Irradiation with 365 nm and 405 nm wavelength shows differences in DNA damage of swine pancreatic islets.

INTRODUCTION: 3D printing is being used more extensively in modern biomedicine. One of the problems is selecting a proper crosslinking method of bioprinted material. Amongst currently used techniques we can distinguish: physical crosslinking (e.g. Ca2+ and Sr2+) and chemical crosslinking-the UV light crosslinking causing the biggest discussion. UV radiation is selectively absorbed by DNA, mainly in the UV-B region but also (to some extent) in UV-A and UV-C regions. DNA excitement results in typical photoproducts. The amount of strand breaks may vary depending on the period of exposition, it can also differ when cells undergo incubation after radiation. AIM: The aim of this study was to show whether and how the time of irradiation with 405 nm and 365 nm wavelengths affect DNA damage in cell lines and micro-organs (pancreatic islets). MATERIALS AND METHODS: The degree of DNA damage caused by different wavelengths of radiation (405 nm and 365 nm) was evaluated by a comet assay. The test was performed on fibroblasts, alpha cells, beta cells and porcine pancreatic islets after 24 hours incubation period. Samples without radiation treatment were selected as a control group. Results analysis consisted of determining the percent of cells with damaged DNA and the tail intensity evaluation. RESULTS: The degree of DNA damage in pancreatic islets after exposure to 405 nm wavelength oscillated between 2% and 6% depending on the tested time period (10 - 300 seconds). However, treating islets using 365 nm wavelength resulted in damage up to 50%. This clearly shows significantly less damage when using 405 nm wavelength. Similar results were obtained for the tested cell lines. CONCLUSIONS: Crosslinking with 405 nm is better for pancreatic islets than crosslinking with 365 nm UV light.

Islet Transplantation - perspective from Poland.

The article describes impact of advanced research in the USA and collaborative approach of US scientists and clinicians on development of the field of islet transplantation in Poland and all over the world. At the same time, it presents negative consequences of islet regulation by FDA as a biological drug leading to decline and extinction of the field in the US, while it is on the rise worldwide.

Endoscopic Islet Autotransplantation Into Gastric Submucosa-1000-Day Follow-up of Patients.

BACKGROUND: Total pancreatectomy and autologous transplantation of pancreatic islets is a treatment option for patients with severe pain due to chronic pancreatitis. In the standard procedure, pancreatic islets are isolated and subsequently administered into the portal vein. In the case of patients with a history of thrombosis or at risk of thrombosis, this route of administration is not viable. Animal studies conducted in our department led to the development of a technique of endoscopic islets transplantation into the gastric submucosa. In 2013 and 2014, the first human autologous transplant procedures were performed. The objective of this study was to present the results of a 3-year follow-up of these patients. METHODS: Two pancreatectomies were performed in our department, the first in 2013 and another in 2014, along with subsequent autologous transplantation of pancreatic islets into the gastric submucosa. RESULTS: Both patients had been diagnosed previously with diabetes, and both had endogenous islet activity detected. Peptide C concentration after pancreatectomy and before pancreatic cell transplantation was 0.1 ng/mL. After the transplantation, peptide C concentrations for the 2 patients were 0.8 and 0.5 ng/mL on day 7, 1.2 and 0.6 ng/mL on day 30, 1.3 and 0.8 ng/mL on day 180, 1.1 and 0.7 ng/mL on day 360, and 3.0 and 0.6 ng/mL at 3 years, respectively, after transplantation. The pain symptoms resolved in both cases. CONCLUSION: Pancreatic islets may survive in the gastric wall. Endoscopic submucosal transplantation may present an alternative for the management of patients who cannot undergo a classic transplantation procedure.

Analysis of Distribution of Expanded- and Standard-Criteria Donors and Complications Among Polish Recipients by Kidney Donor Risk Index Value.

INTRODUCTION: The approach toward transplanting kidneys from expanded-criteria donors (ECDs) in Poland is largely site-dependent. The Kidney Donor Risk Index (KDRI) allows for obtaining a more precise characteristic of ECDs and further stratification into "better" and "worse" quality grafts. METHODS: Comparison of the incidence of delayed graft function (DGF) and biopsy-proven acute rejection (BPAR), median of hospitalization time and median of estimated glomerular filtration rate (eGFR) at 1 year after transplantation among kidney graft recipients (n = 468), divided by donor status (ECD/standard-criteria donor [SCD]) and KDRI value (I: 0.67-1.2, II: 1.21-1.6, III: 1.61-2.0, IV: 2.01-3.48). RESULTS: ECD kidneys have been transplanted to 32.47% of recipients. There were no ECD recipients in KDRI compartment I, 16.55% in compartment II, 79.22% in compartment III, and 100% in IV. In KDRI compartment II, DGF was diagnosed in 34.9% of SCDs and 56% of ECDs (P = .003), BPAR occurred in 7.8% of SCDs and 16% of ECDs (P = .073), median hospital stay was 12 days for SCDs and ECDs (P = 1), and eGFR was 50.7 mL/min for SCDs and 49.4 mL/min for ECDs (P = .734). In KDRI compartment III, DGF was diagnosed in 43.8% of SCDs and 49.2% of ECDs (P = .139), BPAR occurred in 6.3% of SCDs and 31.7% of ECDs (P = .001), median hospital stay was 10 days for SCDs and 12 days for ECDs (P = .634), and eGFR was 49.5 mL/min for SCDs and 45.2 mL/min for ECDs (P = .382). Among ECD recipients, DGF was diagnosed in 56.0%, 49.2%, and 47.7% of patients for KDRI compartments II, III, and IV respectively (P = .776); BPAR occurred in 16% (compartment II), 31.7% (compartment III), and 23.1% (compartment IV) (P = .273); the median hospital stay was 12 days (compartment II), 12 days (compartment III), and 12.5 days (compartment IV) (P = 1); and eGFR was 49.5 mL/min (compartment II), 45.4 mL/min (compartment III), and 36.1 mL/min (compartment IV) (P = .002). CONCLUSION: Assessment using both the ECD and KDRI systems allows for a more precise evaluation of prognosis and predicting complications among recipients.

Islets Allotransplantation Into Gastric Submucosa in a Patient with Portal Hypertension: 4-year Follow-up.

BACKGROUND: Islets transplantation is an established treatment method for patients suffering from brittle diabetes with hypoglycemia unawareness. The standard implantation technique is through the portal vein into the liver. In case of liver diseases or portal hypertension, finding an extra-hepatic site is recommended. There have been attempts to perform islets transplantations into muscles and into the gastric submucosa. OBJECTIVE: The aim of this study is to show a 4-year follow-up of allotransplantation into gastric submucosa in a case of portal hypertension observed during the procedure of islets infusion. PATIENTS AND METHODS: A 36-year-old woman with complicated diabetes for over 30 years was selected to receive simultaneous islets and kidney transplantation. The patient underwent an unsuccessful simultaneous pancreas and kidney transplantation 2 years earlier in another transplantation center. The patient's daily insulin requirement was 60 IU, which corresponded to 1.15 IU/kg of body weight. The HbA1c level was 7.4%. C-peptide levels, both fasting and stimulated, were 0.01 ng/mL. On December 7, 2013, the patient received transplanted kidney and islets procured from the same donor. Only 124,000 islets equivalents (IEQ) were isolated (2400 IEQ/kg body weight). Islets were suspended in 300 mL of Ringer's solution along with albumin, antibiotics, and heparin. After infusing 100 mL of the islets suspension into the portal vein, pressure in portal vein increased from 5 mm Hg to 23 mm Hg. Despite stopping the infusion, pressure did not drop after 30 minutes. The decision was made to transplant the reminder of the islets (200 mL) into the gastric wall. RESULTS: No complications were observed after the procedure. Serum creatinine level was 1.6 mg/dL on day 10 and 1.5 mg/dL 4 years after the transplantation. Fasting C-peptide levels were 1.7, 0.65, 0.55, 0.69, 0.68, and 0.2 ng/mL at 1, 3, 6, 12, 18, and 36 months after the transplantation, respectively. HbA1c levels were 5.2, 6.4, 4.7, 5.2, and 5.9% at 3, 6, 12, 18, and 36 months, respectively. The patient's insulin requirement dropped to 15 U/day immediately after transplantation and equaled 20 and 27 U/day at 18 and 48 months after the simultaneous islet and kidney transplantation, respectively. CONCLUSION: Allotransplantation of islets into the gastric wall may be a safe alternative in cases of contraindications for transplantation into the portal vein.

Renal Transplantation Using Kidneys Procured From Elderly Donors Older Than 70 Years.

AIM: A major problem for the transplant society is a shortage of organs for transplantation compared with the number of patients on the waiting list. This study aimed to assess the results of the transplantation of kidneys procured from older donors. PATIENTS AND METHODS: A total of 27 kidneys procured from donors age 70 years or older were transplanted between January 1, 2010, and April 25, 2015. These represented only 4.1% of the 657 kidneys transplanted from deceased donors during this period at the same center. RESULTS: Delayed graft function (DGF) in the recipients of kidneys procured from donors age 70 or older occurred in 46.1% of patients, whereas the recipients of kidneys from younger donors showed DGF at a frequency of 32.7% (P = NS). The annual and 3-year survival rates of kidneys in the study group were 85% and 80%, respectively, and in the control group were 92.5% and 88.6%, respectively (P = NS). According to the Polish National Organ Procurement Organization (Poltransplant), the annual survival rate of a transplanted kidney in Poland stands at 89%, whereas the 3-year survival rate is 82%. We detected no significant posttransplantation differences in the serum creatinine concentration and in the estimated glomerular filtration rate between the study and control groups. The donor age and donor creatinine were the variables independently associated with DGF. CONCLUSIONS: The results of transplantation of kidneys from elderly donors were comparable to those of transplantation from younger donors. Kidneys harvested from elderly donors should be used for a transplant after a preliminary assessment.

Nephropathy Evolving Within the First Two Posttransplant Months With No Typical Cytopathic Lesions: Two Cases Presentation.

BACKGROUND: We report 2 cases of polyomavirus-associated nephropathy (PyVAN) emerging within the initial 8 posttransplant weeks. These cases were characterized by intraepithelial BK virus replication without typical nuclear inclusions in epithelial cells. METHODS AND RESULTS: A 70-year-old male recipient of a cadaveric kidney transplant had experienced unsatisfying graft function since the time of transplantation (Tx). One month after Tx, results of a graft biopsy revealed mild tubulointerstitial inflammation. No intraepithelial nuclear inclusions suggestive of viral infection were present at that time. The patient received intravenous methylprednisolone, and the dosage of tacrolimus was increased. Due to a further drop in the glomerular filtration rate, a subsequent kidney biopsy was performed during posttransplant week 10, which revealed lesions typical of PyVAN. Retrospectively performed SV40 staining revealed that intragraft polyomavirus replication was already present on posttransplant day 30. Basic immunosuppression reduction and ciprofloxacin administration were followed by BK viremia elimination, stabilization of graft function, and resolution of PyVAN. In another patient, a 62-year-old male recipient of a cadaveric renal graft, BK viremia was monitored from the time of Tx. Two months after Tx, the patient was found to have a BK viral load of 6 × 4 log(10)/mL. Results of the graft biopsy revealed fully preserved tubular epithelium, but SV40 staining was positive in some of these cells. After basic immunosuppression reduction and introduction of ciprofloxacin, the BK viral load dropped to 1 × log(10)/mL with graft function stabilization. CONCLUSIONS: PyVAN may emerge as early as 4 weeks after Tx, with near-normal or acute rejection-like graft morphology. The early monitoring of plasma BK viral load, as well as SV40 staining, avoids misdiagnosis of this severe posttransplant complication.

Renal Tumor in Allogeneic Kidney Transplant Recipient.

BACKGROUND: Malignancies will be a leading cause of mortality in renal transplant recipients in the next 20 years. Renal cell cancer (RCC) is the most common urologic cancer in kidney transplant recipients. The risk of RCC development in kidney transplant recipients is 15-100 times higher than in the general population. The purpose of the current retrospective study was to assess the frequency of nephrectomies performed because of renal tumors in the native kidneys in kidney transplant recipients in the Department of General and Transplantation Surgery at the Medical University of Warsaw between 2010 and 2014 year; the identification of kidney recipients diagnosed with RCC; and epidemiologic, clinical, and histopathological aspects associated with RCC. PATIENTS AND METHODS: A total of 319 nephrectomies were performed in the Department of General and Transplantation Surgery at the Medical University of Warsaw between 2010 and 2014 year. Renal tumors were diagnosed in 25 renal transplant recipients. RESULTS: Among malignant tumors, 13 cases of RCC and 1 case of post-transplant lymphoproliferative disorder (PTLD) were observed. There was no significant difference between age and duration of pretransplantation dialysis in patients with RCC and patients with benign tumors (P = .14 and P = .91, respectively). Body mass index was significantly higher in patients with RCC than in patients with benign tumors (P = .04). CONCLUSIONS: Renal cell cancer is more common among male kidney recipients. There is a good Polish screening system allowing detection of kidney cancer in native kidney. We recommend performing periodic screening for kidney cancers to obtain an early diagnosis.

Glomerular filtration rate estimation in prospective living kidney donors: preliminary study.

INTRODUCTION: Kidney transplantation prolongs life expectancy in end-stage renal disease patients at a lesser cost than dialysis. Estimation of kidney function is crucial in the evaluation of prospective living kidney donors. Although unsurpassed in their precision methods of glomerular filtration rate (GFR) measurement with exogenous substances are invasive, expensive, and carry a risk for anaphylactic reactions. Alternatively, kidney function can also be assessed by GFR estimation formulas based on serum creatinine or novel markers such as cystatin C or ß-trace protein (BTP). The aim of this study was to compare the performance of GFR estimation methods with reference scintigraphy-measured GFR in population of living kidney donor candidates. METHODS: We included 25 prospective kidney donors (aged 28-64 years) and measured GFR with the following equations: Cockcroft-Gault, Modification of Diet in Renal Disease (MDRD), Mayo Clinic, Nankivell, Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI; including cystatin C), and BTP based. GFR were assessed by (99)mTc-DTPA for reference. All estimation methods were compared with a reference by general linear models. RESULTS: The precision of GFR estimation by all methods is unsatisfactory (30% margin of reference held in <50% of cases). Direction of regression coefficients is negative for some of the methods even when adjusted for body mass index (BMI). Of the study subjects, 64% were overweight/obese. BMI value is significantly correlated with measured GFR (P < .01). CKD-EPI estimation equations are the most precise methods of GFR estimation in this analysis; in addition, CKD-EPI cystatin C and combined creatinine/cystatin C estimators are robust to overweight/obesity. CONCLUSIONS: The precision of GFR estimation is unsatisfactory, in part because of overweight, which adversely influences measured GFR, but also renders estimation methods unusable, except for CKD-EPI cystatin C and combined creatinine/cystatin C formulae. GFR measurement with exogenous substances remains the method of choice in the assessment of kidney function in prospective kidney donors. In addition, it provides useful information on differential (split) renal function.

Impact of pretransplant body mass index on early kidney graft function.

BACKGROUND: An increase in the number of obese patients on transplantation waiting lists can be observed. There are conflicting results regarding the influence of body mass index (BMI) on graft function. METHODS: We performed a single-center, retrospective study of 859 adult patients who received a renal graft from deceased donors. BMI (kg/m(2)) was calculated from patients' height and weight at the time of transplantation. Kidney recipients were subgrouped into 4 groups, according to their BMI: Groups A (<18.5; n = 57), B (18.6-24.9; n = 565), C (25-29.9; n = 198) and D (>30; n = 39). Primary or delayed graft function (DGF), acute rejection (AR) episodes, and number of reoperations, graft function expressed by glomerular filtration rate (GFR) and serum creatinine concentration and number of graft loss as well as the recipient's death were analyzed. The follow-up period was 1 year. RESULTS: Obese patients' grafts do not develop any function more frequently in comparison with their nonobese counterparts (P < .0001; odds ratio [OR], 32.364; 95% CI, 2.174-941.422). Other aspects of the procedure were analyzed to confirm that thesis: Cold ischemia time and number of HLA mismatches affect the frequency of AR (OR, 1.0182 [P = .0029] and OR, 1.1496 [P = .0147], respectively); moreover, donor median creatinine serum concentration (P = .00004) and cold ischemia time (P = .00019) are related to delayed graft function. BMI did not influence the incidence of DGF (P = .08, OR; 1.167; 95% CI, 0.562-2.409), the number of AR episodes (P > .1; OR, 1.745; 95% CI, 0.846-3.575), number of reoperations, GFR (P = .22-.92), or creatinine concentration (P = .09). Number of graft losses (P = .12; OR, 1.8; 95% CI, 0.770-4.184) or patient deaths (P = .216; OR, 3.69; 95% CI, 0.153-36.444) were not influenced. CONCLUSION: Greater recipient BMI at the time of transplantation has a significant influence on the incidence of primary graft failure.

Bacterial and fungal infections in the early post-transplant period after kidney transplantation: etiological agents and their susceptibility.

BACKGROUND: Infections remain serious complications in solid-organ transplant recipients, despite professional medical care, the introduction of new immunosuppressive drugs, and treatment that decreases the risk of infections. METHODS: The study covered 295 adult patients undergoing kidney transplantation (KTx) between September 2001 and December 2007. All the patients were followed prospectively for infections from the KTx date and during the first 4 weeks after surgery. Samples of clinical materials were investigated for microbiological cultures. The microorganisms were cultured and identified in accordance with standard bacteriological procedures. Susceptibility testing was carried out through the use of Clinical and Laboratory Standards Institute procedures. RESULTS: From 295 KTx recipients, 1073 clinical samples were taken for microbiological examination. Positive cultures were 26.9% (n = 289) of all samples tested; 525 strains were collected. Gram-positive bacteria were isolated in 52.2% (n = 274), Gram-negative bacteria were isolated in 40.8% (n = 214), and fungal strains were isolated in 7% (n = 37). Urine specimens (n = 582) were obtained from 84.5% of 245 recipients during the first month after transplantation. Among the isolated bacterial strains (n = 291), the most common were Gram-negative bacteria (56.4%). Gram-positive bacteria comprised 35.7%; fungal strains were found in 23 cases (7.9%). In surgical site specimens (n = 309), Gram-positive bacteria (72.1%) were the most common. Gram-negative bacteria comprised 24.4%. In blood specimens (n = 138), Gram-positive bacteria (81.6%) were the most common. Gram-negative bacteria comprised 15.8%; fungi were isolated in 2.6%. In respiratory tract specimens (n = 13), among the isolated bacterial strains (n = 8), the most common were Gram-positive bacteria (57.1%). Gram-negative bacteria comprised 14.3%; fungi were isolated in 28.6%. CONCLUSIONS: Urine samples were predominantly positive after KTx. Our study showed Gram-positive bacteria in 52.2% after kidney transplantation. The proportion of isolates of multi-drug-resistant bacterial strains (MRCNS, vancomycin-resistant strains, high-level aminoglycoside-resistant strains, extended-spectrum beta-lactamase producers, and high-level aminoglycoside-resistant strains) was increased. These data indicate the need for strict adherence to infection control procedures in these patients.

Characteristics of potential living kidney donors and recipients: donor disqualification reasons--experience of a Polish center.

INTRODUCTION: Kidney transplantation is efficacious as a renal replacement, particularly pre-emptive living donation. In Poland, the rate of transplantation of living donor kidneys is only 3%. The aim of the study was to identify the most common reasons to disqualify a potential living kidney donor. METHODS: We evaluated 124 kidney donor candidates for 111 potential recipients at 1 medical center for genders and ages of donor and recipient; thus relation, donor disqualification reasons, number of potential donors for a particular recipient, prior transplantations, and kidney vasculature. RESULTS: The 111 recipients of ages 2-62 years had, 1, 2, or 3 potential donors were tested in 101, 1, and 7, cases respectively. We had 18.9% recipients referred for pre-emptive transplantation; 59.5% were on haemodialysis and 21.6% on peritoneal dialysis. In all, 89% recipients sought first kidney transplantations. Kidneys were procured from 49/124 (39.5%) of the initially evaluated donors. The full examination was completed by 92 potential donors with 68/124 donors disqualified early. Single and multiple renal arteries were detected in 56 and 36 potential donors, respectively. Donor disqualification was due to medical contraindications (39.7%), earlier transplantation from a deceased donor (25%), immunologic constraints (23.5%), donor consent withdrawn (6%) or psychological and social reasons (4.4%). CONCLUSIONS: A considerable number of donor candidates are disqualified for medical reasons.

Assessment of kidneys procured from expanded criteria donors before transplantation.

BACKGROUND: Using expanded criteria donor (ECD) organs is 1 strategy to make more organs available for transplant. To reduce the number of posttransplant complications and failures, there is a need to create a comprehensive system of evaluation before transplantation, especially for kidneys harvested from ECD. The aim of this study was to assess the results of kidneys procured from ECD seeking to discover the most useful factors for kidney evaluation before transplantation. PATIENTS AND METHODS: One hundred seventy-two patients received cadaveric renal transplants between January 1, 2006, and August 31, 2008. We collected data on donors, recipients, and perfusion parameters. We analyzed patient and graft survivals, as well as immediate, delayed, and slow graft function. Kidney recipient function was assessed by serum creatinine concentrations and by creatinine clearance calculated according to the Cockroft-Gault formula. Renal biopsy specimens were obtained in the perioperative periods 147 cases. RESULTS: The overall 1-year graft survival was 86.9%. More than 25% of transplanted kidneys were harvested from ECD. There were no significant differences in patient survival between recipients of standard criteria donor kidneys (RSCDK) versus of expanded criteria donor kidneys (RECDK). One-year graft survival was higher among the RSCDK group than the RECDK group, namely, 94.4% versus 62.5%, (P = .004). There were no differences in the incidence of primary nonfunction or in delayed graft function between the groups. RECDK were more likely to show slow graft function (69.2% vs 37.8%; P = .033). A lower graft survival at 6 months after transplantation was observed among organs harvested from ECD compared with standard criteria donor (SCD) kidneys who showed histologic lesions or a flow at the fourth hour of machine perfusion below 0.4 mL/g. Using a logistic regression model, chronic histologic changes were shown to influence kidney survival at 6 months after transplantation. CONCLUSION: There was no significant difference in patient survival between recipients of kidneys harvested from expanded versus standard criteria donors. ECD kidneys displayed lower graft survival rates. There was no significant difference in the incidence of delayed graft function between recipients of kidneys harvested from expanded versus standard criteria donors. Pretransplant evaluation of ECD kidneys should include 3 variables: donor parameters, histologic findings, and machine perfusion parameters.

Complications of transplantation of kidneys from expanded-criteria donors.

BACKGROUND: Organ shortage is the primary barrier to kidney transplantation. To maximize organ use, organs from expanded-criteria donors (ECDs) have been used increasingly. Expanded-criteria donors are defined as individuals older than 60 years or older than 50 years with at least 2 of the following risk factors: hypertension, stroke as the cause of death, or serum creatinine concentration greater than 1.5 mg/dL. OBJECTIVE: To assess the incidence of complications posttransplantation in ECD kidneys compared with kidneys from standard-criteria cadaveric donors (SCDs). PATIENTS AND METHODS: One hundred seventy-two patients received cadaveric renal transplants between January 1, 2006, and August 31, 2008. Donor and recipient data were collected, as well as patient and graft survival and immediate, delayed, or slow graft function. Complication rates for lymphocele, urinary leak, thrombosis, hematoma, urinary tract infection, and cytomegalovirus infection were recorded. Follow-up was for 3 to 35 months, ending on November 30, 2008. RESULTS: Overall, mean 1-year graft survival was 86.9%, and mean creatinine concentration was 1.58 mg/dL. One incidence of primary nonfunction (0.6%) was observed. More than 25% of transplanted kidneys were from ECDs. No significant differences were noted in postoperative complications between recipients of ECD or SCD organs. CONCLUSION: The rate of complications in recipients of ECD and SCD kidneys is comparable.

Surgical site infections in the early posttransplant period after simultaneous pancreas-kidney transplantation.

OBJECTIVE: This study evaluated the frequency of microbial isolates and their susceptibility profiles among cultures from the "surgical site" of 26 simultaneous pancreas-kidney (SPKT) recipients in the early posttransplant period. PATIENTS AND METHODS: Data on microbiologic cultures of 26 adult patients undergoing SPKT were collected prospectively from 2001 to the end of 2006. The isolation and identification of cultured micro-organisms was performed according to standard microbiological procedures and commercially available tests. Susceptibility of the strains to antibacterial agents was made by the Clinical and Laboratory Standards Institute (CLSI) guidelines. RESULTS: All patients were followed prospectively for the first 4 weeks after surgery yielding 168 microbial isolates from the surgical site. The most commonly isolated organisms were Gram-positive bacteria (65.5%) with domination of staphylococci (52.7%) as methicillin-resistant S aureus and methicillin-resistant coagulase-negative staphylococci. The second most common were enterococci (33.6%) with the presence of an high level aminoglycoside-resistant strains (64.9%) and vancomycin-resistant strains (2.7%). Gram-negative bacteria comprised 19% of positive cultures; among them were isolated extended spectrum beta-lactamase producers and carbapenem-resistant strains. Yeast-like fungi comprised 15.5% of positive cultures. In conclusion, we observed predominantly Gram-positive bacteria, comprising 65.5% of isolates. The increased proportion of multi-drug-resistant bacterial isolates may be due to the frequent prophylaxis of bacterial infections in patients.

Urinary tract infections in the early posttransplant period after simultaneous pancreas-kidney transplantation.

OBJECTIVE: Urinary tract infection (UTI) is among the common infection in simultaneous pancreas-kidney transplantation (SPKT). PATIENTS AND METHODS: The study included 26 adult patients undergoing SPKT between September 2001 and December 2006. All the patients were followed prospectively for UTI during the first 4 weeks after surgery. Urine samples were investigated for bacteriologic cultures. The micro-organisms were identified in accordance with standard bacteriologic procedures. Susceptibility testing was carried out using Clinical and Laboratory Standards Institute (CLSI) procedures. RESULTS: Among 77 urine specimens obtained from all recipients during the first month, there were 30 isolated bacterial strains. The most common were Gram-positive bacteria (53.3%) with predominance of enterococci (75%) associated with high levels of aminoglycoside resistant strains (HLAR; 58.3%) and vancomycin-resistant strains (VRE; 25%). Gram-negative bacteria were detected in 46.7% of positive cultures. CONCLUSIONS: In our study, enterococci predominated as 75% of Gram-positive isolates. The increased proportion of multi-drug-resistant bacteria, which can caused severe UTI in patients after SPKT, may be due to the frequent use of prophylaxis of bacterial infections in patients.

Etiologic agents of bacteremia in the early period after simultaneous pancreas-kidney transplantation.

BACKGROUND: Bacteremia is among the known complications in simultaneous pancreas-kidney transplantation (SPKT). This study evaluated the frequency of microbial isolates and their susceptibility profiles among cultures of clinical samples obtained from blood and from the tips of blood vessel catheters of 26 SPKT recipients suspected of bacteremia in the early posttransplant period. PATIENTS AND METHODS: Data on microbiologic blood cultures of 26 adult patients undergoing SPKT were collected prospectively from 2001 to the end of 2006. The isolation and identification of cultured microorganisms were performed according to standard microbiological procedures and commercially available tests. The susceptibility of the strains to antibacterial agents was established by the Clinical and Laboratory Standards Institute guidelines. RESULTS: All patients were followed prospectively for the first 4 weeks after surgery. Among 66 clinical samples, there were 23 microbial isolates from blood samples of 17 recipients and catheter tips of 12 recipients. The most common isolates were Gram-positive bacteria (73.9%) with domination of staphylococci (64.7%) and MRCNS strains (81.8%). Gram-negative bacteria comprised 17.4% of positive cultures, whereas yeast-like fungi, 8.7% with a predominance of Candida glabrata. CONCLUSION: Our study showed predominately Gram-positive bacteria in 73.9% of isolates. The increased proportion of multi-drug-resistant bacteria and fungi to antimicrobial agents may be due to the frequent use of these agents for prophylaxis of bacterial infections in patients.

Chlamydia pneumoniae infection in patients after kidney transplantation treated with spiramycin.

INTRODUCTION: Previous research has pointed to a role of Chlamydia pneumoniae infection in the development of chronic renal allograft dysfunction, chronic liver rejection, and vasculopathy in the transplanted heart. The aim of this study was to evaluate the presence of C. pneumoniae prior to and after kidney transplantation as well as to determine the role of spiramycin therapy among kidney transplant recipients. MATERIALS AND METHODS: The study group consisted of 50 patients (25 pairs) who received kidney transplants from cadaveric donors. One of the 2 kidneys from a donor was transplanted to a patient randomized to spiramycin (2 x 3 million U/d orally for 3 months; group S) and the other to a patient assigned as control (group C). Markers of infection were assessed on day 1 posttransplantation and 3 months later (average, 94 days). All 50 patients were examined for the presence of bacterial DNA in peripheral blood leukocytes using real-time polymerase chain reaction (PCR) and for titers of serum anti C. pneumoniae immunoglobulin (IgG) and IgA antibodies using microimmunofluorescence (MIF). C. pneumoniae infection was diagnosed by the presence of C. pneumoniae DNA in peripheral blood leukocytes or positive antibodies of both classes. RESULTS: C. pneumoniae infection was initially diagnosed in 14 patients among group S and 8 patients among group C (P = not significant [ns]) and after 3 months in 12 and 9 patients, respectively (P = ns). Conversion from positive to negative C. pneumoniae status occured in 7 patients among group S and 1 patient among group C (P = .04). Conversion from negative to positive C. pneumoniae status occured in 5 patients from group S and 2 patients from group C (P = ns). CONCLUSIONS: These results suggest a possible role for spiramycin treatment of C pneumoniae infection in kidney allograft recipients. C. pneumoniae infection diagnosis and treatment should be considered to be routine for every patient awaiting transplantation.

The role of orexin A in the control of prolactin and growth hormone secretions in sheep--in vitro study.

Orexin A may play a special role in animals' sensitivity to the day length changes such as sheep. The localization of mRNA for prepro-orexin in the ovine hypothalamus was found to correspond to the pattern described in rodents. The results of that research also showed that the expression of the orexin gene depends on the length of a day and is higher during short days. Other study revealed that mRNA for orexin receptors (OxR)1 and OxR2 shows strong expression in the anterior, intermediate and posterior pituitary lobes of the rat. In addition, it was also found that in the anterior pituitary, OxR1 is more strongly expressed than OxR2. These observations indicate that the pituitary gland is capable of receiving the orexin signal. The aim of the study was to determine the interaction of season and orexin A on PRL and GH secretion by pituitary explants in short-term culture. Studies were carried out on pituitaries explants collected from lactating Polish Longwool sheep during the long (LD, May, n=5) and short day (SD, December, n=5). Glands were transected saggitally into halves, with each incubated in 2.5 ml of M-199 for 180-min in medium containing either 0 or 1000 ng/ml of orexin A. Treatment with orexin during LD increased significantly the secretion of PRL (P < 0.01) and GH (P < 0.05), compared to controls. In cultures from glands collected during SD, orexin significantly (P < 0.05) decreased the secretion of both hormones, compared to controls. We conclude that the secretion of PRL and GH from the ovine pituitary gland is negatively responsive to orexin A during SD; whereas orexin may stimulate PRL and GH secretion during LD.

Long-term results of kidney transplantation from HCV-positive donors.

BACKGROUND: Due to the shortage of organs for transplantation, procurement of kidneys from marginal donors is inevitable. Not infrequently, these donors are infected with hepatitis C virus (HCV). AIM: We sought to determine the effect of transplanting kidneys from anti-HCV-positive donors to anti-HCV-positive recipients. PATIENTS AND METHODS: Among 765 procedures between 1994 and 2006, 259 kidney recipients were anti-HCV-positive, including 60 who received kidneys from anti-HCV-positive donors (HCV(+)/HCV(+) group) and the others, from seronegative donors (HCV(-)/HCV(+) group). The control group of 506 seronegative recipients received kidneys from seronegative donors (HCV(-)/HCV(-) group). All kidneys from anti-HCV-positive donors were preserved with machine perfusion. We investigated recipient liver function tests (LFTs; alanine aminotrasferase, aspartate aminotransferase; alkaline phosphatase, and bilirubin), graft survival, and patient survival. RESULTS: No significant difference was observed between the groups among the biochemistry results (LFTs, creatinine at 5 years). No significant differences, were observed in patient survival, graft survival, or number of patients returning to dialysis. CONCLUSION: Transplantation of kidneys from HCV-positive donors to HCV-positive recipients did not influence long-term liver function, or long-term renal allograft function. This strategy enhances the availability of transplantation as means of end-stage renal disease treatment.