Crotamine, one of the main toxic components of
Crotalus durissus terrificus venom, is a small non-enzymatic basic polypeptide, which causes hind limb paralysis and necrosis of muscle cells. It is ...well-known that several toxins penetrate into the cytosol through endocytosis, although in many cases the mechanism by which this occurs has not been fully investigated. Recently, using low concentrations of crotamine, we demonstrated the uptake of this toxin into actively proliferative cells via endocytosis, an event that ensues crotamine binding to cell membrane heparan sulfate proteoglycans. Thus, crotamine can be regarded as a cell-penetrating peptide that, additionally, has been shown to be able of delivering some biologically active molecules into various cells. Herein, we investigate one of the mechanisms by which crotamine exerts its cytotoxic effects by following its uptake into highly proliferative cells, as CHO-K1 cells. Crotamine accumulation in the acidic endosomal/lysosomal vesicles was observed within 5
min after treatment of these cells with a cytotoxic concentration of this toxin, a value determined here by classical MTT assay. This accumulation caused disruption of lysosomal vesicles accompanied by the leakage of these vesicles contents into the cytosol. This lysosomal lysis also promoted the release of cysteine cathepsin and an increase of caspase activity in the cytoplasm. This chain of events seems to trigger a cell death process. Overall, our data suggest that lysosomes are the primary targets for crotamine cytotoxicity, a proposal corroborated by the correlation between both the kinetics and concentration-dependence of crotamine accumulation in lysosome compartments and the cytotoxic effects of this protein in CHO-K1 cells. Although crotamine is usually regarded as a myotoxin, we observed that intraperitoneal injection of fluorescently labeled crotamine in living mice led to significant and rapid accumulation of this toxin in the cell cytoplasm of several tissues, suggesting that crotamine cytotoxicity might not be restricted to muscle cells.
Campylobacteriosis is one of the most common types of bacterial gastroenteritis affecting humans, and poultry is considered a major source of the causative organism, Campylobacter spp. Broilers may ...arrive contaminated at slaughterhouses, and transport crates could be considered a potential source of contamination. Thus, cleaning and disinfection procedures are crucial to avoid cross-contamination among flocks. Despite its public health importance in Latin American countries, virulence factors of Campylobacter jejuni remain poorly studied in this region. Thus, this study aimed to: 1) determine the occurrence of contaminated crates at a poultry slaughterhouse, 2) compare the contamination before and after the cleaning and disinfection procedures, and 3) detect virulence-associated genes in C. jejuni strains by PCR. Campylobacter spp. were recovered from 8 of the 10 flocks evaluated, and C. jejuni was detected as the main species. There was no significant difference in the Campylobacter detection or quantification between crates at the reception platform and crates after the cleaning/disinfection processes. However, crates after 24 h of natural drying, presented a significant (P < 0.05) lower amount of Campylobacter cells than before the cleaning and disinfection processes. A negative relationship (R2 = 0.210, P = 0.045) between environmental conditions and Campylobacter quantification was found for transport crates after 24 h of natural drying. There was no significant difference (P > 0.05) in the detection of two C. jejuni virulence genes, flaA (encode a major flagellin protein) and cadF (encode an adhesion and fibronectin-binding protein), among various stages of the cleaning and disinfection processes. Our results demonstrate the high contamination levels of Campylobacter strains in broiler flocks and the potential involvement of poultry transport crates in transmitting these bacteria. This study also suggests that ineffective cleaning and disinfection procedures can increase Campylobacter contamination and facilitate the spread of bacteria in poultry establishments.
The trivalent lanthanide isonicotinates were synthesized to obtain stoichiometry Lu(IN)
3
and Ln(IN)
3
·2H
2
O (Ln = Tb to Lu, and Y; IN = isonicotinate). A deep study of the thermal behavior in ...oxidant (air) and inert (N
2
) atmospheres was carried out using the following thermoanalytical techniques: simultaneous thermogravimetry and differential scanning calorimetry (TG–DSC), differential scanning calorimetry (DSC) and evolved gas analysis (EGA by TG–DSC–FTIR). From these results, it was possible to determine that dehydration occurs in a single step and the thermal decomposition of the anhydrous compounds occurs in one or two (air), and two or three steps (N
2
). The final residues of thermal decomposition were Tb
4
O
7
and Ln
2
O
3
(Ln = Dy to Lu, and Y) in air atmosphere, while in N
2
atmosphere the mass loss is still being observed up to 1000 °C. The identified gaseous products evolved during the thermal decomposition in dynamic dry air and nitrogen atmospheres were water, CO
2
, pyridine and CO. From these thermoanalytical data, it was possible to propose a general equation of thermal decomposition of these compounds in the N
2
atmosphere. DSC curves of Tb and Ho compounds presented endothermic peaks corresponding to a reversible phase transition, confirmed by powder X-ray diffractometry (XRD), not previously reported. In addition, infrared vibrational spectroscopy (IR) suggests the coordination through carboxylate as bridging bidentate ligand toward the heavy trivalent lanthanides metals. This paper is complementary to our previous study involving the series of light trivalent lanthanides isonicotinates.
Despite jellyfish’s ecological and socioeconomic impacts, they are still an understudied component of marine ecosystems. Even with its conspicuous size, reaching ~ 1 m in bell diameter,
Drymonema ...gorgo
has been rarely observed, with only a few occurrences in the literature, suggesting that it is not a common species. Here, we gathered historical records from literature, personal communication, and novel data from the World Wide Web (WWW) to obtain observations of
D. gorgo
along the Brazilian coast. A total of 63 observations from 1857 to 2022 were compiled, of which 57 were extracted from the web. Observations on this jellyfish concentrated between 22° S and 27° S, especially in the Cabo Frio region in Rio de Janeiro. They were reported throughout the year, except in August and October, with the highest concentrations in March (late summer). Based on the notable complement on information about its occurrence in the last ten years, we hypothesize that
D. gorgo
is probably not as rare as suggested by the near absence of previous literature records. In addition, biological associations of
D. gorgo
with other species were also discovered such as (1) predation on the jellyfish
Aurelia
sp. and
Chrysaora lactea
; (2) food item for the green turtle
Chelonia mydas
; and (3) association with fishes. This study provides evidence of the potential use of the WWW to obtain ecological data about conspicuous marine species occurrence, such as
D. gorgo
, helping to fill knowledge gaps and overcome difficulties in its detection using traditional methodologies.
Brazil's mangroves: Natural carbon storage da S Bezerra, Denilson; de Lima Santos, Adriano; Bezerra, Janaina Santos ...
Science (American Association for the Advancement of Science),
2022-Mar-18, 2022-03-18, 20220318, Letnik:
375, Številka:
6586
Journal Article
Solid-state Ln(Mef)
3
compounds, where Ln stands for some trivalent lanthanides (La, Ce, Pr and Nd) and Mef is the mefenamate (C
15
H
14
NO
2
−
) ligand, were synthesized. The characterization and ...thermal and spectroscopic study of the compounds were performed using elemental analysis, complexometry, image analysis by field emission gun, simultaneous thermogravimetry and differential scanning calorimetry coupled to Fourier transform infrared spectroscopy, X-ray diffractometry, near-infrared and mid-infrared spectroscopy. All the analyses showed that the compounds were obtained in the anhydrous state. The thermoanalytical results showed that the stability and thermal behavior were markedly influenced by the atmosphere used. Moreover, the curves show that the thermal decomposition takes place in two or three steps, with the formation of lanthanide oxide (La
2
O
3
, CeO
2
, Pr
6
O
11
and Nd
2
O
3
), as the final residue. Furthermore, the theoretical and experimental spectroscopic data suggested the possible modes of coordination of the ligand with the metals. The DR spectra provided information about the ligand absorption bands and the
f
–
f
transitions of Nd
3+
and Pr
3+
ions.
The majority of current approaches for bias and fairness identification or mitigation in machine learning models are applications for a particular issue that fails to account for the connection ...between the application context and its associated sensitive attributes, which contributes to the recognition of consistent patterns in the application of bias and fairness metrics. This can be used to drive the development of future models, with the sensitive attribute acting as a connecting element to these metrics. Hence, this study aims to analyze patterns in several metrics for identifying bias and fairness, applying the gender-sensitive attribute as a case study, for three different areas of applications in machine learning models: computer vision, natural language processing, and recommendation systems. The gender attribute case study has been used in computer vision, natural language processing, and recommendation systems. The method entailed creating use cases for facial recognition in the FairFace dataset, message toxicity in the Jigsaw dataset, and movie recommendations in the MovieLens100K dataset, then developing models based on the VGG19, BERT, and Wide Deep architectures and evaluating them using the accuracy, precision, recall, and F1-score classification metrics, as well as assessing their outcomes using fourteen fairness metrics. Certain metrics disclosed bias and fairness, while others did not, revealing a consistent pattern for the same sensitive attribute across different application domains, and similarities for the statistical parity, PPR disparity, and error disparity metrics across domains, indicating fairness related to the studied sensitive attribute. Some attributes, on the other hand, did not follow this pattern. As a result, we conclude that the sensitive attribute may play a crucial role in defining the fairness metrics for a specific context.
Antioxidant compounds protect plants against oxidative stress caused by environmental conditions. Different light qualities, such as UV‐A radiation and blue light, have shown positive effects on the ...production of phenols in plants. Kalanchoe pinnata (Lamarck) Persoon (Crassulaceae) is used for treating wounds and inflammations. Some of these beneficial effects are attributed to the antioxidant activity of plant components. We investigated the effects of blue light and UV‐A radiation supplementation on the total phenol content, antioxidant activity and chromatographic profile of aqueous extracts from leaves of K. pinnata. Monoclonal plants were grown under white light, white plus blue light and white plus UV‐A radiation. Supplemental blue light improved the antioxidant activity and changed the phenolic profile of the extracts. Analysis by HPLC of supplemental blue‐light plant extracts revealed a higher proportion of the major flavonoid quercetin 3‐O‐α‐l‐arabinopyranosyl (1→2) α‐l‐rhamnopyranoside, as well as the presence of a wide variety of other phenolic substances. These findings may explain the higher antioxidant activity observed for this extract. Blue light is proposed as a supplemental light source in the cultivation of K. pinnata, to improve its antioxidant activity.
This study reports a potential use of blue light as a supplemental light source in cultivation of Kalanchoe pinnata, an important medicinal species. We investigated the effects of blue and UV‐A light supplementation on the total phenol content, antioxidant activity and chromatographic profile of K. pinnata extracts. Supplemental blue light improved the antioxidant activity and changed the phenolic profile. Analysis by HPLC of supplemental blue‐light plant extracts (A) revealed a higher proportion of the major flavonoid in K. pinnata, as well as the presence of quercitrin and a wide variety of other phenolic substances, when compared with control (B).
Human H-kininogen (120 kDa) plays a role in many pathophysiological processes and interacts with the cell surface through protein receptors and proteoglycans, which mediate H-kininogen endocytosis. ...In the present work we demonstrate that H-kininogen containing bradykinin domain is internalized and different endogenous kininogenases are present in CHO-K1 cells. We used CHO-K1 (wild type) and CHO-745 (mutant deficient in proteoglycans biosynthesis) cell lines. H-kininogen endocytosis was studied using confocal microscopy, and its hydrolysis by cell lysate fraction was determined by immunoblotting. Bradykinin release was also measured by radioimmunoassay. H-kininogen interaction with the cell surface of CHO-745 cells resulted in bradykinin release by serine proteases. In CHO-K1 cells, which produce heparan and chondroitin sulfate proteoglycans, internalization of H-kininogen through its bradykinin domain can occur on lipid raft domains/caveolae. Nevertheless bradykinin-free H-kininogen was not internalized by CHO-K1 cells. The H-kininogen present in acidic endosomal vesicles in CHO-K1 was approximately 10-fold higher than the levels in CHO-745. CHO-K1 lysate fractions were assayed at pH 5.5 and intact H-kininogen was totally hydrolyzed into a 62 kDa fragment. By contrast, at an assay pH 7.4, the remained fragments were 115 kDa, 83 kDa, 62 kDa and 48 kDa in size. The antipain-Sepharose chromatography separated endogenous kininogenases from CHO-K1 lysate fraction. No difference was detected in the assays at pH 5.5 or 7.4, but the proteins in the fraction bound to the resin released bradykinin from H-kininogen. However, the proteins in the unbound fraction cleaved intact H-kininogen at other sites but did not release bradykinin. H-kininogen can interact with extravascular cells, and is internalized dependent on its bradykinin domain and cell surface proteoglycans. After internalization, H-kininogen is proteolytically processed by intracellular kininogenases. The present data also demonstrates that serine or cysteine proteases in lipid raft domains/caveolae on the CHO cell can hydrolyze H-kininogen, thus releasing kinins.
The aim of this work was to evaluate the role of human plasma prekallikrein assembly and processing in cells and to determine whether proteoglycans, along with high molecular weight kininogen ...(H-kininogen), influence this interaction.
We used the endothelial cell line ECV304 and the epithelial cell lines CHO-K1 (wild type) and CHO-745 (deficient in proteoglycans). Prekallikrein endocytosis was studied using confocal microscopy, and prekallikrein cleavage/activation was determined by immunoblotting using an antibody directed to the prekallikrein sequence C364TTKTSTR371 and an antibody directed to the entire H-kininogen molecule.
At 37°C, prekallikrein endocytosis was assessed in the absence and presence of exogenously applied H-kininogen and found to be 1,418.4±0.010 and 1,070.3±0.001 pixels/cell, respectively, for ECV304 and 1,319.1±0.003 and 631.3±0.001 pixels/cell, respectively, for CHO-K1. No prekallikrein internalization was observed in CHO-745 in either condition. Prekallikrein colocalized with LysoTracker in the absence and presence of exogenous H-kininogen at levels of 76.0% and 88.5%, respectively, for ECV304 and at levels of 40.7% and 57.0%, respectively, for CHO-K1. After assembly on the cell surface, a plasma kallikrein fragment of 53 kDa was predominant in the incubation buffer of all the cell lines studied, indicating specific proteolysis; plasma kallikrein fragments of 48-44 kDa and 34-32 kDa were also detected in the incubation buffer, indicating non-specific cleavage. Bradykinin free H-kininogen internalization was not detected in CHO-K1 or CHO-745 cells at 37°C.
The prekallikrein interaction with the cell surface is temperature-dependent and independent of exogenously applied H-kininogen, which results in prekallikrein endocytosis promoted by proteoglycans. Prekallikrein proteolysis/activation is influenced by H-kininogen/glycosaminoglycans assembly and controls plasma kallikrein activity.
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