Estrogen- and Satiety State-Dependent Metabolic Lateralization in the Hypothalamus of Female Rats.

Hypothalamus is the highest center and the main crossroad of numerous homeostatic regulatory pathways including reproduction and energy metabolism. Previous reports indicate that some of these functions may be driven by the synchronized but distinct functioning of the left and right hypothalamic sides. However, the nature of interplay between the hemispheres with regard to distinct hypothalamic functions is still unclear. Here we investigated the metabolic asymmetry between the left and right hypothalamic sides of ovariectomized female rats by measuring mitochondrial respiration rates, a parameter that reflects the intensity of cell and tissue metabolism. Ovariectomized (saline injected) and ovariectomized+estrogen injected animals were fed ad libitum or fasted to determine 1) the contribution of estrogen to metabolic asymmetry of hypothalamus; and 2) whether the hypothalamic asymmetry is modulated by the satiety state. Results show that estrogen-priming significantly increased both the proportion of animals with detected hypothalamic lateralization and the degree of metabolic difference between the hypothalamic sides causing a right-sided dominance during state 3 mitochondrial respiration (St3) in ad libitum fed animals. After 24 hours of fasting, lateralization in St3 values was clearly maintained; however, instead of the observed right-sided dominance that was detected in ad libitum fed animals here appeared in form of either right- or left-sidedness. In conclusion, our results revealed estrogen- and satiety state-dependent metabolic differences between the two hypothalamic hemispheres in female rats showing that the hypothalamic hemispheres drive the reproductive and satiety state related functions in an asymmetric manner.

Molecular pathogenesis of post-transplant acute kidney injury: assessment of whole-genome mRNA and miRNA profiles.

Acute kidney injury (AKI) affects roughly 25% of all recipients of deceased donor organs. The prevention of post-transplant AKI is still an unmet clinical need. We prospectively collected zero-hour, indication as well as protocol kidney biopsies from 166 allografts between 2011 and 2013. In this cohort eight cases with AKI and ten matched allografts without pathology serving as control group were identified with a follow-up biopsy within the first twelve days after engraftment. For this set the zero-hour and follow-up biopsies were subjected to genome wide microRNA and mRNA profiling and analysis, followed by validation in independent expression profiles of 42 AKI and 21 protocol biopsies for strictly controlling the false discovery rate. Follow-up biopsies of AKI allografts compared to time-matched protocol biopsies, further baseline adjustment for zero-hour biopsy expression level and validation in independent datasets, revealed a molecular AKI signature holding 20 mRNAs and two miRNAs (miR-182-5p and miR-21-3p). Next to several established biomarkers such as lipocalin-2 also novel candidates of interest were identified in the signature. In further experimental evaluation the elevated transcript expression level of the secretory leukocyte peptidase inhibitor (SLPI) in AKI allografts was confirmed in plasma and urine on the protein level (p<0.001 and p = 0.003, respectively). miR-182-5p was identified as a molecular regulator of post-transplant AKI, strongly correlated with global gene expression changes during AKI. In summary, we identified an AKI-specific molecular signature providing the ground for novel biomarkers and target candidates such as SLPI and miR-182-5p in addressing AKI.

MARS therapy, the bridging to liver retransplantation - Three cases from the Hungarian liver transplant program.

Besides orthotopic liver transplantation (OLT) there is no long-term and effective replacement therapy for severe liver failure. Artificial extracorporeal liver supply devices are able to reduce blood toxin levels, but do not replace any synthetic function of the liver. Molecular adsorbent recirculating system (MARS) is one of the methods that can be used to treat fulminant acute liver failure (ALF) or acute on chronic liver failure (AoCLF). The primary non-function (PNF) of the newly transplanted liver manifests in the clinical settings exactly like acute liver failure. MARS treatment can reduce the severity of complications by eliminating blood toxins, so that it can help hepatic encephalopathy (HE), hepatorenal syndrome (HRS), and the high rate mortality of cerebral herniation. This might serve as a bridging therapy before orthotopic liver retransplantation (reOLT). Three patients after a first liver transplantation became candidate for urgent MARS treatment as a bridging solution prior to reOLT in our center. Authors report these three cases, fo-cusing on indications, MARS sessions, clinical courses, and final outcomes.

Detection and management of renal cell carcinoma in the renal allograft.

PURPOSE: Tumours of the transplanted kidney represent a rare form of post-transplantation malignancies. An important aspect of the treatment option is whether the transplanted kidney can be saved or not. Aim of our study was the analysis of our allograft tumours. METHODS: In the Budapest Centre, 3,530 kidney transplantations were performed between 1973 and 2012. Retrospective analysis of 9 patients who developed renal cell carcinoma (RCC) in the transplanted kidney was done. RESULTS: Mean age of recipients was 45.3 ± 13.4 years at the time of transplantation and 57.0 ± 11.6 years at the time of tumour detection. Mean age of their donors was 43.5 ± 11.5 years. Mean time from transplantation to tumour diagnosis was 134.6 ± 40.8 months. Seven RCC were stage pT1a, 1 was stage pT1b and 1 was pT3a. Eight patients had stage I. (pT1a-b, N0, M0) and 1 patient had stage IV. (pT3a, N1, M1) disease. Histological types were clear cell (n = 6), papillary (n = 2) and sarcomatoid (n = 1) carcinomas. The tumour growth rate of RCC was 16.7 ± 13.5 mm/year. In 4 cases, transplant nephrectomy was performed; 5 cases had percutaneous radiofrequency ablation (RFA). Ablative therapy had no influence on renal graft function. Six patients (including 5 patients who were treated with RFA) are still alive and tumour-free; 3 patients died. CONCLUSIONS: According to our observation, we can state that RCC of the kidney allograft diagnosed at an early stage can be successfully treated with RFA instead of graft removal. A longer follow-up is needed to assess the effectivity of the RFA treatment in these cases.

Periodic limb movements in sleep are associated with stroke and cardiovascular risk factors in patients with renal failure.

Periodic limb movements in sleep (PLMS) is prevalent among dialysed patients and is associated with increased risk of mortality. Our study aimed to determine the prevalence of this disease in a sample of transplanted and waiting-list haemodialysed patients. One hundred transplanted and 50 waiting-list patients underwent polysomnography. Moderate and severe diseases were defined as periodic limb movements in sleep index (PLMSI) higher than 15 and 25 events h(-1), respectively. The 10-year coronary heart disease risk was estimated for all patients using the Framingham Score. Moreover, the 10-year estimated risk of stroke was calculated according to the modified version of the Framingham Stroke Risk Profile. PLMS was present in 27% of the transplanted and 42% of the waiting-list group (P = 0.094); the proportion of severe disease was twice as high in waiting-list versus transplanted patients (32 versus 16%, P = 0.024). Patients with severe disease had a higher 10-year estimated risk of stroke in the transplanted group [10 (7-17) versus 5 (4-10); P = 0.002] and a higher 10-year coronary heart disease risk in both the transplanted [18 (8-22) versus 7 (4-14); P = 0.002], and the waiting-list groups [11 (5-18) versus 4 (1-9); P = 0.032]. In multivariable linear regression models the PLMSI was associated independently with the Framingham cardiovascular and cerebrovascular scores after adjusting for important covariables. Higher PLMSI is an independent predictor of higher cardiovascular and cerebrovascular risk score in patients with chronic kidney disease. Severe PLMS is less frequent in kidney transplant recipients compared to waiting-list dialysis patients.

[Incidence of thrombophilia and risk of renal vessel thrombosis in kidney transplant recipients]. / A thrombophilia elofordulása és a veseerek thrombosisának veszélye veseátültetett betegeknél.

INTRODUCTION: Kidney transplantation is the optimal treatment of end stage kidney disease. The most common vascular complication in the early postoperative period is thrombosis of the renal artery and vein. These complications usually lead to the loss of the transplanted kidney. AIM: of our study was to identify those factors which represent an increased risk for thrombotic complication and determine whether routine screening for thrombophilia is justifiable before transplantation. As an illustration to this problem we report a case of successful renal vein recanalisation after thrombosis. METHODS: We give an overview of the literature about incidence of renal graft thrombosis, hypercoagulable states, predictive value of factor V. Leiden and prothrombin G20210A mutations in venous thromboembolism. We discuss those publications that suggest a preoperative screening of transplant candidates for hypercoagulable states and thrombophilia and those that do not think that such screening is reasonable. In our case a 28 year old male patient received a cadaveric kidney. Thrombosis of the renal vein was diagnosed 8 hours after transplantation. Reoperation was performed immediately: venous anastomosis was opened, the thrombus removed. After reoperation the circulation of the kidney recovered, intravenous heparin treatment was introduced immediately. RESULTS: 24 months later the kidney is still functioning well. Postoperative thrombophilia screening showed heterozygosity for factor V Leiden. CONCLUSION: There are only few publications reporting on successful recanalisation after renal vein thrombosis. In our case rapid diagnosis and immediate operative treatment saved the graft. There is no uniform proposal in the literature whether preoperative screening for thrombophilia is justifiable or not. In our view screening for thrombophilia and thromboprophylaxis is mandatory. Extensive prospective studies should be undertaken to refine the risks and establish the associations of thrombophilia and thromboembolism after kidney transplantation.

[Evaluation of patients' readiness for surgery when called from the waiting list for kidney transplantation: experience of the Budapest Centre]. / Veserecipiensek alkalmassága transzplantációra való behíváskor. A budapesti centrum tapasztalatai.

UNLABELLED: The aim of this study was to assess the impact of the establishment of waiting list committee on recipient evaluation for kidney transplantation. Studies on this issue have not been previously reported. METHODS: Data of 714 patients were collected between September 1, 2007 and April 20, 2010. Of the 714 patients 354 were transplanted. Data from the first 16 months period were compared to those obtained during the second 16 months period. RESULTS: During the first period 171 patients (47.9%) were unfit for transplantation [129 patients (36.1%) were refused after telephone information and 42 patients (11.8%) after clinical investigation]. During the second period 141 (39.5%) patients were found to be unsuitable [80 patients (22.4%) after telephone information and 61 patients (17.1)] after clinical examination. During the two periods of the study 44% of patients were unfit for transplantation. A considerable number of patients were refused because of cardiologic reasons. The percent of fit patients was 52.1% in the first period and 60.5% in the second period. In addition, the percent of unfit patients who were unsuitable for kidney transplantation after telephone information decreased from 36.1% to 22.4%, while the percent of unfit patients after clinical evaluation increased from 11.% to 17.1%. CONCLUSIONS: Authors conclude that waiting list committee made an effective work.

Primary rat hepatocyte culture on 3D nanofibrous polymer scaffolds for toxicology and pharmaceutical research.

Primary rat hepatocytes are a widely used experimental model to estimate drug metabolism and toxicity. In currently used two-dimensional (2D) cell culture systems, typical problems like morphological changes and the loss of liver cell-specific functions occur. We hypothesize that the use of polymer scaffolds could overcome these problems and support the establishment of three-dimensional (3D) culture systems in pharmaceutical research. Isolated primary rat hepatocytes were cultured on collagen-coated nanofibrous scaffolds for 7 days. Cell loading efficiency was quantified via DNA content measurement. Cell viability and presence of liver-cell-specific functions (albumin secretion, glycogen storage capacity) were evaluated. The activity of liver-specific factors was analyzed by immunofluorescent staining. RNA was isolated to establish quantitative real-time PCR. Our results indicate that primary rat hepatocytes cultured on nanofibrous scaffolds revealed high viability and well-preserved glycogen storage. Albumin secretion was existent during the entire culture period. Hepatocytes remain HNF-4 positive, indicating highly preserved cell differentiation. Aggregated hepatocytes re-established positive signaling for Connexin 32, a marker for differentiated hepatocyte interaction. ZO-1-positive hepatocytes were detected indicating formation of tight junctions. Expression of cytochrome isoenzymes was inducible. Altogether the data suggest that nanofibrous scaffolds provide a good in vitro microenvironment for neo tissue regeneration of primary rat hepatocytes.

[Cytomegalovirus infection after solid-organ transplantation, its risk factors, direct and indirect effects and prevention strategies]. / A cytomegalovírus-fertozés rizikófaktorai, hatásai és a megelozés lehetoségei transzplantációt követoen.

The human cytomegalovirus is widely prevalent among human population and it is the most common viral pathogen that affects both the graft's and solid-organ transplant recipient's survival. The risk is highest in donor-seropositive, recipient-seronegative pairing transplantation. These recipients carry increased risk of developing symptomatic primary CMV infection; however, other risk factors may have an impact on cytomegalovirus activation as well: intensity of immunosuppression, type of organ transplanted, rejection and/or treatment for rejection, HLA-mismatch between recipient and donor, certain HLA-types of the recipient, female sex etc. Cytomegalovirus infection in transplant patients has been associated with both direct (symptoms) and indirect effects which are derived from the immunomodulating impact of the virus such as cellular effects and cytokine expression or systemic immune suppression leading to other opportunistic infections. Prevention of the direct and indirect effects of cytomegalovirus infection is the therapeutic goal in transplanted patients. Most transplant centers use either universal prophylaxis or preemptive therapy to prevent the infection. The advantages and disadvantages of these two preventive strategies and current evidence-based recommendations for preventing cytomegalovirus disease in solid-organ transplant recipients are discussed according to others' and the authors' own observations. According to recommendations of the American and Canadian Societies of Transplantation, most of the centers--after analyzing of the CMV-infection risk factors of the recipients--divide them into three groups: high-, moderate- and low-risk groups. The preventive strategy is attached to the risk-group type. In the high-risk group (R-/D+ and lung transplant patients) the use of the universal prophylaxis is necessary. The patients administered anti-lymphocyte antibodies (ATG, ALG or OKT3) need selective (subtype of universal) prophylaxis. Among the moderate-risk patients (R+/D+ or R+/D-) the doctors may choose either universal prophylaxis or preemptive therapy. Selection of a strategy requires consideration of patient-specific factors as well as practical considerations such as available resources. For avoidance of the indirect effects of CMV infection universal prophylaxis is preferred. The use of preventive proceedings in low-risk patients is the matter of the center's decision.

HLA-DQ3 is a probable risk factor for CMV infection in high-risk kidney transplant patients.

BACKGROUND: Cytomegalovirus (CMV) infection in transplant patients with special risk factors remains a major hazard. CMV-seronegative recipients with seropositive donors have the highest risk of developing acute CMV disease. We suggest that the HLA-type may influence the occurrence and the severity of primary CMV infection of these recipients and the measurement of the special HLA-types may be useful in the prediction of acute infection. METHODS: Since 1999 1213 cadaver kidney transplantations have been performed in our clinic. 163 of 1213 recipients were CMV-seronegative (13%) and 129 of them received the kidney from seropositive donors. All 129 patients received CMV infection prophylaxis. Of 129 CMV-seronegative patients 49 developed acute CMV infection (38%) during the first posttransplant year. CMV infection was diagnosed by CMV antigenemia test and serologic measurements (ELISA). The particular HLA-genotypes of the recipients were studied before the transplantation. The occurrence and the severity of CMV infection was investigated in association with HLA-types. RESULTS: We found different acute CMV infection distribution in the careers and non-careers of investigated HLA-types: HLA-A2, HLA-B12, HLA-Cw7, HLA-DR6 and HLA-DR11, but the differences were not significant in these HLA-types (P = 0.26, P = 0.37, P = 0.83, P = 0.07 and P = 0.37). While investigating HLA-DQ3, we found that of 68 DQ3-positive patients 32 (47%), of 61 DQ3-negative patients 17 (28%) had acute CMV infection and this difference was found to be significant. This result was confirmed by univariate and multivariate Cox Regression (P = 0.001) and the appropriate significance level was considered by Bonferroni correction. CONCLUSIONS: HLA-DQ3 was found to be an independent predictor of CMV infection. Our data suggest that patients positive for HLA-DQ3 are more susceptible to CMV infection than a comparable group of patients negative for HLA-DQ3. This result was not due to rejection and/or treatment for rejection and was not influenced by induction therapy. Although we found more symptomatic infections among DQ3+ patients the difference was not significant (P = 0.19). Comparing the gender proportion among all 1213 kidney recipients and among CMV-seronegative recipients we found that the proportion of males is significantly higher among CMV-seronegative recipients (P < 0.001).

[Role of apoptosis in the kidney after reperfusion]. / Az apoptosis szerepe a vesében reperfúzió során.

UNLABELLED: Organ transplantation is one of the most important achievements of medicine in the 20th century. Ischaemia-reperfusion has a great impact on both: immediate organ function and long-term survival. Organ transplantation can be regarded as a clinical model of ischaemia-reperfusion phenomenon. AIM: The prevention of ischaemia-reperfusion damages. Apoptosis plays a key-role in these processes. METHODS: Apoptosis was investigated in the course of human kidney transplantation. In animal experiments the characteristics of apoptosis were analysed after short ischaemia. Calcium antagonists: verapamil, nifedipine, bepridil, fendiline and (-)-deprenyl, an irreversible selective inhibitor of monoamino oxidase type B, (-)-deprenyl, were administered to prevent apoptosis. RESULTS: The observations showed that in the course of human kidney transplantation necrotic, apoptotic and proliferating renal tubular cells can be observed. All calcium channel blockers and (-)-deprenyl decreased the occurrence and degree of apoptosis in rat kidney. CONCLUSIONS: The functional capacity of tubular cells is a significant factor in the adequate kidney function. The reduction of the apoptosis of tubular cells possibly could improve the function of transplanted kidneys.

Malignancies after renal transplantation during 33 years at a single center.

This study provides an analysis of incidence and characteristics of malignant tumors of 2535 patients who underwent renal transplantation between 1973 and 2007 at the Transplantation Center in Budapest. One hundred ninety-three malignant diseases were found in 188 patients (7.6%). The incidence of thyroid-, renal- hepatic-, skin- and gastric cancers as well as of Kaposi sarcoma and lymphomas increased in our transplant patient cohort compared to the figures of the general population based on the data of our Cancer Registry. On the other hand, colorectal-, oralprostate and lung cancers were underrepresented in our patient cohort. The mean time of diagnosis of malignancies following kidney transplantation was 58.5+/-44.8 months. One fifth of the tumors were detected within the first year. Patients with malignancies were distributed into four groups based on the immunosuppressive regimen: group I (8.5%), azathioprine + prednisone; group II (59.0%), cyclosporine + prednisone; group III (26.6%), cyclosporine + mycophenolate mofetil + prednisone; group IV (5.9%), tacrolimus + mycophenolate mofetil + prednisone. The mean age of patients was 47.3, 53.5, 55.5 and 58.1 years in group I, II, III and IV, respectively. Oncologic and immunosuppressive therapy was decided individually. Immunosuppression was switched to rapamycin-containing regimens in 63 cases. We lost 92 patients (48.9%) with a mean survival time of 25.8+/-39.4 months. Cumulative 1- and 5-year survivals were 69.5% and 52%, respectively. The increasing number of cancers seen early after transplantation and the increased risk of developing a cancer due to the older age of recipients draw attention to the importance of regular oncologic screening in patients on the waiting list and after transplantation.

Antiapoptotic effect of (-)-deprenyl in rat kidney after ischemia-reperfusion.

BACKGROUND: Since apoptosis of renal tubular cells is the basis of the damage caused by ischaemia-reperfusion, the antiapoptotic effect on kidney tubular epithelial cells of the monoamino oxidase-B (MAO-B) inhibitor (-) deprenyl (selegiline), known neuroprotective agent, with antiapoptotic properties, was studied in a rat model. MATERIAL/METHODS: Warm renal ischaemia was caused by clamping the left renal artery of rats for 30 minutes. With the start of reperfusion 0.015 mg/kg, 0.15 mg/kg and 1.5 mg/kg of (-)-deprenyl was injected simultaneously into the carotid artery of the animals, respectively. Five rats served as control, in which renal artery clamping was performed, but the rats were only treated with the solvent (physiological saline). After 6 hours of reperfusion the rats were exsanguinated and the kidneys were histologically examined. RESULTS: Severe tubular damage characterised by apoptosis was found in the kidneys of the untreated rats. Apoptosis was verified on the basis of morphological features, methylgreen-pyronin staining and TUNEL reaction. (-)-Deprenyl diminished dose-dependently the apoptotic damage, 0.15 mg/kg being the most effective dose. The same dose of (-)-Deprenyl is used in the therapy of human Parkinson's disease. CONCLUSIONS: Our findings suggest, that (-)-deprenyl might have an impact on decreasing renal injury also in case of human cadaveric renal transplantation.