Use of elderly living kidney donors: twenty years' experience in the Balkans.

The Balkan region has dramatically changed during the last 20 years. Despite transplantation efforts, dialysis remains the usual way to treat end stage renal diseases. Living donor renal transplantation is still the predominant transplant activity. Seeking to solve the problem, we decided to accept expanded criteria living donors, including elderly, marginal, unrelated, and ABO-incompatible individuals. Herein we have presented our 20 years' experience with 230 living donor renal transplantations using elderly individuals, including 90 older than 65 years (mean age 68 ± 4.5; range = 65-86; ED group). The predominantly haploidentical recipients had a mean age of 45 ± 6 years (range = 18-66). Sequential immunosuppressive protocols were used in all cases including induction with anti-thymocyte-globulin or interleukin-2 receptor antagonists. We analyzed the 5-year Kaplan-Meier graft survival rate, rejection episodes, delayed graft function, and renal function for comparison with these outcomes of 110 kidneys from younger donors (mean age = 53.4 years; range = 25-62; YD group) and haploidethical recipients (mean age = 32.2, range = 16-42), performed within the same period. The 3- and 5-year cumulative graft survival rates in the ED group were 81% and 72% compared with 85% and 81% in the YD group respectively (P > .9; NS). The incidences of acute rejection episodes were also comparable for both groups (19% and 17%, respectively). Delayed graft function occurred in 15% of the ED group but only 8% of the YD group. The serum creatinine value at the end of 60 months' follow-up was 146.04 µmol/L in the ED group versus 123.38 µmol/L in the YD group (P < .001). There were no major surgical complications in either group. We recommend the use of elderly living donors as a valuable source of kidneys, especially in countries wherein deceased donor transplantation is not yet established.

Biventricular pacing in the early postoperative period after cardiac surgery.

Cardiac resynchronization therapy is not commonly used in the early postoperative period in patients undergoing cardiac surgery who have left ventricular (LV) dysfunction and a history of heart failure. We performed a prospective randomized clinical trial to compare atrial synchronous right ventricular (DDD RV) and biventricular (DDD BIV) pacing within 72 hours after cardiac surgery in patients with an EF ≤35 %, a QRS interval longer than 120 msec and who had LV dyssynchrony detected by real-time three-dimensional echocardiography (RT3DE). Epicardial pacing was provided by a modified Medtronic INSYNC III pacemaker. An LV epicardial pacing lead was implanted on the latest activated segment of the LV based on RT3DE. The study included 18 patients with ischemic heart disease, with or without valvular heart disease (14 men, 4 women, average age 71 years). Patients undergoing DDD BIV pacing had a statistically significant greater CO and CI (CO 6.7±1.8 l/min, CI 3.4±0.7 l/min/m(2)) than patients undergoing DDD RV pacing (CO 5.5±1.4 l/min, CI 2.8±0.7 l/min/m(2)), p<0.001. DDD BIV pacing in the early postoperative period after cardiac surgery corrects LV dyssynchrony and has better hemodynamic results than DDD RV pacing.

Tissue-engineered heart valves.

Currently-used mechanical and biological heart valve prostheses have several disadvantages. Mechanical prostheses, based on carbon, metallic and polymeric components, require permanent anticoagulation treatment, and their usage often leads to adverse reactions, e.g. thromboembolic complications and endocarditis. Xenogenous and allogenous biological prostheses are associated with immune reaction, thrombosis and degeneration, and thus they have a high rate of reoperation. Biological prostheses of autologous origin, such as pulmonary autografts, often burden the patient with a complicated surgery and the risk of reoperation. Therefore, efforts are being made to prepare bioartificial heart valves with an autologous biological component by methods of tissue engineering. They should be biocompatible, durable, endowed with appropriate mechanical properties and able to grow with a child. For this purpose, scaffolds composed of synthetic materials, such as poly(lactic acid), poly(caprolactone), poly(4-hydroxybutyrate), hydrogels or natural polymers, e.g. collagen, elastin, fibrin or hyaluronic acid, have been seeded with autologous differentiated, progenitor or stem cells. Promising results have been obtained with nanostructured scaffolds, and also with cultivation in special dynamic bioreactors prior to implantation of the bioartificial grafts into an animal organism.

Living-unrelated (paid) renal transplantation--ten years later.

Due to the increase of organ shortage and still inadequate development of cadaver transplantation, many end-stage patients from the Balkan region travel mostly to India to buy a kidney. Despite all the ethical dilemmas and discussions, organ sales is present nowdays in Third-World countries. Sixteen patients (13 from Macedonia and 3 from Kosovo, SCG) were observed clinically during a period of 10 years. Recipients of mean age 36.5 years (range 10 to 58) displayed the following underlying diseases: chronic glomerulonephritis (n = 5), urethral valves with reflux (n = 2), ADPKD (n = 1), hypertensive nephropathy (n = 4), lithiasis (n = 1), and unknown cause of ESRD (n = 3). The donor population was young (22 to 29 years). Most patient records did not include data on HLA, cross-match, MLC, kind of surgery, or usual pretransplant workup. The immunosuppressive protocol included CyA, PRED, and AZA or MMF. All transplanted patients were followed on an outpatient basis in our department; patients with complications were hospitalized. The 1, 3, 5, and 10 year Kaplan Meier graft survival rates were 78.6%, 50.2%, 33.3%, and 18.8%, respectively. Seven patients were lost (43.7%), two during the first month after transplantation, two at the end of the first year, and three at 5, 6, and 8 years thereafter. The main reasons for death were severe pulmonary infections with sepsis, hepatitis B with liver cirrhosis, Kala Azar, CMV, and cancer of the colon. Five grafts were lost due to repeated rejection episodes and chronic graft nephropathy. The last three cases remained with good renal function and actual serum creatinine values of 135 +/- 9. In view of this experience, the authors cannot recommend this type of transplantation, not only from the ethical point of view, but also from frequent medical and surgical complications which are sometimes life threatening.

[Mechanical cardiac support--the first use in Czech Republic]. / Pouzití mechanické srdecní podpory--první zkusenosti v Ceské republice.

BACKGROUND: Implantation of mechanical assist device is widely accepted modality of treatment of patients with refractory heart failure. In the present study we evaluated our first one-year experiences with this method for bridging patients to cardiac transplantation. METHODS AND RESULTS: Between April 2003 and May 2004, the Thoratec VAD (Thoratec, Pleasanton, CA, USA) was implanted in 6 patients ( males; age 28-61 years) as a bridge-to-transplant procedure after having received maximum inotropic support and who were at imminent risk of death. In all patients was performed VAD as biventricular device (BiVAD). During a week after placement was observed recovery of organs function in all patients. Five patients survived to heart transplantation. One patient died 21 days after BiVAD placement due to massive bleeding to the respiratory tract. In post-transplantation period 1 patient died second day from acute graft failure and other patient died 34 days after from intracranial bleeding. Three patients has been discharged from the hospital and they are surviving more than I year. CONCLUSIONS: Analysis of our first experiences with the Thoratec BiVAD implantation as bridging to heart transplantation suggests that it is well suited method with respect to long- term prognosis of this group of patients.

Living emotionally related renal transplantation (LERT)--single center experience in the Balkans.

BACKGROUND: As elsewhere, the growing organ shortage is a main problem for organ transplantation. To solve the problem, we started accepting genetically unrelated, but emotionally related living donors. METHODS: In the period of 1998-2002, 14 LERT are performed in the University Clinical Centre in Skopje, Republic of Macedonia. As suitable donors are used predominantly spouses, but also mother and brother in law. The immunosuppression included a quadruple protocol with Interleukin-2R antagonists, late cyclosporin A, MMF and steroids. The two-year graft and patients survival of LERT was compared with 22 living genetically related donor transplantation (LRT) performed in the same time. RESULTS: The two years graft survival was 100% in LERT and 92% in LRT. There are not any significant difference among the medical and surgical complications between the two groups of pts. The actual serum creatinin was 101+22 in LERT compared with 142+34 in LRT. CONCLUSION: The authors recommend the LERT as a valid alternative especially in the countries where the regular cadaver transplantation is not yet established.

Chaperone activity of tobacco HSP18, a small heat-shock protein, is inhibited by ATP.

NtHSP18P (HSP18), a cytosolic class I small heat-shock protein from tobacco pollen grains, was expressed in Escherichia coli. The viability of these cells was improved by 50% at 50 degrees C, demonstrating its functionality in vivo. Purified recombinant protein formed 240 kDa HSP18 oligomers, irrespective of temperature. These oligomers interacted with the model substrate citrate synthase (CS) to form large complexes in a temperature-dependent manner. Furthermore, HSP18 prevented thermally induced aggregation of CS at 45 degrees C. The fluorescence probe bis-ANS revealed the exposure of HSP18 hydrophobic surfaces at this temperature. Reactivation of chemically denatured CS was also significantly enhanced by HSP18. Surprisingly, HSP18 function was inhibited (in contrast to the related chaperone alphabeta-crystallin and plant sHSPs studied so far) by the presence of ATP in a concentration-dependent manner. The conformational changes of HSP18 imposed by ATP binding were indicated by the difference in the quenching of intrinsic tryptophan fluorescence, and implied more compact structure with ATP. Fluorescence measurements with bis-ANS showed that the conformational shift of HSP18 is suppressed in the presence of ATP. Decreased chaperone activity of HSP18 in the presence of ATP is caused by the lower affinity of conformationally blocked HSP18 for the substrate, as demonstrated by a higher susceptibility of model substrate, malate dehydrogenase, to proteolytic cleavage. Our results suggest that the chaperone activity of some plant sHSPs could be regulated by the availability of ATP in the cytoplasm, which would provide a mechanism to monitor the cell environment, control biological activity of sHSPs, and coordinate it with other ATP-dependent chaperones such as HSP70.

[Kidney transplantation using living donors over age 65]. / Transplantation rénale utilisant des donneurs vivants de plus de 65 ans.

Efforts to increase the donor pool of available organs have resulted in some unconventional kidney transplantation procedures. One of these is the use of elderly donors for both living and cadaver kidney transplantations. The aim of this study was to review our experience with kidney transplants from living elderly donors. During a period of 10 years, 70 living renal transplantations were performed. In 32 transplants the age of the donor was above 65 years (mean 69 +/- 4 years, range: 65 to 81 years) and in 10 of these 32 transplants the age of the donor was over 70 years. The survival rate was compared with that of 38 transplants from younger donors (mean age 51 +/- 6 years, range: 24 to 59 years). The time to cold and warm ischemia, the preservation procedure and time to anastomosis of blood vessels were comparable in both groups of donors. Immunosuppression included a sequential quadruple protocol, using thymoglobulin (ATG), prednisolone (PRED), azathioprin (AZA) and cyclosporin A (CsA), which replaced ATG/PRED after day seven. A triple drug maintenance therapy (AZA, PRED, CsA) was used in all recipients. Kaplan-Meier survival curves at 1, 3 and 5 years showed that graft survival was 88%, 79% and 64% respectively for grafts from the advanced age donor group and 92%, 82% and 68% respectively for grafts from the younger donor group. The difference was slightly statistically significant (p < 0.05). Functioning of the graft was delayed in six patients who had received grafts from elderly donors and in one patient who had received a graft from a young donor. Despite worse results in transplantation with grafts from elderly donors, we consider this population as an important source of kidneys, which might help solve the present organ shortage, especially in our region.

Changes of plasma fibronectin levels in pregnancy induced hypertension.

A number of laboratory tests are available for evaluation of hypertension in pregnancy. These tests can be used to either predict and/or prognosticate preeclampsia and other hypertensive disorders of pregnancy. The aim of this study was to evaluate alterations in fibronectin homeostasis in normotensive pregnancy and in hypertensive disorders of pregnancy subclassified into chronic hypertension, preeclampsia superimposed on chronic hypertension, and pregnancy induced hypertension. A prospective, longitudinal study was conducted in 115 pregnant women aged 20-39 years, divided into four groups: normotensive (n = 40), chronic hypertension (n = 18), preeclampsia superimposed on chronic hypertension (n = 20), and pregnancy induced hypertension (n = 37). Plasma concentrations of fibronectin were measured by using single radial immunodiffusion assay (RIA) in the 8th, 18th, 23rd, 28th, 32nd and 36th week of gestation. Plasma fibronectin concentration showed no significant changes in normotensive pregnancy, but was significantly elevated in the third trimester in women destined to become preeclamptic or with preeclampsia in whom it reached a mean (+/- SD) of 0.40 +/- 0.09 g/L in the 36th week of gestation. In the groups with preeclampsia superimposed on chronic hypertension and with pregnancy induced hypertension, there was a significant difference between plasma fibronectin concentrations in 32nd (p < 0.01) and 36th (p < 0.001) week of gestation compared with either other levels in the respective group (in the 8th, 18th, 23rd and 28th week of gestation) or those recorded in other groups in the same period of pregnancy. These results suggested that the measurement of plasma fibronectin might be of diagnostic value in preeclampsia but could not be considered a useful predictor for preeclampsia.

Complications of rhytidectomy in an otolaryngology training program.

OBJECTIVES/HYPOTHESIS: Complications of rhytidectomy have been widely reported in the literature. This study examines the incidence of complications after rhytidectomy in the hands of chief residents under appropriate attending supervision in an otolaryngology-head and neck surgery training program. MATERIALS AND METHODS: The charts of 96 consecutive SMAS rhytidectomy patients were retrospectively reviewed. Patients were selected for surgery from a clinic designed exclusively for cosmetic facial surgery patients. This clinic was run by the otolaryngology chief resident and was supervised by an attending staff surgeon. Most patients elected local anesthesia and sedation administered by the surgical team. Submental liposuction was performed followed by SMAS plication rhytidectomy. RESULTS: Follow-up ranged from 1 to 60 months. Complications included expanding hematoma (1%), temporary facial nerve weakness (3%), pretragal/mastoid skin slough (4.2%), permanent ear numbness (1%), hypertrophic scar (3.1%), wound infection (1%), and dissatisfaction with result (4%). There were no cases of permanent facial nerve injury. CONCLUSION: These complication rates compare favorably with reported rates of larger studies over the past 30 years. These data support the conclusion that rhytidectomy can be performed safely by otolaryngology residents with little morbidity and good patient satisfaction.

Otitis media following tympanostomy tube placement in children with IgG2 deficiency.

Children with IgG2 deficiency commonly develop recurrent acute otitis media. It is believed that these infections are secondary to impaired antibody response rather than eustachian tube dysfunction and are therefore less responsive to treatment with tympanostomy tubes. The authors compared the incidence of acute otitis media in IgG2-deficient patients following tympanostomy tube placement with controls in a retrospective cohort study. The charts of 20 patients (10 with IgG2 deficiency and 10 controls) were reviewed. Episodes of otitis media were recorded for 12 months. IgG2-deficient patients experienced three times as many occurrences of otitis media as did controls. This suggests that otitis media is much more common in these patients following tympanostomy tube placement. We believe that an immunodeficiency workup should be considered in patients with multiple episodes of otitis media following placement of tympanostomy tubes.

Topoisomerase I activity in squamous cell carcinoma of the head and neck.

In recent preclinical and clinical trials, topoisomerase I inhibitors have shown great promise as antitumor agents. These agents are most effective against tumors with high topoisomerase I activity. Therefore, determining topoisomerase I activity in advance may predict response to topoisomerase I inhibitors. Squamous cell carcinoma of the head and neck and normal tissue samples were obtained from 12 patients. Cellular extracts were prepared, and topoisomerase I activity assays were performed. The results suggest that topoisomerase I activity in squamous cell carcinoma of the head and neck is increased approximately sixtyfold compared to normal tissue. Increased activity often correlates with clinical responsiveness; these results predict that topoisomerase I inhibitors should be effective and selective against squamous cell carcinoma of the head and neck.

Contactless semi-implantable electromagnetic middle ear device for the treatment of sensorineural hearing loss. Short-term and long-term animal experiments.

A contactless electromagnetic hearing device has been designed following basic science experiments, improvement of electronics, and precision micromechanics. Different prototypes have been developed and tested in the laboratory, fresh human temporal bones, and acute and chronic animal experimentation. A conductive hearing loss model was first developed in the cat using samarium cobalt as the target magnet. Later, a highly efficient electromagnetic air-core coil was selected to vibrate a neodymium iron boron magnet cemented to the body of the incus and tested in acute and chronic experiments using the cat as the model. In this group of animals, the ossicular chain was left intact. There was no failure of the target magnet, driving coil, or implanted electronics. The only problem encountered in this evaluation was a malfunction of the receiving antenna that had to be redesigned and retrofitted into the implanted units. This system would be suitable for the treatment of moderate to severe sensorineural hearing loss. Planning to begin human clinical trials is ongoing.

Two nicking enzyme systems specific for mismatch-containing DNA in nuclear extracts from human cells.

We have identified two novel enzyme systems in human HeLa nuclear extracts that can nick at specific sites of DNA molecules with base mismatches, in addition to the T/G mismatch-specific nicking enzyme system (Wiebauer, K., and Jiricny, J. (1989) Nature 339, 234-236). One enzyme (called all-type) can nick all eight base mismatches with different efficiencies. The other (A/G-specific) nicks only DNA containing an A/G mismatch. The all-type enzyme can be separated from the T/G-specific and A/G-specific nicking enzymes by Bio-Rex 70 chromatography. Further purification on a DEAE-5PW column separated the A/G-specific nicking enzyme from the T/G-specific nicking enzyme. Therefore, at least three different enzyme systems are able to cleave mismatched DNA in HeLa nuclear extracts. The all-type and A/G-specific enzymes work at different optimal salt concentrations and cleave at different sites within the mismatched DNA. The all-type enzyme can only cleave at the first phosphodiester bond 5' to the mispaired bases. This enzyme shows nick disparity to only one DNA strand and may be involved in genetic recombination. The A/G-specific enzyme simultaneously makes incisions at the first phosphodiester bond both 5' and 3' to the mispaired adenine but not the guanine base. This enzyme may be involved in an A/G mismatch-specific repair similar to the Escherichia coli mutY (or micA)-dependent pathway.