Results of Renal Transplant From Deceased Anti-hepatitis C Virus Donors, Poland 1998-2012.

Kidney transplant (KTx) is the best method of renal insufficiency treatment. In dialyzed patients, mortality rises with the time on dialysis. There is a continuing shortage of organs for transplantation, hence a propensity to expand the donor pool with expanded-criteria donors, anti-hepatitis C virus-positive included. In the above case a transmission of hepatitis C virus (HCV) genotype to recipient is present. It has been proven that contamination with more than 1 HCV genotype did not worsen KTx outcomes. There are 2.6% anti-HCV(+) donors in Poland. Use is only possible in cases of anti-HCV(+) and anti-HCV RNA(+) recipients. METHODS: Retrospective analysis covered 8675 deceased donors (1998-2012 Polish data from Poltransplant). The early (after 12 months) and late (after 60 months) graft and patient survival was assessed in KTx recipients, with documented recipient and donor data spanning at least 1 year after KTx. In comprehensive analysis, 7016 KTx recipients with known anti-HCV status were included according to anti-HCV profile of recipient and donor. The results are in absolute and percentage values and P < .05 assessed with χ2 test. RESULTS: Twelve-month survival: recipient (R) (95%), graft (G) (89%), total; R (95% vs 89%, P < .001), G (88 vs 79, P < .001) in HCV(-) to HCV(+/-) vs HCV(+) to HCV(+); R (95 vs 94, P = .2), G (88 vs 83, P < .001), HCV(-) to HCV(-) vs HCV(-) to HCV(+); R (93 vs 95, P = .004), G (82 vs 89, P < .001) in HCV(+/-) to HCV(+) vs HCV(-) to HCV(-); R (95 vs 89, P < .001), G (88 vs 79, P < .001) in HCV(-) to HCV(-) vs HCV(+) vs HCV (+). Sixty-month survival: R (86%), G (75%), total; R (84 vs 88, P = .01), G (63 vs 71, P = .001) in HCV(+/-) to HCV(+) vs HCV(-) to HCV(-); R (88 vs 80, P = .003) in HCV(-) to HCV(-) vs HCV(+) to HCV(+). CONCLUSIONS: The worst anti-HCV serological profile was HCV(+) to HCV(+), although transplanting HCV(+) to HCV(+) did not worsen outcomes in that group. Worse KTx outcomes of HCV(+) over HCV(-) donors can be attributed to HCV(+) status of the recipient.

Sex-related differences of bone properties of pelvic limb and bone metabolism indices in 14-month-old ostriches (Struthio camelus).

1. Sex-related differences of long pelvic limb bones and serum bone metabolism indices were evaluated in 14-month-old female (N = 7) and male (N = 7) ostriches of similar body weights. 2. Densitometric parameters of femur, tibia and tarsometatarsus were determined using quantitative computed tomography (volumetric bone mineral density, calcium hydroxyapatite density and mean volumetric bone mineral density) and dual energy X-ray absorptiometry (bone mineral density and bone mineral content) methods. Geometrical parameters such as cortical bone area, cross-sectional area, second moment of inertia, mean relative wall thickness and cortical index were determined in the midshaft of bones. Mechanical properties of bones (maximum elastic strength and ultimate strength) were evaluated using three-point bending test. Serum concentrations of free amino acids, osteocalcin, N-terminal propeptide of type I procollagen, C-terminal telopeptides of type II collagen and total antioxidative capacity were also determined. 3. Bone weight and relative bone weight of all bones were significantly higher in males than in females. Significantly lower values of trabecular bone mineral density and calcium hydroxyapatite density were found in the trabecular bone of tibia in males. The highest number of the sex-related differences was observed in the tarsometatarsus where bone length, bone mineral content, cortical bone area, cross-sectional area and ultimate strength were higher in males. Serum concentrations of taurine, hydroxyproline, valine and isoleucine were significantly higher in males. 4. Higher loading of the tarsometatarsus in comparison to femur and tibia may be an important factor interacting with sex hormones in regulation of bone formation and mineralisation processes. Sex-related differences of bone properties were associated with increased serum concentration of selected amino acids in males.

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.

Influence of Double-J Catheters on Urinary Infections After Kidney Transplantation.

BACKGROUND: One of the most important problem in kidney transplantation is risk of the urinary anastomosis stricture. In uncertain cases the use of a double-J (or JJ) catheter is a standard solution. In case of urinary leak or fistula after the reanastomosis operation, transuretheral JJ implantation is used. A number of patients after JJ use present urinary infection. METHODS: Between 2012 and 2014, 283 kidney transplantations were performed in our center. In 77 cases (27.2%), a JJ was used at the time of operation, in 10 other cases (3.5%) during postoperative period. RESULTS: Urinary infection was observed in 95 patients (33.6%), with Proteus, Klebsiella, Enterobacter, Escherichia, Enterococcus, Pseudomonas, Morganella, and Staphylococcus cultures. In the group of patients without JJ catheters, infections were found in 27.6% of cases and in group with JJ, in 47.1% (46.8% in intraoperative use of catheters and 50% in postoperative curse). CONCLUSIONS: The analysis shows that use of JJ catheters gives urinary infections almost twice more frequent (47.1% vs 27.6%; P = .001) with no difference if the implantation took place during or after the operation.

Pterostilbene induces accumulation of autophagic vacuoles followed by cell death in HL60 human leukemia cells.

Pterostilbene, a naturally occurring structural analog of resveratrol, has been reported to exert antiproliferative and proapoptotic effects in various cancer types. Recently, it has been demonstrated to induce both autophagy and apoptosis in human bladder and breast cancer cell lines. The aim of this study was to evaluate the effects of pterostilbene on HL60 human leukemia cells. Cell morphology was examined using confocal and electron microscopy. Cell viability was determined by MTT, neutral red uptake and trypan blue exclusion assays. LC3 processing was studied based on Western blotting and immunofluorescence analyses. Flow cytometry was used to study cell cycle distribution, phosphatidylserine externalization, caspase activation, disruption of mitochondrial membrane potential and intracellular production of reactive oxygen species. DNA degradation was examined by gel electrophoresis. We found that treatment of HL60 cells with pterostilbene at the IC90 concentration resulted in the G0/G1 cell cycle arrest. Pterostilbene induced conversion of cytosolic LC3-I to membrane-bound LC3-II and accumulation of large LC3-positive vacuolar structures. Pterostilbene also led to phosphatidylserine externalization, internucleosomal DNA fragmentation, caspase activation and disruption of mitochondrial membrane potential. Moreover, it did not induce oxidative stress. Our results suggest that pterostilbene induces accumulation of autophagic vacuoles followed by cell death in HL60 cells.

A note on the organization and expression of ß-defensin genes in Polish goats.

Nucleotide (cds) and amino acids sequences of the caprine ß2-defensin genes were in silico compared to search for the sequence variation and for the LAP gene sequences in the goat genome and for the presence of LAP gene transcripts in goat tissues. The comparison of the exon sequences revealed that the first 64 amino acids are identical in both LAP and ß1-defensin. However, the GBD-1 prepropeptide is shorter by 18 amino acids due to the presence of the stop codon UAA at position 209-211 in GBD-1 mRNA. The LAP gene, which was found, so far, only in Indian goat breeds, is absent in the genome of Polish dairy goats. The introns of the caprine ß1- and ß2-defensin genes were, for the first time, sequenced; their sequences showed 99.6 % identity, differing in six nucleotide positions.

Twelve-year cohort study on the influence of caprine arthritis-encephalitis virus infection on milk yield and composition.

This long-term observational cohort study was carried out to evaluate the effect of caprine arthritis-encephalitis virus (CAEV) infection on the quantitative and qualitative characteristics of milk production in dairy goats. For this purpose, a dairy herd comprising both CAEV-infected and uninfected female goats was observed for 12 consecutive years. Records on daily milk yield, somatic cell count (SCC), and contents of the major milk components (fat, protein and lactose) were collected every month. In total, 3,042 records (1,114 from CAEV-positive and 1,928 from CAEV-negative animals) from 177 female goats were used for statistical analysis. The multi-trait repeatability test-day animal model using the derivative-free multivariate analysis package with the average information-REML method was applied to eliminate the influence of factors other than CAEV infection on milk production in goats. The statistical significance of the differences between estimates for seropositive and seronegative goats was evaluated using Student's t-test. The effect of age of goats (parity) on their serological status was also estimated with the one-trait repeatability test-day model. The serological status of goats was linked to parity: the higher the parity, the greater the probability of CAEV infection. No significant differences between infected and uninfected goats with respect to daily milk yield and SCC were found. On the other hand, the milk of uninfected goats contained more total protein (3.40% vs. 3.35%), fat (3.69% vs. 3.54%), and lactose (4.30% vs. 4.25%) than the milk of infected goats. Even though these differences were highly significant, they were small when expressed numerically.

Vaccination against porcine parvovirus protects against disease, but does not prevent infection and virus shedding after challenge infection with a heterologous virus strain.

The demonstration of field isolates of porcine parvovirus (PPV) that differ genetically and antigenically from vaccine strains of PPV raises the question of whether the broadly used inactivated vaccines can still protect sows against the novel viruses. Ten specific-pathogen-free primiparous sows were assigned to three groups and were vaccinated with one of two vaccines based on the old vaccine strains, or served as non-vaccinated controls. After insemination, all sows were challenged with the prototype genotype 2 virus, PPV-27a, on gestation day 41; fetuses were delivered on gestation day 90 and examined for virus infection. The fetuses of the vaccinated sows were protected against disease, but both the vaccinated and the non-vaccinated sows showed a marked increase in antibody titres after challenge infection, indicating replication of the challenge virus. All sows (vaccinated and non-vaccinated) shed the challenge virus for at least 10 days after infection, with no difference in the pattern or duration of virus shedding.

Chemical composition, cholesterol content, and fatty acid profile of pigeon meat as influenced by meat-type breeds.

Two muscles, breast and thigh, were excised from 3 pigeon meat-type breeds (Europigeon, Wroclawski, King) and were analyzed for proximate analysis, cholesterol content, and fatty acid (FA) profile. Among the breeds considered, the lowest protein content of breast muscle was found in Kings (21.73%), whereas the highest fat (7.07%) and ash (1.11%) content of breast muscle (P

Evaluation of matrix metalloproteinases in serum of patients with amyotrophic lateral sclerosis with pattern recognition methods.

Matrix metalloproteinases (MMPs) are zinc-dependent endopeptidases, which are present in central and peripheral nervous system. They are considered to be involved in the pathogenesis of several neurological diseases, as multiple sclerosis, Alzheimer disease, and amyotrophic lateral sclerosis (ALS). The aim of the present study was to evaluate the application of the pattern recognition methods for the assessment of MMPs in serum of patients with ALS. Thirty patients with amyotrophic lateral sclerosis (ALS), in two subgroups: (i) with mild and (ii) severe progressing ALS, and 15 control healthy subjects were studied. The metalloproteinases MT-MMP-1, MMP-2, MMP-9 were examined. Additional variables (age of subjects and disease duration) were also analyzed by using a standard, parallel and hierarchical classifiers. Our results indicate that: (i) MMP-2 in serum may be an important marker for the evaluation of ALS progress; (ii) the set of two features {MT-MMP-1, MMP-9} may be helpful in differentiation between ALS and healthy subjects; (iii) the error rates obtained for the pair-wise linear classifier were similar to those received for the classifiers (standard, parallel, and hierarchical) based on k-NN rule. We conclude that the pattern recognition methods may be useful for the evaluation of significance MMPs as markers in neurodegenerative diseases, such as ALS.

Different distribution of CD4 and CD8 T cells in synovial membrane and peripheral blood of rheumatoid arthritis and osteoarthritis patients.

Rheumatoid arthritis (RA) and osteoarthritis (OA) are chronic diseases associated with morphological joint changes. Synovial membrane (SM) involvement was established for RA, but the data for OA are limited, because OA is usually regarded as noninflammatory disease. Changes in immune system in RA are not limited to joints, and the significant role of T cells of peripheral blood (PB) is not disputable. However, there is still an open debate about PB immunological profile in OA. Therefore, we decided to measure the distribution of CD4+ and CD8+ T cells, regarding CD28 expression, both in PB and SM of RA and OA patients, on the same day. Altogether, eleven RA patients, 11 OA patients and similar numbers of age-matched healthy controls were included into the study. Flow cytometry was used for T cells subpopulation distinguishing and quantification; monoclonal antibodies against CD3, CD4, CD8 and CD28 with different fluorochromes were used for stainings. The RA patients had significantly higher percentage of CD3+4+ cells in PB as compared to OA patients and relevant control group. Both within the CD4+ and CD8+ compartments, significantly lower percentages of cells bearing the CD28 marker were found in the PB of OA as compared to RA patients. The proportion of CD3+CD4+ cells in SM was dependent on age of OA patients, older OA patients had significantly higher value of their SM/blood ratio than RA patients. Older OA subjects were also characterized by higher values of the SM/blood ratio of both CD4+CD28+ and CD8+CD28+ subpopulations than RA or younger OA patients. In conclusion, in contrast to the traditional view of OA disease, our results give support to the hypothesis that OA may also (like RA) be a disease with a local immunological involvement.

Cytoprotective effect of lithium against spontaneous and induced apoptosis of lymphoid cell line MOLT-4.

Lithium (Li) is still useful in the treatment of bipolar disorder. Cellular mechanisms of Li action are not fully understood and include some cytoprotective properties. Data concerning Li effect on the apoptotic mechanisms in cells other than neurons are fragmentary and contradictory. We have investigated anti-apoptotic activity of Li in a lymphoid derived MOLT-4 cell line. Spontaneous and camptothecin-induced apoptosis was analyzed in cells treated with 0-20 mM Li carbonate. Early apoptosis was identified as significant mitochondrial depolarization (JC-1 staining). Later stages of apoptosis were estimated with annexin V binding and by the proportion of cells containing sub-G1 amounts of DNA (PI staining). We have observed a biphasic effect of Li on the proportion of spontaneously apoptotic cells;namely, low (therapeutic) concentrations of Li had a significant effect stabilizing the mitochondrial membrane polarization, while 10 and 20mM Li increased apoptosis. The latter could be seen both as mitochondrial depolarization as well as an increased proportion of sub-G1 cells, accompanied by reduced proportion of S phase cells. Li at concentrations above 2 mM had a significant, dose-dependent, anti-apoptotic effect on the cells undergoing camptothecin induced apoptosis. In conclusion, demonstrated cytoprotective effect of Li is at least partially related to stabilization of mitochondrial membrane potential and to the reduction of DNA damaging effects in proliferating cells; both may form part of the mechanism through which Li is useful in therapy of bipolar disorder, but may have more general consequences.

Flow cytometry as a rapid test for detection of penicillin resistance directly in bacterial cells in Enterococcus faecalis and Staphylococcus aureus.

We studied the usefulness of flow cytometry for detection of penicillin resistance in E. faecalis and S. aureus by direct binding of commercially available fluorescent penicillin, Bocillin FL, to cells obtained from culture. There were significantly lower percentages of fluorescent cells and median and mean fluorescence values per particle in penicillin-resistant than in penicillin-sensitive strains of both species observed. The method allows rapid detection of penicillin resistance in S. aureus and E. faecalis. The results encourage further investigations on the detection of antibiotic resistance in bacteria using flow cytometry.

Recognition of gait disturbances in patients with normal pressure hydrocephalus using a computer dynography system.

Gait and body balance disturbances are important clinical problems in patients with normal pressure hydrocephalus (NPH). They affect patients' locomotion and lead to a higher risk of falls. The gait pattern may be described as durations of the single and double support and of a stance phase. The aim of the present study was to apply the pattern recognition methods for the evaluation of gait disturbances in patients with NPH before and after neurosurgical treatment (shunt implantation). The results indicate that the parameters measured with a Computer DynoGraphy (CDG) system may effectively differentiate changes of gait in patients with NPH.

Induced sputum in patients with interstitial lung disease: a non-invasive surrogate for certain parameters in bronchoalveolar lavage fluid.

Induced sputum is a useful non-invasive method for the assessment of airway and parenchymal lung diseases. This study aimed to compare induced sputum and bronchoalveolar lavage fluid (BALF) cellular composition and T-lymphocyte subpopulations in patients with interstitial lung disease. We evaluated 33 patients: 15 with sarcoidosis, 11 with hypersensitivity pneumonitis, and 7 with idiopathic pulmonary fibrosis. The percentage of macrophages was significantly lower in induced sputum than in BALF in sarcoidosis (P=0.005), and the percentage of neutrophils was higher in induced sputum than in BALF in sarcoidosis (P=0.001) and hypersensitivity pneumonitis (P=0.006). A significant correlation was found between the BALF and induced sputum CD4+, CD8+ subsets and the CD4+/CD8+ ratio in both the whole patient group (r(s)=0.80, r(s)=0.88, r(s)=0.88, P<0.001, respectively) and in the 3 subgroups. A strong correlation of the T-lymphocyte subsets in induced sputum and BALF in patients with interstitial lung disease shows that induced sputum may be a non-invasive surrogate for certain parameters in BALF in these patients.

Respiratory responses to acute intermittent hypoxia and hypercapnia in awake rats.

This article deals with the recognition of early changes in the breathing pattern, in response to acute intermittent stimuli in awake rats. Two different types of stimuli were given: 9% hypoxia in N(2) and 10% hypercapnia in O(2). Animals were exposed to 3 consecutive cycles consisting of 3-min stimulus period separated by 8-min normoxic recovery intervals. Features of the breathing pattern, such as respiratory frequency, tidal volume, minute ventilation, inspiration and expiration times, peak inspiratory and expiratory flows, were measured by whole body plethysmography. The data were analyzed with the use of pattern recognition methods. We conclude that the overall respiratory changes were rather slight. However, computerized analysis using a k-nearest neighbor decision rule (k-NN) allowed for a good recognition of the respiratory responses to the stimuli. The misclassification rate (E(r)) varied from 5 to 10%. After feature selection, E(r) decreased below 1%. The k-NN classifier differentiated correctly also the type of intermittent stimulus. Our experimental results demonstrate usefulness of pattern recognition algorithms in studying respiratory effects in biological models.

The polymorphism in the beta4-defensin gene and its association with production and somatic cell count in Holstein-Friesian cows.

The aim of this study was to find a polymorphism of the bovine beta4-defensin gene and search for its association with milk yield and composition and with the somatic cell count in milk. The data were from the years 1999 to 2004 on 212 Holstein-Friesian (HF) dairy cows, descended from 70 sires. Based on the sequence of the bovine beta4-defensin gene (GenBank no. AF008307) the primers were designed for the amplification of the 924-bp or 393-bp long fragments. The 924-bp long fragment was sequenced and the sequence was compared with that available in the GenBank. Ten putative nucleotide sequence polymorphisms were found in the intron of the bovine beta4-defensin gene. One of them, a C-->T transition at position 2239, that creates a new NlaIII (Hin1II) restriction site, was genotyped with polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in a cohort of 212 HF cows. The CC genotype was the most common (72%). The heterozygous CT genotype was found in 26% of the genotyped cows and four cows (2%) were TT homozygotes. In order to determine the relationship between the polymorphism of the beta4-defensin gene and milk production traits a multi-trait repeatability test-day animal model was used. The Derivative-free Multivariate analysis program was used for computation. The differences between estimates for genotypes were checked using Student's t-test. The model included the animal genotype, year-season of calving and parity as fixed effects and the animal additive genetic effect and permanent environmental effect of individual cows as well as dates of the tests as random effects. Significant associations were found between the RFLP-NlaIII and milk fat, protein and lactose contents. Also, a significant effect was shown of the defensin genotype on the somatic cell count in the milk.

Distinguishing the strength of hypoxic stimulus in intermittent hypoxia.

The intermittent hypoxia (IH) phenomenon is a subject of intensive examinations. In this study we examined whether it could be possible to distinguish the strength of the hypoxic stimulus given in IH cycles on the basis of observed changes in the breathing pattern. We investigated the ventilatory responses to five hypoxic-normoxic cycles (1 min hypoxia/3 min normoxia) in rabbits. Two different hypoxic stimuli were given: gas mixtures of 14% and 11% O2 in N2, each one in a separate run of IH. Ventilatory features: frequency (f), tidal volume (VT) and minute ventilation (VE) were analyzed using the algorithms of the pattern recognition theory. The probability of wrong classification was used as a criterion for the recognition quality evaluation. This probability can be estimated experimentally by calculating the percentage of misclassifications, i.e., an error rate (Er). When the features were analyzed alone, the VT offered the lowest misclassification rate of 19.3% and 10.3% in the stimulus and normoxic periods, respectively. However, using the single features measured during the stimulus and recovery phases allowed to decrease the error rate more than 2-fold, achieving 4.3% for VT. The best results were obtained for both phases associated with single cycles of IH. Suitable feature selection procedures enabled reducing the global misclassification rate to 0.7%. In conclusion, the pattern recognition approach may potentially be useful for controlling the stimulus strength in intermittent hypoxic training.

Statistical evaluation of ventilatory patterns in response to intermittent hypoxia in the rabbit.

In the present study we investigated whether classical or non-classical statistical methods might be useful in the diagnosis of early changes evoked by intermittent hypoxia (IH) in an experimental model. The experiments were carried out in anesthetized, spontaneously breathing rabbits. IH consisted of 5 cycles of breathing 14% O(2) in N(2) for 1 min interspersed with 3 min normoxic intervals. The following ventilatory variables were evaluated: frequency breathing, tidal volume, and minute ventilation. The results indicate that IH had a progressively stimulatory effect on the baseline ventilation and on the hypoxic ventilatory responses. Further, the algorithms of the pattern recognition theory might be a suitable tool for the recognition of early ventilatory effects of recurrent hypoxic events in the IH model. The recognition of IH states may be useful in clinical and sports medicine.

Comparison of the immunofluorescence assay with RT-PCR and nested PCR in the diagnosis of canine distemper.

Two pairs of primers were prepared, both localized within the sequences of the nucleoprotein gene (NP) of canine distemper virus (CDV). A number of experiments were done to optimize the conditions of RT-PCR and nested PCR methods. The nucleic acids of the Onderstepoort, Rockborn, Snyder Hill and Lederle strains of CDV could be detected with these primers. However, they did not react with the sequences of the Edmonston strain of the measles virus. The detection limit for RT-PCR was 10 TCID50 and for nested PCR 0.1 TCID50 of CDV. The RT-PCR was able to demonstrate the nucleic acid of CDV in the blood of all seven puppies vaccinated with a modified live virus. Blood samples of 23 dogs clinically suspected of distemper were examined by RT-PCR combined with nested PCR, and the results were compared with the detection of the CDV antigen in the smears from the mucous membranes by the direct immunofluorescence (IF) test. Of the 23 dogs, 12 were positive in nested PCR, six in the IF assay, and only two in single RT-PCR. It is concluded that nested PCR seems to be the most sensitive method for ante-mortem diagnosis of canine distemper, especially in its subacute or chronic forms.