Pharmacogenomics of Methotrexate Pathway in Rheumatoid Arthritis Patients: Approach toward Personalized Medicine.

Background: Methotrexate (MTX) is one of the most common medications used for rheumatoid arthritis (RA) treatment. Single-nucleotide polymorphisms (SNPs) could potentially predict variability in therapeutic outcomes. Aim: This study aims to assess the impact of SNPs in genes encoding for the MTX pathway for predicting clinical and therapeutic responses to MTX in a cohort of Egyptian patients with RA. Subjects and Methods: Data from 107 Egyptian RA patients (aged 44.4 ± 11.4 years) treated with MTX monotherapy, for a duration of 3.7 ± 3.3 years, were collected. Genotypes of 10 SNPs from four different genes were analyzed using the allelic discrimination PCR technique. Results: The ATIC rs3821353 G/T (p = 0.034) and the C/T and C/C of SLC19A1 rs7279445 (p = 0.0018) were associated with a non-response to MTX, while DHFR rs10072026 C/T and C/C were associated with a good response (p < 0.001). Carriers of the ATIC rs382135 3 G (p = 0.001) and ATIC rs4673990 G (p < 0.001) alleles were more likely to develop RA, while the SLC19A1 rs11702425 T (p < 0.001) and GGH rs12681874 T (p = 0.003) allele carriers were more likely to be protected against RA. Carriers of the ATIC rs4673990 A/G genotype (p < 0.001) were at risk of developing RA, while carriers of the following genotypes were mostly protected against RA: ATIC rs3821353 T/T (p < 0.001), ATIC rs3821353 G/G (p = 0.004), SLC19A1 rs11702425 T/T (p = 0.001), SLC19A1 rs11702425 C/T (p = 0.003), GGH rs12681874 C/T (p = 0.004) and GGH rs12681874 T/T (0.002). Conclusion: The genotyping of genes involved in the MTX pathway may be helpful to predict which RA patients will/will not benefit from MTX, and thus, may help to apply a personalized medicine approach in RA.

The association of CD40 polymorphism (rs1883832C/T) and soluble CD40 with the risk of systemic lupus erythematosus among Egyptian patients.

BACKGROUND: Systemic lupus erythematosus (SLE) is a complex autoimmune disorder of unknown etiology. Considerable evidence supports a genetic basis for susceptibility to SLE. Genetic and functional data suggested the CD40 receptor (CD40) and CD40 ligand (CD40L) as strong candidate genes for SLE. AIM: To investigate whether CD40 gene rs1883832 C/T single-nucleotide polymorphism (SNP) and/or soluble CD40 (sCD40) are associated with SLE in the Egyptian population. SUBJECTS AND METHODS: The study included a hundred SLE patients, and a fifty age- and gender-matched healthy control subjects. CD40 gene rs1883832 C/T genotyping was carried out using restriction fragment length polymorphism (RFLP), while sCD40 levels were measured by ELISA. RESULTS: CD40 rs1883832C/T genotypes (CC, TT, and CT) as well as CD40 alleles (C and T) did not differ between SLE patients and normal control (p = 0.63, 0.37, and 0.31 respectively). Though did not reach statistical significance, carriers of genotype CT had 1.5 times more chance to develop SLE compared to wild homozygous CC genotype carriers (OR 1.44), while carriers of genotype TT had ~ 2 times more chance to have SLE than CC carries (OR 1.96). Accordingly, the carriers of the T allele had ¬ 1.5 times more chance to get SLE compared to the carriers of the C allele (OR 1.4). The serum sCD40 level was significantly higher in SLE patients compared to healthy control (3.4 vs. 0.8 ng/mL, p < 0.001). In SLE patients, using CC as the reference genotype, serum sCD40 level was significantly higher in the carriers of the homozygous genotype TT (3.8 ± 1.3 vs. 2.9 ± 1.9, p = 0.0001), and T allele (3.6 ± 1.4 vs. 3.0 ± 1.5, p = 0.003). Moreover, sCD40 could discriminate SLE patients from normal subjects at a cutoff value of 0.885 ng/mL with 98% sensitivity and 96% specificity (AUC = 0.999, p < 0.001). CONCLUSIONS: The study did not prove CD40 gene (rs1883832 C/T) polymorphism as a clear risk factor of SLE in this cohort of Egyptian patients, though it was highly likely associated with the carriers of T allele. In the same context, significant high sCD40 levels were observed in the T allele carriers.

Homocysteine, insulin-like growth factor one and oestrogen levels in patients with erectile dysfunction-associated chronic hepatitis C virus infection.

We assessed the change of homocysteine (Hcy), insulin-like growth factor one (IGF-Ι) and oestrogen (E2) levels in patients with erectile dysfunction (ED) associated with chronic hepatitis C virus (HCV) infection. Eighty-five male patients with chronic HCV and/or ED were enrolled in this study. Seventy-five men were assigned to three equal groups (n = 25/each); Group A: patients who had chronic HCV and ED. Group B: patients who had chronic HCV and had no ED complaint. Group C: patients who had ED with no chronic HCV. In addition to 10 control patients with no ED or chronic HCV (Group D). All patients were subjected to: detailed medical and sexual history, complete physical examination, laboratory assessment including measurement of serum Hcy, IGF-1 and E2. The means of international index of erectile function scores were 8 and 16 in groups A and C respectively. There were significant differences in Hcy, IGF-I and E2 among study groups (p < 0.05 for each). There were significant differences in Hcy between patients with Child B and Child C. A strong association between severity of ED and chronic HCV was demonstrated. There was statistically significant increase of Hcy and E2 levels and reduction in IGF-I level in patients with ED associated with chronic HCV infection.

Serum interleukin-8 and insulin like growth factor-1 in Egyptian bladder cancer patients.

BACKGROUND: Bladder cancer is a major health problem in Egypt as it represents the most common malignancy. AIM: Evaluation of the potential usefulness of serum IL-8 and IGF-1 in Egyptian bladder cancer patients. METHODS: Serum levels of IL-8 and IGF-1 were determined in 51 bladder cancer patients and 45 controls using a chemiluminescence enzyme immunometric assay. RESULTS: Serum IL-8 was significantly higher in cancer patients than in controls (P < 0.0001). It was significantly higher in patients with invasive cancer than those with superficial cancer (P < 0.01). Also, IL-8 showed a significant elevation in relation to schistosomal infection (P = 0.02) however, it did not differ in relation to either pathological type of tumor or its grade (P > 0.05). Receiver operating characteristic (ROC) curve indicated that IL-8 cut-off value of 35 pg/mL yielded the best combination of sensitivity (71%) and specificity (98%) for differentiating patients with bladder cancer from control subjects. Serum IGF-1 levels showed no significant difference between bladder cancer patients and controls (P > 0.05). There was no relationship between IGF-1 levels and clinicopathological parameters. CONCLUSIONS: In Egyptian patients with bladder cancer, serum IL-8 is significantly elevated and its level is related to tumor invasion and associated schistosomal infection. Moreover, serum IGF-1 level does not help as a serum tumor marker in these patients.

Impact of hepatitis C virus infection on neutrophil oxidative burst function in hemodialysis patients.

Polymorphonuclear leukocyte (PMN) functions have been studied extensively in hemodialysis (HD) patients; however, results are contradictory and the mechanisms that modulate phagocytosis and oxidative burst are not completely understood. Hepatitis C virus (HCV) is a frequent complication of HD that may be associated with disturbed PMN function; however, the impact of HCV infection on neutrophil oxidative burst function in HD patients is unknown. We investigated Neutrophil oxidative burst function in 24 HD patients (15 HCV-positive and, 9 HCV-negative patients) before and after dialysis. HCV-RNA was detected by RT-nested PCR while, quantitative measurement of oxidative burst function was assessed by flowcytometry. Neutrophil Oxidised burst function was significantly diminished in HD patients as comapred to controls (P = 0.001, oxidised PMN (%); P = 0.02 mean flueresnce intensity, MFI), and in pre-dialysis as compared to post-dialysis samples (oxidised PMNs (%): 60.5 +/- 3.2 vs. 72.1 +/- 3.9, P = 0.02); (MFI: 352 +/- 42 vs. 500 +/- 50, P = 0.03). Alteration in Neutrophil oxidative burst function in the pre-dialysis samples was significant in HCV-positive patients as compared to HCV-negative patients (oxidized PMNs (%): 50 +/- 2.9 vs. 63 +/- 5.1, P = 0.02); (MFI: 291 +/- 31 vs. 438 +/- 64, P = 0.006). Marked reduction in E. coli induced burst in pre-dialysis samples compared to post-dialysis was found in HCV-positive when compared to HCV-negative patients (oxidized PMNs (%): 50 +/- 2.9 vs. 74.8 +/- 4.7, P = 0.001), (MFI: 291 +/- 31 vs. 493 +/- 63, P = 0.002). In conclusion, a possible role of concomitant HCV infection in alteration of Neutrophil oxidative burst function is highly suggested.

Age related testosterone depletion in patients with erectile dysfunction.

PURPOSE: We assessed the pattern of age related testosterone depletion in patients with erectile dysfunction. MATERIALS AND METHODS: A total of 305 patients with erectile dysfunction who had a normal testosterone level at baseline visit and who completed the study were candidates for analysis. Erectile function was assessed using the International Index of Erectile Function. Patients underwent routine laboratory investigations plus total testosterone and prolactin assessment at the baseline visit and on a yearly basis for 4 years. RESULTS: The mean age +/- SD was significantly higher in 210 patients with decreased testosterone (55.3 +/- 7.3 years) than in 95 patients with steady testosterone (remaining within the normal range) (50.8 +/- 10.2 years). There was a significant decrease in yearly mean testosterone level throughout the study in all the age groups (determined by decades) older than 30 years. Of the study population 68.9% had decreases in testosterone levels throughout the 4 years of visits. Hypogonadism (testosterone lower than normal range) developed in 7.6% of the study population. There was a significant decrease in mean testosterone at any visit in comparison to previous visits. There were significant associations between decreased levels of testosterone and increased severity of erectile dysfunction at baseline visit, longer duration and poor metabolic control of diabetes, ischemic heart disease, hyperprolactinemia and low desire. CONCLUSIONS: This study clearly demonstrated a decrease in testosterone level throughout the 4-year followup in patients with erectile dysfunction. Patients with decreasing testosterone were older than patients with a steady testosterone level.

Pattern of endocrinal changes in patients with sexual dysfunction.

INTRODUCTION: Many patients with endocrinal changes (endocrinopathy) have some degrees of sexual dysfunction that necessitate assessment and treatment. AIM: To assess the prevalence, and identify the pattern, of endocrinopathy in patients with sexual dysfunction in our community. METHODS: A total of 1,248 male patients with sexual dysfunction were enrolled in this study. Patients were screened for erectile dysfunction (ED) and sexual desire by the erectile function and the sexual desire domains of the International Index of Erectile Function (IEEF). Patients underwent routine laboratory investigations as well as total testosterone and prolactin assessment. All patients were referred to an endocrinologist for clinical and biochemical assessment of their endocrine function. The evaluation consisted of comprehensive history taking, physical examination, and, as needed, laboratory investigations. RESULTS: Mean ages+/-SD were 51.9+/-12.2 and 52.3+/-11.7 years for patients with and without endocrinopathy, respectively. Of the study population, 23.8% had endocrinopathy. The most frequent endocrinal changes were low testosterone level (15%), hyperprolactinemia (13.7%), and hypothyroidism (3.1%). There were significant associations between endocrinopathy and obesity, smoking, low desire, and premature ejaculation (P<0.05 for each). Also, significant associations were found between low desire and low testosterone level, hyperprolactinemia, and hypothyroidism (P<0.05 for each). Hyperprolactinemia was significantly associated with premature ejaculation (P<0.05) but not with low testosterone level (P>0.05). There was no significant association between endocrinopathy and age, cigarette smoking (number and duration), and ED (duration, severity, type of onset, and progression) (P>0.05 for each). CONCLUSION: Endocrinopathy is not a rare condition among ambulatory patients with sexual dysfunction. This study provides a quantitative estimate of endocrinopathy in ambulatory patients with sexual dysfunction.

Prostatic specific antigen in patients with hypogonadism: effect of testosterone replacement.

INTRODUCTION: The effect of parenteral testosterone replacement therapy on prostatic specific antigen (PSA) level or the development or growth of prostate cancer is unclear. AIM: To assess the effect of testosterone replacement on PSA level in patients with hypogonadism associated with erectile dysfunction (ED). METHODS: A total of 187 male patients above the age of 45 with hypogonadism associated with ED were enrolled in this study. Patients were screened for ED by the erectile function domain of the International Index of Erectile Function (IIEF). Patients underwent routine laboratory investigations, plus total testosterone, and PSA assessment. Replacement treatment with parenteral testosterone every 2-4 weeks for 1 year was instituted. Total testosterone and PSA serum levels were assessed every 3 months during the treatment course. RESULTS: Mean age +/- SD was 62.8 +/- 11.4. Of the patients 87.7% were sexually active. Of the patients 10.2% had mild, 40.6% had moderate and 49.2% had severe ED. Of the study population, 62.5% had ED complaints for less than 5 years and 84.5% had gradual onset of their complaint. The majority of the patients (91.4%) had either progressive or stationary course while the minority reported regressive course and improvement of the condition. There was a significant increase of the post-treatment testosterone level in comparison to pretreatment level (P < 0.05). No significant increase in the post-treatment PSA level in comparison to pretreatment (P > 0.05). No significant difference between pre- and post-treatment categories of PSA level (normal, borderline, high) in relation to the severity of ED (P > 0.05). There was no significant association between PSA level and the duration of testosterone replacement therapy in the study population (P > 0.05). CONCLUSION: The current study demonstrated that the level of PSA was not significantly changed after 1 year of testosterone replacement therapy in patients with hypogonadism associated with ED.

Soluble ICAM-1 in patients with chronic hepatitis C infection: a prognostic marker of disease activity.

Soluble intercellular adhesion molecule-I (sICAM-1) is an important early marker of response to inflammatory mediators and immune activation released from a variety of cells including hepatocytes. At present, the most reliable determination of severity and prognosis in chronic viral hepatitis is the histological staging of the disease which is an invasive procedure and is often not well accepted by patients. The search for alternative non-invasive methods is mandatory especially in follow ups after initial assessment by biopsy. Serum sICAM-1 level was measured in 19 patients with chronic HCV, 19 patients with non-B, non-C chronic liver diseases (NBNC-CLD) and in 19 healthy control subjects using ELISA. Serum sICAM-1 levels were significantly higher in patients with chronic HCV and in NBNC-CLD patients compared to normal subjects (mean +/- SD, [1003 +/- 453 vs. 232 +/- 177, p<0.001], and [881 +/- 328 vs. 232 +/- 177, p<0.001]), respectively. Furthermore, serum levels of sICAM-1 were significantly higher in HCV-RNA positive patients than in HCV-RNA negative patients (p<0.001). Positive correlations were detected between serum levels of sICAM-1 and serum alanine aminotranseferase (ALT) (p<0.001), aspartate aminotranseferase (AST) (p<0.001), prothrompin time (p<0.001), and alkaline phosphatase (p<0.001), while, a negative correlation with albumin was found (p<0.001). Also, there was a significant correlation between clinical, ultrasonic findings and the level of sICAM-1 in chronic HCV patients as regards hepatomegaly, splenomegaly and normal liver echogenecity. High knodell score was significantly associated with high sICAM-1 level (p<0.001) in both patient groups. while no association between sICAM-1 and fibrosis was found. In conclusion, the measurement of sICAM-1 serum levels in chronic hepatitis C and NBNC-CLD patients is a useful non-invasive marker for monitoring liver disease activity that could replace follow up liver biopsies that are mostly not welcomed by the patients.