Alloreactive memory B cell detection by flow cytometric cross match using polyclonally activated memory B cell culture supernatants.

In addition to alloantibodies, alloreactive memory B cell (mBC) evaluation has a potential for immunological risk assessment during transplantation processes. For the alloreactive mBCs evaluation currently, direct Flow Cytometric (FC) analysis using the HLA tetramer staining is an option. Evaluation of alloantibodies produced by the polyclonally stimulated alloreactive mBCs in in vitro culture system seems to be another useful approach, but this needs further downstream applications. In this study, we investigated the usefulness of the Flow Cytometric Cross Match (FCXM-supernatant) in which in vitro polyclonally activated mBCs culture supernatants and potential donor's lymphocytes being used for the mBC detection. FCXM-supernatant assays were performed between culture supernatants of polyclonally activated mBCs obtained from 4 allosensitized multiparous women and 14 renal transplant patients, and their non-alloimmunized spouses' or donors' lymphocytes, and vice versa. HLA typing was performed by SSP method. Anti-HLA antibodies produced by in vitro activated alloreactive mBCs were also evaluated by the Luminex assays. The success of in vitro polyclonal activation of mBCs was evaluated by a total IgG ELISA test and antibody secreting cell analyses by FC. Donor specific alloreactive mBCs were detected by FCXM-supernatant in 45% of the 18 allosensitized cases. Detection rate was 85% (6 out of 7) in the strongly allosensitized cases. No alloreactive mBCs was detected in control cases without allosensitization. FCXM-supernatant negative results of the allosensitized cases were related to low level of allosensitization and insufficient polyclonal stimulation evaluated by total IgG antibody tests of the supernatants. We herein report a practical methodology for alloreactive mBC detection as a donor specific manner using the FCXM-supernatant assay so that this would easily be transformed into a routine test performed in tissue typing laboratories.

Comparison of Cytotoxic Flow Cytometric Cross Match With Complement Dependent Lymphocytotoxicity and Flow Cytometric Cross Match in Renal Transplant Patients.

Cytotoxic flow cytometric crossmatch (cFCXM), identified by detecting complement-mediated cytotoxic cell death in addition to the capability of showing the alloantibodies binding onto lymphocytes at the same time, can reduce the necessary time and workload in evaluating alloantibodies. More data from clinical samples are needed for cFCXM to be accepted by tissue typing laboratories. In this study, we compared cFCXM with complement-dependent lymphocytotoxicity and standard flow cytometric crossmatch in 41 renal pretransplant patients. A comparison of the obtained data was performed using Spearman's correlation test. We found that cFCXM showed no statistically significant differences with complement-dependent lymphocytotoxicity and flow cytometric crossmatch. We believe that cFCXM can be used in clinical laboratories in the near future following intra-laboratory validation.

Effects of Benign Prostate Hyperplasia on Allografts in Kidney Transplant.

OBJECTIVES: Benign prostate hyperplasia is frequently diagnosed in elderly chronic renal failure patients. Although it is asymptomatic because urine flow declines in those with chronic renal failure, it may become symptomatic when urine flow increases after renal transplant. Untreated benign prostate hyperplasia may lead to several complications that adversely affect the kidney allograft. MATERIALS AND METHODS: Our retrospective study investigated data from 286 male transplant patients. Data obtained included medical history, clinical examination, prostate-specific antigen, prostate volume, and residual urine volume results. Patients had completed an International Prostate Symptom Score questionnaire to assess the low urinary symptoms. Creatinine level, residual urine volume, International Prostate Symptom Score, and uroflow-metry results were reviewed. RESULTS: The average age of patients in our group was 54.8 years. Seven patients were diagnosed with benign prostate hyperplasia, and transurethral resection of the prostate was performed. The average creatinine levels before and after transurethral resection were 2.53 ± 0.76 mg/dL and 1.66 ± 1.12 mg/dL, respectively. Creatinine levels measured 6 months after transurethral resection versus before resection were statistically significant (P < .018). Furthermore, there was a statistically significant difference (P < .017) in the postmicturition residual urine volume between the preoperative and postoperative values, which were 132.14 ± 19.33 and 47.6 ± 18.6 mL, respectively. Maximum urine flow rates before and after transurethral resection were also significantly different (P < .017). CONCLUSIONS: Although the main reasons for graft function loss after renal transplant are rejection and infections, obstructive pathologies should also be considered. Especially for elderly patients, anamnesis, uroflowmetry, and digital rectal examination play critical roles in the evaluation of benign prostate hyperplasia before and after transplant.

Evaluation of Pre-Transplant Panel Reactive Antibody Levels and Sensitization: A Single-Center Study.

BACKGROUND Sensitization is one of the most important barriers against transplantation. Our aim was to evaluate the sensitization status of our patients awaiting cadaveric transplantation and to identify factors causing sensitization. MATERIAL AND METHODS A total of 140 patients on the cadaveric waiting list during January 2014 were included in this retrospective cross-sectional study. The parametric t-test and the non-parametric chi-square test were used to detect differences between PRA-positive and -negative patients. Multivariate analysis was used to identify factors associated with PRA positivity. One-way analysis of variance was used to compare PRA-negative and -positive results. RESULTS Anti-HCV positivity (p=0.040), history of transfusion (p=0.041), and mean number of blood product transfused (p=0.047) were significantly related to class 1 PRA positivity. History of transfusion (p=0.038) and mean number of blood product transfused (p=0.044) were related to class 2 PRA positivity. The multivariate analysis indicated that transfusion and more than 5 units of blood product transfused were related to either class 1 or class 2 PRA positivity. No associations were found between PRA positivity and pregnancy, transplantation, age, sex, infection, abortion, cardiovascular disease, diabetes mellitus, hepatitis B, or time spent on dialysis and being on the transplantation waiting list. CONCLUSIONS Anti-HCV positivity and transfusion are risk factors for sensitization. Particular emphasis should be given to sensitization and its prevention to reduce waiting time for transplantation.

Brown Tumors: A Case Report and Review of the Literature.

Brown tumors are focal bone lesions, encountered in patients with uncontrolled hyperparathyroidism. They can be located in any part of the skeleton. Clinically significant lesions in the craniofacial bones are rare. Craniofacial involvement may cause facial disfiguration and compromise social ease of the patient and normal functions, such as chewing, talking, and breathing. In this case report, we present a patient with a brown tumor of the craniofacial bones provoked by secondary hyperparathyroidism and review the last 10 years of craniofacial brown tumors associated with secondary hyperparathyroidism in the English literature.

Controversial issues in biliary pancreatitis: when should we perform MRCP and ERCP?

OBJECTIVES: The majority of bile duct stones (BDS) that cause acute biliary pancreatitis (ABP) pass spontaneously into the duodenum. If not passed, they worsen the prognosis or cause recurrence. Therefore, they must be treated. The purpose of this study was to assess the number and timing of spontaneous passage of BDS using magnetic resonance cholangiopancreatography (MRCP) and to determine the effect of this approach on endoscopic retrograde cholangiopancreatography (ERCP). METHODS: Sixty patients diagnosed with ABP were evaluated prospectively. MRCP was performed between the 1st and 4th days of an acute attack in all the patients. A control MRCP was performed after 7 days in patients with MRCP-identified choledocholithiasis. Patients in whom BDS were visible on imaging or who showed no decrease in bilirubin or cholestasis enzymes underwent ERCP. RESULTS: MRCP revealed choledocholithiasis in 20 (33%) of the 60 patients. In the control MRCP imaging, choledocholithiasis was detected in 16 of 20 (80% of those who had stone initially) patients. ERCP was performed in these patients and in 2 patients who did not have BDS on the control MRCP but whose bilirubin values and cholestatic enzyme levels had not decreased. ERCP verified choledocholithiasis in 16 of the 18 patients. The positive predictive value of MRCP was 93.7% (15/16). CONCLUSIONS: MRCP performed in the second week in ABP patients with a nonworsening prognosis and a suspicion of choledocholithiasis will give more specific results. This will avoid unnecessary ERCP and the potential morbidity and mortality that can develop with this invasive procedure.

Elective and emergency surgery in chronic hemodialysis patients.

PURPOSE: Aim of this study was to report our experience in elective and emergency surgery on chronic hemodialysis (CH) patients for end-stage renal disease (ESRD). METHODS: All patients on CH for ESRD who underwent various surgical procedures in our unit within the past 9-year period (2001-2010) were included in this study. These patients were divided into two groups according to the type of surgery performed: elective or emergency. Demographic data, indications for surgery, primary causes of ESRD, surgical procedures, postoperative complications, and mortality rates were studied. RESULTS: Of 130 patients, 121 underwent elective surgery while 10 were addressed for emergency operation. In the elective surgery group, the most common diseases were secondary hyperparathyroidism, kidney diseases, cholelithiasis, and diabetic foot gangrene. Complications occurred in nine patients (morbidity rate, 7%) and only one patient died (mortality rate, 0.8%). In the emergency surgery group, the most common diseases were diabetic foot gangrene and obstructed sigmoid colon cancer. In this group, complications occurred in seven patients (total morbidity rate, 70%) and two patients died (mortality rate, 20%). CONCLUSIONS: Elective surgery in patients on CH for ESRD can be performed with acceptable surgical risks provided careful preoperative preparation, intraoperative, and postoperative precautions are taken.

Predictors of hypocalcemia occurring after a total/near total thyroidectomy.

PURPOSE: The aim of this study was to identify the predictors of early postoperative hypocalcemia after a total/near total thyroidectomy in order to select patients for prompt treatment to prevent symptomatic hypocalcemia. METHODS: Patients with hypocalcemia within 24 h of surgery were identified as Group I and normocalcemic patients as Group II. The perioperative serum total calcium (tCa, ionized calcium (iCa) and intact parathormone (iPTH) were measured perioperatively. Skin closure (SC) was accepted as the reference time point. Data are expressed as the mean +/- SEM. RESULTS: The study included 73 patients. Hypocalcemia (Group I) was detected in 40 patients (54%) within the first 24 h postoperatively. Symptomatic hypocalcemia was detected in 40% of the patients in Group I. Intact parathormone values at 10 min of SC were significantly lower in Group I (P = 0.001). IPTH measurement at 10 min of SC showing a >/=30% decrease had a 92.3% sensitivity and 92.6% specificity in predicting hypocalcemia after a total/near total thyroidectomy. The postoperative day 15 mean tCa, iCa, and iPTH values were similar in both groups of patients. The mean iPTH level was 16.79 +/- 2.5 pg/dl at 10 min after SC in patients who developed symptomatic hypocalcemia. CONCLUSIONS: Intact parathormone measurement 10 min after SC is helpful to predict early postoperative hypocalcemia. An IPTH decrease >/=30% at this time point estimates the risk of postoperative hypocalcemia.