and mortality in CKD patients. METHODS: The study included 75 CKD patients classified according to eGFR into three groups; group-1 included 15 patients with stage-1 CKD, group-2 included 30 patients with stage-2 and stage-3 CKD, and group-3 included 30 patients with stage-4 and stage-5 CKD. The three groups were compared to 20 matched controls. Interleukin-6 (IL-6) and sTWEAK were measured using ELISA and chemiluminescent techniques respectively. Carotid intima-media thickness (IMT) was also measured. RESULTS: We found that IL-6 showed significant difference between patient groups and controls, being highest in stage 4 and 5 CKD patients and lowest in controls. Soluble TWEAK showed significant difference between patient groups and controls, being lowest in stage 4 and 5 CKD patients and highest in controls. Soluble TWEAK level showed significant negative correlation with IL-6 (r = -0.68; P < 0.01) and carotid IMT (r = -0.95; P < 0.01). After two years follow up, nine out of 75 CKD patients developed ischemic heart disease (IHD). Two patients developed cerebrovascular stroke and another patient developed peripheral arterial disease. These patients had significantly lower levels of sTWEAK at baseline compared to other patients (160.5 ± 60.2 versus 274.8 ± 90 pg/mL; P < 0.05). CONCLUSION: Soluble TWEAK can be a novel biomarker of atherosclerosis and endothelial dysfunction as well as cardiovascular outcome in CKD patients.