Comparison of different transient gene expression systems for the production of a new humanized anti-HER2 monoclonal antibody (Hersintuzumab).

BACKGROUND: Producing therapeutic proteins can be done quickly and on a large scale through Transient Gene Expression (TGE). Chinese hamster ovary (CHO) cell lines are commonly used to achieve this. Although there are few comparative studies, TGE has been observed in suspension-adapted CHO cells. OBJECTIVES: We tested TGE's effectiveness in DG-44, CHO-S, and ExpiCHO-S cell lines with four transfection reagents. METHODS: A design of experiments (DoE) was followed to optimize transfection using a recombinant monoclonal antibody (mAb) construct. To evaluate the efficacy, flow cytometry and ELISA were used. Feeding strategies and temperature shifts were implemented to enhance transfection effectiveness. The quality of the mAb was assessed through ELISA, SDS-PAGE, and proliferation inhibition assays. RESULTS: We adapted all cell lines to grow in suspension using a serum-free medium. Our findings from flow cytometry and ELISA tests indicate that PEI and Pmax reagents had a higher rate of transfection and mAb production than the ExpiCHO commercial transfection reagent. While DG-44 cells had better transfection efficiency than CHO-S and ExpiCHO-S, there was no significant difference between CHO-S and ExpiCHO-S. Our TGE system was more productive at 32 °C than at 37 °C. In the optimized TGE of Pmax-based transfection in DG-44 at 37 and 32 °C, the production level of mAb was more than half of the amount of the commercial ExpiCHO-S expression system. Still, the number of transfected cells was three times higher, making it more efficient. The purified mAb from all transfected cell lines had similar structural and functional properties under different conditions. CONCLUSION: Our research shows that using Pmax and DG-44 cells in the TGE system is a cost-effective and efficient way to produce humanized monoclonal antibodies. We discovered that this method outperforms the ExpiCHO-S kit.

Urinary Beta-2Microglobulin: An Indicator of Renal Tubular Damage after Extracorporeal Shock Wave Lithotripsy.

PURPOSE: This study aims to determine extracorporeal shock wave lithotripsy (ESWL)-induced renal tubular damageand the affecting factors by measuring urinary beta2microglobulin (ß2M) excretion. MATERIALS AND METHODS: This is a cross-sectional study conducted on 91 patients with renal stones who underwentESWL during 2012. Urinary beta2microglobulin was measured immediately before and after the procedure foreach patient and analyzed based on different variables to evaluate factors affecting ESWL-induced renal tubularinjury. RESULTS: Mean ± SD urinary beta2-microglobulin values, before and after ESWL were 0.08 ± 0.07 and 0.22 ± 0.71mg/dL respectively, the average difference between which was equal to 0.14 ± 0.07 mg/dL. These figures exhibiteda 166.66% rise in the urinary ß2M concentration after ESWL which was statistically significant (P < .001). Multivariateanalysis showed that hypertension (P = .05) and the history of ESWL (P = .02) were predictive factors ofhigher post-ESWL urinary beta2-microglobulin excretion. CONCLUSION: Urinary excretion of beta2-microglobulin increased significantly immediately after ESWL. Thesechanges could indicate that ESWL is a contributing factor to renal tubular damage. It also seems that in patientswith hypertension and a previous history of ESWL the likelihood of this injury is higher than others.

Effects of isosorbide dinitrate on the urinary flow rate in patients with benign prostatic hyperplasia.

PURPOSE: To compare the immediate effects of a systemic nitric oxide (NO) donor with placebo on the uroflowmetric parameters in patients with benign prostatic hyperplasia (BPH). MATERIALS AND METHODS: Eighty patients with the mean age of 61.5 years (range, 49 to 74 years) who suffered from BPH were enrolled in the study. We examined peak flow rate, average flow rate, and residual urine in all the patients. Then, patients were randomized to receive either 20 mg sublingual isosorbide dinitrate (ISDN) (n = 40) or placebo (n = 40) 20 minutes prior to the second uroflowmetry, which was performed one day after the first test. RESULTS: The mean peak flow rate increased from 7.6 ± 0.41 mL/s to 10.2 ± 0.54 mL/s (P = .013) in the ISDN group, while it increased +0.40 mL/s in the placebo group (P > .05). Mean residual urine volume decreased significantly from 51 ± 3.1 mL to 29 ± 2.9 mL and from 56 ± 4.1 to 51 ± 2.6 in the ISDN (P = .02) and the placebo groups (P > .05), respectively. At baseline, the mean arterial pressure was 95 ± 2.1 mmHg and under the influence of the NO-donor, it decreased to 83 ± 1.9 mmHg, which was significant (P < .001). No significant changes of micturition parameters were found in the placebo group. CONCLUSION: Organic nitrates influence micturition parameters in patients with BPH. This new approach could offer a potential pharmacological option to treat obstructive lower urinary tract symptoms.

Assessment of clinical efficacy of intranasal desmopressin spray and diclofenac sodium suppository in treatment of renal colic versus diclofenac sodium alone.

OBJECTIVES: To determine the effect of the combination of intranasal desmopressin spray and diclofenac sodium suppository on acute renal colic and compare it with diclofenac sodium suppository alone. METHODS: A total of 150 patients aged 15-65 years referred to our hospital with acute renal colic were included in a double-blind controlled clinical trial study. Patients in group 1 received desmopressin, 40 microg intranasally plus diclofenac sodium suppository 100 mg, and patients in group 2 received diclofenac sodium suppository 100 mg plus a placebo spray consisting of normal saline 0.9%. RESULTS: Significant differences were found in the pain scores at 15 and 30 minutes between the 2 groups (P < .05). Also, significant differences were found in the mean pain scores in the first 15 and first 30 minutes after treatment between the 2 groups (P < .05). Of the patients in group 1, 37.3% had no pain relief and required pethidine. However, this rate in group 2 was 69.3%. In 17 cases, we prescribed pethidine within 20 minutes after treatment, and these patients were excluded from our study. CONCLUSIONS: According to our results, intranasal desmopressin plus diclofenac sodium suppository caused prompt pain relief with significant decreases in pain scores after 15 and 30 minutes. We suggest that intranasal desmopressin spray is a useful supplemental therapy for renal colic in combination with nonsteroidal anti-inflammatory drugs, especially to reduce the use of opioids.

Tubeless percutaneous nephrolithotomy for staghorn stones.

PURPOSE: To assess the outcome and safety of tubeless percutaneous nephrolithotomy (PCNL) to treat patients with staghorn stones. PATIENTS AND METHODS: Between March 2006 and March 2007, 75 patients underwent 82 PCNLs at our hospital. Of that total, 42 procedures were done without a nephrostomy tube (tubeless group) and 40 procedures were done with insertion of a nephrostomy tube at the end of the procedure (standard group). RESULTS: The mean age was 41.55 +/- 15.15 years and 50.29 +/- 9.42 years in the tubeless group and the standard group, respectively. Seven patients had bilateral stones. None of our patients had organ trauma or any other significant complication. The average operative time was shorter in the tubeless group than in the standard group (93.76 v 109.98 minutes, respectively; P = 0.03). The mean analgesic doses given postoperatively were 79.17 +/- 62.2 and 93.75 +/- 40.7 mg of meperidine in the tubeless group and the standard group, respectively, with no statistically significant difference. The mean postoperative hospital stay was 1.7 +/- 0.6 days in the tubeless group and 4 +/- 1.6 days in the standard group (P < 0.0001). The stone-free rate was 37 units (88.09%) and 35 units (87.5%) units in the tubeless and standard groups, respectively. CONCLUSIONS: Complications, operative time, and the length of hospitalization in selected patients undergoing tubeless PCNL were all lower than those seen in the standard group. Tubeless PCNL was thus found to be safe and effective, even in patients with staghorn stones.

Vasal irrigation with sterile water and saline solution for acceleration of postvasectomy azoospermia.

INTRODUCTION: Vasectomy is the safest and most reliable method of all the contraception methods, but azoospermia is not achieved immediately by this method. We decided to determine whether irrigation of the vas deferens with sterile water or hypertonic saline solution irrigation during vasectomy would reduce the time needed to obtain azoospermia. MATERIALS AND METHODS: A total of 126 fertile men presented for vasectomy were divided in 3 groups. No-scalpel vasectomy was done for all of the participants and irrigation of the vas deferens was carried out during the procedure in 2 groups with either sterile water or hypertonic saline solution (9 g/L sodium chloride solution). Forty-two participants underwent vasectomy without irrigation. Semen analysis was performed at 4, 8, 12, and 16 weeks after vasectomy. RESULTS: Azoospermia was achieved in all of the men with sterile water after 12 weeks, while at the end of the study (16 weeks) it was achieved in 37 (88.1%) of those with saline solution and in 11 (26.2%) of those without irrigation. There were significant differences in the rates of azoospermia between the participant with sterile water and saline solution at 8 weeks (38.1% versus zero; P < .001), 12 weeks (100% versus 30.9%; P < .001), and 16 weeks (100% versus 88.1%; P = .02). No pregnancy developed during the follow-up and no complication was reported. CONCLUSION: Vasal irrigation with sterile water and hypertonic saline solution during vasectomy were effective in removing sperm from the distal vas and increasing the rate at which men achieved azoospermia. Sterile water was a promising option with no complications.

The risk factors of prostate cancer: a multicentric case-control study in Iran.

Prostate cancer (PC), in Iran, is the third most frequently diagnosed visceral cancer among men and the seventh most common underlying cause of cancer mortality. We evaluated the relation between speculated factors and PC risk using data from a multicentric case-control study conducted in Iran from 2005 to 2007 on 130 cases of incident, clinicopathologically confirmed PC, and 75 controls admitted to the same network of hospitals without any malignant disease. Odds ratios (OR) and corresponding 95% confidence intervals (CIs) were estimated using conditional logistic regression models. The risk of PC was increased with aging (OR: 5.35, 95% CI: 2.17-13.19; P<0.0001), and with the number of sexual intercourse >or=2 times/week (OR: 3.14, 95% CI: 1.2-8.2; P=0.02). One unit elevation in serum estradiol and testosterone concentration was related to increase (OR: 1.04, 95% CI: 1.01-1.06; P=0.006) and decrease (OR: 0.79; 95% CI: 0.64-0.96; P=0.02) of PC risk, respectively. Cases were less likely to have a history of diabetes (OR: 0.34, 95% CI: 0.12-0.98; P=0.04). Increasing in dietary consumption of lycopene and fat was associated with declined (OR: 0.45, 95% CI: 0.09-2.12) and increased (OR: 2.38, 95% CI: 0.29-19.4) PC development, respectively. Other factors including educational level, marriage status, dietary meat consumption, vasectomy and smoking have not been shown to affect PC risk in the Iranian population. Our study adds further information on the potential risk factors of PC and is the first epidemiologic report from Iran. However, justification of these results requires more well-designed studies with a larger number of participants.

Diagnostic accuracy of C-reactive protein and erythrocyte sedimentation rate in patients with acute scrotum.

INTRODUCTION: Our aim was to investigate the diagnostic accuracy of C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR) in patients with acute scrotum. MATERIALS AND METHODS: One hundred and twenty patients with acute scrotum were evaluated and divided into 3 groups: 46 with acute epididymitis (group 1), 23 with spermatic cord torsion (group 2), and 51 with other noninflammatory causes of acute scrotum (group 3). Serum levels CRP and ESR were measured at the time of admission. RESULTS: Of the patients in group 1, 44 (95.6%) had elevated serum levels of CRP (mean, 67.77 +/- 47.80 mg/L). In contrast, only 1 patient in group 2 had a significant increase in the serum level of CRP (mean, 9.0 +/- 4.90 mg/L), and the patients in group 3 did not have any significant increase in the CRP levels (mean, 7.0 +/- 2.2 mg/L) (P < .001). The mean ESR values were 45.9 +/- 21.4 mm/h, 14.2 +/- 11.2 mm/h, and 8.8 +/- 7.5 mm/h, in groups 1 to 3, respectively (P < .001). The cutoff points for distinguishing between epididymitis and noninflammatory causes of acute scrotum were 24 mg/L for CRP and 15.5 mm/h for ESR. The sensitivity and specificity values were 93.4% and 100% for CRP and 95.6% and 85.1% for ESR, respectively. CONCLUSION: Based on our findings, serum levels of CRP and ESR can provide helpful information for differentiation between epididymitis and other causes of acute scrotum. We recommend CRP and ESR measurements before making a decision of surgical intervention.