Analytical method for detection and quantification of new emerging drug etaqualone in human blood and urine by gas chromatography tandem mass spectrometry

Analytical procedure for detection and quantification of etaqualone in human blood and urine using GC–MS/MS was established and applied to authentic human samples obtained from volunteers. A liquid–liquid extraction method was employed. Each 1.0 mL of blood or urine was alkalized and extracted with diethyl ether. The solvent layer was evaporated to dryness and reconstituted with methanol then analyzed by GC–MS/MS. linear relationships within the concentration range of 1–100 ng/mL were obtained in calibrators for both blood and urine, demonstrating correlation coefficients values being>0.999. For blood and urine samples, the intra-day assay precision and accuracy values are each less than 3.65%, 7.13%, and 6.02%, 9.12%; those values of the inter-day assay are each less than 1.82%, 6.74%, and 3.99%, 7.41%. The extraction recovery rates for etaqualone ranged from 98.7% to 106%. The lower limit of quantifications was 1.0 ng/mL in both blood and urine. Stabilities of etaqualone in blood and urine were satisfactory under various temperatures within 15 days. 8.51 and 2.06 ng/mL of etaqualone in blood and urine were detected at 4 h later oral ingestion; 6.91 and 3.94 ng/mL of etaqualone were also detected 30 min and 2 h later smoking from blood and urine.     

Incidence and predictors of post-reperfusion syndrome in living donor liver transplantation

A characteristic pattern of hemodynamic changes that may occur in reperfusion phase of liver transplantation (LT) is known as post-reperfusion syndrome (PRS). In this study, we determined the frequency of PRS and evaluated possible predictors of PRS. The medical records of 152 patients who underwent living donor LT were reviewed. PRS was defined as a decrease in mean arterial pressure of more than 30% from the baseline value for more than one min during the first five min after reperfusion. The frequency of PRS was determined, and patients were divided into two groups: PRS group and non-PRS group. Donor factors, preoperative and intraoperative recipient factors, and postoperative outcomes were compared between the two groups. PRS occurred in 58 recipients (34.2%). Preoperative model for end-stage liver disease scores of recipients and percentage of graft steatotic changes were higher in PRS group. PRS group showed higher heart rates and lower hemoglobin values preoperatively. Before reperfusion, PRS group received more transfusion and their urine output was less than that of non-PRS group. Postoperatively, peak bilirubin during the first five d after LT was higher in PRS group. In conclusion, both severity of liver disease and graft steatosis may increase risk for PRS in LT. Further prospective studies of PRS in its relationship to outcome are indicated.     

Worldwide variability in deceased organ donation registries

The variability in deceased organ donation registries worldwide has received little attention. We considered all operating registries, where individual wishes about organ donation were recorded in a computerized database. We included registries which recorded an individual's decision to be a donor (donor registry), and registries which only recorded an individual's objection (non-donor registry). We collected information on 15 characteristics including history, design, use and number of registrants for 27 registries (68%). Most registries are nationally operated and government-owned. Registrations in five nations expire and require renewal. Some registries provide the option to make specific organ selections in the donation decision. Just over half of donor registries provide legally binding authorization to donation. In all national donor registries, except one, the proportion of adults (15+) registered is modest (<40%). These proportions can be even lower when only affirmative decisions are considered. One nation provides priority status on the transplant waiting list as an incentive to affirmative registration, while another nation makes registering a donation decision mandatory to obtain a driver's license. Registered objections in non-donor registries are rare (<0.5%). The variation in organ donor registries worldwide necessitates public discourse and quality improvement initiatives, to identify and support leading practices in registry use.     

Significance of low-level DSA detected by solid-phase assay in association with acute and chronic antibody-mediated rejection

We sought to clarify the controversial issue of whether detecting low‐level anti‐donor‐specific HLA antibody (HLA‐DSA) by single‐antigen flow‐bead assay (SAFB) may have a potential role in reducing acute and chronic antibody‐mediated rejection (AMR). We retrospectively studied the preoperative serum of ABO‐compatible living kidney transplantation recipients transplanted between 2001 and 2004 by SAFB using a Luminex platform. HLA‐DSA was detected only by SAFB in 24 patients, although all of them showed negative T‐cell and B‐cell complement‐dependent cytotoxicity (CDC) crossmatches. The HLA‐DSA patients went on to have surprisingly high levels of acute and chronic AMR despite being only weakly sensitized (acute AMR, 33.3%; chronic AMR, 41.7%). After 2005, we implemented SAFB routinely and any patient having a positive HLA‐DSA was considered to be a desensitization candidate. The 52 patients found to have HLA‐DSA underwent kidney transplantation after prior treatment with a single dose of rituximab (RIT) and three or four sessions of double‐filtration plasmapheresis (DFPP) in addition to regimens commonly used between 2001 and 2004. After 2005, there was a significant reduction in the occurrence of acute and chronic AMR (acute AMR, 4.7%, P < 0.001; chronic AMR, 4.7%, P < 0.001). The 5‐year graft survival rate also improved after implementing SAFB (83.3–98.1%, P = 0.032). The RIT/DFPP‐induction protocol may improve graft survival even in patients with low‐level DSA.     

Preemptive deceased donor kidney transplant not associated with patient survival benefit in minority kidney transplant recipients

Previous studies have shown an inverse association between pre-transplant dialysis exposure and post-kidney transplant outcomes. Socioeconomic and allocation factors, in contrast to medical factors, play a greater role in dialysis exposure among minorities, and medical causes for delay may impact post-transplant outcomes. This study sought to test whether minorities behaved similarly to Caucasians with regard to the effect of duration of dialysis on post-transplant outcomes. All primary deceased donor kidney transplants between 1997 and 2004 (n = 54,162) were analyzed from the Organ Procurement and Transplant Network database and were categorized as either Caucasian or minority. Adjusted patient and graft survivals were determined in each subgroup based on the duration of pre-transplant dialysis. Caucasians recipients show a clear stepwise increase in risk of graft failure and death with increasing duration of dialysis. The risk of graft failure among minorities increased less without a clear stepwise pattern. The risk of death, however, showed a U-shaped risk profile with the highest risk of death among preemptive transplants and recipients with more than five yr of dialysis. The disparate effect of dialysis on minorities suggests that a selection bias and not a biologic effect may explain the association between dialysis duration and outcomes after kidney transplantation previously reported.     

Laparoscopic live donor nephrectomy: Current practice and results of renal transplantation

Objective: In 2009, 1659 patients with end‐stage renal failure in Hong Kong were waiting for a renal transplant. The overall number of renal transplants carried out locally remains low, with an even lower number being live donor donations. Yet, live donor kidney transplantation yields results that are consistently superior to those of deceased donor kidney transplantation, and laparoscopic donor nephrectomy (LDN) is increasingly accepted worldwide as a safe and preferred surgical option. We aim to evaluate the outcome of LDN in our setting, and to compare with that of deceased donors in this retrospective review. Patients and Methods: A total of 12 patients received LDN over the study period of 2006–2009. Standard left transperitoneal LDN was carried out. Grafts including three with double vessels were prepared using the bench technique. The postoperative outcomes up to 1 year for both the donors and the recipients were studied. Contemporary results for the 47 deceased donor kidneys were studied and compared. Results: All donors had an eventful recovery. The operating time was 225.0 ± 67.4 min. The hospital stay was 5.6 ± 2.3 days. The recipient outcomes including hospital stay and creatinine levels at discharge and 1 year were 11 days, 121 umol/L and 116 umol/L, respectively. Specifically, no ureteric stricture or graft loss was noted at the 1‐year follow up. Recipient complications included haematoma (1 patient), renal artery stenosis (1 patient) and redo of vascular anastomosis (1 patient). In contrast, the deceased donor graft recipients had a hospital stay of 11 days, and creatinine levels of 205 umol/L on discharge and 205 umol/L at 1 year, respectively. The delayed graft function rates for the live donor and deceased donors group were 0% and 14.9%, whereas the 1‐year graft survival rates were 100% and 87.2% respectively. Conclusion: The results showed that the donor morbidity rate was low, as reflected by the short hospital stay. Also, the overall parameters of recipients were good. In particular, no ureteric stricture was noted, and graft survival was 100% at 1 year. Living donor kidney transplant program using the laparoscopic technique is a viable option to improve the pool of kidneys for transplantation.     

Donor‐Derived Fungal Infections in Organ Transplant Recipients: Guidelines of the American Society of Transplantation,Infectious Diseases Community of Practice†

Donor‐derived fungal infections can be associated with serious complications in transplant recipients. Most cases of donor‐derived candidiasis have occurred in kidney transplant recipients in whom contaminated preservation fluid is a commonly proposed source. Donors with cryptococcal disease, including those with unrecognized cryptococcal meningoencephalitis may transmit the infection with the allograft. Active histoplasmosis or undiagnosed and presumably asymptomatic infection in the donor that had not resolved by the time of death can result in donor‐derived histoplasmosis in the recipient. Potential donors from an endemic area with either active or occult infection can also transmit coccidioidomycosis. Rare instances of aspergillosis and other mycoses, including agents of mucormycosis may also be transmitted from infected donors. Appropriate diagnostic evaluation and prompt initiation of appropriate antifungal therapy are warranted if donor‐derived fungal infections are a consideration. This document discusses the characteristics, evaluation and approach to the management of donor‐derived fungal infections in organ transplant recipients.     

Pulsatile Pump Decreases Risk of Delayed Graft Function in Kidneys Donated After Cardiac Death

Organ storage techniques have been under scrutiny to determine the best preservation method, particularly in donation after cardiac death (DCD) kidneys. Conflicting literature on the benefit of pulsatile perfusion (PP) over cold storage (CS) warrants further investigation. We analyzed the risk of developing delayed graft function (DGF) in recipients of DCD and donation after brain death (DBD) kidneys undergoing PP or CS. We stratified on basis of cold ischemic time (CIT) to determine the interaction of preservation techniques, CIT and DCD kidneys on developing DGF. Of 54 136 recipients, 4923 received DCD kidneys of which 3330 (67%) underwent PP. Of 49 213 DBD recipients, 7531 (15%) underwent PP. DCD had a higher risk of DGF versus DBD (adjusted odds ratio, AOR 3.2; 3.0–3.5). PP kidneys had less DGF (AOR 0.59; 0.56–0.63) compared to CS. Interaction models of method by donor type referenced to PP/DBD revealed CS/DBD kidneys had higher DGF (AOR 1.8; 1.7–1.9), whereas CS/DCD kidneys had the highest risk of DGF (AOR 5.01; 4.43–5.67). Even though suggestive for a benefit of PP on DGF, this retrospective analysis cannot address whether this is an intrinsic effect of PP or is associated with the logistics of PP such as discard of DCD kidneys based on pump parameters.     

RT-nPCR Assays for Amplification and Sequencing of VP1 Genes in Human Enterovirus A-D from Clinical Specimens

ObjectiveTo develop RT-nPCR assays for amplifying partial and complete VP1 genes of human enteroviruses (HEVs) from clinical samples and to contribute to etiological surveillance of HEV-related diseases.MethodsA panel of RT-nPCR assays, consisting of published combined primer pairs for VP1 genes of HEV A–C and in-house designed primers for HEV-D, was established in this study. The sensitivity of each RT-nPCR assay was evaluated with serially diluted virus stocks of five serotypes expressed as CCID₅₀ per μL and copies per μL, and the newly established methods were tested in clinical specimens collected in recent years.ResultsThe sensitivity of RT-nPCR assays for amplifying partial VP1 gene of HEVs was 0.1 CCID₅₀ per μL and 10 virus copies per μL, and for the complete VP1 gene was 1 CCID₅₀ per μL and 100 virus copies per μL, using serially-diluted virus stocks of five serotypes. As a proof-of-concept, 25 serotypes were identified and complete VP1 sequences of 23 serotypes were obtained by this system among 858 clinical specimens positive for HEVs during the past eight surveillance seasons.ConclusionThis RT-nPCR system is capable of amplifying the partial and complete VP1 gene of HEV A–D, providing rapid, sensitive, and reliable options for molecular typing and molecular epidemiology of HEVs in clinical specimens.     

3D后腹腔镜下活体供肾切取术的临床分析

目的  总结3D后腹腔镜下活体供肾切取术的经验,并探讨其临床效果和安全性。 方法  收集19例3D后腹腔镜下活体供肾切取术的临床资料。记录手术时间、术中失血量、肾脏热缺血时间、肾动脉长度、肾静脉长度、输尿管长度、切口长度、手术并发症。观察供、受者术后肾功能的情况等。 结果  19例活体供肾切取术均在3D后腹腔镜下顺利完成,无术中改为常规腹腔镜和中转开放者。3D后腹腔镜下活体供肾切取术手术时间80.5～125.2(平均102.3)min;术中出血量40.6～90.4(60.8)ml;肾脏热缺血时间100～230(161)s。供肾动脉长度2.6～3.2(2.9)cm;供肾静脉长度2.2～3.0(2.6)cm;供肾输尿管长度8～13(10)cm;切口长度约5～6 cm,伤口愈合良好;供者术后24 h尿量2 000～2 500 ml;术后3 d查血清肌酐轻度增高1例,术后7 d和1个月复查血清肌酐恢复正常。术后住院时间5～7(6)d。移植手术均获成功,未发生移植肾功能延迟恢复。 结论  3D腹腔镜手术系统可有效提高术中操作的精准性,3D后腹腔镜下活体供肾切取术安全可行。