Extracellular vesicles (EVs) have been identified within different body fluids and cell culture media. However, there is very little information on the secretion of these vesicles during early ...embryonic development. The aims of this work were first to demonstrate the secretion of extracellular vesicles by pre-implantation bovine embryos and second to identify and characterize the population of EVs secreted by bovine blastocysts during the period from day seven to nine of embryo culture and its correlation with further embryo development up to day 11. Bovine embryos were produced by in vitro fertilization (IVF) or parthenogenetic activation (PA) and cultured until blastocyst stage. Blastocyst selection was performed at day 7 post IVF/PA considering two variables: stage of development and quality of embryos. Selected blastocysts were cultured in vitro for 48 hours in groups (exp. 1) or individually (exp. 2) in SOF media depleted of exosomes. At day 9 post IVF/PA the media was collected and EVs isolated by ultracentrifugation. Transmission electron microscopy revealed the presence of heterogeneous vesicles of different sizes and population: microvesicles (MVs) and exosomes (EXs) of rounded shape, enclosed by a lipid bi-layer and ranging from 30 to 385 nm of diameter. Flow cytometry analysis allowed identifying CD63 and CD9 proteins as exosome markers. Nanoparticle tracking analysis generated a large number of variables, which required the use of multivariate statistics. The results indicated that the concentration of vesicles is higher in those blastocysts with arrested development from day 9 up to day 11 of in vitro development (6.7 x 108 particles/ml) derived from IVF (p <0.05), compared to PA blastocysts (4.7 x 108 particles/ml). Likewise, the profile (concentration and diameter) of particles secreted by embryos derived from IVF were different from those secreted by PA embryos. In conclusion, we demonstrated that bovine blastocysts secrete MVs/EXs to the culture media. Data suggest that characteristics of the population of EVs vary depending on embryo competence.
Tumor suppressor gene TUSC2/FUS1 (TUSC2) is frequently inactivated early in lung cancer development. TUSC2 mediates apoptosis in cancer cells but not normal cells by upregulation of the intrinsic ...apoptotic pathway. No drug strategies currently exist targeting loss-of-function genetic abnormalities. We report the first in-human systemic gene therapy clinical trial of tumor suppressor gene TUSC2.
Patients with recurrent and/or metastatic lung cancer previously treated with platinum-based chemotherapy were treated with escalating doses of intravenous N-1-(2,3-dioleoyloxy)propyl-N,N,N-trimethylammonium chloride (DOTAP):cholesterol nanoparticles encapsulating a TUSC2 expression plasmid (DOTAP:chol-TUSC2) every 3 weeks.
Thirty-one patients were treated at 6 dose levels (range 0.01 to 0.09 milligrams per kilogram). The MTD was determined to be 0.06 mg/kg. Five patients achieved stable disease (2.6-10.8 months, including 2 minor responses). One patient had a metabolic response on positron emission tomography (PET) imaging. RT-PCR analysis detected TUSC2 plasmid expression in 7 of 8 post-treatment tumor specimens but not in pretreatment specimens and peripheral blood lymphocyte controls. Proximity ligation assay, performed on paired biopsies from 3 patients, demonstrated low background TUSC2 protein staining in pretreatment tissues compared with intense (10-25 fold increase) TUSC2 protein staining in post-treatment tissues. RT-PCR gene expression profiling analysis of apoptotic pathway genes in two patients with high post-treatment levels of TUSC2 mRNA and protein showed significant post-treatment changes in the intrinsic apoptotic pathway. Twenty-nine genes of the 82 tested in the apoptosis array were identified by Igenuity Pathway Analysis to be significantly altered post-treatment in both patients (Pearson correlation coefficient 0.519; p<0.01).
DOTAP:chol-TUSC2 can be safely administered intravenously in lung cancer patients and results in uptake of the gene by human primary and metastatic tumors, transgene and gene product expression, specific alterations in TUSC2-regulated pathways, and anti-tumor effects (to our knowledge for the first time for systemic DOTAP:cholesterol nanoparticle gene therapy).
ClinicalTrials.gov NCT00059605.
The influence of technological factors (decaffeination, brew volume, coffee species, and roast degree) on antiradical activity and phenolics content of espresso coffee is described. The screenings of ...phenolics profile and other compounds (caffeine and trigonelline), as well as the quantification of hydroxymethylfurfural, were performed by LC-DAD-ESI-MS. Significantly lower (p < 0.05) scavenging activities and phenolics contents were found in decaffeinated espressos when compared with regular ones (32 vs 38% and 324 vs 410 mg/30 mL cup, respectively). A long espresso (70 mL) offers more than twice the phenolics amount of a short one (20 mL). Robusta brews showed higher (p < 0.05) antiradical activity and phenolic contents than arabica ones, for all roast degrees (light, medium, and dark). No significant differences (p > 0.05) were observed for scavenging activities of differently roasted robusta brews, whereas an increase in medium-dark brews was observed for arabica samples. Total phenolics in robusta espressos decreased (p < 0.05) with the increase of roast degree, but no significant differences (p > 0.05) were found between arabica espressos from different roasts. By LC-DAD-ESI-MS, 23 hydroxycinnamic derivatives were found, including chlorogenic acids, lactones, and cinnamoyl−amino acid conjugates. The amount of each compound was differently affected by species and roast. Robusta brews presented superior levels of caffeine and chlorogenic acids, whereas arabica ones contained more trigonelline. Hydroxymethylfurfural contents in the brew (30 mL) varied from 2.60 to 0.84 mg for light- and dark-roasted arabicas and from 1.29 to 0.68 mg for light- and dark-roasted robustas, respectively.
Despite myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) affecting millions of people worldwide, many clinicians lack the knowledge to appropriately diagnose or manage ME/CFS. ...Unfortunately, clinical guidance has been scarce, obsolete, or potentially harmful. Consequently, up to 91% of patients in the United States remain undiagnosed, and those diagnosed often receive inappropriate treatment. These problems are of increasing importance because after acute COVID-19, a significant percentage of people remain ill for many months with an illness similar to ME/CFS. In 2015, the US National Academy of Medicine published new evidence-based clinical diagnostic criteria that have been adopted by the US Centers for Disease Control and Prevention. Furthermore, the United States and other governments as well as major health care organizations have recently withdrawn graded exercise and cognitive-behavioral therapy as the treatment of choice for patients with ME/CFS. Recently, 21 clinicians specializing in ME/CFS convened to discuss best clinical practices for adults affected by ME/CFS. This article summarizes their top recommendations for generalist and specialist health care providers based on recent scientific progress and decades of clinical experience. There are many steps that clinicians can take to improve the health, function, and quality of life of those with ME/CFS, including those in whom ME/CFS develops after COVID-19. Patients with a lingering illness that follows acute COVID-19 who do not fully meet criteria for ME/CFS may also benefit from these approaches.
Abstract
Aims
Chitosan‐based biomaterials exhibit several properties of biological interest for endodontic treatment. Herein, a low molecular weight chitosan (CH) solution was tested for its ...antimicrobial activity against
Enterococcus faecalis
(
E. faecalis
) and effects on dentine structure.
Methodology
The root canal of 27 extracted uniradicular teeth were biomechanically prepared, inoculated with a suspension of
E. faecalis
and randomly assigned to be irrigated with either 5.25% sodium hypochlorite (NaClO), 0.2% CH or sterile ultrapure water (W). Bacteriologic samples were collected from root canals and quantified for of
E. faecalis
colony‐forming units (CFUs). The effectiveness of CH over
E. faecalis
biofilms was further measured using the MBEC Assay®. Additionally, dentine beams and dentine powder were obtained, respectively, from crowns and roots of 20 extracted third molars. Dentine samples were treated or not with 17% EDTA and immersed in either CH or W for 1 min. The effects of CH on dentine structure were evaluated by assessment of the modulus of elasticity, endogenous proteolytic activity and biochemical modifications.
Results
The number of
E. faecalis
CFUs was significantly lower for samples irrigated with CH and NaClO. No significant differences were found between CH and NaClO treatments. Higher modulus of elasticity and lower proteolytic activity were reported for dentine CH‐treated specimens. Chemical interaction between CH and dentine was observed for samples treated or not with EDTA.
Conclusions
Present findings suggest that CH could be used as an irrigant during root canal treatment with the triple benefit of reducing bacterial activity, mechanically reinforcing dentine and inhibiting dentine proteolytic activity.
Vapor contact is an alternative when essential oils (EO's) and microorganisms are placed separately in some sealed environment. The aim of this study was to compare the antifungal efficacy of orange ...peel EO at selected concentrations, applied either by vapor exposure or direct addition on the growth of Aspergillus flavus. Orange peel EO was obtained from fresh oranges (Citrus sinensis var. Valencia). EO was obtained by vapor distillation, analyzed by means of GC–MS chromatography, and applied to potato-dextrose agar inoculated with A. flavus, using the techniques of direct addition to the agar or by generating EO vapors in airtight containers. Radial growth rate and lag phase were calculated using the Gompertz equation. Main compounds identified in the orange peel EO were: limonene, β-myrcene, β-pinene, α-pinene, as well as citral Z and E; of which, limonene represented 96.62%. The minimum inhibitory concentration for the growth of A. flavus by direct addition was 16,000 mg l−1, while for the vapor contact was 8000 mg of EO l−1 of air. For both studied methods A. flavus growth decreased when increasing EO concentration. Although the effect of orange peel EO direct addition was faster, orange peel EO vapors were more effective, since lower concentrations were required to achieve the same antifungal effect.
The regulation of gastric ghrelin secretion Nunez‐Salces, Maria; Li, Hui; Feinle‐Bisset, Christine ...
Acta Physiologica,
March 2021, 2021-03-00, 20210301, Letnik:
231, Številka:
3
Journal Article
Recenzirano
Ghrelin is a gastric hormone with multiple physiological functions, including the stimulation of food intake and adiposity. It is well established that circulating ghrelin levels are closely ...associated with feeding patterns, rising strongly before a meal and lowering upon food intake. However, the mechanisms underlying the modulation of ghrelin secretion are not fully understood. The purpose of this review is to discuss current knowledge on the circadian oscillation of circulating ghrelin levels, the neural mechanisms stimulating fasting ghrelin levels and peripheral mechanisms modulating postprandial ghrelin levels. Furthermore, the therapeutic potential of targeting the ghrelin pathway is discussed in the context of the treatment of various metabolic disorders, including obesity, type 2 diabetes, diabetic gastroparesis and Prader‐Willi syndrome. Moreover, eating disorders including anorexia nervosa, bulimia nervosa and binge‐eating disorder are also discussed.
The photolysis of iodide anions promotes the reaction of carbon dioxide with hydrogen sulfide or thiols to quantitatively yield formic acid and sulfur or disulfides. The reaction proceeds in ...acetonitrile and aqueous solutions, at atmospheric pressure and room temperature by irradiation using a low‐pressure mercury lamp. This transition‐metal‐free photocatalytic process for CO2 capture coupled with H2S removal may have been relevant as a prebiotic carbon dioxide fixation.
Connecting the dots: The electron photodetachment from iodide anions, the hydrogen transfer from thiols to carbon dioxide radical anions, and the oxidation of thiols by iodine define a robust and transition‐metal‐free photocatalytic method for the reduction of carbon dioxide to formic acid by using thiols and hydrogen sulfide at atmospheric pressure in aqueous and acetonitrile solution.
Streptococcus gallolyticus subspecies gallolyticus (Sgg) has a strong clinical association with colorectal cancer (CRC) and actively promotes the development of colon tumors. However, the molecular ...determinants involved in Sgg pathogenicity in the gut are unknown. Bacterial type VII secretion systems (T7SS) mediate pathogen interactions with their host and are important for virulence in pathogenic mycobacteria and Staphylococcus aureus. Through genome analysis, we identified a locus in Sgg strain TX20005 that encodes a putative type VII secretion system (designated as SggT7SST05). We showed that core genes within the SggT7SST05 locus are expressed in vitro and in the colon of mice. Western blot analysis showed that SggEsxA, a protein predicted to be a T7SS secretion substrate, is detected in the bacterial culture supernatant, indicating that this SggT7SST05 is functional. Deletion of SggT7SST05 (TX20005Δesx) resulted in impaired bacterial adherence to HT29 cells and abolished the ability of Sgg to stimulate HT29 cell proliferation. Analysis of bacterial culture supernatants suggest that SggT7SST05-secreted factors are responsible for the pro-proliferative activity of Sgg, whereas Sgg adherence to host cells requires both SggT7SST05-secreted and bacterial surface-associated factors. In a murine gut colonization model, TX20005Δesx showed significantly reduced colonization compared to the parent strain. Furthermore, in a mouse model of CRC, mice exposed to TX20005 had a significantly higher tumor burden compared to saline-treated mice, whereas those exposed to TX20005Δesx did not. Examination of the Sgg load in the colon in the CRC model suggests that SggT7SST05-mediated activities are directly involved in the promotion of colon tumors. Taken together, these results reveal SggT7SST05 as a previously unrecognized pathogenicity determinant for Sgg colonization of the colon and promotion of colon tumors.
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