Molecular Assessment of Kidney Allografts: Are We Closer to a Daily Routine?

Kidney allograft pathology assessment has been traditionally based on clinical and histological criteria. Despite improvements in Banff histological classification, the diagnostics in particular cases is problematic reflecting a complex pathogenesis of graft injuries. With the advent of molecular techniques, polymerase-chain reaction, oligo- and microarray technologies allowed to study molecular phenotypes of graft injuries, especially acute and chronic rejections. Moreover, development of the molecular microscope diagnostic system (MMDx) to assess kidney graft biopsies, represents the first clinical application of a microarray-based method in transplantation. Whether MMDx may replace conventional pathology is the subject of ongoing research, however this platform is particularly useful in complex histological findings and may help clinicians to guide the therapy.

Significance of the Pathologic Findings in Implantation Biopsies for Kidney Allografts Survival.

BACKGROUND: There is no consistent association between individual histological lesions and composite scores in donor kidney biopsy and transplant outcomes. OBJECTIVE: To evaluate which acute or chronic individual histological lesions and composite scores in donor kidney were associated with graft survival in the recipient. METHODS: We investigated the association of individual histological lesions and 8 composite scoring systems in implantation biopsies of cadaveric (n=101) and living (n=29) kidneys with 5-year death-censored graft survival. RESULTS: We found a high frequency of chronic lesions in donor kidneys, mostly associated with arteriosclerosis, and less dependent from donor age. Acute, chronic, and total Banff scores for post-transplant biopsies, chronic and total Banff scores for pre-implant biopsies, donor damage score and chronic damage score predicted death-censored graft loss. However, only chronic and total Banff-scores had significant effects in multivariate model. Chronic pre-implant and total post-transplant Banff scores demonstrated the highest area under the curve (AUC) of 0.722 and 0.717, respectively. Among individual lesions, glomerulosclerosis ≥20%, interstitial inflammation >0, arteriosclerosis =3, arteriolar hyalinosis >0, and interstitial fibrosis >0, assessed with Banff-grading criteria, were associated with lower allograft survival. We created the Donor Kidney Damage Index (DKDI), by summing regression coefficients for these lesions, which yielded the AUC of 0.747. When combined with retransplantation, cold ischemia time and acute rejection, DKDI, chronic pre-implant and total post-transplant Banff scores further improved their predictive accuracy, yielding AUCs of 0.842, 0.807, and 0.802, respectively. CONCLUSION: DKDI, chronic pre-implant and total post-transplant Banff scores alone and combined with clinical variables may facilitate decision making in post-transplant period.

[URINE UNDER CHRONIC DYSFUNCTION OF RENAL ALLO-TRANSPLANT].

The study was organized to provide additional characteristic of chronic dysfunction of renal allo-transplant using such biomarkers of serum and urine as enzymes (alanine aminotransferase), aspartate aminotransferase, gamma- glutamiltransferase, alkaline phosphatase, N-acetyl-ß-D-glucosaminidase, interleukins (IL-2, IL-8, IL-10), beta-2- microglobulin. The chronic dysfunction of renal allo-transplant is characterized by increasing of concentration of IL-10 and beta-2-microglobulin in serum and increasing of concentration of beta-2-microglobulin, IL-2, IL-8 in urine and increasing of activity of N-acetyl-ß-D-glucosaminidase, alkaline phosphatase, aspartate aminotransferase, gamma-glutamiltransferase as compared with patients with satisfactory function of renal allo-transplant. The multivariant logistic regression analysis established that only activity of N-acetyl-ß-D-glucosaminidase in urine was reliably independently related to chronic dysfunction of renal allo-transplant. It is assumed that increasing of concentration of beta-2-microglobulin in serum testifies glomerular dysfunction and in urine--tubular dysfunction of renal allo-transplant. The enzymeuria indicates continuing damage of epithelium of proximal tubules of nephron. The classification of patients with satisfactory function and chronic dysfunction of renal allo-transplant established that the highest indicators of square under ROC-curves had concentration of beta-2-microglobulin in serum (0.858 ± 0.061) and urine (0.733 ± 0.079) and activity of N-acetyl-ß-D-glucosaminidase in urine (0.701 ± 0.061). To specify diagnosis of chronic dysfunction of renal allo-transplant the most useful (ratio of likelihood of positive result 10 and 11 correspondingly) are tests of beta-2- microglobulin in serum (more than 8.55 mkg/ml) and N-acetyl-ß-D-glucosaminidase/creatinine in urine (more than 34 nmol/(sl)/ mmol/l). These discoveries require further validation and confirmation by implementation of morphological analysis of bioptat of renal allo-transplant.

[P300 potential parameters at the stages of formation of the subcortical vascular dementia in elderly].

Auditory P300 latency and amplitude values were assessed in 11 elderly persons without cognitive impairments (WCI), mean age--(70.73 +/- 4.24) years, 20 patients with subcortical vascular mild cognitive impairment (SCMCI), mean age--(75.35 +/- 5.48) years and 20 patients with subcortical vascular dementia (SCVaD), mean age (75.80 +/- 6.51) years. Obtained data suggest that P300 latency value increase at the stages of the SCVaD progression. Mean values of P300 latency were (341.09 +/- 107.70) ms in WCI group, (655.70 +/- 87.08) ms in SCMCI patients, (732.45 +/- 74.64) ms in SCVaD patients. There were significant differences between the groups. This parameter allows differentiating patients with SCMCI from elderly persons WCI, and patients with SCVaD from SCMCI. Increasing P300 latency has sufficient relationship to severity and specificity of cognitive deficit of SCVaD stages development reflecting progressive deterioration of attention, working memory, kinetic and regulator praxis.

[Serum level of S100B as a marker of progression of vascular mild cognitive impairment into subcortical vascular dementia and therapy effectiveness].

We evaluated serum level of S100B in 11 patients with subcortical vascular dementia (SVD) and 19 patients with subcortical vascular mild cognitive impairment (SVMCI). Comparable groups were age-matched (79.18 +/- 7.76 in SVD group, 77.84 +/- 3.83 in SVMCI; P = 0.53). 22 patients were assessed after 1 month therapy. It was shown that the serum S100B level significantly increased--(0.065 +/- 0.020) micro/l (P = 0.0005) in SVD patients comparing to SVMCI ones - (0.043 +/- 0.010) microg/l. S100B level was significantly correlated with the clinical parameters: MMSE performance (r(s) = -0.61), CDR (r(s) = 0.58), attention task (r(s) = -0.46), pseudobulbar syndrome severity (r(s) = 0.37) and walking alteration (r(s)= 0.37). In patients with reduction of S100B level due to therapy (positive dynamics, n = 12) we registered significant improvement of some clinical parameters: MMSE, attention level, walking. In patients with increasing of S100B level (negative dynamics, n = 10) we didn't registered improvement of any clinical parameters. We made the conclusion that the serum level of S100B could be used as marker of progression SVMCI into SVD and therapy effectiveness.

[Up-to-date techniques of diagnostics, prognosis and prophylaxis of dysfunction of the renal allotransplant].

On the basis of their experience with 530 kidney transplantations and literature data the authors argue the necessity of careful assessment of the kidney allograft state at all stages of its life. An accent is thus done on the necessity of the complex diagnostics with using noninvasive and invasive methods that will facilitate the kidney allograft dysfunction prediction and prevention, and longer time of the kidney allograft survival.

[The state of lipid peroxidation and antioxidant defense system in recipients of renal allotransplant].

The peroxidal oxidation of lipids (POL) and antioxidant defense (AOD) system state in the patients blood before renal transplantation performing and on the fourth day after it were studied, depending on kind of familial or potential agonal with beating (DBH) or nonbeating (DNBH) heart donor applied and the presence of complications--delayed renal autotransplant (RAT) function and acute reaction of rejection (ARR). In chronic renal insufficiency, RAT transplantation from cadaver, DNBH and ARR the POL processes intensification and AOD system activity lowering were noted.

[Estimation of soluble serum CD30 in the diagnosis of early renal allograft dysfunction].

We aimed to reveal factors influencing serum soluble CD30 level in the recipients of kidney allograft and to estimate its pathogenetic significance. We tested the sCD30 level in the serum before and the 4th day after operation by ELISA. It was established, thats CD30 levels before transplantation were virtually the same in patients who experienced rejection and in non-rejecting patients. However, there was a significant decrease in the level of sCD30 after transplantation in non-rejecting patients, contrary to rejecting patients. A significant decrease of sCD30 level was detected on the day 4th after the transplantation independently of dialysis requirement. The decrease of sCD30 on the day 4th after operation in the patients with delayed graft function and its stability in the patients with acute rejection may be used distinguish these complications.

[Study of apoptosis manifestations in streptozotocin diabetes mellitus]. / Vyvchennia proiaviv apoptozu pry streptozototsynovomu tsukrovomu diabeti.

The aim of this work was to study manifestations of apoptosis in pancreas, brain and myocardium of rats at the type I diabetes mellitus. To reveal the apoptotic changes in cells, specific damages of DNA were identified by the TUNEL-method. Specific for apoptosis changes in chromatin and cell membranes were found after counterstaining of a specimen by methyl green. In addition, the apoptotic and non-apoptotic cells were differentiated automatically with the help of image analyzing computer system by an index of optical density of cellular nucleus. This method allowed to find out for sure that apoptotic cells were available in all the studied tissues of healthy animals. However the possibility of their revealing by the TUNEL-method depends on the way of tissue fixation. Development of the type I diabetes mellitus was accompanied with an increase in the number of such cells, which gives evidence for an important role of apoptosis in diabetes pathogenesis.