Кількісний ПЛР-аналіз кишкової мікробіоти у підсисних поросят різних вікових груп за діареї

The article presents the results of quantitative PCR detection of intestinal intestinal microbiota in suckling piglets of different age groups with diarrhea. To determine the etiology of diarrhea in piglets, 4 groups of animals were formed – two experimental and two control aged 3–5 and 17–21 days of life. Each group consisted of 6 piglets. The piglets in the experimental groups showed signs of diarrhea, and the pigs in the control groups were clinically healthy. Rectal smears were taken from piglets and immediately frozen at a temperature of -18 °C – -22 °C and stored in this form until laboratory examination. Isolation of genetic material from rectal smears was performed using a set of reagents for DNA/RNA extraction manufactured by Biosellal (France) on an automatic device “KingFisher Duo Prime” (USA). Performed mechanical lysis of biological material on the device for homogenization “FastPrep-24” (France). Specific regions of rotavirus RNA types A and C (PRV A, PRV C), the causative agent of swine diarrhea (PEDV) and Cl DNA fragments were determined in the purified nucleic acid solution. perfringens, E. coli, Cryptosporidium spp, Eimeria spr, Cystoisospora suis, as well as toxin-producing genes A and B Cl. difficile. Determinations of pathogenic E. coli was performed by toxin genes – STa, STb, LT, STX1, STX2 and EAE, as well as for by  coding genes – F4, F5, F18 and F41. The obtained results of the study of the genetic material of microorganisms extracted from rectal smears from clinically healthy pigs and with manifestations of diarrhea characterize the state of the microbiome and the distribution of microorganisms in the intestines of suckling piglets. It was found that the species composition of intestinal microorganisms in clinically healthy suckling piglets is represented by two Cl bacteria. perfringens and E. coli with dominance of the latter, in the population of which there are trace amounts of enterotoxigenic forms. At the same time, it was found that diarrhea in piglets aged 3–5 days of age is induced by a pathogenic effect on the body of rotavirus type C, which is complicated by the effect of toxin synthesizing Cl. difficile, and in pigs 17–21 days of age – co-infection of rotavirus type A and enterotoxigenic forms of E. coli.У статті наведено результати кількісного ПЛР-аналізу кишкової мікробіоти у підсисних поросят різних вікових груп за діареї. Для визначення етіології діареї у поросят було сформовано 4 групи тварин – дві дослідні та дві контрольні віком 3–5 та 17–21 доба життя. Кожна група налічувала по 6 поросят. Поросята дослідних груп мали ознаки діареї, а свині контрольних груп були клінічно здоровими. Від поросят відбирали ректальні мазки які одразу заморожувались при температурі -18 °С – -22 °С і у такому вигляді зберігались до лабораторного дослідження. Виділення генетичного матеріалу з ректальних мазків проводили за допомогою комплекту реактивів для екстракції ДНК/РНК виробництва Biosellal (Франція) на автоматичному приладі “KingFisher Duo Prime” (США). Попередньо проводили механічний лізис біологічного матеріалу на приладі для гомогенізації “FastPrep-24” (Франція). У очищеному розчині нуклеїнових кислот визначали специфічні ділянки РНК ротавірусу типів А і С (PRV A, PRV C), збуднику епідемічної діареї свиней (PEDV) та фрагменти ДНК Cl. perfringens, E. coli, Cryptosporidium spp, Eimeria spр, Cystoisospora suis, а також токсин продукуючі гени А і В Cl. difficile. Визначення патогенних E. coli проводили за шістьма токсин кодуючими генами –STа, STb, LT, STX1, STX2 і EAE, а також за адгезин кодуючими генами – F4, F5, F18 і F41. Отримані результати дослідження генетичного матеріалу мікроорганізмів екстрагованого з ректальних мазках від свиней клінічно здорових та з проявами діареї характеризують стан мікробіому та розподіл мікроорганізмів у кишечнику підсисних поросят. Встановлено, що видовий склад мікроорганізмів кишечнику клінічно здорових підсисних поросят представлений двома бактеріями Cl. perfringens та E. coli з домінуванням останньої, у популяції якої присутні слідові кількості ентеротоксигенних форм кишкової палички. Одночасно з цим з’ясовано, що діареї у поросят віком 3–5 діб життя індуковані патогенною дією на організм ротавірусу типу С, що ускладнюється впливом В токсин синтезуючої Cl. difficile, а у свиней 17–21 добового віку – коінфекцією ротавірусу типу А та ентеротоксигенних форм E. coli

Impact of uncertainties in exposure assessment on estimates of thyroid cancer risk among Ukrainian children and adolescents exposed from the chernobyl accident

The 1986 accident at the Chernobyl nuclear power plant remains the most serious nuclear accident in history, and excess thyroid cancers, particularly among those exposed to releases of iodine-131 remain the best-documented sequelae. Failure to take dose-measurement error into account can lead to bias in assessments of dose-response slope. Although risks in the Ukrainian-US thyroid screening study have been previously evaluated, errors in dose assessments have not been addressed hitherto. Dose-response patterns were examined in a thyroid screening prevalence cohort of 13,127 persons aged <18 at the time of the accident who were resident in the most radioactively contaminated regions of Ukraine. We extended earlier analyses in this cohort by adjusting for dose error in the recently developed TD-10 dosimetry. Three methods of statistical correction, via two types of regression calibration, and Monte Carlo maximum-likelihood, were applied to the doses that can be derived from the ratio of thyroid activity to thyroid mass. The two components that make up this ratio have different types of error, Berkson error for thyroid mass and classical error for thyroid activity. The first regression-calibration method yielded estimates of excess odds ratio of 5.78 Gy-1 (95% CI 1.92, 27.04), about 7% higher than estimates unadjusted for dose error. The second regression-calibration method gave an excess odds ratio of 4.78 Gy-1 (95% CI 1.64, 19.69), about 11% lower than unadjusted analysis. The Monte Carlo maximum-likelihood method produced an excess odds ratio of 4.93 Gy-1 (95% CI 1.67, 19.90), about 8% lower than unadjusted analysis. There are borderline-significant (p= 0.101-0.112) indications of downward curvature in the dose response, allowing for which nearly doubled the low-dose linear coefficient. In conclusion, dose-error adjustment has comparatively modest effects on regression parameters, a consequence of the relatively small errors, of a mixture of Berkson and classical form, associated with thyroid dose assessment

Epidemiological, clinical, outcome and antibiotic susceptibility differences between PVL positive and PVL negative Staphylococcus aureus infections in Western Australia: A case control study

Background: Panton Valentine Leukocidin (PVL) has been associated with invasive Staphylococcus aureus soft tissue and pneumonic infections. Methods: From September 2007 to January 2009 at Royal Perth Hospital we tested for the PVL gene in S. aureus isolates from an invasive site, a suspected PVL-related soft tissue infection and all MRSA isolates. We could access medical records for 141 PVL positive (PVL + ve) infections and compared these to a control group comprised of 148 PVL negative (PVL-ve) infections. Results: In the PVL + ve group 62 isolates were MRSA (48 were ST93-MRSA-IV) and 79 isolates were methicillin-sensitive S. aureus, and in the PVL-ve group 56 were MRSA (50 were WA-MRSA strains) and 92 were methicillin-sensitive S. aureus. We found the presence of PVL to be significantly associated with younger age, aboriginality, intravenous drug use, community acquisition, shorter length of hospital stay and lower mortality at 1 year. Overall PVL + ve infections more often required surgical intervention (73.0% versus 44.6%, p &lt; 0.001) and were less often polymicrobial (8.5% versus 41.2%, p &lt; 0.001). PVL + ve isolates were more often susceptible to clindamycin (87.9% versus 73.0%, p = 0.002). Conclusions: This study demonstrates that PVL + ve infections are associated with a distinct clinical picture, predominantly pyogenic skin and soft tissue infections often requiring surgery, disproportionately affecting patients who are younger, indigenous or with fewer health-care risk factors

Фінансова архітектоніка

Навчальний посібник спрямований на обґрунтування теоретичних і прикладних засад фінансових процесів та явищ. Зокрема, розглядаються питання теорії розвитку фінансів, нормативного забезпечення розвитку та становлення фінансів, особливості становлення фінансової архітектоніки та функціонування фінансових систем, державних фінансів, місцевих фінансів, податків і податкової системи, фінансового ринку, міжнародних фінансів, фінансового менеджменту, фінансового аналізу, інформаційних систем і технологій у фінансах, статистичних способів та прийомів у фінансах, фінансів підприємств та фінансів домогосподарств. Видання передбачене для студентів вищих навчальних закладів, керівників та спеціалістів управлінських структур, науковців, фінансових організацій, аспірантів

Antibody responses in furunculosis patients vaccinated with autologous formalin-killed Staphylococcus aureus

Autologous vaccines (short: autovaccines) have been used since the beginning of the 20th century to treat chronic staphylococcal infections, but their mechanisms of action are still obscure. This prospective pilot study involved four patients with furunculosis who were vaccinated with autologous formalin-killed Staphylococcus aureus cells. Vaccines were individually prepared from the infecting S. aureus strain and repeatedly injected subcutaneously in increasing doses over several months. We characterized the virulence gene repertoire and spa genotype of the infecting and colonising S. aureus strains. Serum antibody responses to secreted and surface-bound bacterial antigens were determined by two-dimensional immunoblotting and flow-cytometry based assays (Luminex®). All patients reported clinical improvement. Molecular characterization showed that all strains isolated from one patient over time belonged to the same S. aureus clone. Already before treatment, there was robust antibody binding to a broad range of staphylococcal antigens. Autovaccination moderately boosted the IgG response to extracellular antigens in two patients, while the antibody response of the other two patients was not affected. Similarly, vaccination moderately enhanced the antibody response against some staphylococcal surface proteins, e.g. ClfA, ClfB, SdrD and SdrE. In summary, autovaccination only slightly boosted the pre-existing serum antibody response, predominantly to bacterial surface antigens

Identification and Characterization of Nucleolin as a COUP-TFII Coactivator of Retinoic Acid Receptor β Transcription in Breast Cancer Cells

The orphan nuclear receptor COUP-TFII plays an undefined role in breast cancer. Previously we reported lower COUP-TFII expression in tamoxifen/endocrine-resistant versus sensitive breast cancer cell lines. The identification of COUP-TFII-interacting proteins will help to elucidate its mechanism of action as a transcriptional regulator in breast cancer.FLAG-affinity purification and multidimensional protein identification technology (MudPIT) identified nucleolin among the proteins interacting with COUP-TFII in MCF-7 tamoxifen-sensitive breast cancer cells. Interaction of COUP-TFII and nucleolin was confirmed by coimmunoprecipitation of endogenous proteins in MCF-7 and T47D breast cancer cells. In vitro studies revealed that COUP-TFII interacts with the C-terminal arginine-glycine repeat (RGG) domain of nucleolin. Functional interaction between COUP-TFII and nucleolin was indicated by studies showing that siRNA knockdown of nucleolin and an oligonucleotide aptamer that targets nucleolin, AS1411, inhibited endogenous COUP-TFII-stimulated RARB2 expression in MCF-7 and T47D cells. Chromatin immunoprecipitation revealed COUP-TFII occupancy of the RARB2 promoter was increased by all-trans retinoic acid (atRA). RARβ2 regulated gene RRIG1 was increased by atRA and COUP-TFII transfection and inhibited by siCOUP-TFII. Immunohistochemical staining of breast tumor microarrays showed nuclear COUP-TFII and nucleolin staining was correlated in invasive ductal carcinomas. COUP-TFII staining correlated with ERα, SRC-1, AIB1, Pea3, MMP2, and phospho-Src and was reduced with increased tumor grade.Our data indicate that nucleolin plays a coregulatory role in transcriptional regulation of the tumor suppressor RARB2 by COUP-TFII

Signal One and Two Blockade Are Both Critical for Non-Myeloablative Murine HSCT across a Major Histocompatibility Complex Barrier

Non-myeloablative allogeneic haematopoietic stem cell transplantation (HSCT) is rarely achievable clinically, except where donor cells have selective advantages. Murine non-myeloablative conditioning regimens have limited clinical success, partly through use of clinically unachievable cell doses or strain combinations permitting allograft acceptance using immunosuppression alone. We found that reducing busulfan conditioning in murine syngeneic HSCT, increases bone marrow (BM):blood SDF-1 ratio and total donor cells homing to BM, but reduces the proportion of donor cells engrafting. Despite this, syngeneic engraftment is achievable with non-myeloablative busulfan (25 mg/kg) and higher cell doses induce increased chimerism. Therefore we investigated regimens promoting initial donor cell engraftment in the major histocompatibility complex barrier mismatched CBA to C57BL/6 allo-transplant model. This requires full myeloablation and immunosuppression with non-depleting anti-CD4/CD8 blocking antibodies to achieve engraftment of low cell doses, and rejects with reduced intensity conditioning (≤75 mg/kg busulfan). We compared increased antibody treatment, G-CSF, niche disruption and high cell dose, using reduced intensity busulfan and CD4/8 blockade in this model. Most treatments increased initial donor engraftment, but only addition of co-stimulatory blockade permitted long-term engraftment with reduced intensity or non-myeloablative conditioning, suggesting that signal 1 and 2 T-cell blockade is more important than early BM niche engraftment for transplant success

Innate non-specific immunity in clinicaly healthy standard breed mares after Streptococcus equi infection

Fagocytoza jest podstawową funkcją neutrofilów, monocytów oraz makrofagów tkankowych. Proces fagocytozy i tym samym jej efektywność ograniczane są znacznie w przypadku zakażeń koni Streptococcus equi. Mała efektywność identyfikacji zołz oraz niska skuteczność w leczeniu koni zarażonych S. equi związana jest z trudnościami w rozpoznaniu bezobjawowych nosicieli. Celem pracy było porównanie przebiegu procesu fagocytozy neutrofilów i monocytów krwi kłusaków rasy Standard bread klinicznie zdrowych z kłusakami, które przechorowały zakażenie Streptococcus equi.Aktywność fagocytarną określano wykorzystując PhagoTest® i cytometrię przepływową. Materiałem do badań była krew pobrana na heparynę od 13 klaczy, które przechorowały streptokokozę wywołaną S. equi i od 12 klaczy, u których nigdy nie obserwowano klinicznych objawów tej choroby. Wykazano statystycznie istotnie większą koncentrację monocytów we krwi klaczy zdrowych. Nie wykazano statystycznie istotnych różnic w liczebności limfocytów i neutrofilów. Odnotowano mniejszą aktywność fagocytarną (fluorescencję) zarówno neutrofilów (P = 0,049), jak i monocytów (P = 0,006), a także odsetek eozynofilów we krwi klaczy ozdrowieńców. U tych klaczy zaobserwowano także większą koncentrację komórek E. coli wokół fagocytujących neutrofilów i monocytów. Mniejsza aktywność układu immunologicznego klaczy, które przechorowały zakażenie, może wynikać z ich bezobjawowego nosicielstwa S. equi, a badanie aktywności fagocytarnej neutrofilów i monocytów krwi koni może być pomocne w diagnozowaniu bezobjawowego nosicielstwa S. Equi.Phagocytosis is an elementary function of neutrophils,monocytes and tissuemacrophages. The process on its own as well as its efficiency is significantly limited in case of Streptococcus equi infections in horses. Strangles diagnosis and poor effectiveness of treatment is due to difficulties in identification of asymptomatic hosts. The aim of this study was to compare the course of neutrophil andmonocyte phagocytosis in clinically healthy horses after Streptococcus equi infection versus control group. The phagocytosis activity was determined by means of PhagoTest® and flow cytometry. The bloodwas obtained from13mares recovered fromstreptococcal infection caused by S. equi and 12mares which never demonstrated clinical symptoms of this disease.A statistically significant differencewas shown in bloodmonocytes concentration in control group.Therewere no statistically significant differences between the amount of lymphocytes and neutrophils. Lower phagocytosis activity (fluorescence) was observed among neutrophils (P = 0.049) andmonocytes (P = 0.006) in recovered mares. Moreover, the percentage of eosinophils was also lower in this group, while the chemotaxis was enhanced. Limited immunological response in the recoveredmaresmay be due to asymptomatic carrier state of S. equi. Therefore, the determination of neutrophil and monocyte phagocytic activity may be useful in diagnosis of asymptomatic S. equi infections

SCRI Project “Efficient Vineyard” Progress Report Project Theme 5: Adoption and Outreach

NYS IPM Type: Project ReportYear 1 Objectives: Determine preferred method(s) of learning of grape growers and industry members. Develop a portal of learning and PR through a public “face” of the project. Create, and make readily available, educational presentations on research available to growers