Aims
The goal of this study was to isolate and characterize a lactic acid bacteria (LAB) from donkey milk with potential beneficial properties.
Methods and Results
Lactic acid bacteria were isolated ...from donkey milk and identified based on physiological, biochemical and molecular methods. The isolate that presented highest bacteriocin potential (Lactobacillus plantarum LP08AD) was evaluated for the production of bacteriocin, including stability in the presence of various enzymes, surfactants, salts, pH and temperatures. Bactericidal effect of bacteriocin LP08AD on Listeria monocytogenes, Enterococcus faecium and Lactobacillus curvatus was shown for actively growing and stationary cells. Similar growth and bacteriocin production were observed when strain LP08AD was cultured in MRS broth at 30°C or 37°C. Bacteriocin LP08AD adhered at low levels on the producer cells (200 AU ml−1). The presence of plantaricin W gene on the genomic DNA was recorded based on PCR. Good growth for strain LP08AD was recorded in MRS broth with pH from 5·0 to 9·0 and LP08AD grew well in the absence of oxbile or concentration below 0·8%. Lact. plantarum LP08AD was applied to the small intestinal epithelial polarized monolayers of H4, PSIc1 and CLAB and demonstrated low attachment ability on all cell lines studied, with values with a similar behaviour for cells from human and pig origin.
Conclusions
Bacteriocin‐producing Lact. plantarum LP08AD might be useful in the design of novel functional foods with potential probiotic or biopreservation properties.
Significance and Impact of the Study
To the best of our knowledge, this is the first report on detection and characterization of bacteriocinogenic Lact. plantarum from donkey milk. The strain LP08AD shows to have potential beneficial properties, as demonstrated by the use of noncancerogenic cell lines.
With the aim of screening proteolytic strains of lactic acid bacteria to evaluate their potential for the reduction of allergenicity of the major bovine milk proteins, we isolated a new proteolytic ...strain of Enterococcus faecalis (Ent. faecalis VB63F) from raw bovine milk. The proteases produced by this strain had strong activity against caseins (αS1-, αS2-, and β-casein), in both skim milk and sodium caseinate. However, only partial hydrolysis of whey proteins was observed. Proteolysis of Na-caseinate and whey proteins, observed after sodium dodecyl sulfate-PAGE, was confirmed by analysis of peptide profiles by reversed-phase HPLC. Inhibition of proteolysis with EDTA indicated that the proteases produced by Ent. faecalis VB63F belonged to the group of metalloproteases. The optimal conditions for their activity were 42°C and pH 6.5. The majority of assessed virulence genes were absent in Ent. faecalis VB63F. The obtained results suggest that Ent. faecalis VB63F could be efficient in reducing the immunoreactivity of bovine milk proteins.
Natural killer (NK) cells mediate spontaneous cell-mediated cytotoxicity and antibody-dependent cell-mediated cytotoxicity. This dual functionality could enable their participation in chronic active ...antibody-mediated rejection (CA-ABMR). Earlier microarray profiling studies have not subcategorized antibody-mediated rejection into CA-ABMR and active-ABMR, and the gene expression pattern of CA-ABMR has not been compared with that of T cell-mediated rejection (TCMR). To fill these gaps, we RNA sequenced human kidney allograft biopsies categorized as CA-ABMR, active-ABMR, TCMR, or No Rejection (NR). Among the 15,910 genes identified in the biopsies, 60, 114, and 231 genes were uniquely overexpressed in CA-ABMR, TCMR, and active-ABMR, respectively; compared to NR, 50 genes were shared between CA-ABMR and active-ABMR, and 164 genes between CA-ABMR and TCMR. The overexpressed genes were annotated to NK cells and T cells in CA-ABMR and TCMR, and to neutrophils and monocytes in active-ABMR. The NK cell cytotoxicity and allograft rejection pathways were enriched in CA-ABMR. Genes encoding perforin, granzymes, and death receptor were overexpressed in CA-ABMR versus active-ABMR but not compared to TCMR. NK cell cytotoxicity pathway gene set variation analysis score was higher in CA-ABMR compared to active-ABMR but not in TCMR. Principal component analysis of the deconvolved immune cellular transcriptomes separated CA-ABMR and TCMR from active-ABMR and NR. Immunohistochemistry of kidney allograft biopsies validated a higher proportion of CD56+ NK cells in CA-ABMR than in active-ABMR. Thus, CA-ABMR was exemplified by the overexpression of the NK cell cytotoxicity pathway gene set and, surprisingly, molecularly more like TCMR than active-ABMR.
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•Solid state actuation in CuAlMn SMAs are reported at extreme low temperatures for the first time.•Compositional effects on martensitic transformation in CuAlMn SMAs are presented below −200 °C.•The ...alloys exhibited 2.9% actuation strain under 250 MPa at subzero temperatures.•The results are significant because well-known NiTi SMAs do not demonstrate actuation below −60 °C.
Solid state actuation characteristics of CuAlMn shape memory alloys (SMAs) are reported at extreme low temperatures (<-100 °C). The effects of composition on the martensitic transformation (MT) characteristics are demonstrated at low temperatures (below −200 °C). The thermomechanical experiments revealed that the alloys exhibited 2.9% actuation strain under 250 MPa at subzero temperatures. CuAlMn SMAs with different Mn concentrations displayed 2% superelasticity at temperatures lower than −100 °C. The results illustrate that CuAlMn SMAs are promising candidates for cryogenic actuation and superelastic applications. The results are significant because none of the well-known NiTi SMAs demonstrate reversible thermally-induced MT below −60 °C. These findings are important for enabling solid state actuation in space and subsea environments.
Delineation of T-cell genes, gene sets, pathways, and T-cell subtypes associated with acute T cell-mediated rejection (TCMR) may improve its management.
We performed bulk RNA-sequencing of 34 kidney ...allograft biopsies (16 Banff TCMR and 18 no rejection NR biopsies) from 34 adult recipients of human kidneys. Computational analysis was performed to determine the differential intragraft expression of T-cell genes at the level of single-gene, gene set, and pathways.
T-cell signaling pathway gene sets for plenary T-cell activation were overrepresented in TCMR biopsies compared with NR biopsies. Heightened expression of T-cell signaling genes was validated using external TCMR biopsies. Pro- and anti-inflammatory immune gene sets were enriched, and metabolism gene sets were depleted in TCMR biopsies compared with NR biopsies. Gene signatures of regulatory T cells, Th1 cells, Th2 cells, Th17 cells, T follicular helper cells, CD4 tissue-resident memory T cells, and CD8 tissue-resident memory T cells were enriched in TCMR biopsies compared with NR biopsies. T-cell exhaustion and anergy were also molecular attributes of TCMR. Gene sets associated with antigen processing and presentation, and leukocyte transendothelial migration were overexpressed in TCMR biopsies compared with NR biopsies. Cellular deconvolution of graft infiltrating cells by gene expression patterns identified CD8 T cell to be the most abundant T-cell subtype infiltrating the allograft during TCMR.
Our delineation of intragraft T-cell gene expression patterns, in addition to yielding new biological insights, may help prioritize T-cell genes and T-cell subtypes for therapeutic targeting.
The lineage restriction of prospectively isolated hematopoietic progenitors has been traditionally assessed by bulk in vitro culture and transplantation of large number of cells in vivo. These ...methods, however, cannot distinguish between homogenous multipotent or heterogeneous lineage-restricted populations. Using clonal assays of 1 or 5 cells in vitro, single-cell quantitative gene expression analyses, and transplantation of mice with low numbers of cells, we show that a common myeloid progenitor (CMP) is Sca-1lolin−c-Kit+CD27+Flk-2− (SL-CMP; Sca-1lo CMP) and a granulocyte/macrophage progenitor (GMP) is Sca-1lolin−c-Kit+CD27+Flk-2+CD150−/lo (SL-GMP; Sca-1lo GMP). We found that mast cell progenitor potential is present in the SL-CMP fraction, but not in the more differentiated SL-GMP population, and is more closely related to megakaryocyte/erythrocyte specification. Our data provide criteria for the prospective isolation of SL-CMP and SL-GMP and support the conclusion that mast cells are specified during hematopoiesis earlier than and independently from granulocytes.
► Single-cell analysis resolves hematopoietic progenitor lineage potential ► Distinct Sca-1lo common myeloid and granulocyte/macrophage progenitors ► The mast cell lineage is independent from granulocyte differentiation ► Mast cell and megakaryocyte/erythrocyte lineages may be closely related
Cathepsin Z (CTSZ) is a cysteine protease responsible for the adhesion and migration of both immune and tumor cells. Due to its dual role, we hypothesized that the site of CTSZ expression could be ...determinant of the pro- or anti-tumorigenic effects of this enzyme. To test this hypothesis, we analyzed CTSZ expression data in healthy and tumor tissues by bioinformatics and evaluated the expression levels of CTSZ mRNA in the blood cells of prostate cancer (PCa) patients by qRT-PCR compared with healthy subjects, evaluating its diagnostic and prognostic implications for this type of cancer. Immune cells present in the blood of healthy patients overexpress CTSZ. In PCa, we found decreased CTSZ mRNA levels in blood cells, 75% lower than in healthy subjects, that diminished even more during biochemical relapse. CTSZ mRNA in the blood cells had an area under the curve for PCa diagnosis of 0.832, with a 93.3% specificity, and a positive likelihood ratio of 9.4. The site of CTSZ mRNA expression is fundamental to determine its final role as a protective determinant in PCa, such as CTSZ mRNA in the blood cells, or a malignant determinant, such as found for CTSZ expressed in high levels by different types of primary and metastatic tumors. Low CTSZ mRNA expression in the total blood is a possible PCa marker complementary to prostate-specific antigen (PSA) for biopsy decisions, with the potential to eliminate unnecessary biopsies.
The abundance of chlorine in the Earth's atmosphere increased considerably during the 1970s to 1990s, following large emissions of anthropogenic long-lived chlorine-containing source gases, notably ...the chlorofluorocarbons. The chemical inertness of chlorofluorocarbons allows their transport and mixing throughout the troposphere on a global scale, before they reach the stratosphere where they release chlorine atoms that cause ozone depletion. The large ozone loss over Antarctica was the key observation that stimulated the definition and signing in 1987 of the Montreal Protocol, an international treaty establishing a schedule to reduce the production of the major chlorine- and bromine-containing halocarbons. Owing to its implementation, the near-surface total chlorine concentration showed a maximum in 1993, followed by a decrease of half a per cent to one per cent per year, in line with expectations. Remote-sensing data have revealed a peak in stratospheric chlorine after 1996, then a decrease of close to one per cent per year, in agreement with the surface observations of the chlorine source gases and model calculations. Here we present ground-based and satellite data that show a recent and significant increase, at the 2σ level, in hydrogen chloride (HCl), the main stratospheric chlorine reservoir, starting around 2007 in the lower stratosphere of the Northern Hemisphere, in contrast with the ongoing monotonic decrease of near-surface source gases. Using model simulations, we attribute this trend anomaly to a slowdown in the Northern Hemisphere atmospheric circulation, occurring over several consecutive years, transporting more aged air to the lower stratosphere, and characterized by a larger relative conversion of source gases to HCl. This short-term dynamical variability will also affect other stratospheric tracers and needs to be accounted for when studying the evolution of the stratospheric ozone layer.
Celotno besedilo
Dostopno za:
DOBA, IJS, IZUM, KILJ, KISLJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Abstract Background There is a concern about the accuracy of the available prognostic indexes when applying them to the emergent population of polypathological patients (PP). Methods To develop a ...1-year mortality predictive index on PP, we developed a multicenter prospective cohort-study recruiting 1.632 PP after hospital discharge, outpatient clinics, or home hospitalization, from 33 hospitals. Potential risk factors were obtained in the 1.525 PP who completed follow-up. Each factor independently associated with mortality in the derivation cohort (757 PP from western hospitals) was assigned a weight, and risk scores were calculated by adding the points of each factor. Accuracy was assessed in the validation cohort (768 PP from eastern hospitals) by risk quartiles calibration, and discrimination power, by ROC curves. Finally, accuracy of the index was compared with that of the Charlson index. Results Mortality in the derivation/validation cohorts was 35%/39.5%, respectively. Nine independent mortality predictors were identified to create the index (age ≥ 85 years, 3points; No caregiver or caregiver other than spouse, 2points; active neoplasia, 6points; dementia, 3points; III–IV functional class on NYHA and/or MRC, 3points; delirium during last hospital admission, 3points; hemoglobinemia < 10 g/dl, 3points; Barthel index < 60 points, 4points; ≥ 4 hospital admissions in last 12 months, 3points). Mortality in the derivation/validation cohorts was 12.1%/14.6% for patients with 0–2points; 21.5%/31.5% for those with 3–6 points; 45%/50% for those with 7–10 points; and 68%/61.3% for those with ≥ 11points, respectively. Calibration was good in derivation/validation cohorts, and discrimination power by area under the curve was 0.77/0.7. Calibration of the Charlson index was good, but discrimination power was suboptimal (area under the curve, 0.59). Conclusions This prognostic index provides an accurate and transportable method of stratifying 1-year death risk in PP.
The aim of this study was to explore the biochemical and genetic features of the two-peptide bacteriocin produced by a Lactobacillus plantarum strain isolated from Italian type salami produced in ...Brazil (Lb. plantarum MBSa4). Identification of bacteriocinogenic Lb. plantarum MBSa4 was performed by 16S rRNA sequencing. Expressed bacteriocin was evaluated for spectrum of activity, heat and pH stability, mechanism of action, and molecular mass. Partial purification was achieved by cation-exchange, and reversed phase - HPLC. Total DNA of Lb. plantarum MBSa4 was extracted and tested for presence of previously described bacteriocin genes. Bacteriocin MBSa4 was heat-stable, unaffected by pH 2.0 to 6.0 and active against all tested Listeria monocytogenes strains and most of tested fungi. Maximal production (1600 AU/ml) in MRS broth occurred after 22 h at 25 °C, presenting bacteriostatic activity as result of combined action of two components. The molecular mass determined by SDS-PAGE was 2.3 kDa. PCR-amplified DNA indicated the same nucleotide sequence of plantaricin W. Results indicate that Lb. plantarum MBSa4 produces plantaricin W, a two-peptide lantibiotic with remarkable anti-Listeria activity.
•Mode of action of bacteriocin produced by Lactobacillus plantarum MBSa4 was determined against Listeria spp.•Produced bacteriocin/s was active against several Listeria monocytogenes from different serotypes.•Lb. plantarum MBSa4 produces antifungal agent.•Lb. plantarum MBSa4 produces two component bacteriocin.•Bacteriocin was partially purified and gene related to plantaricin W detected.
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