Transfer of mesenchymal stem cell mitochondria to CD4+ T cells contributes to repress Th1 differentiation by downregulating T-bet expression.

BACKGROUND: Mesenchymal stem/stromal cells (MSCs) are multipotent cells with strong tissue repair and immunomodulatory properties. Due to their ability to repress pathogenic immune responses, and in particular T cell responses, they show therapeutic potential for the treatment of autoimmune diseases, organ rejection and graft versus host disease. MSCs have the remarkable ability to export their own mitochondria to neighboring cells in response to injury and inflammation. However, whether mitochondrial transfer occurs and has any role in the repression of CD4+ Th1 responses is unknown. METHODS AND RESULTS: In this report we have utilized CD4+ T cells from HNT TCR transgenic mice that develop Th1-like responses upon antigenic stimulation in vitro and in vivo. Allogeneic bone marrow-derived MSCs reduced the diabetogenic potential of HNT CD4+ T cells in vivo in a transgenic mouse model of disease. In co-culture experiments, we have shown that MSCs were able to reduce HNT CD4+ T cell expansion, expression of key effector markers and production of the effector cytokine IFNγ after activation. This was associated with the ability of CD4+ T cells to acquire mitochondria from MSCs as evidenced by FACS and confocal microscopy. Remarkably, transfer of isolated MSC mitochondria to CD4+ T cells resulted in decreased T cell proliferation and IFNγ production. These effects were additive with those of prostaglandin E2 secreted by MSCs. Finally, we demonstrated that both co-culture with MSCs and transfer of isolated MSC mitochondria prevent the upregulation of T-bet, the master Th1 transcription factor, on activated CD4+ T cells. CONCLUSION: The present study demonstrates that transfer of MSC mitochondria to activated CD4+ T cells results in the suppression of Th1 responses in part by downregulating T-bet expression. Furthermore, our studies suggest that MSC mitochondrial transfer might represent a general mechanism of MSC-dependent immunosuppression.

L-carnitine as primary or adjuvant treatment in infertile patients with varicocele. A systematic review.

BACKGROUND: Varicocele has been found to impair the function of the epididymis resulting in subfertility whereas the varicocelectomy can resolve the phenomenon. L-carnitine is regarded as a biomarker for the function of the epididymis and has been found in reduced concentrations in infertile patients of various causes, including infertile men with varicocele. It seems that Lcarnitine and varicocele share clinical significance and the area of research looks promising. OBJECTIVE: To identify the role of L-carnitine in the treatment of varicocele. MATERIALS AND METHODS: A systematic search was performed in Pubmed/Medline with the terms (L-carnitine) and (varicocele) and (L-carnitine) and (varicocelectomy). Inclusion criteria were studies reported outcomes of L-carnitine administration alone or in duet, as primary or adjuvant treatment to varicocele. Exclusion criteria were non-English language and animal studies. Studies using L-carnitine as part of a panel of therapeutic agents were avoided. RESULTS: Only four suitable studies were identified for discussion. In one randomized study, the combination of L-carnitine and cinnoxicam improved semen parameters in patients with non-high-grade varicocele compared to L-carnitine alone and had a favourable effect on pregnancy rates but the effect of grade is unknown. In another study, as an adjuvant treatment to varicocelectomy, L-carnitine showed no clear benefit. Finally, in comparison to surgery, the results are inconclusive; two studies showed some benefit might be expected in low-grade or subclinical varicocele, but surgery appears superior. CONCLUSIONS: The evidence regarding the role of L-carnitine as a primary or adjuvant treatment of varicocele is sparse. The pathophysiological significance of L-carnitine implicates a potential role of the molecule in the management of varicocele, but the evidence so far is controversial for any recommendations. L-carnitine might be taken into consideration in selected cases; however, further search is needed in order the optimal role of L-carnitine in infertile patients with varicocele to be clarified.

Quality of leadership in multidisciplinary cancer tumor boards: development and evaluation of a leadership assessment instrument (ATLAS).

BACKGROUND: High-quality leadership and chairing skills are vital for good performance in multidisciplinary tumor boards (MTBs), but no instruments currently exist for assessing and improving these skills. OBJECTIVE: To construct and validate a robust instrument for assessment of MTB leading and chairing skills. DESIGN AND SETTING: We developed an observational MTB leadership assessment instrument (ATLAS). ATLAS includes 12 domains that assess the leadership and chairing skills of the MTB chairperson. ATLAS has gone through a rigorous process of refinement and content validation prior to use to assess the MTB lead by two urological surgeons (blinded to each other) in 7 real-live (n = 286 cases) and 10 video-recorded (n = 131 cases) MTBs. OUTCOME MEASURES AND STATISTICAL ANALYSIS: ATLAS domains were analyzed via descriptive statistics. Instrument content was evaluated for validity using the content validation index (CVI). Intraclass correlation coefficients (ICCs) were used to assess inter-observer reliability. RESULTS: Instrument refining resulted in ATLAS including the following 12 domains: time management, communication, encouraging contribution, ability to summarize, ensuring all patients have treatment plan, case prioritization, keeping meeting focused, facilitate discussion, conflict management, leadership, creating good working atmosphere, and recruitment for clinical trials. CVI was acceptable and inter-rater agreement adequate to high for all domains. Agreement was somewhat higher in real-time MTBs compared to video ratings. Concurrent validation evidence was derived via positive and significant correlations between ATLAS and an established validated brief MTB leadership assessment scale. CONCLUSION: ATLAS is an observational assessment instrument that can be reliably used for assessing leadership and chairing skills in cancer MTBs (both live and video-recorded). The ability to assess and feedback on team leader performance provides the ground for promotion of good practice and continuing professional development of tumor board leaders.

Magnetic resonance cholangiopancreatography in diagnosis of biliary disorders in children - Sharing our experience.

OBJECTIVE: To evaluate the role of Magnetic Resonance Cholangio-pancreatography in the diagnosis of biliary disorders in children. METHODS: The retrospective study was conducted at Aga Khan University Hospital, Karachi, and comprised data related to the period between August 2005 and December 2013. All children from age of 1 day to 17 years who had undergone Magnetic Rasonance Cholangio-pancreatography examination for suspected pancreaticobiliary disorders were included. Clinical presentation, Magnetic Rasonance Cholangio-pancreatography findings, operative findings and histopathological results were recorded. Sensitivity and specificity of Magnetic Rasonance Cholangio-pancreatography for different diseases was worked out. RESULTS: Of the 50 patients in the study, 12(24%) showed findings of choledochal cyst. Of these patients, 11(91.6%) underwent surgery and operative findings were consistent with choledochal cyst. Only 1(8.3%) who was assessed as biliary atresia was found to have choledochal cyst on surgery Magnetic Resonance Cholangio-pancreatography was found to be 91% sensitive and 100% specific for diagnosis of choledochal cyst with diagnostic accuracy of 98%. CONCLUSIONS: Magnetic Resonance Cholangio-pancreatography is a very accurate non-invasive investigation for the diagnosis of biliary disorders.

Transient Corneal Edema After Phacoemulsification.

OBJECTIVE: To determine the association between pre-operative and intra-operative factors leading to transient corneal edema after phacoemulsification. STUDY DESIGN: Cohort study. PLACE AND DURATION OF STUDY: Department of Ophthalmology, Shifa Foundation Community Health Centre, Islamabad, from October 2011 to September 2012. METHODOLOGY: Patients undergoing phacoemulsification and Intraocular Lens (IOL) implantation were enrolled in the study using consecutive non-probability sampling. Pre-operative risk factors including peripheral corneal degenerations, the type and density of cataract were documented. Surgical risk factors included the incision site, the type of intraocular lens, the phacotechnique and the phacopower time. Postoperatively the patients were assessed for corneal clarity and the degree of striate keratopathy. Statistical analysis was done using SPSS version 17. RESULTS: There were 43% male and 57% female patients (n = 182). Mean age was 58.92 ± 13.00 years (median and mode-60 years). Factors which increased the risk of transient corneal edema after phacoemulsification included hypertension (p = 0.022), dense nuclear cataracts (p=0.006), divide and conquer technique (p = 0.008), duration of phacopower use (p < 0.001) and peripheral corneal degenerations (p < 0.001). CONCLUSION: Patients with peripheral corneal degenerations and dense nuclear cataracts had significantly higher rates of postoperative corneal edema. Use of phaco-chop technique and less phaco-power time helps in decreasing corneal edema.

Nephroureterectomy for emphysematous pyelonephritis: An aggressive approach is sometimes necessary. A case report and literature review.

INTRODUCTION: Emphysematous pyelonephritis (EPN) is a life-threatening urological emergency. A high index of suspicion is required for diagnosis as such patients may present to physicians with typical features of pyelonephritis. PRESENTATION OF CASE: A 67 year old lady presented atypically to the Emergency Department with symptoms of renal colic. The diagnosis of emphysematous pyelonephritis was established on prompt CT scanning. She did not respond to conservative management. Due to acute, critical deterioration, she underwent a radical right nephroureterectomy. The resected kidney involved a long segment of necrotic, gangrenous ureter. The patient had a smooth post-operative recovery and was successfully discharged. She remains well on follow-up after one year. DISCUSSION: Early radiological diagnosis is imperative for risk stratification of EPN. Current evidence recommends percutaneous catheter drainage with interval nephrectomy as the gold standard treatment. We review the literature for pathophysiology and clinical prognostic factors. This case adds onto the limited evidence base on ureteric involvement in EPN, suggesting a revision of EPN classification. CONCLUSION: Further research on ureteric involvement and treatment outcomes in EPN is required. Even in the current era of minimally invasive surgery and renal preservation therapies, early open nephrectomy still has a role in the management of EPN.

Laparoscopic varicocelectomy in the management of chronic scrotal pain.

BACKGROUND AND OBJECTIVES: To evaluate the usefulness of laparoscopic varicocelectomy in the management of chronic scrotal pain. METHODS: Between 2009 and 2011, 48 patients in total were treated with laparoscopic varicocelectomy for dull scrotal pain that worsened with physical activity and was attributed to varicoceles. All patients were followed up at 3 and 6 months and biannually thereafter with a physical examination, visual analog scale score, and ultrasonographic scan in selected cases. RESULTS: The mean age was 38.2 years (range, 23-54 years). The mean follow-up period was 19.6 months (range, 6-26 months). Bilateral varicoceles were present in 7 patients (14.6%), and a unilateral varicocele was present in 41 (85.4%). The varicocele was grade 3 in 27 patients (56.3%), grade 2 in 20 (41.6%), and grade 1 in 1 (2.1%). The mean preoperative visual analog scale score was 4.8 on a scale from 0 to 10. The mean postoperative visual analog scale score at 3 months was 0.8. After the procedure, 42 patients (87.5%) had a significant improvement in the visual analog scale score (P<.001); 5 (10.4%) had symptom improvement, although it was not statistically significant; and 1 (2.1%) remained unchanged. During follow-up, we observed 5 recurrences (10.4%) whereas de novo hydrocele formation was identified in 4 individuals (8.3%). CONCLUSION: Laparoscopic varicocelectomy is efficient in the treatment of symptomatic varicoceles with a low complication rate. However, careful patient selection is necessary because it appears that individuals presenting with sharp, radiating testicular pain and/or a low-grade varicocele are less likely to benefit from this procedure.

Reducing postoperative pterygium recurrence: comparison of free conjunctival auto-graft and conjunctival rotation flap techniques.

OBJECTIVE: To compare the recurrence of pterygium between free conjunctival auto-graft and conjunctival rotation flap following simple surgical excision of pterygium. STUDY DESIGN: Quasi-experimental study. PLACE AND DURATION OF STUDY: Shifa Foundation Community Health Clinic, Shifa College of Medicine, Islamabad, from January to November 2012. METHODOLOGY: Fifty seven cases aged above 18 years, with a pterygium corneal encroachment of ³ 2 mm which was responsible for visual disability or was cosmetically undesirable were recruited for the study and randomly assigned to conjunctival auto-graft group and conjunctival rotation flap group. Cases with a history of glaucoma or glaucoma suspect, prior pterygium surgery, pterygium with concurrent ocular surface and lid disease, conjunctival inflammation and scarring, pseudo-pterygium or collagen vascular disease were excluded. After simple pterygium excision conjunctival auto-graft group (n=26) cases received a free conjunctival flap was transplanted, while conjunctival rotation flap group (n=31) cases received a conjunctival rotation flap. All cases were followed-up for 6 months after surgery for recurrence and complications. Frequency distribution and significance of association of recurrence using Fisher's exact test and Mann-Whitney U-test was carried out using Statistical Package for Social Sciences (SPSS) version 20. RESULTS: The median (and inter-quartile range) age and surgery duration in conjunctival auto-graft group and conjunctival rotation flap group were 60 (51.50-63.00) and 57 (45.00-60.00) years, 28.50 (27.00-30.50) and 16.00 (15.00-17.00) minutes respectively. Recurrence was seen in 2 (7.96%) and 3 (9.76%) cases in auto-graft and rotation flap groups respectively. No significant difference was seen in postoperative complications between the two groups (p=0.60). CONCLUSION: The surgical time for conjunctival rotation flap procedure is less as compared to free auto-graft, while their recurrence and complications are comparable.

Validation of team performance assessment of multidisciplinary tumor boards.

PURPOSE: We construct validated the instrument to evaluate assessor learning curves and the feasibility and interrater reliability of MTB-MODe for assessing the decision making process using video recorded multidisciplinary tumor board meetings. MATERIALS AND METHODS: Multidisciplinary tumor boards are becoming standard practice for managing cancer internationally but no standards have been agreed on to assess the efficacy of such teams. The MTB-MODe tool assesses the process of multidisciplinary tumor board decision making by standardized observation (1 to 5 anchored scales) of the quality of information presented at the multidisciplinary tumor board as well as board member contributions to the case review. We assessed 683 multidisciplinary tumor board case discussions using MTB-MODe in a multiphase study, including 332 cases (9 urology boards) by 1 urologist in vivo and 224 cases (6 urology boards) by 2 urologists in vivo. The instrument was refined and subsequently used to rate 127 video recorded case discussions (5 tumor types) by a total of 8 multidisciplinary tumor boards. RESULTS: Good interrater reliability was achieved in vivo and at the video recorded multidisciplinary tumor board meetings (ICC ≥0.70). MTB-MODe scores were higher in cases that resulted in a decision than in cases in which no decision was made (mean ± SD 2.54 ± 0.47 vs 2.02 ± 0.65, p ≤0.001). CONCLUSIONS: A standardized method to assess the quality of multidisciplinary tumor board discussions can enhance the quality of cancer care and the ability of the boards to self-evaluate performance, thus, promoting good practice. Video recordings offer a feasible, reliable method of assessing how multidisciplinary tumor boards work.

Should every patient with hematospermia be investigated? A critical review.

Hematospermia or hemospermia is defined as the presence of blood in ejaculate. It often invokes considerable anxiety and is frightening for the patient. Mostly, it is due to infectious causes and regarded as a benign and self-limiting condition particularly in younger patients. Patients above 40 years of age and those with high risk factors require thorough evaluation. Detailed examination is mandatory, and should include: blood pressure measurement and abdominal palpation to identify hepatosplenomegaly or renal enlargement. Genital examination must also be performed to assess for the presence of testicular lumps and urethral discharge, as well as a rectal examination to assess the prostate. Further investigations include cystoscopy, transrectal ultrasound, and prostate biopsy. Diagnosing prostatic pathologies is made easier by performing transrectal ultrasound. It is useful in diagnosing calculi, cysts, prostatic varices, and inflammatory changes, as well as therapeutic in certain cases where cyst or abscess is drained and is found to be the cause of hematospermia. Complex investigations depend on history and examination. A role of MRI is emerging to rule out rare causes of hematospermia. Evidence based evaluation of hematospermia is not only useful in definitive diagnosis, but it can also be cost effective. Therefore, we suggest that patients with high risk factors should be investigated thoroughly. However, younger patients with one episode can be monitored closely and investigated only if deemed necessary.

Magnetic resonance spectroscopy in focal brain lesions.

OBJECTIVE: To describe the spectrum of Magnetic Resonance spectroscopy in focal brain lesions and determine its diagnostic accuracy using histopathology as gold standard in differentiating neoplastic and non-neoplastic focal brain lesions. METHODS: The study was conducted in Department of Radiology, Aga Khan University Hospital, Karachi from Dec 2006 till Jan 2009. Fifty three patients found to have focal brain lesions on magnetic resonance imaging (MRI) were included in the study. Magnetic Resonance spectroscopy (MRS) was performed in these focal lesions. These lesions were deemed neoplastic and non-neoplastic on MR Spectrum findings. Correlation of all these findings was done with histopathology obtained in all these patients. Sensitivity, specificity, positive, negative predictive values and diagnostic accuracy of MR Spectroscopy was calculated. Percentage agreement between spectroscopy and histopathology was also calculated using kappa statistics. RESULTS: Increase Choline/creatine and Choline/NAA ratio noted in neoplastic lesions compared to nonneoplastic lesion with significant p-value. MR Spectroscopy has a sensitivity of 93.02%, specificity of 70%, positive predictive value of 93.02%, negative predictive value of 70% and diagnostic accuracy of 88.67% in differentiating neoplastic and non-neoplastic brain lesions. Kappa statistics shows a good agreement between MR Spectroscopy and histopathology (k = 0.630). CONCLUSION: Magnetic Resonance spectroscopy (MRS) is non-invasive sensitive, however, relatively nonspecific modality in differentiating neoplastic and non-neoplastic brain lesions. This modality should be considered as an adjunct to conventional imaging rather than replacement for histopathological evaluation.

Triphasic computed tomography (CT) scan in focal tumoral liver lesions.

OBJECTIVE: To assess the diagnostic accuracy of triphasic spiral CT in differentiating benign from malignant focal tumoral liver lesions. METHODS: The study was conducted in Department of Radiology of Aga Khan University Hospital and Sind Institute of Urology and Transplantation, Karachi from Feb 2006 to Feb 2007. By convenient sampling, 45 patients found to have focal tumoral liver lesions were recruited for one year period and their triphasic CT scans findings were evaluated and later correlated with histopathology. Sensitivity, specificity, positive predictive value, negative predictive value and diagnostic accuracy of triphasic CT scan were calculated. RESULTS: Among 45 patients, 136 liver lesions (11 benign and 125 malignant) were detected with the help of different enhancement patterns. Out of these, 37(82.2%) patients had malignant while 8 (17.8%) had benign lesions. On later histopathological examination, 35 (77.8%) of the total 45 cases had malignant lesions while 10 (22.2%) were diagnosed as benign lesions. Based on these results, it could be assessed that triphasic CT Scan has a sensitivity of 100%, specificity of 80%, positive predictive value of 94.5%, negative predictive value of 100% and diagnostic accuracy of 95.5% in differentiating benign from malignant liver lesions. CONCLUSION: Triphasic CT Scan is a good non-invasive tool in characterizing and differentiating benign from malignant liver lesions.

Effect of post-hypoxic reoxygenation on DNA fragmentation in cortical neuronal nuclei of newborn piglets.

Previous studies have shown an increased fragmentation of genomic DNA following hypoxia in cortical neuronal nuclei of newborn piglets. The present study tests the hypothesis that DNA fragmentation following hypoxia persists during reoxygenation in cortical neuronal nuclei of newborn piglets. To test this hypothesis, DNA fragmentation was assessed in 36 newborn piglets divided into six groups: normoxic (Nx), hypoxic (Hx) and hypoxic/reoxygenated for 6, 12, 24h and 7 days. The Hx groups were exposed to 7% oxygen for 1h followed by reoxygenation to room air for 6, 12, 24h and 7 days. Cerebral tissue hypoxia was confirmed biochemically by ATP and phosphocreatine (PCr) levels. Nuclei were isolated and purified using discontinuous sucrose gradient. DNA was isolated by phenol/chloroform/isoamyl-alcohol extraction method. ATP/PCr (micromol/g brain) were 4.11+/-0.15/3.67+/-0.30 for Nx, 1.31+/-0.68/0.74+/-0.30 for Hx, 3.81+/-0.11/3.24+/-0.14 for 6h reoxygenation, 4.21+/-0.12/3.27+/-0.09 for 12h reoxygenation and 4.63+/-0.09/3.75+/-0.27 for 24h reoxygenation and 4.31+/-0.12/3.70+/-0.21 for 7 days reoxygenation. There was a significant difference in the ATP and PCr values between Nx and Hx groups (p<0.05) and between Hx and hypoxic reoxygenated groups (p<0.05). DNA fragments (OD/mm(2)) increased from 1776+/-267 in the Nx group to 3211+/-285 in the Hx group (p<0.05). In the reoxygenation groups, DNA fragments (OD/mm(2)) decreased to 2018+/-249 after 6h (p<0.05 versus Hx) but increased to 3408+/-206, 2782+/-406 and 3256+/-302 after 12, 24h and 7 days, respectively. The data show a decrease in DNA fragmentation in the early phase (6h) of reoxygenation but is comparable to acute hypoxia during the later phases (12, 24h and 7 days) of reoxygenation. We propose that the biphasic pattern of DNA fragmentation during reoxygenation occurs by an initial oxidative DNA injury followed by an enzymatic cleavage of DNA by endonucleases activation.

Percutaneous radiologic gastrostomy: results and complications.

BACKGROUND: Percutaneous endoscopic gastrostomy is usually difficult in patients with malignant involvement of oral cavity, pharynx and esophagus. Flouroscopic guided insertion of Gastrostomy catheter with Gastropexy have gained acceptance because it is easy and less time consuming as well as less invasive as compare to surgical procedure. This study was done to evaluate the safety and efficacy of percutaneous placement of gastrostomy with gastropexy using imaging guidance in patients with oropharyngeal and esophageal cancers. METHODS: Over five years, 105 patients were referred to our department for percutaneous radiologic gastrostomy. In five patients the procedure was not performed because of overlying viscera and high position of stomach. We performed 100 gastrostomies with gastropexy procedures using seldinger technique. RESULTS: Success rate for percutaneous radiologic gastrostomy was 100%. No major complication had occurred. There were 11 minor complications occurred including 4 stomal infection, 3 catheter obstruction, one peritonism and three were extensive pneumoperitoneum. Stomal infection and catheter obstruction were not related to procedure. So, our true minor complications were only 4 (4%) which is comparable to literature. CONCLUSION: Percutaneous radiologic gastrostomy is an effective and safe procedure for enteric access of nutrition in patients with oral, pharyngeal and esophageal cancer where percutaneous endoscopic gastrostomy is difficult.

Hypoxia-induced Ca2+-influx in cerebral cortical neuronal nuclei of newborn piglets.

The present study was designed to investigate the effect of hypoxia on nuclear calcium-influx in the cerebral cortex of newborn piglets. Anesthetized and ventilated newborn piglets divided into normoxic (n=4) and hypoxic groups with varying degrees of tissue hypoxia (n=10) were studied. Nuclear Ca(2+)-influx was determined using (45)Ca(2+) and plotted against ATP and phosphocreatine levels. The plots were analyzed by non-linear regression (exponential) analysis that showed a curvilinear relationship (r=0.92 for ATP and r=0.88 for phosphocreatine). These data suggest a threshold at which there is a sudden increase in the nuclear calcium-influx that then continues to increase with further decrease in the ATP and phosphocreatine levels. The results demonstrate an increase in the nuclear Ca(2+)-influx during hypoxia in newborn piglets and that this increase correlates in a curvilinear fashion with the increase in the degree of cerebral tissue hypoxia. We propose that the hypoxia-induced increase in intranuclear Ca(2+) is due to altered nuclear membrane Ca(2+)-influx mechanisms and will lead to Ca(2+)-mediated alteration of apoptotic gene expression as well as Ca(2+)-dependent activation of endonucleases that result in DNA fragmentation and subsequent programmed neuronal cell death.