Alternatives to zinc oxide in pig production.

Zinc oxide (ZnO) has been applied for many years in the production of pigs to reduce the number of diarrhoea in weaned piglets. In June 2022, the European Union banned the use of zinc oxide (ZnO) in pig feed. According to scientific reports, the may reason was the accumulation of this microelement in the environment of pig production. It has been shown that frequent application of ZnO can lead to increased antibiotic resistance in pathogenic swine microflora. The main alternatives to ZnO are probiotics, prebiotics, organic acids, essential oils, and liquid feeding systems. Alternatives to ZnO can be successfully used in pig production to reduce the number of diarrhoea among piglets during the postweaning period. Additional reports indicated that bacteriophage supplementation has a positive effect on the health of pigs. The article provides an overview of current ZnO substitutes that can be used in pig farming.

Effects of the microencapsulated feed additive of lactic acid bacteria on production parameters and post-vaccinal immune response in pigs.

The aim of the study was to determine the effects of feed addition of LAVIPAN PL5 probiotic preparation containing compositions of microencapsulated lactic acid bacteria (Leuconostoc mesenteroides, Lactobacillus casei, Lactobacillus plantarum, Pediococcus pentosaceus) on production parameters and post-vaccinal immune response in pigs under field condition. The study was performed on 400 pigs in total and 60 pigs from this group were used to evaluate the effect of the product tested on the post-vaccinal response. The animals were divided into two groups: control group, fed without additive of LAVIPAN PL5 and the study group, receiving LAVIPAN PL5 at doses recommended by manufacturer from weaning to the end of fattening. The following parameters were recorded: main production parameters, including weight gains, fattening time (slaughter age) and animal health status during the study (mortality), and specific humoral post-vaccinal response after vaccination against swine erysipelas. The results indicate that the application of LAVIPAN PL5 had positive influence on the animals` productivity and did not significantly affect the post-vaccinal antibody levels and the development and maintenance of the post-vaccinal response, albeit the levels of antibodies were slightly higher in the animal receiving the test preparation. The higher average daily weight gains (by over 3%) which resulted in a 2 kg higher average weight at slaughter and a reduction of the fattening period by 5 days, undoubtedly contributed to significant economic benefits.

Novel statistical approach for evaluating flow cytometric in vitro micronucleus data.

Statistical methods currently recommended for analysis of in vitro micronucleus data are based on small sample sizes. The tests are designed to evaluate linear trends and differences between treated and control samples. When using flow cytometric analysis, >5 times the number of cells are easily evaluated, and the variance estimates from these large samples are small. Application of these recommended tests to large samples resulted in statistically significant outcomes which were not considered to be biologically meaningful. Alternative statistical methods for testing trends and differences among treatments that were either widely used, or sample-size independent, were investigated. Using data from 95 experiments (from 2011-2013) where 19% of the experiments were considered positive, results for the various statistical methods were compared. When using either the recommended or alternate methods, 42-68% of the experiments resulted in statistically significant results (p < 0.05). A new concept was then tested using the same data sets: the "z' factor", designed to identify 'hits' during high throughput screening. Using this simple-to-compute statistic the number of significant calls was reduced to 27%. Then, when combined with a biological criterion based on historical vehicle control data, there was restoration of the original positive frequency (19%). Given the larger sample sizes evaluated using flow cytometry, we have demonstrated that traditional statistical tests may be overly sensitive to small changes in micronucleus induction, and that a simple-to-compute index of separation (z') may be a better tool for analysis, provided that the response is first determined to be biologically meaningful. Environ. Mol. Mutagen. 57:589-604, 2016. © 2016 Wiley Periodicals, Inc.

Interactive Pharmacometric Applications Using R and the Shiny Package.

Interactive applications, developed using Shiny for the R programming language, have the potential to revolutionize the sharing and communication of pharmacometric model simulations. Shiny allows customization of the application's user-interface to provide an elegant environment for displaying user-input controls and simulation output-where the latter simultaneously updates with changing input. The flexible nature of the R language makes simulations of population variability possible thus promoting the combination of Shiny with R in model visualization.

Thoracic endovascular aortic repair with left subclavian artery coverage without prophylactic revascularisation-early and midterm results.

BACKGROUND: The management of the left subclavian artery when coverage is necessary during thoracic aorta endografting remains a matter of debate. MATERIALS AND METHODS: A retrospective analysis of a single-centre experience with thoracic endovascular aorta repair (TEVAR) was performed. Between April 2004 and October 2012, 125 cases of TEVAR were performed. The analysis focused on patients who required coverage of the left subclavian artery (LSA). We analysed mortality and morbidity with special attention to the rates of cerebrovascular accidents (CVAs) and spinal cord ischaemia (SCI) in the early and midterm. RESULTS: Of the 125 patients, 53 (42 %, group A) required an intentional coverage of the LSA to obtain an adequate proximal seal for the endograft; the remaining patients constituted group B. None of the patients in group A had protective LSA revascularisation prior to TEVAR. The primary technical success rate was 79.2 vs. 90.3 % (group A vs. group B, p = 0.08), and the primary clinical success rate was 77.4 vs. 82 % (group A vs. group B, p = 0.53). The 30-day mortality rate was 11.3 vs. 11.1 % (group A vs. group B, p = 0.97). The 30-day morbidity was 7.5 vs. 13.9 % (group A vs. group B, p = 0.4). CVA occurred in 1.9 % of group A patients, compared to 1.4 % of patients from group B (p = 0.82). The SCI incidence rate was 0 vs. 1.4 % (p = 0.39). The mean follow-up of group A was 24.1 months (range 2-64.6 months, SD = 19). Additionally, the 1-year estimated survival was 85.5 %, and the 3-year estimated survival was 78 %. There were no midterm CVAs; one event of SCI occurred in the seventh post-operative month in group A. CONCLUSION: Our analysis, although retrospective and based on one institution experience, shows a realistic population of TEVAR patients. We prove that TEVAR with coverage of LSA origin can be accomplished with minimal neurological morbidity in this patient population. The study shows that LSA revascularisation is not mandatory before endograft deployment, especially in emergency settings. We also prove that although zone 2 TEVAR extends the proximal landing zone, it does not prevent type IA endoleaks from appearing. A multicentre randomised control trial with higher number of patients is necessary for proper, robust conclusion to be established.

Association of hypothyroid disease in Doberman Pinscher dogs with a rare major histocompatibility complex DLA class II haplotype.

Canine hypothyroid disease is similar to Hashimoto's disease in humans, which has been shown to be associated with human major histocompatibility complex (MHC) genes. We have collected 27 Doberman Pinschers affected with primary hypothyroid disease and compared their MHC class II haplotypes with 129 unaffected Doberman Pinschers. Three dog-leucocyte antigen (DLA) genes, DLA-DRB1, DQA1 and DQB1, were characterized by sequence-based typing and assigned to haplotypes for each dog. One rare haplotype was found at an increased frequency in the affected dogs compared to the unaffected dogs (Odds ratio = 2.43, P < 0.02). This haplotype has only been found in Doberman Pinschers and Labradors to date.

Role of p53 tumor suppressor in ageing: regulation of transient cell cycle arrest and terminal senescence.

In this study we investigated the function of p53 as a regulator of cell cycle progression in cycling and senescent cells. Using the conditional temperature-sensitive (ts) mutant we could prevent the detrimental effect of constitutive expression of high levels of wt p53 protein. High levels of wt p53 inhibited cell proliferation by blocking the cells to progress from G1 to S phase of the cell cycle. Flow cytometric analysis revelaed a maintenance of G1 cell population for a longer time depending on the prolonged expression of wt p53 protein. The p53 mediated inhibition of cell proliferation and of the cycle was reversible. However, a spontaneous increase of wt p53 occurring in ageing normal human MRC-5 fibroblasts was associated with irreversible reduction of proliferative potential. The accumulation of G1 cells was detected by flow cytometry. By the measurement of DNA content it is not possible to discriminate between cells arrested in G1 and G0 phase, therefore, the expression of G1 markers was determined. Analysis of the expression of distinct cell cycle regulators revealed that quiescent MRC-5 cells were in G0 phase. Our results indicate that cell cycle arrest occurring in senescent cells is associated with the G0 transition.

Cell cycle arrest induced in human breast cancer cells by cyclin-dependent kinase inhibitors: a comparison of the effects exerted by roscovitine and olomoucine.

Cyclin-dependent kinases (CDKs) are serine/threonine kinases that play a key role in the regulation of the cell cycle progression. In proliferating cells, distinct CDKs activated upon complexing with specific cyclins and upon site-specific phosphorylation coordinate in an orchestrated way the appropriate transition between consecutive phases of the cell cycle. Aberrant expression or altered activity of distinct CDK complexes results in escape of cells from the cell cycle control and leads to malignant transformation. Therefore, the inhibition of CDKs in malignant cells provides a new strategy in the fight against cancer. Recently, selective CDK inhibitors targeting distinct CDKs were developed. They represent promising anti-cancer drugs due to their strong anti-proliferative efficacy combined with a relative low direct cytotoxicity. The aim of this study was to compare the effect of two related CDK inhibitors: roscovitine (ROSC) and olomoucine (OLO) on the cell cycle progression in human breast cancer MCF-7 cells. Both examined CDK inhibitors differentially affected the cell cycle progression in MCF-7 cels. Whereas ROSC arrested cells in G(2)/M, OLO inhibited cells at S to G(2) transition and increased the number of cells residing in the S-phase. Moreover, both CDK inhibitors modulated the cell cycle progression with distinct kinetics. Accumulation of G(2)/M-arrested cells beginning 6 h after exposure of cells to ROSC coincided with a strong up-regulation of the p53. Interestingly, ROSC triggered apoptosis in MCF-7 cells by activation of mitochondrial pathway. Loss of the integrity of mitochondrial membrane observed after exposure of cells to ROSC for 6 h led to release of distinct mitochondrial proteins, e.g. apoptosis inducing factor (AIF). In contrast to ROSC, OLO-induced cell cycle changes could be detected after 12 h of the treatment. OLO did not up-regulate p53 protein. It indicates that both examined CDK inhibitors are selective and block the cell cycle progression of human breast carcinoma cells at different phases.

Activation of p53 protein in normal and in tumor cells by a novel anticancer agent CHS 828.

CHS 828, a novel cyanoguanidine, represents a new class of drugs for cancer therapy, with an unknown primary mechanism of action. It is generally known that anticancer drugs induce p53 response thereby triggering cell cycle arrest or apoptosis. We investigated the effect of CHS 828 on p53 response in normal and tumor cells and compared this effect with that exerted by conventional anticancer drugs. After 24 h of treatment with CHS 828, we observed a dose-dependent up-regulation of wild type (WT) p53 protein in human breast carcinoma MCF-7 cells as well as in normal human and mouse fibroblasts. The highest p53 increase was observed at 300 nM to 1 microM CHS 828. CHS 828 induced phosphorylation of p53 protein at Ser-15 in normal cells. However, the drug failed to induce p53 protein in mouse cells in which the poly(ADP-ribose)-1 gene (PARP-1) was disrupted even at a 30-fold higher dose and after prolonged treatment. Combined treatment of PARP-1 -/- cells by multidrug resistance modulators did not alter p53 expression. CHS 828 inhibited cell proliferation and DNA replication in the tested cells. Interestingly, DNA synthesis as well as proliferation of PARP-1 deficient cells was inhibited by drug concentrations that were approximately 3-fold lower than their conventional counterparts. Treatment of cells with CHS 828 for 48 h did not induce apoptosis.

DLA-DRB1 histocompatibility genotyping using RT-nested PCR and cycle sequencing.

Class-II histocompatibility genes are associated with predisposition to autoimmune diseases in many mammal species. We have developed a technique using reverse transcriptase and nested-PCR for amplification from blood samples of expressed sequences encoded by canine DLA-DRB1 loci. In the first polymerase chain reaction (PCR), we utilize primers DR-SP and DR-STOP as developed by Sarmiento et al. (1990). In the nested PCR, we utilize two additional primers, namely primer 57 [5'-TCTTGGAGGCTCCTGGATGACAGC-3'] and primer 367 [5'-CACAACTACGGGGTGATTGAGAGC-3'] to produce a 334 bp amplified product. After digestion with restriction endonucleases, some of the alleles can be identified by restriction fragment length polymorphism (RFLP). The increasing information on new DLA-DRB1 alleles over the last two years renders the DLA-DRB1 too diverse for convenient use of RFLP. However, the expressed sequences amplified by our protocol can be conveniently identified by cycle sequencing. This RT n-PCR protocol will suffice for the genotyping of individual dogs at the DLA-DRB1 locus.

Expanded indications for the treatment of postcatheterization femoral pseudoaneurysms with ultrasound-guided compression.

BACKGROUND: The purpose of this study was to define the factors that predict successful ultrasound-guided compression repair (UGCR) of postcatheterization femoral pseudoaneurysms (PA) and to determine risks for recurrence, the most appropriate follow-up, and the optimal management of compression failures and recurrences. METHODS: A retrospective chart review was made. RESULTS: UGCR thrombosed 52 of 60 PA (87%). Predictors of compression failure were PA size of 8 cm and an associated arteriovenous fistula (AVF). AVF was the only predictor of recurrence. All seven recurrences (13%) were discovered on the first follow-up scan. Four were thrombosed with additional UGCR. Late rescanning after a mean of 264 days identified no recurrences. Four anticoagulated patients failed initial UGCR but were thrombosed in another session when their anticoagulation was briefly reversed. CONCLUSIONS: UGCR should be the initial management of PA because it is safe, effective, and durable. Temporary discontinuation of anticoagulation and multiple prolonged compression sessions may help treat recalcitrant cases. One follow-up scan is adequate for most patients. Recurrences should be initially treated with repeat UGCR.

D-dimer and thrombin/antithrombin III complex--diagnostic tools in deep venous thrombosis?

In order to evaluate the usefulness of determining D-dimer and thrombin-antithrombin III complex (TAT) in the diagnosis of deep venous thrombosis (DVT), three D-dimer assays were tried: one ELISA and one latex test from Diagnostica Stago and one new latex method from Biopool. TAT was assessed using an ELISA (Behringwerke). We studied 96 consecutive outpatients with suspected DVT, of whom 36 had phlebographically confirmed DVT. Statistical calculations showed high sensitivity and a negative predictive value for the D-dimer ELISA (97% for both), confirming the results obtained by others. The new latex method (Biopool) showed similar figures (96% for both). The latex method from Diagnostica Stago and TAT showed lower sensitivity and negative predictive values. No differences in the D-dimer results were found with or without antifibrinolytics in the tubes for blood sampling. Our data suggest that negative results when using the new simple and cheap latex method (Biopool) may reduce the number of phlebographic examinations.

Biologic markers in hospital workers exposed to low levels of ethylene oxide.

Operators of hospital sterilizers that use ethylene oxide were studied to determine if there was a relationship between exposure and a battery of biological markers. A total of 73 workers from nine hospitals in the United States (U.S.) and one hospital in Mexico City was evaluated for ethylene oxide exposure during four months prior to collection of peripheral blood. The frequency of hemoglobin adducts (p = 0.0006) and sister-chromatid exchanges (SCEs) (p = 0.002) increased with cumulative exposure to ethylene oxide in U.S. subjects when controlling by regression analysis for various confounding factors, including cigarette smoking. Hemoglobin adducts, but not SCEs, were also increased in Mexican subjects (p = 0.0012). Chromosomal micronuclei showed no consistent relationship with exposure. The U.S. study participants were classified by four-month cumulative exposure levels of 10 ppm-h (n = 8), greater than 0 to 32 ppm-h (n = 32) and greater than 32 ppm-h (n = 11) of ethylene oxide exposure. The group with an exposure of greater than 32 ppm-h had an increased frequency of hemoglobin adducts (p = 0.002) and SCEs (p = 0.0001) compared to the nonexposed group. The estimated mean of the 8-h time-weighted average (8-h TWA) exposure levels for the highest U.S. exposure group (greater than 32 ppm-h) was 0.16 +/- 0.007 ppm (mean +/- SD). A similar exposure-related differential was observed in the Mexican subjects for hemoglobin adducts (p = 0.04) but not for SCEs. The latter finding may have been due to longer shipping times for the specimens in the cytogenetic assays. The estimated mean of the 8-h TWA exposure levels for the highest Mexican exposure group (greater than 32 ppm-h) was 0.48 +/- 0.08 ppm. This study is the third to suggest that exposures less than the U.S. OSHA standard of 1 ppm 8-h TWA result in biochemical and biologic changes. It is not known whether these changes may be indicative of increased risk of disease; however, they do appear to reflect exposure to relatively low levels of ethylene oxide. The exact meaning of these changes is unknown.

Comparison of single-, double- or triple-exposure protocols for the rodent bone marrow/peripheral blood micronucleus assay using 4-aminobiphenyl and treosulphan.

Two human carcinogens, 4-aminobiphenyl (4AB) and treosulphan (Treo), were tested in male B6C3F1 mice for the induction of micronuclei in bone marrow and peripheral blood cells by 1-, 2- and 3-exposure protocols. Both compounds tested positive. The magnitude of response with respect to the incidence of micronucleated polychromatic erythrocytes by 2- and 3-exposure protocols was considerably higher than by the single-exposure protocol. The peripheral blood results for Treo were as typically seen with a 24-h delay when compared to the bone marrow. The peripheral blood results for 4AB, however, differed from those expected. The incidence of MN-PCE in peripheral blood of animals exposed to 4AB was significantly greater than seen in the bone marrow in 2- and 3-exposure protocols. There was also an increase in the % PCE at the 60 mg/kg dose level as a function of time. Based on these studies, it is concluded that a step-wise scoring scheme may be the best protocol for rodent micronucleus assay, involving a 3-exposure protocol with single sampling of bone marrow (24 h after the last treatment) and two samplings of peripheral blood (24 h and 48 h after the first treatment). This approach is cost-effective, it limits the number of animals required and provides maximum sensitivity.

Double contrast barium enema and flexible rectosigmoidoscopy: a reliable diagnostic combination for detection of colorectal neoplasm.

The efficiency of double contrast barium enema and flexible rectosigmoidoscopy (to 60 cm) in the assessment of patients with a positive Hemoccult II test in a randomized screening study for colorectal neoplasms was evaluated. A positive test was present in 625 patients, of whom 530 had a complete enema and rectosigmoidoscopic assessment. A carcinoma was diagnosed in 26 and an adenoma greater than or equal to 1 cm in diameter in 71. As a control, 323 patients with a negative assessment repeated the Hemoccult II test and of these 67 had a positive second test, of whom 55 underwent colonoscopy. One carcinoma (Dukes' A) and two adenomas greater than or equal to 1 cm in diameter were diagnosed. The efficiency of the assessment was also checked by rescreening the whole group 1-2 years after the first study and by continuing follow-up. It was found that two more carcinomas and one adenoma greater than or equal to 1 cm in diameter had been overlooked at the primary assessment. The sensitivity for neoplasms greater than or equal to 1 cm in diameter at the primary assessment was 72 per cent for double contrast barium enema and 86 per cent for rectosigmoidoscopy. The sensitivity for the combined methods was 94 per cent and the specificity was 99 per cent. The combination of double contrast barium enema and rectosigmoidoscopy in the primary assessment of patients with a positive Hemoccult II test gives an acceptable result and immediate retesting of those with a negative assessment is not necessary.

Synoviorthesis with 198Au colloid gold in haemophilia patients. A preliminary report.

In 1988-1989 fifteen patients with severe haemophilia A and recurrent bleedings into the knee joint, aged from 19 to 44 years were treated by an intraarticular injection of 198Au colloid gold. So far 10 of them were assessed after 6 months follow-up. In 6 cases cesseation and in 2 cases reduction in number and volume of bleedings were observed. Only in 2 patients the frequency of haemarthroses remained unchanged. No significant difference in tracer uptake was observed between pretreatment 99mTc-pertechnate gamma scans of the knee joints and controls completed 6 months after the radiogold injections. It is worthy to stress the lower costs of the 198Au synoviorthesis as compared with surgical synovectomy of the knee joint. The radioisotope method is also much less traumatic to the patient than the surgical one.

[Functional capacity of pancreatic B cells in patients with diabetes mellitus type 2 with late true ineffectiveness of sulfonylurea derivatives]. / Sprawnosc czynnosciowa komórek B wysp trzustkowych u chorych na cukrzyce typu 2 z pózna rzeczywista nieskutecznoscia pochodynych sulfonylomocznika.

Fasting concentration of the C peptide in serum was estimated in 150 patients with type 2 diabetes treated with insulin because of the late, true ineffectiveness of the sulphonylurea derivatives. In 36 patients selected out of the total group at random the secretion of that peptide was measured after i.v. injection of 1 mg of glucagon. Only 9 patients showed trace amounts of that peptide at morning fast (Group A--0.17 +/- 0.08 nmol/l), in 69 the secretion was normal (Sub-Group B1--0.80 +/- 0.25 nmol/l), in 48 moderately elevated (Sub-Group B2--1.67 +/- 0.10 nmol/l) and in 24 markedly elevated (Sub-Group B3--4.54 +/- 2.57 nmol/l). The increments of the peptide C concentration after glucagon stimulation were parallel to its fasting concentration, which indicated a proper reactivity of the pancreatic beta-cells in patients with normal or increased basal secretion. The patients with only trace secretion of the peptide C differed from the other by their small, normal body mass and by a longer duration of insulin treatment. Very similar insulin needs must be stressed in the patients of the Groups A and B as well as within the Sub-Groups B. In patients with hyperactivity of the beta-cells (Sub-Group B2 and B3) no differences were found, as compared with the other patients, in the prevalence of chronic diabetes complications of the micro- or macroangiopathy type, also prevalence of hypertension was equal. The results presented show that in the most patients with type 2 diabetes, with the late, true ineffectiveness of the sulphonylurea derivatives the secretory function of the pancreatic islets beta-cells remains normal or is even increased.