Successful application of modified crude DNA extraction from muscle tissues for various types of PCR amplifications.

BACKGROUND: One of the most challenging aspects of nucleic acid amplification tests is the extraction of genomic DNA. However, achieving satisfactory quality and quantity of genomic DNA is not always easy, while the demand for rapid, low-cost and less laborious DNA isolation methods is ever-increasing. METHODS AND RESULTS: We have developed a rapid (⁓2 min) crude DNA extraction method leading to direct-PCR that requires minimum reagents and laboratory equipment. It was developed by eliminating the time-consuming purification steps of DNA extraction, by processing the sample in optimized amounts of Taq KCl PCR buffer and DNARelease Additive/Proteinase K in only two minutes and carrying out amplification using conventional Taq DNA polymerase. The DNA preparation method was validated on muscle tissue samples from 12 different species as well as 48 cooked meat samples. Its compatibility was also successfully tested with different types of PCR amplification platforms extensively used for genetic analysis, such as simplex PCR, PCR-RFLP (Restriction Fragment Length Polymorphism), multiplex PCR, isothermal amplification, real-time PCR and DNA sequencing. CONCLUSIONS: The developed protocol provides sufficient amount of crude DNA from muscle tissues of different species for PCR amplifications to identify species-of-origin via different techniques coupled with PCR. The simplicity and robustness of this protocol make nucleic acid amplification assays more accessible and affordable to researchers and authorities for both laboratory and point-of-care tests.

Does male factor infertility impact on self-esteem and sexual relationship?

Nonsomatic factors play a significant role in erectile dysfunction. We assessed the impact on men suffering from male factor infertility by employing the validated Self-Esteem and Relationship questionnaire (SEAR) and compared the scores with controls. Men with primary infertility were asked to fill SEAR questionnaire. Normal fertile men visiting clinic for unrelated problem or normal male accompanying infertile patients constitute the control group. Transformed score of each domain and total score of SEAR questionnaire and means were compared. Data were analysed by R version 5.2. Univariate and multivariate analyses were done to determine factors predicting self-esteem and total relationship score. There were 45 men each in the study and control groups. The mean transformed self-esteem score and total score of infertile men were significantly lower as compared to controls (74.44 versus 95.83) and (73.54 versus 95.86) p-value of <.0001. Longer duration of infertility, advanced age, diabetes and higher education were factors significantly lowering the scores on univariate analysis, and on multivariate analysis, diabetes was the only significant predictor of total SEAR score in infertile men. Infertility leads to lower self-esteem, sexual performance and confidence among infertile men as compared to controls. The relationship score worsens with increasing duration of infertility.

The impact of post PCNL tube type on blood loss and postoperative pain.

OBJECTIVE: To evaluate the impact of nephrostomy tube type on postoperative pain and blood loss following percutaneous nephrolithotomy (PCNL). METHODS: This is a prospective non-randomized study performed at Aga Khan University Hospital from July 2017 to June 2018. In this study we prospectively studied adult patients (16 to 65 years) who underwent unilateral PCNL. Patients who had nephrostomy with balloon (12Fr Foley's catheter) were compared with patients who had nephrostomy without balloon (12Fr Nelaton™ catheter). STONE Nephrolithometry score was used to assess the stone complexity. Mean pain score at six and 24 hours and mean hemoglobin drop at 24 hours was compared between two groups using independent sample t-test, p-value of <0.05 was considered significant. RESULTS: Over one year, 198 PCNL were performed out of which 119 were included for analysis. Sixty-six had nephrostomy tube with balloon and 53 had nephrostomy tube without balloon. Mean STONE score (9.66±1.4 vs. 9.64±1.24) and operative time (72.84±28.34 vs. 86.05±32.1 minutes) was comparable. Mean postoperative pain score at 6 hours and 24 hours postoperative was significantly lower in balloon group as compared to without balloon group. Mean Hemoglobin drop was similar in both groups (p=0.60). CONCLUSION: The use of nephrostomy tube with balloon after PCNL as this is associated with less pain and comparable hemoglobin drop as compare to nephrostomy tube without balloon.

Foreign Body in Kidney Presenting as Renal Stone.

Retained foreign bodies in the urinary system are commonly found in the urinary bladder but are a rare finding in the renal pelvis. Foreign objects can reach the renal pelvis via different pathways. The presence of a foreign body could be iatrogenic, or via direct penetration from the gastrointestinal tract. Presentation could mimic tumor, abscess, hematoma, or even renal stone. We report an unusual case of intrarenal foreign body presenting as renal stone. The patient presented with flank pain and symptoms suggestive of renal stone and a non-contrast CT scan was also indicative of stone, however on endoscopy a foreign body was identified and retrieved.

Efficacy and Safety of Semi-rigid Ureteroscopic Lithotripsy (URS) for Proximal Ureteral Stone ≥10 mm.

OBJECTIVE: To evaluate the efficacy and safety of the semi-rigid URS with pneumatic lithotripsy for the treatment of large (>10 mm) proximal ureteral calculi. STUDY DESIGN: Cohort study. PLACE AND DURATION OF STUDY: Section of Urology, Department of Surgery, The Aga Khan University Hospital, Karachi, Pakistan, from January 2010 to December 2018. METHODOLOGY: All patients with single, radio-opaque unilateral proximal ureteral stones >10 mm were included in the study. Patients with urosepsis, pregnancy, solitary kidney, and indwelling JJ stent or prior percutaneous nephrostomy drainage were excluded. Patients without any visualisation of stone on abdominal X-Ray done at 4 weeks of the procedure were labelled as stone-free. RESULTS: One hundred and three cases with stones >10 mm and fulfilling the selection criteria were included. The mean age was 40.83 + 14.92 years. The mean stones size was 13.33 + 4.64 mm. The mean procedure time was 41.50 + 15.60 minutes. Indwelling JJ stents were placed in 49 (47.6 %) cases. The stone-free rate was 83.5% at four weeks with calculated efficiency quotient (EQ) of 0.72, using a standard equation. The overall complication rate was 27% with the majority being minor (Clavian-Dindo grade 1). Only one patient had urosepsis (MCG IV). Ancillary procedures were performed in 17 (16.5%) cases, most commonly the extracorporeal shock wave lithotripsy (ESWL) in 14 (13.5%) cases, followed by the secondary URS in 3 (2.9%) cases. CONCLUSION: Semi-rigid URS is an effective and safe procedure for the large proximal ureteral stones, with limited access to flexible instruments. Key Words: Ureteroscopy, Semi-rigid, Stone, Proximal, Clearance, Safety.