Simultaneous analysis of drugs administered to lung-transplanted patients using liquid chromatography-tandem mass spectrometry for therapeutic drug monitoring.

Several drugs are administered to lung-transplanted patients, which are monitored using therapeutic drug monitoring (TDM). Therefore, we developed and validated a liquid chromatography-tandem mass spectrometry method to simultaneously analyze immunosuppressive drugs such as mycophenolic acid, antifungal drugs such as voriconazole and itraconazole, and its metabolite hydroxyitraconazole. Chromatographic separation was achieved using a C18 column and gradient flow of mobile phase comprising 20 mM aqueous ammonium formate and 20 mM ammonium formate-methanol solution. A simple protein precipitation treatment was performed using acetonitrile/methanol and mycophenolic acid-2 H3 , voriconazole-2 H3 , itraconazole-2 H4 , and hydroxyitraconazole-2 H4 as internal standards. The linearity ranges of mycophenolic acid, voriconazole, itraconazole, and hydroxyitraconazole were 100-20,000, 50-10,000, 5-1000, and 5-1000 ng/mL, respectively. The retention time of each target was less than 2 min. The relative errors in intra- and inter-day were within ±7.6%, the coefficient of variation was 8.9% or less for quality control low, medium, and high, and it was 15.8% or less for lower limit of quantitation. Moreover, the patient samples were successfully quantified, and they were within the linear range of measurements. Therefore, our new method may be useful for TDM in lung-transplanted patients.

Plasma mycophenolic acid concentration and the clinical outcome after lung transplantation.

BACKGROUND: The therapeutic drug monitoring of mycophenolic acid (MPA) has been investigated for renal and heart transplantations; however, its usefulness in lung transplantation is unclear. METHODS: The MPA area under the plasma concentration-time curve (AUC) was calculated in 59 adult lung transplant recipients. The MPA AUC0-12 s were compared among the three groups determined by the presence of adverse events (no events, infection, and chronic lung allograft dysfunction [CLAD]). Next, MPA AUC0-12 thresholds for the adverse events were identified by receiver operating characteristic analysis. Cumulative occurrence rate of the adverse events was compared between two groups (adequate and inadequate groups) according to the thresholds. RESULTS: The MPA AUC0-12 s in the no event, infection, and CLAD groups were 30.3 ± 6.5, 36.8 ± 10.7, and 20.6 ± 9.6 µg·h/mL, respectively (P = .0027), while the tacrolimus trough levels were similarly controlled in the groups. The thresholds of MPA AUC0-12 for the occurrence of infection and CLAD were 40.5 and 22.8 µg·h/mL, respectively. The cumulative occurrence rate of adverse events of adequate group (15.3%) was significantly lower than that of inadequate group (56.0%) (P = .0050). CONCLUSIONS: The MPA AUC0-12 may affect the occurrence of adverse events in lung transplant recipients.

Improvement of native pulmonary alveolar proteinosis after contralateral single living-donor lobar lung transplantation: A case report.

PAP is a rare disease characterized by the accumulation of surfactant materials in the alveolar spaces due to the imbalance of surfactant homeostasis (production and clearance). We herein report a case of an 8-year-old girl who developed PAP after BMT from her mother for the treatment of DBA. The anemia was improved by BMT; however, respiratory dysfunction due to graft-versus-host disease gradually progressed. She eventually underwent right single LDLLT from her mother when she was 14 years old. A pathological examination of the excised lung confirmed the finding of diffuse bronchiolitis obliterans and unexpectedly revealed widespread alveolar proteinosis. Interestingly, the GGO of her native left lung on chest X-ray was improved after LDLLT. We present the very unique clinical course of this patient and discuss the mechanisms underlying the development of PAP after BMT and its improvement after LDLLT from the same donor.

Development of a precise quantitative method for monitoring sirolimus in whole blood using LC/ESI-MS/MS.

Sirolimus is used on patients after solid organ transplantation and on lymphangioleiomyomatosis (LAM) patients, and therapeutic drug monitoring is required in clinical practice. We have previously reported an accurate method for quantitative determination of sirolimus, but its sample preparation step was complicated. In this study, we developed a modified liquid chromatography/electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS) method for sirolimus quantification. A supported liquid extraction cartridge was used to purify sirolimus from whole blood and ion suppression was mostly prevented. The validation results met the acceptance criteria. This method was compared with the antigen conjugated magnetic immunoassay (ACMIA) and our previously reported method, using whole blood samples from LAM patients. Comparison of the Bland-Altman plots of the currently developed method and the previous method revealed no significant difference between the two methods (mean bias, -2.02%; 95% CI, -7.81-3.78). The values obtained using ACMIA were significantly higher than those obtained using the current method by 13.87% (95% CI, 6.49-21.25) owing to cross-reactivity. The degrees of cross reactivities in LAM patients and in organ transplant patients were similar, and our LC/ESI-MS/MS method precisely measured the blood concentrations of sirolimus.

Estimation of Blood Sirolimus Concentration Based on Tacrolimus Concentration/Dose Normalized by Body Weight Ratio in Lung Transplant Patients.

BACKGROUND: Sirolimus and tacrolimus require accurate drug dosing based on their target blood levels to produce better clinical outcomes, specifically, the avoidance of drug-induced adverse effects and the maintenance of efficacy. However, because the ideal dose of sirolimus and the schedule for measuring its blood levels are unclear in lung transplant patients, an index is required for estimating sirolimus blood concentrations. The aim of this work is to study the correlation between the trough concentration/dose normalized by body weight (C0/D) ratios of sirolimus and tacrolimus in lung transplant patients. METHODS: Thirteen lymphangiomyomatosis patients who underwent lung transplantation and were treated with sirolimus and tacrolimus from February 2015 to July 2018 were divided into 2 groups, one receiving twice-daily (TD, n = 6) and the other once-daily (OD, n = 7) tacrolimus formulations. The correlation between the C0/D ratio of sirolimus and patient background was evaluated using Spearman's rank correlation coefficient. Correlations between sirolimus and tacrolimus C0/D ratios or doses were analyzed by single regression analysis. RESULTS: Significant correlations were found between the C0/D ratios of sirolimus and tacrolimus. The regression equations from the initial data of TD and OD groups at steady state were y = 1.880x + 32.636 (adjusted R = 0.743, P = 0.017) and y = 1.684x + 38.816 (adjusted R = 0.919, P < 0.001), respectively. In addition, the regression equations from all data of TD and OD groups were y = 1.883x + 4.170 (adjusted R = 0.546, P < 0.001) and y = 1.950x + 43.188 (adjusted R = 0.898, P < 0.001), respectively. A significant correlation between the dosage of sirolimus and tacrolimus was observed only in the OD group, with relatively low accuracy. CONCLUSIONS: Blood sirolimus concentrations can be estimated using the C0/D ratio of tacrolimus, suggesting that the C0/D ratio of tacrolimus is an index of required sirolimus dosage and the frequency of blood sirolimus concentration measurements.

Limited Sampling Strategy for the Estimation of Mycophenolic Acid and its Acyl Glucuronide Metabolite Area under the Concentration-Time Curve in Japanese Lung Transplant Recipients.

PURPOSE: The dose of mycophenolate mofetil (MMF) used to prevent rejection after lung transplantation is often adjusted based on the 12-hour area under the concentration-time curve (AUC0-12) of mycophenolic acid (MPA). A limited sampling strategy (LSS) is useful to define the pharmacokinetic (PK) profiles of MPA and mycophenolic acid acyl glucuronide (AcMPAG). Therefore, this study aimed to design a LSS based on multiple linear regression for estimating the AUC0-12 of MPA and AcMPAG at the minimum blood sampling points in Japanese lung transplant patients with concomitant tacrolimus. METHODS: Forty-five lung transplantation recipients were enrolled in a PK study of MPA, mycophenolic acid glucuronide (MPAG), and AcMPAG. The plasma MPA, MPAG, and AcMPAG concentrations were determined just before and at 0.5, 1, 2, 4, 8, and 12 hours after dosing. The AUC0-12 of MPA and AcMPAG was calculated using a linear trapezoidal rule from the plasma concentration of each blood sampling time. LSS was used to develop models for estimated AUC in the model group (n = 23) and was evaluated in the validation group (n = 22). RESULTS: The best three time-point equation was 4.04 + 1.64·C1 + 3.08·C4 + 5.17·C8 for MPA, and -0.13 + 3.01·C1 + 3.51·C4 + 5.74·C8 for AcMPAG. The prediction errors (PE) and the absolute prediction errors (APE) were within the clinically acceptable ± 5% and 15% range, respectively (MPA: PE = 2.00%, APE = 11.66%, AcMPAG: PE = 0.98%, APE = 14.69%). The percentage of estimated AUC0-12 within ± 15% of the observed AUC0-12 was 77.27% for MPA and 81.82% for AcMPAG. CONCLUSION: LSS using three time-point (C1, C4, and C8) provides the most reliable and accurate simultaneous estimation of the AUC0-12 of MPA and AcMPAG in Japanese lung transplant patients.

[Successful Multidisciplinary Treatment Accomplished by Collaboration of Multiple Clinical Departments for Synchronous Quadruple Cancer].

A 60-year-old man consulted a clinic complains of sore throat. Squamous cell carcinoma of the hypopharynx and adenocarcinoma of the stomach were pointed out and he was refered to our hospital. As a result of detailed systemic examination, squamous cell carcinoma of the esophagus and squamous cell carcinoma of the right lung were also pointed out, which led to a diagnosis of synchronous quadruple cancer. On the basis of discussions among multiple clinical departments, systemic chemotherapy with cisplatin(CDDP), fluorouracil (5-FU) and docetaxel(DTX) was preceded locolegional therapies. After that, complete thoracoscopic right lower lobectomy and then a laparoscopic distal gastrectomy was performed. Radiation therapy was applied for hypopharyngeal cancer. Finally, endoscopic submucosal dissection for esophageal cancer was performed. Twenty months have passed since the last treatment, the patient is alive with a relapse-free condition.

Usefulness of Inflammation-Based Prognostic Score in Patients Undergoing Lung Metastasectomy for Colorectal Carcinoma.

OBJECTIVES: There is increasing evidence that Glasgow Prognostic Score (GPS), based on systemic inflammatory response and albumin level, is a useful predictor of overall survival in patients with various types of cancer. METHODS: Patients with lung metastasis from colorectal carcinoma who underwent a lung metastasectomy from 2000 to 2015 were retrospectively investigated. Routine laboratory measurements including serum C-reactive protein (CRP), albumin, and the tumor marker carcinoembryonic antigen were performed before the metastasectomy. RESULTS: Ninety-nine patients underwent 132 lung metastasectomy procedures during the study period. Kaplan-Meier analysis revealed that GPS (p = 0.017), number of metastases (p = 0.004), and the presence of liver metastasis (p = 0.010) were associated with overall survival, while univariate analysis selected GPS (p = 0.028), number of metastases (p = 0.005), and liver metastasis (p = 0.014) as predictive factors associated with overall survival. Multivariate analysis also indicated GPS (p = 0.004), number of metastases (p = 0.004), and liver metastasis (p = 0.013) as predictive factors associated with overall survival. CONCLUSION: In addition to number of metastases and liver metastasis, GPS is an important predictor of overall survival in colorectal cancer patients who undergo a lung metastasectomy.

[Infectious Diseases after Lung Transplantation].

Lung transplantation is getting to be a common treatment for the end-stage respiratory failure. Therefore we might encounter the various issues associated with infection under immunosuppression. At 1st, virus infections occur by Cytomegalovirus, Epstein-Barr virus, hepatitis B and C virus and John Cunningham virus. The 2nd, bacterial infections exacerbate graft condition by methicillin-resistant Staphylococcus aureus, Pseudomonas, Burkholderia cepacia and coagulase negative staphylococci. The 3rd, fungal infections are induced by Aspergillus, Candida and Cryptococcus. Lastly there is mycobacterial infection. These opportunistic infections contribute poor prognosis for lung transplant recipients, so that we have to manage these infectious diseases.

[Spindle Cell Sarcoma Originated in the Anterior Mediastinum;Report of a Case].

An anterior mediastinal sarcoma is a very rare type of mediastinal tumor. A 45-year-old female visited our hospital with swelling in the right anterior chest wall. Radiographically, the tumor was found to originate from the thymus and to infiltrate to the major pectoral muscle through the 1st intercostal space. Positron emission tomography revealed fluorodeoxyglucose accumulation at a standardized uptake maximum value of 16.1. Percutaneous needle biopsy showed the pathological findings of sarcoma. The tumor was resected along with the thymus and chest wall tissues including the right hemi-manubrium of the sternum, clavicle head, 1st and second ribs, and major pectoral muscle. After 4 months, tumor relapse occurred at the site of needle biopsy and additional resection was performed. At 10 months after the 2nd surgery, the patient was free from disease. From histologic and immunohistochemical findings, the tumor was diagnosed as spindle cell sarcoma.

Successful bilateral lung transplantation after 16 h of lung preservation with EP-TU solution: report of a case.

We report a case of bilateral lung transplantation (BLT) after preservation of the donor graft for 16 h 5 min with EP-TU, an extracellular phosphate-buffered lung preservation solution. The recipient was a 26-year-old woman with idiopathic pulmonary arterial hypertension and the graft ischemic time was prolonged significantly because of the time required to induce peripheral veno-arterial extracorporeal membrane oxygenation (V-A ECMO) under local anesthesia, and address the severe intrathoracic and pericardial adhesions from past surgery for partial anomalous pulmonary venous return, with concurrent annular plication of the tricuspid valve. After the operation, ECMO and continuous hemodiafiltration were started preemptively to protect the grafts against excessive edema. Postoperative chest X-ray showed diffuse bilateral infiltrates, which improved within a few days and she was weaned off ECMO on day 9. Successful BLT after a graft ischemic time of over 16 h has rarely been described in clinical lung transplantation.

[Two types of diaphragmatic plication; therapeutic plication and prophylactic plication].

Plication of the hemidiaphragm is effective to relieve respiratory symptoms such as exertional dyspnea and orthopnea of the patients with unilateral diaphragmatic paralysis. We have experiences of 2 types of diaphragmatic plication;one for the symptomatic patients with unilateral diaphragmatic paralysis, and the other for the patients who underwent phrenicotomy during extended surgery for a recurrent thymoma. The former plication was performed for a therapeutic purpose to alleviate the patient's symptoms. The latter was performed for a prophylactic purpose to keep the patient free from possible symptoms or postoperative morbidity after phrenicotomy. Case 1:A 65-year-old female with an eventration of the right diaphragm underwent a sternal resection and reconstruction for a large desmoid tumor. After continuous mechanical ventilation for 3 months, she was still under noninvasive positive pressure ventilation( NPPV) for 24 hours a day. Plication of the right hemidiaphragm was performed and the patient was successfully weaned from NPPV. Case 2:A 41-year-old female with myasthenia gravis underwent a surgery for a recurrent thymoma. Left phrenicotomy was necessary to achieve a complete resection of the tumor. Plication of the left diaphragm was performed during the tumor resection to prevent possible respiratory complications. Postoperative course was uneventful, and her vital capacity was well maintained.

[Contribution of surgical margin for surgical outcome of the chest wall tumors].

We present here our institutional review of surgical treatment for the chest wall tumors. Chest wall resections were performed on 80 patients, and subsequent chest wall reconstructions were performed on 45 patients. Primary malignant tumors in the chest wall required more extensive rib resections combined with the neighboring structures such as the sternum and the vertebral bones than benign or metastatic/recurrent tumors did. Postoperative mortality and morbidity occurred in 5 patients who underwent the sternal resection and the rib resection combined with the vertebral bodies. Primary malignant tumors in the chest wall are sarcomas originating from the bones, the cartilage tissues, and the soft tissues of the chest wall. We general thoracic surgeons may not have expertise in treating sarcomas,because primary malignant chest wall tumors are rare and a single institution has limited experiences in surgical treatment of such tumors. We should be aware that a surgical margin of primary malignant chest wall tumors is important to achieve excellent local control and better prognosis. We recommend cooperation with an orthopedic oncologist who is experienced with treating sarcomas. Not only preoperative planning but also intraoperative evaluation for the surgical margin with orthopedic oncologists is necessary for better surgical outcome.

Successful closure of an open-window thoracostomy wound by negative-pressure wound therapy: report of a case.

Negative-pressure wound therapy is a newly developed, noninvasive technique to manage a wide variety of wounds. This novel therapy was successfully used to heal the wound after open-window thoracostomy without surgical closure. A 46-year-old woman was admitted to hospital because of a painful mass on the right side of her chest. Radiological findings revealed an abscess on the right chest wall that had ruptured into the right lung and caused empyema. Antibiotic therapy did not yield sufficient improvement. Open-window thoracostomy was performed to achieve a sterile pleural space, and negative-pressure wound therapy was then applied. The pleural space was reduced, and the patient was discharged home with self-administered wound care. The wound healed completely in 5 months without any need for surgical closure.

Clinical application of an extracellular phosphate-buffered solution (EP-TU) for lung preservation: preliminary results of a Japanese series.

PURPOSE: We evaluated the effect of an extracellular phosphate-buffered lung preservation solution (EP-TU solution) on acute postoperative graft function and recipient survival in a Japanese series of cadaveric lung transplantation. METHODS: The subjects were ten patients who received lung grafts preserved with EP-TU solution at three of four designated lung transplant centers between 2000 and 2005. Three of the patients underwent single lung transplantation and seven underwent bilateral lung transplantation. Postoperative graft function was evaluated by primary graft dysfunction (PGD) grade (0-3), arterial oxygen tension/inspired oxygen fraction (PaO(2)/FiO(2)) ratio, and the previously reported chest roentgenogram score (CRS, 0-4) during the first 48 h after intensive care unit admission. Survival rates were also calculated. RESULTS: The average graft ischemic times for the first and second grafts were 431 and 571 min, respectively. The average PGD grade ranged from 0.5 to 1.0 and the average PaO(2)/FiO(2) ratio ranged from 350 to 400 Torr during the 48 h. The average CRS was always under 1.0. Survival rates at 30 days and 5 years were 90 and 80%, respectively. CONCLUSIONS: EP-TU solution promoted excellent postoperative lung graft function and enhanced recipient survival rates, despite a long average preservation time.

[Thoracoscopic thymectomy for thymoma].

Complete resection of Masaoka stage I and II thymoma leads to good prognosis. In recent years, technological development of endoscopy enables thymectomy by thoracoscopic method instead of median sternotomy approach. We perform thoracoscopic thymectomy for thymoma without myasthenia gravis via 2 trocars introduced in unilateral pleural cavity and a small incision on epigastric region with sternal lifting. Thoracoscopic thymectomy for thymoma with myasthenia gravis is performed by bilateral pleural cavity approach with sternal lifting. We had 18 patients with thymoma resected by thoracoscopic thymectomy between 2005 and 2007. Fourteen patients had thymoma with myasthenia gravis. Nine patients had Masaoka I thymoma, 8 had Masaoka II thymoma and 1 had Masaoka III thymoma. The average follow-up period of the patients is 51 months. One patient had a recurrence tumor 56 months after operation. It was developed on the left phrenic nerve from which the resected thymoma had removed. Thoracoscopic thymectomy can be expected as standard surgical treatment for Masaoka I and II thymoma.

[Clinicopathological features of peripheral small-sized (2 cm or less in diameter) non-small cell lung cancer].

Ninety-three patients with completely resected peripheral non-small cell lung cancer, clinically diagnosed 2 cm or less in diameter, were retrospectively reviewed. Their preoperative computed tomography (CT) and positron emission tomography (PET) findings, carcinoembryonic antigen (CEA) values, clinico-pathological features and postoperative outcomes were analysed. Ground-glass opacity (GGO) ratio( soft tissue density area of the tumor/maximum area of the tumor in diameter) was measured. The overall survival rate at 3 years was 93.3% and the relapse-free survival rate at 3 years was 89.4% with a median follow-up period of 38.5 months. Patients with GGO ratio 0.25 or less had no lymph node (LN) involvement nor lymph vascular invasion. Only 2 of them (8%) had vascular invasion. Fisher's exact probability test revealed CEA ≥ 5 ng/ ml as risk factor for LN involvement( p=0.0400). Multiple logistic regression analysis showed that solid adenocarcinoma and squamous cell carcinoma recurred more frequently than adenocarcinoma with GGO (p=0.0619, odds ratio 4.969, 95%CI 0.9242~37.67).

Living Donor Lobar Lung Transplant for a Patient With Lung Disease Caused by ABCA3 Gene Mutations: A Case Report.

Recessive gene mutations in ABCA3 cause lethal neonatal respiratory distress, and pediatric and adult interstitial lung disease. The effectiveness of medical treatments is limited and a subset of such patients will eventually require lung transplantation. A 20 months old boy developed interstitial lung disease and was treated with hydroxychloroquine, which had a significant effect. Sequence analysis of ABCA3 gene revealed newly discovered compound heterozygous mutations. His respiratory dysfunction gradually progressed over years and he underwent living-donor lobar lung transplantation (LDLLT) at 8 years of age with his parents serving as bilateral lobar donors. The parents had been genetically examined beforehand and found to be carriers who had one allele with an ABCA3 gene mutation and the other with no mutation. The recipient has been well without chronic lung allograft dysfunction and his parents have been enjoying healthy social lives for 7 years since the operations. LDLLT appears to be a valid option for selected children with ABCA3 gene mutations who are too ill to wait for cadaveric lung transplantation. When relatives of the recipient with ABCA3 gene mutation are deemed potential donors for LDLLT, sequence analyses of the donors are indispensable to exclude the possibility that they are late-onset patients of this recessive hereditary disease.

A total pleural covering technique in patients with intractable bilateral secondary spontaneous pneumothorax: Report of five cases.

We herein present five cases of bilateral intractable secondary spontaneous pneumothorax associated with chronic severe lung diseases that were successfully treated with a modified form of a previously reported surgical procedure, the "total pleural covering technique," under video-assisted thoracic surgery. We performed the total pleural covering technique modified with a preceding coverage of air-leak points with polyglycolic acid sheets. In this series, the median length of surgery was 106 min (range: 67-220 min) on the unilateral side (10 sides). No significant surgical complications were observed, but one patient died on day 23 after the operation, due to respiratory insufficiency on the basis of the underlying lung disease. The remaining four patients have been followed up regularly (mean follow-up time: 23 months; range: 1-54 months) and there has been no recurrences of pneumothorax. We believe that the total pleural covering technique is a useful method; however, special attention should be paid to the underlying disease in order to identify patients who would be most likely to benefit from the procedure.

The intensity of bronchiolar epithelial cell injury caused by an alloimmune response is ameliorated by transbronchial human interleukin-10 gene transfer in a rat model of lung transplantation.

The aim of the present study was to determine whether the intensity of bronchiolar epithelial cell injury is ameliorated by transbronchial human IL-10 (hIL-10) gene transfer in a rat model of lung allograft rejection. The left lung was extracted from a donor Brown Norway rat and transbronchially instilled with encoding hIL-10 (IL-10 group) or ß-galactosidase (control group). The lung graft was then transplanted into a Lewis rat and harvested on day 6 after transplantation. The allografts were histologically examined and all bronchioles in the slides were assigned one of three grades according to the intensity of epithelial cell loss. The distribution of the grades was significantly different between the two groups, and the epithelial cell injury was significantly improved in the IL-10 group. The present study demonstrated the effect of transbronchial hIL-10 gene transfer on ameliorating bronchiolar epithelial cell injury in a rat model of lung allograft rejection.