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1.
Benef Microbes ; 15(2): 211-225, 2024 Mar 11.
Article in English | MEDLINE | ID: mdl-38688481

ABSTRACT

Enterococcus faecium SF68 (SF68) is a well-known probiotic with a long history of safe use. Recent changes in the taxonomy of enterococci have shown that a novel species, Enterococcus lactis, is closely related with E. faecium and occurs together with other enterococci in a phylogenetically well-defined E. faecium species group. The close phylogenetic relationship between the species E. faecium and E. lactis prompted a closer investigation into the taxonomic status of E. faecium SF68. Using phylogenomics and ANI, the taxonomic analysis in this study showed that probiotic E. faecium SF68, when compared to other E. faecium and E. lactis type and reference strains, could be re-classified as belonging to the species E. lactis. Further investigations into the functional properties of SF68 showed that it is potentially capable of bacteriocin production, as a bacteriocin gene cluster encoding the leaderless bacteriocin EntK1 together with putative Lactococcus lactis bacteriocins LsbA, and LsbB-like putative immunity peptide (LmrB) were found located in an operon on plasmid pF9. However, bacteriocin expression was not studied. Competitive exclusion experiments in co-culture over 7 days at 37 °C showed that the probiotic SF68 could inhibit the growth of specific E. faecium and Listeria monocytogenes strains, while showing little or no inhibitory activity towards an entero-invasive Escherichia coli and a Salmonella Typhimurium strain, respectively. In cell culture experiments with colon carcinoma HT29 cells, the probiotic SF68 was also able to strain-specifically inhibit adhesion and/or invasion of enterococcal and L. monocytogenes strains, while such adhesion and invasion inhibition effects were less pronounced for E. coli and Salmonella strains. This study therefore provides novel data on the taxonomy and functional properties of SF68, which can be reclassified as Enterococcus lactis SF68, thereby enhancing the understanding of its probiotic nature.


Subject(s)
Bacteriocins , Enterococcus faecium , Phylogeny , Probiotics , Enterococcus faecium/genetics , Enterococcus faecium/classification , Enterococcus faecium/physiology , Bacteriocins/genetics , Bacteriocins/metabolism , Humans , Antibiosis , Plasmids/genetics , Multigene Family , HT29 Cells
2.
Neurology ; 101(19): 859-860, 2023 11 07.
Article in English | MEDLINE | ID: mdl-37580160

Subject(s)
Chorea , Female , Humans , Aged
3.
Clin Pract Cases Emerg Med ; 7(2): 68-72, 2023 May.
Article in English | MEDLINE | ID: mdl-37285497

ABSTRACT

INTRODUCTION: Wellens' syndrome represents an important, at times overlooked, spectrum of left anterior descending (LAD) coronary artery occlusion, spontaneous reperfusion, and impending reocclusion. Once considered pathognomonic for a thromboembolic coronary event, an increasing number of clinical scenarios have been demonstrated to result in pseudo-Wellens' syndrome, each requiring unique forms of assessment and management. CASE REPORT: We describe two clinical presentations in which myocardial bridging (MB) of the LAD led to clinical and electrophysiologic presentations of a pseudo-Wellens' syndrome. CONCLUSION: These reports represent a rare cause of pseudo-Wellens' syndrome attributed to MB of the LAD. Transient ischemia secondary to myocardial compression of the traversing LAD leads to intermittent angina and electrocardiogram changes that are typical in patients presenting with Wellens' syndrome secondary to an occlusive coronary event. As with other previously reported pathophysiologic mechanisms that have been shown to mimic Wellens' syndrome, myocardial bridging should be considered in patients presenting with a pseudo-Wellens' syndrome.

4.
J Dairy Sci ; 105(10): 7951-7958, 2022 Oct.
Article in English | MEDLINE | ID: mdl-35965117

ABSTRACT

Cheese, especially ripened varieties, harbor a very complex and heterogeneous microbiota. In addition to the desired microorganisms (starter cultures) added during cheese production, potentially harmful bacteria may also enter the production chain. Regarding the latter, the focus of this study was on coagulase-negative staphylococci (CNS) and Macrococcuscaseolyticus. Both are known to harbor a variety of genes coding for antibiotic resistance, including mecA, mecB, mecC, and mecD. Coagulase-negative staphylococci or macrococci carrying such genes or other virulence factors should not be present in cheese. Cheese samples (101 in total) were collected from retail sources. Coagulase-negative staphylococci and M. caseolyticus were isolated utilizing selective agars, and species were identified by phenotypical tests and partial sequencing of the sodA gene. The results allowed identification of 53 CNS strains and 19 M. caseolyticus strains. Among the CNS, 11 isolates of Staphylococcus saprophyticus and one Staphylococcus epidermidis isolate were obtained. Both species are potential human pathogens and may thus adversely affect the safety of these food products. Screening for antimicrobial resistance was performed by application of disc diffusion tests, a gradient strip-test, and 14 different PCR tests. Evidence for methicillin resistance (by either positive disc diffusion assay for cefoxitin or by mec PCR) was found in CNS isolates and M. caseolyticus (9 isolates each). Regarding other virulence factors, no genetic determinants for coagulase or the most common staphylococcal enterotoxins sea, seb, sec, sed, and see were detected in any of the CNS or M. caseolyticus isolates by PCR testing. In conclusion, the presence of facultatively pathogenic CNS and carriers of genes for antibiotic resistance in both groups of microorganisms, especially mec genes, and the respective food safety issues need further evaluation and surveillance.


Subject(s)
Anti-Infective Agents , Cheese , Animals , Cefoxitin , Cheese/microbiology , Coagulase/genetics , Enterotoxins/genetics , Humans , Staphylococcus , Virulence Factors/genetics
5.
Science ; 375(6584): 1025-1030, 2022 Mar 04.
Article in English | MEDLINE | ID: mdl-35239388

ABSTRACT

The motion of a spin excitation across topologically nontrivial magnetic order exhibits a deflection that is analogous to the effect of the Lorentz force on an electrically charged particle in an orbital magnetic field. We used polarized inelastic neutron scattering to investigate the propagation of magnons (i.e., bosonic collective spin excitations) in a lattice of skyrmion tubes in manganese silicide. For wave vectors perpendicular to the skyrmion tubes, the magnon spectra are consistent with the formation of finely spaced emergent Landau levels that are characteristic of the fictitious magnetic field used to account for the nontrivial topological winding of the skyrmion lattice. This provides evidence of a topological magnon band structure in reciprocal space, which is borne out of the nontrivial real-space topology of a magnetic order.

6.
J Appl Crystallogr ; 55(Pt 1): 14-20, 2022 Feb 01.
Article in English | MEDLINE | ID: mdl-35145354

ABSTRACT

A method is reported to determine the phase and amplitude of sinusoidally modulated event rates, binned into four bins per oscillation, based on data generated at the resonant neutron spin-echo spectrometer RESEDA at FRM-II. The presented algorithm relies on a reconstruction of the unknown parameters. It omits a calculation-intensive fitting procedure and avoids contrast reduction due to averaging effects. It allows the current data acquisition bottleneck at RESEDA to be relaxed by a factor of four and thus increases the potential time resolution of the detector by the same factor. The approach is explained in detail and compared with the established fitting procedures of time series having four and 16 time bins per oscillation. In addition the empirical estimates of the errors of the three methods are presented and compared with each other. The reconstruction is shown to be unbiased, asymptotic and efficient for estimating the phase. Reconstructing the contrast increases the error bars by roughly 10% as compared with fitting 16 time-binned oscillations. Finally, the paper gives heuristic, analytical equations to estimate the error for phase and contrast as a function of their initial values and counting statistics.

7.
J Nucl Med ; 63(3): 415-423, 2022 03.
Article in English | MEDLINE | ID: mdl-34168013

ABSTRACT

Fibroblast activation protein (FAP) is a promising target for diagnosis and therapy of numerous malignant tumors. FAP-2286 is the conjugate of a FAP-binding peptide, which can be labeled with radionuclides for theranostic applications. We present the first-in-humans results using 177Lu-FAP-2286 for peptide-targeted radionuclide therapy (PTRT). Methods: PTRT using 177Lu-FAP-2286 was performed on 11 patients with advanced adenocarcinomas of the pancreas, breast, rectum, or ovary after prior confirmation of uptake on 68Ga-FAP-2286 or 68Ga-FAPI-04 PET/CT. Results: Administration of 177Lu-FAP-2286 (5.8 ± 2.0 GBq; range, 2.4-9.9 GBq) was well tolerated, with no adverse symptoms or clinically detectable pharmacologic effects being noticed or reported in any of the patients. The whole-body effective dose was 0.07 ± 0.02 Gy/GBq (range, 0.04-0.1 Gy/GBq). The mean absorbed doses for kidneys and red marrow were 1.0 ± 0.6 Gy/GBq (range, 0.4-2.0 Gy/GBq) and 0.05 ± 0.02 Gy/GBq (range, 0.03-0.09 Gy/GBq), respectively. Significant uptake and long tumor retention of 177Lu-FAP-2286 resulted in high absorbed tumor doses, such as 3.0 ± 2.7 Gy/GBq (range, 0.5-10.6 Gy/GBq) in bone metastases. No grade 4 adverse events were observed. Grade 3 events occurred in 3 patients-1 with pancytopenia, 1 with leukocytopenia, and 1 with pain flare-up; 3 patients reported a pain response. Conclusion:177Lu-FAP-2286 PTRT, applied in a broad spectrum of cancers, was relatively well tolerated, with acceptable side effects, and demonstrated long retention of the radiopeptide. Prospective clinical studies are warranted.


Subject(s)
Adenocarcinoma , Positron Emission Tomography Computed Tomography , Feasibility Studies , Female , Gallium Radioisotopes , Humans , Peptides , Prospective Studies , Radioisotopes/therapeutic use , Tissue Distribution
8.
Clin Nucl Med ; 46(5): 409-410, 2021 May 01.
Article in English | MEDLINE | ID: mdl-33630790

ABSTRACT

ABSTRACT: The presence of primary neuroendocrine tumors in the liver is still a matter of controversy. We present a case of a somatostatin-receptor-positive mass of the liver in the 68Ga-DOTATOC PET/CT. No other primary tumor was found after conventional imaging, endoscopically, and after liver-segment resection. Immunohistochemically, a constellation of findings was found to be compatible with a primary neuroendocrine neoplasm of the liver.


Subject(s)
Liver Neoplasms/diagnostic imaging , Neuroendocrine Tumors/diagnostic imaging , Positron Emission Tomography Computed Tomography , Aged , Female , Humans , Liver Neoplasms/pathology , Male , Middle Aged , Neuroendocrine Tumors/pathology , Octreotide/analogs & derivatives , Organometallic Compounds
9.
Int J Colorectal Dis ; 35(7): 1221-1230, 2020 Jul.
Article in English | MEDLINE | ID: mdl-32307587

ABSTRACT

PURPOSE: Anastomotic leakage constitutes a dreaded complication after colorectal surgery, leading to increased morbidity and mortality as well as prolonged hospitalization. Most leakages become clinically apparent about 8 days after surgery; however, early detection is quintessential to reduce complications and to improve patients' outcome. We therefore investigated the significance of specific protein expression profiles as putative biomarkers, indicating anastomotic leakage. METHODS: In this single-center prospective cohort study serum and peritoneal fluid samples-from routinely intraoperatively inserted drainages-of colorectal cancer patients were collected 3 days after colorectal resection. Twenty patients without anastomotic leakage and 18 patients with an anastomotic leakage and without other complications were included. Protein expression of seven inflammatory markers in serum and peritoneal fluid was assessed by multiplex ELISA and correlated with patients' clinical data. RESULTS: Monocyte chemoattractant protein 2 (CCL8/MCP-2), leukemia-inhibiting factor (LIF), and epithelial-derived neutrophil-activating protein (CXCL5/ENA-78) were significantly elevated in peritoneal fluid but not in serum samples from patients subsequently developing anastomotic leakage after colorectal surgery. No expressional differences could be found between grade B and grade C anastomotic leakages. CONCLUSION: Measurement 3 days after surgery revealed altered protein expression patterns of the inflammatory markers CCL8/MCP2, LIF, and CXCL5/ENA-78 in peritoneal fluid from patients developing anastomotic leakage after colorectal surgery. Further studies with a larger patient cohort with inclusion of different variables are needed to evaluate their potential as predictive biomarkers for anastomotic leakage.


Subject(s)
Anastomotic Leak , Colorectal Neoplasms , Anastomosis, Surgical , Anastomotic Leak/diagnosis , Anastomotic Leak/etiology , Biomarkers , Chemokine CCL8 , Chemokine CXCL5 , Colorectal Neoplasms/surgery , Early Detection of Cancer , Humans , Leukemia Inhibitory Factor , Prospective Studies
10.
J Dent ; 84: 89-94, 2019 05.
Article in English | MEDLINE | ID: mdl-30959076

ABSTRACT

OBJECTIVES: The study evaluated the suitability of pure crystalline hydroxyapatite (HA) as a substitute for human (H) and bovine (B) enamel in erosion experiments. METHODS: Human and bovine enamel and hydroxyapatite specimens (n = 18 each) were submitted to demineralisation by HCl at pH values of 3.0, 2.6 and 2.0 (1.0, 2.5, and 10 mmol/l concentration). Specimens were eroded in a superfusion chamber for 7 min (flow rate: 1 µl/s). Multiple fluid samples were taken per specimen throughout the exposure period. The dissolved calcium content was measured using a colorimetric assay with Arsenazo III reagent, to serve as surrogate for erosive substance loss. Additionally, erosive tissue loss was quantified by profilometry. Differences in erosive substance loss were assessed by the Kruskal-Wallis and Conover's posthoc test. RESULTS: Good agreement in erosive substance loss between HA and human specimens was found at pH 2.6, as no statistically significant difference and considerable overlap of their respective interquartile ranges was observed. At pH 2.0 and 3.0, a statistically significant difference between HA and human and bovine enamel was determined. HA consistently tended to have lower mean dissolution rates, with a maximum deviation from human enamel of -16% at pH 3. It displayed lower specimen variability with an average coefficient of variation of 17%, compared to 25% (H) and 25% (B), respectively. CONCLUSIONS: Crystalline hydroxyapatite may not be suitable for full substitution of biologic enamel in erosion experiments focusing on absolute measurement values, but is useful for establishing consistent relative trends between erosive agents due to biosimilar erosion behavior and lower experimental variability. This is especially true for preliminary studies where approval for use of biological samples is restricted. CLINICAL SIGNIFICANCE: Crystalline hydroxyapatite is a useful substitute of human or bovine enamel in experiments studying chemical aspects of dental erosion.


Subject(s)
Durapatite , Tooth Erosion , Animals , Calcium , Cattle , Dental Enamel , Humans , Indicators and Reagents
11.
Appl Microbiol Biotechnol ; 103(6): 2731-2743, 2019 Mar.
Article in English | MEDLINE | ID: mdl-30666364

ABSTRACT

The proteinase-encoding prtB gene of Lactobacillus (Lb.) delbrueckii (d.) subsp. bulgaricus 92059 was cloned and sequenced. Two soluble, secreted, C-terminally His-tagged derivatives were constructed and expressed in Lactococcus lactis by means of the NICE® Expression System. In both obtained derivatives PrtBb and PrtB2, the C-terminal, cell wall-binding domain was deleted. In addition, in derivative PrtB2, the C-terminal part of the B domain was deleted and the signal sequence was replaced by a lactococcal export signal. The affinity-purified derivatives were both proteolytically active. Peptide hydrolysates produced from casein with each of the derivatives showed identical peptide composition, as determined by liquid chromatography-mass spectrometry. Comparison of the peptides generated to those generated with living Lb. d. subsp. bulgaricus 92059 cells (Kliche et al. Appl Microbiol Biotechnol 101:7621-7633, 2017) showed that ß-casein was the casein fraction most susceptible to hydrolysis and that some significant differences were observed between the products obtained by either the derivatives or living Lb. d. subsp. bulgaricus 92059 cells. When tested for biological activity, the hydrolysate obtained with PrtBb showed 50% inhibition of angiotensin-converting enzyme at a concentration of 0.5 mg/ml and immunomodulation/anti-inflammation in an in vitro assay of TNF-α induced NFκB activation at concentrations of 5 and 2.5 mg/ml, respectively. The enzymatically obtained hydrolysate did not show any pro-inflammatory or cytotoxic activity.


Subject(s)
Bacterial Proteins/genetics , Caseins/metabolism , Endopeptidases/genetics , Lactobacillus delbrueckii/enzymology , Peptides/metabolism , Protein Hydrolysates/metabolism , Angiotensin-Converting Enzyme Inhibitors/isolation & purification , Cell Line , Endopeptidases/metabolism , Humans , Immunologic Factors/isolation & purification , Lactobacillus delbrueckii/genetics , Lactococcus lactis/genetics , Peptide Biosynthesis , Peptidyl-Dipeptidase A/metabolism , Protein Sorting Signals , Proteolysis
12.
Rev Sci Instrum ; 90(12): 125101, 2019 Dec 01.
Article in English | MEDLINE | ID: mdl-31893808

ABSTRACT

A time-of-flight modulation of intensity by zero effort spectrometer mode has been developed for the Larmor instrument at the ISIS pulsed neutron source. The instrument utilizes resonant spin flippers that employ electromagnets with pole shoes, allowing the flippers to operate at frequencies up to 3 MHz. Tests were conducted at modulation frequencies of 103 kHz, 413 kHz, 826 kHz, and 1.03 MHz, resulting in a Fourier time range of ∼0.1 ns to 30 ns using a wavelength band of 4 Å-11 Å.

13.
Oncotarget ; 9(24): 16932-16950, 2018 Mar 30.
Article in English | MEDLINE | ID: mdl-29682195

ABSTRACT

INTRODUCTION: Peptide receptor radionuclide therapy (PRRT) of patients with somatostatin receptor expressing neuroendocrine neoplasms has shown promising results in clinical trials and a recently published phase III study. METHODS: In our center, 2294 patients were screened between 2004 and 2014 by 68Ga somatostatin receptor (SSTR) PET/CT. Intention to treat analysis included 1048 patients, who received at least one cycle of 90Yttrium or 177Lutetium-based PRRT. Progression free survival was determined by 68Ga SSTR-PET/CT and EORTC response criteria. Adverse events were determined by CTCAE criteria. RESULTS: Overall survival (95% confidence interval) of all patients was 51 months (47.0-54.9) and differed significantly according to radionuclide, grading, previous therapies, primary site and functionality. Progression free survival (based on PET/CT) of all patients was 19 months (16.9-21), which was significantly influenced by radionuclide, grading, and origin of neuroendocrine neoplasm. Progression free survival after initial progression and first and second resumption of PRRT after therapy-free intervals of more than 6 months were 11 months (9.4-12.5) and 8 months (6.4-9.5), respectively. Myelodysplastic syndrome or leukemia developed in 22 patients (2.1%) and 5 patients required hemodialysis after treatment, other adverse events were rare. CONCLUSION: PRRT is effective and overall survival is favorable in patients with neuroendocrine neoplasms depending on the radionuclide used for therapy, grading and origin of the neuroendocrine neoplasm which is not exactly mirrored in progression free survival as determined by highly sensitive 68Ga somatostatin receptor PET/CT using EORTC criteria for determining response to therapy.

14.
Pol J Vet Sci ; 20(2): 321-327, 2017 Mar 01.
Article in English | MEDLINE | ID: mdl-28865211

ABSTRACT

The study aimed to investigate the possible anti-inflammatory activity of larch sawdust as feed supplement in lactating sows' diet and its possible effect on the prevalence of Postpartum Dysgalactia Syndrome under field conditions. In a Greek farrow-to-finish pig farm, fifteen sows were randomly and equally allocated to a negative control group (NC group), a positive control group (PC group), and a treatment group (LT group). The animals of the first two groups received 99% basic diet and 1% corn starch, while LT group animals received 99% basic diet and 1% larch sawdust. The whole trial period lasted 35 days (7 days prior to farrow - day of weaning). At parturition day, animals of the PC group received 2 ml of an anti-inflammatory drug intramuscularly (meloxicam, Metacam®, Boehringer Ingelheim Vetmedica), while the animals of both other groups, received 2 ml of normal saline. Results showed insignificant differences among experimental groups for parameters such as post-partum rectal temperature and piglets performance. On the contrary, a significant increase of mean milk lactation index was observed in LT and PC groups on the 4th day of lactation period, when compared with NC group (p=0.014). Additionally, mean IL-6 concentrations in blood in the LT group showed a tendency for reduction when compared with those found in NC, and insignificant difference (p>0.05) when compared with those observed in PC group 24 hours postpartum. Moreover, the respective TNFα mean level in the LT group at 24 and 72 hours after parturition was similar to that found in PC group, respectively) and significantly lower than that determined in the NC group (p=0.003, p=0.024. The results suggest a possible anti-inflammatory effect of larch sawdust in sows.


Subject(s)
Animal Feed/analysis , Anti-Inflammatory Agents/pharmacology , Diet/veterinary , Larix/chemistry , Swine/physiology , Animals , Anti-Inflammatory Agents/administration & dosage , Female , Lactation , Wood/chemistry
15.
Theriogenology ; 89: 295-304, 2017 Feb.
Article in English | MEDLINE | ID: mdl-28043366

ABSTRACT

Optimizing cryopreservation protocols for nondomestic felids contributes to the successful development of assisted reproduction techniques and genetic resource banking. In this study, we describe a simple cryopreservation procedure for African lion (Panthera leo) ejaculates, which was tested with different packaging options and different sperm numbers per dose. By applying urethral catheterization and electroejaculation, 17 ejaculates with greater than 20% motile and greater than 5% progressively motile sperm were collected. A lyophilized extender (a modified egg yolk-Tes-Tris-fructose-glycerol medium) was rehydrated and added in one step at ambient temperature (∼25 °C) to semen, which was prediluted in cell culture medium M199. After slow cooling of insulated samples to 15 °C in a refrigerator (4 °C), the samples were fast frozen over the surface of liquid nitrogen or in a dry shipper. Aliquots of 300 µL containing 20 × 106 sperm were frozen in cryovials and in 0.5-mL straws. Differences were observed in the total motility after thawing between vial (31.5 ± 14.1%) and straw freezing (20.1 ± 8.6%). However, the subpopulations of vital (22.7 ± 7.8% for vial and 19.8 ± 8.5% for straw) and progressively motile (10.0 ± 7.9% for vial and 10.0 ± 6.4% for straw) sperm after washing and 1 hour incubation at 38 °C were of similar magnitude, velocity, and linearity for both packaging options. After freezing of five ejaculates with 20, 60, and 100 × 106 sperm per dose, best results were achieved at the lowest concentration. In general, post-thaw results were highly variable (2.2% and 56.5% total motility) and not correlated to motility or morphology of the fresh semen. To further characterize semen quality, we assessed the protective potential of seminal fluid against oxidative stress, which might be challenged on freeze thawing. The capacity of seminal fluid to reduce radicals was measured in 10 semen samples by electron spin resonance spectroscopy and a spin-labeled fatty acid as a radical probe. Moreover, we determined the lysophosphatidylcholines (LPC) as potential lipid oxidation products in the sperm and erythrocytes of the males. Individuals with a high radical reduction capacity in the seminal fluid and a low LPC content in their erythrocytes showed a better cryosurvival of sperm. This is a first indication that seminal fluid may affect the freezing potential of African lion ejaculates.


Subject(s)
Cryopreservation/veterinary , Lions , Semen Preservation/veterinary , Animals , Cryopreservation/methods , Lysophosphatidylcholines/metabolism , Male , Semen/cytology , Semen/metabolism , Semen Analysis/veterinary , Semen Preservation/methods
16.
Rev Sci Instrum ; 87(11): 113902, 2016 Nov.
Article in English | MEDLINE | ID: mdl-27910441

ABSTRACT

We report the development of a versatile material preparation chain for intermetallic compounds, which focuses on the realization of a high-purity growth environment. The preparation chain comprises an argon glovebox, an inductively heated horizontal cold boat furnace, an arc melting furnace, an inductively heated rod casting furnace, an optically heated floating-zone furnace, a resistively heated annealing furnace, and an inductively heated annealing furnace. The cold boat furnace and the arc melting furnace may be loaded from the glovebox by means of a load-lock permitting to synthesize compounds starting with air-sensitive elements while handling the constituents exclusively in an inert gas atmosphere. All furnaces are all-metal sealed, bakeable, and may be pumped to ultra-high vacuum. We find that the latter represents an important prerequisite for handling compounds with high vapor pressure under high-purity argon atmosphere. We illustrate the operational aspects of the preparation chain in terms of the single-crystal growth of the heavy-fermion compound CeNi2Ge2.

17.
Cell Death Dis ; 7(11): e2461, 2016 11 10.
Article in English | MEDLINE | ID: mdl-27831556

ABSTRACT

CD44, a large family of transmembrane glycoproteins, plays decisive roles in physiological and pathological conditions. CD44 isoforms are involved in several signaling pathways essential for life such as growth factor-induced signaling by EGF, HGF or VEGF. CD44 is also the main hyaluronan (HA) receptor and as such is involved in HA-dependent processes. To allow a genetic dissection of CD44 functions in homeostasis and disease, we generated a Cd44 floxed allele allowing tissue- and time-specific inactivation of all CD44 isoforms in vivo. As a proof of principle, we inactivated Cd44 in the skin epidermis using the K14Cre allele. Although the skin of such Cd44Δker mutants appeared morphologically normal, epidermal stiffness was reduced, wound healing delayed and TPA induced epidermal thickening decreased. These phenotypes might be caused by cell autonomous defects in differentiation and HA production as well as impaired adhesion and migration on HA by Cd44Δker keratinocytes. These findings support the usefulness of the conditional Cd44 allele in unraveling essential physiological and pathological functions of CD44 isoforms.


Subject(s)
Epidermis/metabolism , Gene Deletion , Hyaluronan Receptors/metabolism , Keratinocytes/metabolism , Stress, Mechanical , Animals , Cell Adhesion/drug effects , Cell Differentiation/drug effects , Cell Movement/drug effects , Cell Proliferation/drug effects , Homeostasis/drug effects , Hyaluronic Acid/pharmacology , Keratinocytes/drug effects , Keratins/metabolism , Mice, Knockout , Organ Specificity/drug effects , Skin/metabolism , Wound Healing/drug effects
18.
Reprod Domest Anim ; 51(2): 195-203, 2016 Apr.
Article in English | MEDLINE | ID: mdl-26817439

ABSTRACT

To support conservation strategies in wild species, simple but highly reproducible procedures of sperm cryopreservation are required for an application under field conditions. We used epididymal sperm of the domestic cat to optimize a sperm freezing procedure for felid species, particularly questioning the demand for sperm cooling to 4°C. We equilibrated sperm during slow cooling to only 15 or 10°C in a Tes-Tris-fructose extender with final concentrations of 4.7% (v/v) glycerol and 10% (v/v) of the water-soluble fraction of hen's egg yolk (low-density lipoproteins). Subsequently, sperm were frozen over liquid nitrogen. Total and progressive motility (mean ± SD) after thawing was 60.7 ± 8.6% and 53.9 ± 9.6% in samples cooled to 15°C or 61.6 ± 9.5% and 55.3 ± 9.9% in samples cooled to 10°C. Therefore, a one-step addition of glycerol to sperm at room temperature together with the freezing extender, the use of cryovials (loaded with diluted sperm aliquots of 300 µl), an equilibration period of 40 min comprising slow cooling to 15°C at a rate of approximately -0.14 K/min before rapid freezing over liquid nitrogen, yielded satisfying results. Cooling, freezing and thawing rates were exactly characterized as a prerequisite for further optimization and to provide a repeatable protocol to other practitioners.


Subject(s)
Cats/physiology , Cryopreservation/veterinary , Epididymis/physiology , Semen Preservation/veterinary , Spermatozoa/physiology , Animals , Cold Temperature , Cryopreservation/methods , Male , Semen Preservation/methods
19.
Scand J Med Sci Sports ; 26(8): 911-8, 2016 Aug.
Article in English | MEDLINE | ID: mdl-26130046

ABSTRACT

The aims of this prospective school cohort study were to describe the epidemiology of diagnosed back pain in childhood, classified as either nontraumatic or traumatic back injury, and to estimate the association with physical activity in different settings. Over 2.5 years, 1240 children aged 6-12 years were surveyed weekly using mobile text messages to ask about the presence or absence of back pain. Pain was clinically diagnosed and injuries were classified using the International Classification of Diseases version 10. Physical activity data were obtained from text messages and accelerometers. Of the 315 back injuries diagnosed, 186 injuries were nontraumatic and 129 were traumatic. The incidence rate ratio was 1.5 for a nontraumatic back injury compared with a traumatic injury. The overall estimated back injury incidence rate was 0.20 per 1000 physical activity units (95% confidence interval 0.18-0.23). The back injury incidence rates were higher for sports when exposure per 1000 physical activity units was taken into consideration and especially children horse-riding had a 40 times higher risk of sustaining a traumatic back injury compared to the risk during non-organized leisure time physical activity. However, the reasonably low injury incidence rates support the recommendations of children continuously being physically active.


Subject(s)
Back Injuries/epidemiology , Back Injuries/etiology , Back Pain/epidemiology , Exercise , Sports , Accelerometry , Athletic Injuries/epidemiology , Back Pain/etiology , Child , Denmark/epidemiology , Female , Humans , Incidence , Longitudinal Studies , Male , Prospective Studies , Text Messaging
20.
Vet J ; 206(3): 322-6, 2015 Dec.
Article in English | MEDLINE | ID: mdl-26526868

ABSTRACT

When larch (Larix spp.) is processed in the wood industry, the sawdust is currently disposed of as waste or used as combustible material, even though it is rich in biologically active compounds. In this study the effect of larch sawdust supplementation on blood parameters as well as milk composition was examined in healthy mid-lactating dairy cows. Twenty-four multiparous Italian Friesian dairy cows were assigned to groups receiving either 300 g/day/cow of larch sawdust or a control diet, and treatments were continued for a 20 day period. Milk parameters were unaffected by treatment. A lower plasma total protein concentration was observed and can be attributed to a decrease in globulin concentration. A lower plasma urea concentration was also detected in the larch group. Moreover, biomarkers of liver function were influenced by the treatment. Total bilirubin was lower in larch-treated animals, and cholesterol tended to be lower. In addition, an interaction between day and treatment was observed for very low density lipoprotein. The concentration of other parameters, including reactive oxygen metabolites, superoxide dismutase, glutathione peroxidase and nitrotyrosine, did not differ between treatments. The observed benefits, together with the good palatability, make larch sawdust a promising candidate for the development of beneficial feed supplements for livestock. Further studies will be useful, particularly to evaluate its efficacy in different health conditions.


Subject(s)
Animal Feed , Cattle , Industrial Waste , Larix , Animals , Dairying , Dietary Fiber , Female , Food Additives , Larix/chemistry , Milk
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