Insights into Leptocybe invasa resistance in Eucalyptus: phenotyping, genotyping and in silico approaches.

The gall wasp, Leptocybe invasa, poses a significant global threat to Eucalyptus cultivation, by causing substantial economic losses. The objective of this study was to differentiate between resistant and susceptible genotypes by morphological characteristics using image analysis based on the damage caused by the gall wasp. In addition, consensus sequences derived from transposable elements (TEs) and the genome of Eucalyptus spp. Were identified by in silico analysis. Furthermore, another objective was to discriminate Eucalyptus genotypes in response to Leptocybe invasa by conducting molecular analyses involving transposable elements and inter simple sequence markers. For image analysis, the GroundEye ® system was used to collect images of 60 leaves from six genotypes, three of which were resistant and three susceptible. Eucalyptus spp. sequences were obtained from the GenBank database by in silico analysis and pairwise alignments with TE sequences were conducted using BLASTN. Multiple sequence alignment was performed with Clustal Omega, followed by the identification of conserved regions in Jalview. A motif signature was generated using Weblogo. For molecular characterization using ISSR markers and TEs, samples of young leaves were obtained from a total of 80 Eucalyptus seedlings, of which 50 were classified as resistant and 30 as susceptible to L. invasa. It was possible to distinguish gall wasp susceptible and resistant genotypes by image analysis. In silico analysis enabled the identification of conserved regions in the Eucalyptus spp. genome, which were associated with proteins involved in secondary metabolite production, e.g., terpenes, which play a role in the response to L. invasa. The discrimination capacity of TEs and ISSR primers was demonstrated and bands were generated that could be used to identify resistant genotypes. However, increasing the number of markers required to discriminate genotypes in both cases is suggested.

Quercetin inhibits neutrophil extracellular traps release and their cytotoxic effects on A549 cells, as well the release and enzymatic activity of elastase and myeloperoxidase / Quercetina inibe a liberação das armadilhas extracelulares dos neutrófilos e a atividade tóxica delas sobre células A549

Neutrophil extracellular traps (NETs) were first reported as a microbicidal strategy for activated neutrophils. Through an immunologic response against several stimuli, neutrophils release their DNA together with proteins from granules, nucleus, and cytoplasm (e.g., elastase and myeloperoxidase). To date, NETs have been implicated in tissue damage during intense inflammatory processes, mainly when their release is dependent on oxygen radical generation. Flavonoids are antioxidant and anti-inflammatory agents; of these, quercetin is commonly found in our daily diet. Therefore, quercetin could exert some protective activity against tissue damage induced by NETs. In our in vitro assays, quercetin reduced NETs, myeloperoxidase (MPO), and elastase release from neutrophils stimulated with phorbol 12-myristate 13-acetate (PMA). The activity of these enzymes also decreased in the presence of quercetin. Quercetin also reduced the cytotoxic effect of NETs on alveolar cells (A549 cell line). Further, in silico assays indicated favorable interactions between quercetin and NET proteins (MPO and elastase). Overall, our results demonstrate that quercetin decreases deleterious cellular effects of NETs by reducing their release from activated neutrophils, and diminishing the enzymatic activity of MPO and elastase, possibly through direct interaction.

As armadilhas extracelulares de neutrófilos (NETs) foram relatadas pela primeira vez como uma estratégia microbicida para neutrófilos ativados. Por meio de uma resposta imunológica contra vários estímulos, os neutrófilos liberam seu DNA ligado a proteínas de grânulos, núcleo e citoplasma (por exemplo, elastase e mieloperoxidase). Até o momento, os NETs têm sido implicadas em danos aos tecidos durante processos inflamatórios intensos, principalmente quando sua liberação depende da geração de radicais de oxigênio. Os flavonóides são agentes antioxidantes e anti-inflamatórios, e destes, a quercetina é comumente encontrada em nossa dieta diária. Portanto, a quercetina pode exercer alguma atividade protetora contra o dano tecidual induzido por NETs. Em nossos ensaios in vitro, a quercetina reduziu a liberação das NETs, mieloperoxidase (MPO) e elastase a partir de neutrófilos estimulados com forbol 12-miristato 13-acetato (PMA). A atividade dessas enzimas também foi diminuída na presença de quercetina. A quercetina também reduziu o efeito citotóxico dos NETs sobre células alveolares (linha celular A549). Além disso, os ensaios in silico indicaram interações favoráveis entre a quercetina e as proteínas da NET (MPO e elastase). No geral, nossos resultados demonstram que a quercetina diminui os efeitos deletérios das NETs, reduzindo sua liberação à partir de neutrófilos ativados e diminuindo a atividade enzimática de MPO e elastase, possivelmente por meio de interação direta.

Resistance profile and biofilm production of Enterococcus spp., Staphylococcus sp., and Streptococcus spp. from dairy farms in southern Brazil.

Milk is a high nutritional value food that helps in human development and growth. However, it can also harbor microorganisms. Therefore, the objective of this study was to isolate, identify and evaluate the resistance profile and pathogenicity factors of gram-positive cocci isolated from liners in milking rooms in the south of Rio Grande do Sul, Brazil. Biochemical and molecular tests were performed for the identification. The following were isolated: Enterococcus faecalis (10), Enterococcus faecium (4), Staphylococcus intermedius (1), Streptococcus uberis (1), and Streptococcus dysgalactiae (1). The susceptibility of isolated microorganisms to eight antibiotics was evaluated according to CLSI, and the genus that proved to be resistant to most of those was Enterococcus. In addition, all 17 isolates were able to form biofilm, which remained viable after the use of neutral, alkaline and alkaline-chlorinated detergent. The only product that was effective against biofilm of all microorganisms was chlorhexidine 2%. The results obtained highlight the importance of pre- and post-dipping tests on dairy properties, in which chlorhexidine is one of the disinfectants used. As observed, products indicated for cleaning and descaling pipes were not effective on biofilms of the different species tested.

Structural and functional properties of the Kunitz-type and C-terminal domains of Amblyomin-X supporting its antitumor activity.

Amblyomin-X is a Kunitz-type FXa inhibitor identified through the transcriptome analysis of the salivary gland from Amblyomma sculptum tick. This protein consists of two domains of equivalent size, triggers apoptosis in different tumor cell lines, and promotes regression of tumor growth, and reduction of metastasis. To study the structural properties and functional roles of the N-terminal (N-ter) and C-terminal (C-ter) domains of Amblyomin-X, we synthesized them by solid-phase peptide synthesis, solved the X-Ray crystallographic structure of the N-ter domain, confirming its Kunitz-type signature, and studied their biological properties. We show here that the C-ter domain is responsible for the uptake of Amblyomin-X by tumor cells and highlight the ability of this domain to deliver intracellular cargo by the strong enhancement of the intracellular detection of molecules with low cellular-uptake efficiency (p15) after their coupling with the C-ter domain. In contrast, the N-ter Kunitz domain of Amblyomin-X is not capable of crossing through the cell membrane but is associated with tumor cell cytotoxicity when it is microinjected into the cells or fused to TAT cell-penetrating peptide. Additionally, we identify the minimum length C-terminal domain named F2C able to enter in the SK-MEL-28 cells and induces dynein chains gene expression modulation, a molecular motor that plays a role in the uptake and intracellular trafficking of Amblyomin-X.

Treatments for Burning Mouth Syndrome: A Network Meta-analysis.

The aim of this systematic review and network meta-analysis (NMA) of randomized controlled trials was to evaluate the effectiveness of treatments for pain relief of burning mouth syndrome (BMS). Five databases and gray literature were searched. Independent reviewers selected studies, extracted data, and assessed the risk of bias. The primary outcome was pain relief or burning sensation, and the secondary outcomes were side effects, quality of life, salivary flow, and TNF-α and interleukin 6 levels. Four comparable interventions were grouped into different network geometries to ensure the transitivity assumption for pain: photobiomodulation therapy, alpha-lipoic acid, phytotherapics, and anxiolytics/antidepressants. Mean difference (MD) and 95% CI were calculated for continuous outcomes. The minimal important difference to consider a therapy beneficial against placebo was an MD of at least -1 for relief of pain. To interpret the results, the GRADE approach for NMA was used with a minimally contextualized framework and the magnitude of the effect. Forty-four trials were included (24 in the NMA). The anxiolytic (clonazepam) probably reduces the pain of BMS when compared with placebo (MD, -1.88; 95% CI, -2.61 to -1.16; moderate certainty). Photobiomodulation therapy (MD, -1.90; 95% CI, -3.58 to -0.21) and pregabalin (MD, -2.40; 95% CI, -3.49 to -1.32) achieved the minimal important difference of a beneficial effect with low or very low certainty. Among all tested treatments, only clonazepam is likely to reduce the pain of BMS when compared with placebo. The majority of the other treatments had low and very low certainty, mainly due to imprecision, indirectness, and intransitivity. More randomized controlled trials comparing treatments against placebo are encouraged to confirm the evidence and test possible alternative treatments (PROSPERO CRD42021255039).

Free omental graft without vascular microanastomosis in the cutaneous wound healing of rabbits / [Enxerto omental livre sem microanastomose vascular na cicatrização de feridas cutâneas em coelhos]

This study aims to evaluate the influence of free omental graft without vascular microanastomosis (FOGWVA) on experimental skin healing in rabbits. Through celiotomy, a 9cm2 free omental fragment was collected in 36 rabbits, with subsequent production of a deep linear cutaneous wound in the dorsal midline measuring 3cm. In 18 animals from the omental group (OG), the omental fragment collected was fixed subcutaneously with six simple interrupted stitches using a 4-0 polyamide thread. In both treated and control (CG) groups, intradermal dermorrhaphy was performed with 4-0 polyamide thread. Experimental wounds were clinically evaluated every day. Each of the groups was divided into three subgroups of 6 animals each for histopathological evaluation on the 7th, 14th, and 28th days of postoperative. In the OG wounds, the increase in volume (omentum activation) stood out after the second postoperative day. Macroscopy showed an organic reaction to the graft on day 7, with a progressive reduction in addition to neovascularization towards the omental graft. The intense presence of mononuclear cells and collagen deposition on day 7 demonstrated an accelerated process of tissue remodeling and repair. The FOGWVA omental graft remained viable and positively influenced the cutaneous healing of the experimental wounds in rabbits.

Neste estudo, objetiva-se avaliar a influência do enxerto omental livre sem microanastomose vascular (FOGWVA) na cicatrização cutânea experimental em coelhos. Por meio de celiotomia, foi coletado fragmento omental livre de 9cm2 em 36 coelhos, com posterior produção de ferida cutânea profunda linear na linha média dorsal medindo 3cm. Apenas em 18 animais, do grupo omento (GO), o fragmento omental coletado foi fixado no subcutâneo com seis pontos simples interrompidos utilizando fio poliamida 4-0. Em ambos os grupos, tratado e controle (GC), efetuou-se dermorrafia intradérmica com fio poliamida 4-0. As feridas experimentais foram avaliadas clinicamente todos os dias. Cada um dos grupos foi dividido em três subgrupos, com seis animais cada, para avaliação anatomopatológica no sétimo, 14º e 28o dias de pós-operatório. Nas feridas do GO, destacou-se aumento de volume (ativação do omento) a partir do segundo dia pós-operatório. A macroscopia evidenciou reação orgânica ao enxerto no dia sete, com redução progressiva, além de neovascularização em direção ao enxerto omental. Intensa presença de células mononucleares e deposição de colágeno no dia sete demonstraram acelerado processo de remodelamento e reparo tecidual. O FOGWVA manteve-se viável e influenciou positivamente na cicatrização cutânea de feridas experimentais em coelhos.

Age and sex influence the response in lipid metabolism of dehydrated Wistar rats.

Aging is associated a decrease in thirst sensation, which makes old people more susceptible to dehydration. Dehydration produces energy metabolism alterations. Our objective was to determinate the effect of water deprivation (WD) in the lipid metabolism of old male and female rats. Here we show that in the state of WD, aging and sex alters retroperitoneal white adipose tissue (R-WAT) weight of rats, WD old female rats had more lipolysis products than old male rats, a sexual dimorphism in the hormonal response related with metabolism of the adipose tissue of old rats during WD, the expression of P-para mRNA in R-WAT did not present any alteration in animals submitted to WD, the expression of Aqp7 mRNA in R-WAT is altered by WD, age, and sex. Also, WD stimulated an increase in the plasma concentration of oxytocin and the expression of mRNA of the oxytocin receptors in R-WAT.

Effective encapsulation of reuterin-producing Limosilactobacillus reuteri in alginate beads prepared with different mucilages/gums.

The mainly aim of this study was to use mucilaginous solutions obtained from tamarind, mutamba, cassia tora, psyllium and konjac powdered to encapsulate reuterin-producing Limosilactobacillus reuteri in alginate beads by extrusion technique. In the particles were determined the bacterial encapsulation efficiency, cell viability during storage and survival under simulated gastric and intestinal conditions. Moreover, the reuterin production, its entrapment into the beads and the influence on viability of encapsulated microorganism were evaluated. Scanning electron microscopy and Fourier Transform Infrared spectroscopy were employed to characterize the produced particles. The beads showed a relatively spherical shape with homogenous distribution of L. reuteri. The use of gums and mucilages combined with alginate improved the encapsulation efficiency (from 93.2 to 97.4%), the viability of encapsulated bacteria during refrigerated storage (especially in prolonged storage of 20, 30 and 60 days) and the survival after exposure to gastric and enteric environments (from 67.7 to 76.6%). The L. reuteri was able to produce reuterin via bioconversion of glycerol in the film-forming solutions, and the entrapment of the metabolite was improved using konjac, mutamba and tamarind mucilaginous solutions in the encapsulation process (45, 44.57 and 41.25%, respectively). Thus, our findings confirm the great potential of these hydrocolloids to different further purposes, enabling its application as support material for delivery of chemical or biological compounds.

Effect of resistance training combined with ß-glucan ingestion on bone of ovariectomized mice.

OBJECTIVE: This study aimed to evaluate the effects of resistance training (RT) combined with prebiotic ß-glucan (BG) ingestion (Saccharomyces cerevisiae) in ovariectomized (OVX) mice. METHODS: Forty-eight mice were divided into six treatments: SHAM group and five OVX groups treated with saline (SAL), alendronate (ALE) (0.1 mg/kg), BG (62.5 mg/kg), RT (climbing a ladder with load) and the combination of BG + RT. All treatments were performed three times per week for 8 weeks starting 38 days after ovariectomy. RESULTS: OVX effectiveness was confirmed by a reduction in uterus mass (p < 0.05). Femur weight was higher in the SHAM group, followed by the ALE and RT groups with intermediate values and the BG + RT, BG and SAL groups with the lowest values (p < 0.05). The bone mineral density (BMDDEXA) results were higher for the SHAM and RT groups compared to the other groups (p < 0.05). RT provided similar bone mass and resistance to fracture to ALE and SHAM. We found a similar number of osteocytes in the RT and SHAM groups, which was significantly higher than that found in animals receiving BG (p < 0.05). The BG + RT combination increased calcium bone concentration. CONCLUSION: RT promoted benefits on bone health in the OVX model, which were not complemented by the consumption of BG.