The effect of HA330 hemoperfusion adsorbent method on inflammatory markers and end-organ damage levels in sepsis: a retrospective single center study.

OBJECTIVE: In this study, we aimed at evaluating the impact of HA330 hemoperfusion adsorbent application on inflammatory markers and end-organ damage markers in patients with sepsis/septic shock. PATIENTS AND METHODS: Patients who were diagnosed with sepsis/septic shock and treated with HA330 hemoperfusion adsorbent in addition to the standard treatment were included in this retrospective study conducted at Inonu University Turgut Ozal Medical Center between January 1, 2019 and January 1, 2021. RESULTS: A total of 150 patients were included in the study. The mean±SD age of the patients was 51.9±17.7 years. 102 patients (68%) were in septic shock. Mean±SD APACHE II scores were 15.3±4.8. The need for mechanical ventilation was noted in 64 patients (42.7%). WBC, neutrophil count, hemoglobin, platelet count, BUN, creatinine, AST, ALT, CRP and procalcitonin levels were measured before and after the procedure. Overall, 104 patients (69.3%) died median (min-max) 2.5 (1-114) days after the cytokine adsorption, while 46 patients (30.7%) recovered from sepsis and were discharged. The increase in BUN levels and decrease in platelet count after the procedure were statistically significant (p≤0.001, 0.041, respectively) in the overall study population. The laboratory findings in 46 survivors indicated significantly decreased AST and ALT levels after cytokine adsorption compared to baseline pre-treatment levels. WBC, neutrophil count, CRP, procalcitonin, BUN and creatinine values were also decreased after cytokine adsorption in survivors, whereas the change was not statistically significant. There was also a non-significant tendency for an increase in platelet count and hemoglobin levels after cytokine adsorption compared to pre-treatment values in these patients. CONCLUSIONS: Although no effect of HA330 hemoperfusion application on inflammatory markers and end-organ damage markers was demonstrated in our study, we used the HA330 hemoperfusion adsorbent method as a last resort in terminal patients with a mortality rate of approximately 90% and for whom antibiotic treatment did not benefit. Therefore, multicenter, prospective studies are needed to clarify the effect of early HA330 hemoperfusion use in the treatment of sepsis.

Incidentally Detected Gastric Gastrointestinal Stromal Tumor during Living Donor Liver Transplant Surgery for Hepatocellular Carcinoma: The First Two Cases.

Coexistence of hepatocellular carcinoma and gastrointestinal stromal tumor is rare. In this case series, we aimed to present an unusual coincidence of a gastrointestinal stromal tumor and hepatocellular carcinoma in patients who underwent living donor liver transplantation for hepatocellular carcinoma who had an incidental gastric gastrointestinal tumor which was detected intraoperatively.

Multidrug-Resistant Bacterial Sepsis and Inferior Vena Cava Thrombosis in Liver Transplant Recipients Used Synthetic Vascular Graft: Three Fatal Cases.

Synthetic vascular grafts are commonly used in liver transplantation. Thrombosis is a possible complication of using expanded polytetrafluoroethylene (e-PTFE) grafts. Herein, we report on 3 cases of liver recipients who died of intermittent sepsis episodes emerged concurrently with the thrombosis in synthetic vascular grafts and inferior vena cava (IVC) vein. Right lobe liver transplantation from living donors was performed for 3 patients by using e-PTFE grafts between the liver and IVC. Although heparin had been administered, thrombosis was developed in vascular graft and IVC extending to the right atrium; it was developed within 1-4 months of transplantations. All 3 patients suffered from recurrent sepsis episodes (4, 5, and 6 attacks for each patient) by different multidrug-resistant bacterial species. Treatment attempts including thrombolytic and antimicrobial drugs made, and surgical, endoscopic and radiological interventions could not resolve the clinical situation. The patients died of septic complications. We concluded that severe recurrent sepsis attacks may develop in liver transplant recipients when IVC and synthetic vascular graft were thrombosed. Removing the e-PTFE graft may be benefit for the treatment.

Endoscopic treatment of biliary complications after living donor liver transplantation in a high volume transplant center in Turkey; a single-center experience.

BACKGROUND AND AIM: Biliary complications are an important cause of mortality and morbidity after living donor liver transplantation (LDLT). We present our endoscopic treatment results after LDLT as a single center with high volume. METHODS: Patients who underwent endoscopic retrograde cholangiopancreatography (ERCP) after LDLT between 2005 and 2015 were included. Clinical data included patient demographics, ERCP indications (stricture or leak), and treatment outcomes, including need for percutaneous and surgical interventions. RESULTS: ERCP was performed in 446 (39.2%) patients with duct-to-duct anastomosis of 1136 LDLT patients. The most common biliary complication was stricture ± stone (70.6%, 315/446). Stricture and leak occurred in 60 (13.4%) patients. Only biliary leak was found in 40 (8.9%) patients. Our endoscopic treatment success rate in patients with biliary stricture after LDLT was 65.1%. Overall endoscopic success rates in our patients were 55.0% in patients with both leak and stricture, and only leak. In all, our percutaneous transhepatic biliary interventions (PTBI) and ERCP success rate was 90.6% in patients with biliary complications after LDLT. CONCLUSIONS: Endoscopic treatments are highly effective for biliary complications after LDLT. Effective use of percutaneous interventions in collaboration with endoscopic treatments significantly reduces the need for surgical treatment.

Living Donor Re-transplantation for Repeated Acute Liver Failure.

Emergency liver transplantation (LT) for acute liver failure (ALF) is a life-saving treatment. Occurrence of this situation in the same patient twice is very rare. Herein, we describe a patient who underwent two emergency LTs for ALF, both from living donors. When she was 26 years old, she underwent a right lobe living donor LT (LDLT) from her sister for ALF due to use of herbal weight loss medications. The next 3 years were uneventful but another ALF developed during a terminal stage pregnancy (37th week). Despite medical treatment, her liver functions worsened, and the baby was delivered by caesarean section. The second time, her brother was the donor and she recovered after the emergency right lobe re-LDLT. Both patient and baby were well at the 2-month follow-up. As far as we know, there is no reported similar case, and we concluded that LDLT is a paramount treatment option for both primary and secondary ALFs.

Pregnancy After Liver Transplantation: Risks and Outcomes.

OBJECTIVE: The aim of this study was to evaluate the outcomes of liver transplant recipients who became pregnant after transplantation. METHODS: The clinical data of all patients who underwent liver transplantation between January 2007 and December 2016 in our liver transplantation institute were reviewed. The following data were analyzed: indications for transplantation, recipient age at the beginning of pregnancy, the interval between transplantation and pregnancy, maternal and fetal complications, type of delivery, the health condition of neonates, and modifications in immunosuppressive therapy. RESULTS: During the study period, 1890 patients underwent liver transplantation. There were 185 women (9.8%) in childbearing age (15-45 years old), and 18 (9.7%) of them became pregnant during the study period. There were a total of 26 pregnancies. The mean age of patients at the time of operation was 25.3 ± 5.2 years, and the mean interval between operation and conception was 32.7 ± 15.3 months. Seventeen pregnancies (65.4%) ended in a live birth in the study. Six pregnancies (23%) resulted with no maternal or fetal complications. The most frequent maternal complication during pregnancy was pregnancy-induced hypertension (n = 3; 16.6%). CONCLUSIONS: Despite advances in immunosuppressive therapy and increasing experience in the management of these patients, pregnancies in liver transplant recipients are still more risky than in the general population for both the mother and the fetus. Thus, the issues related to fertility should be comprehensively discussed with the patients and their partners, preferably before transplantation, and pregnancies in liver transplant recipients should be followed up more carefully by a multidisciplinary team.

Telescopic Biliary Reconstruction in Patients Undergoing Liver Transplantation With 1-Year Follow-up.

BACKGROUND: Biliary complications are important during liver transplantation because of their effect on recipient and graft survival, incidence, and the long treatment period. These complications are associated with 50% morbidity and 30% mortality rates in recent studies. One of the most important reasons for biliary anastomosis complications is arterial ischemia. We present the results of our telescopic biliary anastomosis technique performed on the mucosa of the main biliary duct. PATIENTS AND METHODS: Fifty-six cases of telescopic biliary reconstruction were performed in 203 patients during 2015. Fifty cases and 52 patients who underwent standard reconstruction were chosen and compared. All patients had been scanned retrospectively. Statistical analyses were conducted with χ2 and Mann-Whitney U tests for the complications that occurred during the first 3 months. A P value <.05 was considered significant. RESULTS: No clinical or demographic differences were detected between the groups. About 90% of both groups were living donor liver transplantation cases. Five (10%) anastomotic leaks occurred in telescopic reconstruction group (n = 50), and 13 (25%) occurred in the standard reconstruction group (n = 52; P < .05). CONCLUSION: The arterial blood supply is better if the biliary anastomosis is made on the mucosal side of the main biliary duct. Early period anastomotic leaks may decrease significantly.

Can Patients Who Develop Cerebral Death in Fulminant Liver Failure Despite Liver Transplantation Be Previously Forseen?

BACKGROUND: The outcome of medical treatment is worse in fulminant liver failure (FLF) developing on acute or chronic ground. Recently, liver transplantations with the use of living and cadaveric donors have been performed in these diseases and good results obtained. In this study, we aimed to present the factors affecting the recovery of cerebral functions after liver transplantation in hepatic encephalopathy (HE) developing in FLF, to identify irreversible patient groups and to prevent unnecessary liver transplantation. METHODS: In Inonu University's Liver Transplant Institute, 69 patients who made an emergency notice to the National Coordination Center for liver transplantation owing to FLF from January 2012 to December 2015 were included in the study. Patients were divided into 2 groups. Group 1 consisted of 52 patients who underwent liver transplantation and recovered normal brain function, and group 2 had 17 patients who underwent liver transplantation and did not recover normal brain function and had cerebral death. All patients were evaluated before surgery for clinical encephalopathy stage, light reflex, and convulsions. Groups were compared and assessed according to age (>40, 10-40 and <10 years), body mass index, etiologic factor, preoperative laboratory values, transplantation type, mortality, and encephalopathy level. Multivariate analysis was done for specific parameters. RESULTS: Prothrombin time (PT), international normalized ratio (INR), and total bilirubin values were significantly different between the groups. There was no significant difference between the groups regarding ammonia and lactate levels. There was a statistically significant difference between the groups regarding sodium and potassium levels from serum electrolytes. However, the averages of both groups were within normal limits. pH and total bilirubin levels were meaningful for multivariate analysis. CONCLUSIONS: HE reversibility, mortality, and morbidity are important in patients with HE who undergo liver transplantation. Therefore, West Haven clinical staging and serum INR, PT, and total bilirubin level may be helpful in predicting the reversibility of FLF patients with HE before liver transplantation. It was determined that West Haven encephalopathy grading is important in determining the reversibility of HE after transplantation in FLF; especially the probability of reversibility of stage 4 HE decreases significantly. High PT and INR levels, hyperbilirubinemia, and serum sodium and potassium concentrations were risk factors for the reversibility of HE in this study.

Surgical Treatment of Portal Vein Thrombosis With the Use of Cadaveric Venous Patch After Donor Hepatectomy: A Case Report.

Live donors should be the priority of transplant professionals to prevent surgery-related morbidity and mortality during living-donor liver transplantation. Portal vein thrombosis after donor hepatectomy is an important complication which can be prevented by careful preoperative as well as perioperative evaluation. If portal vein thrombus occurs after donor hepatectomy, anticoagulation and surgical thrombectomy and even portal vein reconstruction should be kept in mind. Cadaveric venous patches can be used for the reconstruction of narrowed and angulated portal veins. Here we report the surgical treatment of a donor with a cadaveric venous patch who developed portal vein thrombosis after donor hepatectomy.

Rapid Detection of Bloodstream Pathogens in Liver Transplantation Patients With FilmArray Multiplex Polymerase Chain Reaction Assays: Comparison With Conventional Methods.

BACKGROUND: Bloodstream infection (BSI) is an important concern in transplant patients. Early intervention with appropriate antimicrobial therapy is critical to better clinical outcome; however, there is significant delay when conventional identification methods are used. METHODS: We aimed to determine the diagnostic performance of the FilmArray Blood Culture Identification Panel, a recently approved multiplex polymerase chain reaction assay detecting 24 BSI pathogens and 3 resistance genes, in comparison with the performances of conventional identification methods in liver transplant (LT) patients. A total of 52 defined sepsis episodes (signal-positive by blood culture systems) from 45 LT patients were prospectively studied. RESULTS: The FilmArray successfully identified 37 of 39 (94.8%) bacterial and 3 of 3 (100%) yeast pathogens in a total of 42 samples with microbial growth, failing to detect only 2 of 39 (5.1%) bacterial pathogens that were not covered by the test panel. The FilmArray could also detect additional pathogens in 3 samples that had been reported as having monomicrobial growth, and it could detect Acinetobacter baumannii in 2 samples suspected of skin flora contamination. The remaining 8 blood cultures showing a positive signal but yielding no growth were also negative by this assay. Results of MecA, KPC, and VanA/B gene detection were in high accordance. The FilmArray produced results with significantly shorter turnaround times (1.33 versus 36.2, 23.6, and 19.5 h; P < .05) than standard identification methods, Vitek II, and Vitek MS, respectively. CONCLUSIONS: This study showed that the FilmArray appeared as a reliable alternative diagnostic method with the potential to mitigate problems with protracted diagnosis of the BSI pathogens in LT patients.

Progressive Multifocal Leukoencephalopathy after Three Consecutive Liver Transplantations.

Progressive multifocal leukoencephalopathy (PML) is a lytic infection of the central nervous system caused by the reactivation of John Cunningham Virus (JCV) in severely immunosuppressed patients. Occurrence of PML after solid organ transplantations, especially after liver transplantation, is rare. If a patient has poor prognostic factors such as atypical radiological involvements or high viral load in cerebrospinal fluid (CSF), overall survival rates could be poor. Herein, we report on a patients who underwent liver transplantation three times and developed PML with unexpected radiological findings; he was also positive for JCV DNA with a high viral load. Although there are limited data about efficacy of cytarabine against JCV, it was given to the patient for five days. Despite the initiation of cytarabine and complete cessation of the immunosuppressive therapy, we lost the patient, unfortunately.

Living Donor Liver Transplantation With Vena Cava Replacement.

OBJECTIVES: This study sought to evaluate the indications, techniques, and results of inferior vena cava (IVC) replacement at living donor liver transplantation (LDLT). MATERIALS AND METHODS: We performed 821 LDLTs and 11 (1.3%) patients required concomitant IVC replacement. We analyzed the indications, replacement materials, and outcomes. RESULTS: Right, left, and left lateral liver lobes were transplanted in 7, 2, and 2 patients, respectively. The indications for IVC replacement were thrombosis/fibrosis in 7 patients (Budd-Chiari 4, hereditary tyrosinemia 1, congenital hepatic fibrosis 1, cryptogenic 1), involvement with mass in 3 patients (Echinococcus alveolaris 2, hepatoblastoma 1) and iatrogenic narrowing at IVC in 1 patient. Cryopreserved grafts (aorta n = 5, IVC n = 4, iliac vein n = 1) or synthetic graft (n = 1) were used for replacements. In 1 patient, hepatic outflow obstruction developed at 39 days and was treated successfully by interventional radiology. There was only 1 hospital mortality (8.9%) that was unrelated to caval replacement (subarachnoid hemorrhage). Of the remaining patients, the caval grafts were patent after a mean 7.7 months of follow-up (range 1 to 17 months). CONCLUSIONS: Although rare, IVC replacement can be necessary at LDLT. Budd-Chiari and E. alveolaris are the main underlying diseases for replacement requirements. Caval replacement with cryopreserved vascular grafts can provide successful short-term and long-term patency.

Circumferential Fence With the Use of Polyethylene Terephthalate (Dacron) Vascular Graft for All-in-One Hepatic Venous Reconstruction in Right-Lobe Living-Donor Liver Transplantation.

Integration of hepatic vein tributaries with a diameter ≥ 5 mm into the drainage system in right-lobe living-donor liver transplantation (LDLT) is of vital importance for graft function. Recently, the most commonly emphasized hepatic venous reconstruction model is the all-in-one reconstruction model. In the final stage of this model that aims to form a common large opening, allogeneic vascular grafts are almost always used to construct a circumferential fence. To date, no other study has reported the use of polyethylene terephthalate (Dacron) vascular graft as a circumferential fence in LDLT. We aimed to present the 1st 4 cases of circumferential fences created with Dacron vascular graft. Four right-lobe grafts weighing 522-1,040 g were used. A polytetrafluoroethylene vascular graft was used for the integration of segment 5 vein and segment 8 vein into the drainage model, whereas a Dacron graft was used to creating a circumferential fence. The patency of hepatic outflow evaluated with the use of multidetector computerized tomography at postoperative day 7. Venous outflow obstruction was not detected in any cases. This study suggested that owing to its flexible structure the polyethylene terephthalate vascular graft can be an alternative to allogeneic vascular grafts in forming circumferential fence.

Influence of Liver Transplantation on Neuropsychiatric Manifestations of Wilson Disease.

OBJECTIVES: This study sought to evaluate the effect of liver transplantation on the neuropsychological manifestations of Wilson disease. MATERIALS AND METHODS: Nine of 42 Wilson disease patients had neuropsychological symptoms before liver transplantation. They were 7 male and 2 female subjects with a median age of 19 years (range 10 to 25). They were analyzed for their preoperative and postoperative hepatic, neurological, and psychological scores described by the Unified Wilson Disease Rating Scale after a mean 36.6 months of follow-up. RESULTS: Preoperative mean Model for End-Stage Liver Disease and Child-Pugh scores were 18.3 (range 15 to 26) and 8.9 (range 6 to 12), respectively. One patient had acute postoperative ischemic stroke unrelated to Wilson disease and was excluded from the statistical analysis. Preoperative and postoperative hepatic, neurological, and psychological scores of the remaining 8 patients were 7.4 ± 2.3 vs 2.4 ± 1.3 (P = .0005), 17.7 ± 11.7 vs 12.7 ± 12.5 (P = .055), and 9.0 ± 1.7 vs 7.0 ± 2.1 (P = .033). CONCLUSIONS: Liver transplantation for Wilson disease can provide some improvement of the neuropsychological symptoms in addition to the hepatic recovery.

Living-Donor Liver Transplantation for Budd-Chiari Syndrome--Resection and Reconstruction of the Suprahepatic Inferior Vena Cava With the Use of Cadaveric Aortic Allograft: Case Report.

BACKGROUND: Living-donor liver transplantation with inferior vena cava resection and reconstruction is rarely indicated for Budd-Chiari syndrome. The aim of this case presentation was to present and discuss the inferior vena cava reconstruction with the use of cadaveric aortic allograft after resection of the suprahepatic inferior vena cava in a patient with Budd-Chiari syndrome who was treated with living-donor liver transplantation. CASE REPORT: A 29-year-old male patient with end-stage liver disease and suprahepatic inferior vena cava obstruction was referred to our center. He was scheduled for living-donor liver transplantation. The suprahepatic inferior vena cava was resected and reconstruction was achieved by means of interposition of the cadaveric aortic allograft between the right atrium and inferior vena cava. Postoperative course was uneventful. DISCUSSION: Liver transplantation and vena cava reconstruction is indicated in some patients with end-stage liver disease and Budd-Chiari syndrome. Limitations in cadaveric organ donation may be compensated for with the use of living-donor liver. In this condition, various aspects of inferior vena cava reconstruction may be discussed. CONCLUSIONS: Budd-Chiari syndrome due to suprahepatic inferior vena cava obstruction close to the right atrium may be treated with vascular reconstruction with the use of a cadaveric aortic allograft.

Living related donor liver transplantation with atrio-caval anastomosis of inferior vena cava graft stored in deep-freeze for budd-Chiari syndrome.

We have previously reported our experience in inferior vena cava resection and reconstruction techniques during liver transplantation for Budd-Chiari syndrome. Herein, we present on a case that demonstrates the importance of experience in complex vascular reconstruction techniques for living donor liver transplantation. A 15-year-old boy was scheduled for living donor liver transplantation for Budd-Chiari syndrome. Venous occlusion was extended up to the right atrial orifice of the supra-hepatic vena cava. Retro- and supra-hepatic segments of the vena cava was resected. Inferior vena cava graft stored in deep-freeze was available. Venous reconstruction was performed with end-to-end atrio-caval anastomosis. Surgical treatment was completed with the implantation of the right liver lobe donated by the patient's mother. Post-surgical course was uneventful.

Temporary Parenteral Tacrolimus Requirement due to Unexplained Low Through Levels in a Liver Transplant Patient with Short Bowel Syndrome.

An adequate level of tacrolimus in serum should be obtained to prevent acute rejection following liver transplantation. Because of good gastrointestinal absorption of oral tacrolimus, adequate trough levels can be achieved even in patients with short bowel syndrome. Rarely, adequate through levels cannot be obtained by oral administration of the drug for several reasons such as inadequate absorption, having a discordant patient, laboratory error, and/or interactions with other drugs and foods. Here, we described a 16-year-old patient who had undergone massive intestinal resection due to mesenteric torsion 5 years previously and required liver transplantation for cryptogenic cirrhosis. Her remnant small bowel length was 90 cm. After a successful living donor liver transplantation, oral tacrolimus administration resulted in inadequate through levels in some parts of the postoperative period. We checked up all the potential reasons but could not identify any cause. An intravenous tacrolimus including immunosuppressive regimen was temporarily required. She maintained adequate blood levels of tacrolimus by parenteral route for a while; thereafter, oral administration resulted in enough blood drug levels. She was discharged with oral tacrolimus therapy. We concluded that very rarely, adequate blood levels of tacrolimus cannot be achieved by oral administration for unexplained reasons. In such cases, temporary administration of parenteral tacrolimus can be life-saving.

Right-lobe living-donor liver transplantation in adult patients with acute liver failure.

BACKGROUND: Right-lobe living-donor liver transplantation (RLDLT) is an excellent option to reduce donor shortages for adult patients with acute liver failure (ALF). The aim of this study was to evaluate the etiologies and outcomes of 30 consecutive adult patients who underwent emergency RLDLT for ALF. METHODS: Between January 2007 and September 2011, we examined data from medical records of patients with ALF who underwent RLDLT. RESULTS: Their mean age was 32.2 ± 13.05 years. The etiologies of ALF were acute hepatitis B (n = 11; 36.6%), hepatitis A (n = 4; 13.3%), drug intoxication (n = 4; 13.3%), pregnancy (n = 2; 6.7%), hepatitis B with pregnancy (n = 1; 3.3%), mushroom intoxication (n = 1; 3.3%), and unknown (n = 7; 23.3%). The mean hepatic coma grade (Model for End-Stage Liver Disease score) was 34.13 ± 8.72. The 43 (48.7%) postoperative complications were minor (grades I-II) and 44 (51.3%) were major (grades III-V). Reoperation was required in 14 of 30 (47%) recipients (grades IIIb-IVa). Deaths occurred owing to pulmonary (n = 2), cardiac (n = 1), septic (n = 2), or encephalopathic (n = 4) complications. The mean durations of intensive care unit stay and postoperative hospitalization were 3.2 ± 2.3 and 29.5 ± 23 days, respectively. The survival rate was 70%. The mean follow-up duration was 305 days (range, 1-1582). CONCLUSION: Liver transplantation is potentially the only curative modality, markedly improving the prognosis of patients with ALF. The interval between ALF onset and death is short and crucial because of the rapid, progressive multiorgan failure. Thus, RLDLT should be considered to be a life-saving procedure for adult patients with ALF, requiring quicker access to a deceased-donor liver graft and a short ischemia time.

Comparison of plasmapheresis and molecular adsorbent recirculating system efficacy in graft failure after living donor liver transplantation.

INTRODUCTION: Liver transplantation may result in graft failure, requiring time and supportive treatment for regeneration of the graft. The aim of this study was to compare the laboratory parameter changes after single-session molecular adsorbent recirculating system (MARS) and plasmapheresis procedures among living donor liver transplantation patients experiencing graft failure. PATIENTS AND METHOD: We analyzed retrospectively the results in 45 liver transplantation patients treated with plasmapheresis and/or MARS between June 2011 and July 2012: (plasmapheresis, n = 17; MARS, n = 15; MARS + plasmapheresis, n = 13). When cadaveric donor cases (n = 11) were excluded, the remaining 34 included patients, underwent. MARS (n = 18) or plasmapheresis (n = 16) at the first session. FINDINGS: Both groups were similar in age, sex, and body mass index features. The MARS group displayed significantly higher levels of international normalized ratio, blood urea nitrogen, and Model for End-stage Liver Disease score. The plasmapheresis cohort, displayed significantly higher levels of initial direct bilirubin and gamma glutamyl transferase (P < .05). The plasmapheresis group showed a significant decrease in GGT after treatment (P < .05). RESULTS: An initial MARS session provided significantly greater decrease in renal function associated with graft failure after living donor liver transplantation.