Antifungal potential of the new copper(II)-theophylline/1,10-phenanthroline complex against drug-resistant Candida species.

Candida spp. are the commonest fungal pathogens worldwide. Antifungal resistance is a problem that has prompted the discovery of novel anti-Candida drugs. Herein, 25 compounds, some of them containing copper(II), cobalt(II) and manganese(II) ions, were initially evaluated for inhibiting the growth of reference strains of Candida albicans and Candida tropicalis. Eight (32%) of the compounds inhibited the proliferation of these yeasts, displaying minimum inhibitory concentrations (MICs) ranging from 31.25 to 250 µg/mL and minimum fungicidal concentration (MFCs) from 62.5 to 250 µg/mL. Drug-likeness/pharmacokinetic calculated by SwissADME indicated that the 8 selected compounds were suitable for use as topical drugs. The complex CTP, Cu(theo)2phen(H2O).5H2O (theo = theophylline; phen = 1,10-phenanthroline), was chosen for further testing against 10 medically relevant Candida species that were resistant to fluconazole/amphotericin B. CTP demonstrated a broad spectrum of action, inhibiting the growth of all 20 clinical fungal isolates, with MICs from 7.81 to 62.5 µg/mL and MFCs from 15.62 to 62.5 µg/mL. Conversely, CTP did not cause lysis in erythrocytes. The toxicity of CTP was evaluated in vivo using Galleria mellonella and Tenebrio molitor. CTP had no or low levels of toxicity at doses ranging from 31.25 to 250 µg/mL for 5 days. After 24 h of treatment, G. mellonella larvae exhibited high survival rates even when exposed to high doses of CTP (600 µg/mL), with the 50% cytotoxic concentration calculated as 776.2 µg/mL, generating selectivity indexes varying from 12.4 to 99.4 depending on each Candida species. These findings suggest that CTP could serve as a potential drug to treat infections caused by Candida species resistant to clinically available antifungals.

Unveiling the antifungal mechanisms of CTP, a new copper(II)-theophylline/1,10-phenanthroline complex, on drug-resistant non-albicans Candida species.

Candida species undeniably rank as the most prevalent opportunistic human fungal pathogens worldwide, with Candida albicans as the predominant representative. However, the emergence of non-albicans Candida species (NACs) has marked a significant shift, accompanied by rising incidence rates and concerning trends of antifungal resistance. The search for new strategies to combat antifungal-resistant Candida strains is of paramount importance. Recently, our research group reported the anti-Candida activity of a coordination compound containing copper(II) complexed with theophylline (theo) and 1,10-phenanthroline (phen), known as "CTP" - Cu(theo)2phen(H2O).5H2O. In the present work, we investigated the mechanisms of action of CTP against six medically relevant, antifungal-resistant NACs, including C. auris, C. glabrata, C. haemulonii, C. krusei, C. parapsilosis and C. tropicalis. CTP demonstrated significant efficacy in inhibiting mitochondrial dehydrogenases, leading to heightened intracellular reactive oxygen species production. CTP treatment resulted in substantial damage to the plasma membrane, as evidenced by the passive incorporation of propidium iodide, and induced DNA fragmentation as revealed by the TUNEL assay. Scanning electron microscopy images of post-CTP treatment NACs further illustrated profound alterations in the fungal surface morphology, including invaginations, cavitations and lysis. These surface modifications significantly impacted the ability of Candida cells to adhere to a polystyrene surface and to form robust biofilm structures. Moreover, CTP was effective in disassembling mature biofilms formed by these NACs. In conclusion, CTP represents a promising avenue for the development of novel antifungals with innovative mechanisms of action against clinically relevant NACs that are resistant to antifungals commonly used in clinical settings.

Interventions to Change Movement Behaviors After Stroke: A Systematic Review and Meta-analysis.

OBJECTIVE: This systematic review aimed to determine which interventions increase physical activity (PA) and decrease sedentary behavior (SB) based on objective measures of movement behavior in individuals with stroke. DATA SOURCES: The PubMed (Medline), EMBASE, Scopus, CINAHL (EBSCO), and Web of Science databases were searched for articles published up to January 3, 2023. STUDY SELECTION: The StArt 3.0.3 BETA software was used to screen titles, abstracts, and full texts for studies with randomized controlled trial designs; individuals with stroke (≥18 years of age); interventions aimed at increasing PA or decreasing SB; and objective measurement instruments. DATA EXTRACTION: Data extraction was standardized, considering participants and assessments of interest. The risk of bias and quality of evidence of the included studies were assessed. DATA SYNTHESIS: Twenty-eight studies involving 1855 patients were included. Meta-analyses revealed that in the post-stroke acute/subacute phase, exercise interventions combined with behavior change techniques (BCTs) increased both daily steps (standardized mean difference [SMD]=0.65, P=.0002) and time spent on moderate-to-vigorous intensity physical activities (MVPAs) duration of PA (SMD=0.68, P=.0004) with moderate-quality evidence. In addition, interventions based only on BCTs increased PA levels with very low-quality evidence (SMD (low-intensity physical activity)=0.36, P=.02; SMD (MVPA)=0.56, P=.0004) and decreased SB with low-quality evidence (SMD=0.48, P=.03). In the post-stroke chronic phase, there is statistical significance in favor of exercise-only interventions in PA frequency (steps/day) with moderate-quality evidence (SMD=0.68, P=.002). In general, the risk of bias in the included studies was low. CONCLUSIONS: In the acute/subacute phase after stroke, the use of BCTs combined with exercise can increase the number of daily steps and time spent on MVPA. In contrast, in the post-stroke chronic phase, exercise-only interventions resulted in a significant increase in daily steps.

Evaluation of postprandial glycemic response in rats (Wistar) fed with different starch sources.

The consumption of readily digestible starch sources, such as corn, can negatively impact endocrine disorders related to hyperglycaemia in predisposing animals. In this sense, starch sources containing slowly digestible and resistant fractions can assist glycemic control. The present study analyzed the postprandial glycemic response and blood variables of rats fed with four extruded diets containing corn, brown rice, sweet potato and pea as the main starch source. Thirty-two male Wistar rats (90 days old) were divided into groups of eight animals each. The rats received one of the experimental diets for 30 days according to a completely randomised design. The glycemia was measured on the 29th and 30th days. The glycemia measured on the 29th day was analyzed at 0, 30, 60, 120 and 240 min after oral administration of 50% glucose solution. On the 30th day, the same protocol was repeated after providing 3 g of the experimental diet for each animal to obtain the glycemic curve. After the euthanasia on the 30th day, 7 ml of blood was collected via cardiac puncture for glycated haemoglobin (HBA1c), triglycerides, cholesterol, and aspartate and alanine aminotransferases analysis. Diets with pea and sweet potato provided lower glycemic index, average and maximum glycemia, and glycemic increment in relation to the other starch sources (p < 0.05). Animals fed with the corn diet had higher serum concentrations of triglycerides and HBA1c than the other treatments (p < 0.05). Results demonstrated that pea and sweet potato are interesting starch sources for the control of metabolic disorders related to glycemia.

Diet supplemented with Saccharomyces cerevisiae from different fermentation media modulates the faecal microbiota and the intestinal fermentative products in dogs.

This study aimed at evaluating the coefficients of total tract apparent digestibility (CTTAD) of nutrients, metabolisable energy (ME), diet palatability, faecal fermentative products and microbiota of dogs fed yeasts from different fermentation media and its fractions. Four diets were evaluated: control, without yeast (CO); diet with 10 g/kg brewer's yeast (BY); diet with 10 g/kg brewer's yeast + corn yeast (BCY); and diet with 10 g/kg BCY + cell wall fractions (BCYF). Twelve adult dogs were distributed in a randomized block design (periods). Each of the four diets was fed to a group of three dogs per period of 20 days, totalling two periods and six repetitions per treatment. Sixteen adult dogs were used for the palatability test, which compared the CO diet versus each one of the yeast diets. Data with normal distribution were subjected to analysis of variance (p < 0.05). Means were compared by orthogonal contrasts (p < 0.05): (A) CO diet versus BY, BCY and BCYF diets; (B) BY diet versus BCY and BCYF diets; (C) BCY diet versus BCYF diet. There was no difference in the CTTAD and ME of the diets (p > 0.05). Yeast diets reduced faecal odour and indole peak area (p < 0.05). Faecal short-chain fatty acids concentration was greater in dogs fed yeast diets compared to those fed the CO (p < 0.05). Yeast diets showed a higher intake ratio compared to the CO (p < 0.05). The BCY and BCYF diets resulted in a greater abundance of Bacteroides, Faecalibacterium, Coprococcus, and Phascolarctobacterium in relation to the CO (p < 0.05). Our results suggest that dietary yeast supplementation results in beneficial changes in intestinal functionality indicators, mainly with the combination of yeasts from brewers and corn fermentation media. In addition, yeast supplementation improves diet palatability without compromising nutrient digestibility.

Fine cassava fibre utilization as a dietary fibre source for dogs: Effects on kibble characteristics, diet digestibility and palatability, faecal metabolites and microbiota.

The objective was to evaluate through three experiments the effects of a fine cassava fibre (CA: 106 µm) on kibble characteristics, coefficients of total tract apparent digestibility (CTTAD) of macronutrients, diet palatability and faecal metabolites and microbiota of dogs. Dietary treatments consisted of a control diet (CO), without an added fibre source and with 4.3% total dietary fibre (TDF), and a diet with 9.6% CA (106 µm), with 8.4% TDF. Experiment I evaluated the physical characteristics of the kibbles. The palatability test was evaluated in experiment II, which compared the diets CO versus CA. In experiment III, 12 adult dogs were randomly assigned to one of the two dietary treatments for 15 days, totalling six replicates/treatment, to assess the CTTAD of macronutrients; faecal characteristics, faecal metabolites and microbiota. The expansion index, kibble size and friability of diets with CA were higher than the CO (p < 0.05). Additionally, the CA diet presented higher palatability than the CO (p < 0.05) but did not affect CTTAD except for those of fibre (p > 0.05). Moreover, a greater faecal concentration of acetate, butyrate and total short-chain fatty acids (SCFA) and a lower faecal concentration of phenol, indole and isobutyrate were observed in dogs fed the CA diet (p < 0.05). Dogs fed with the CA diet presented a greater bacterial diversity and richness and a greater abundance of genera considered to be beneficial for gut health, such as Blautia, Faecalibacterium and Fusobacterium when compared to the CO group (p < 0.05). The inclusion of 9.6% of a fine CA improves the expansion of kibbles and diet palatability without affecting most of the CTTAD of nutrients. Besides, it improves the production of some SCFA and modulates the faecal microbiota of dogs.

Decoding the antifungal resistance mechanisms in biofilms of emerging, ubiquitous and multidrug-resistant species belonging to the Scedosporium/Lomentospora genera.

The opportunistic filamentous fungi belonging to the Scedosporium and Lomentospora genera are highly tolerant to all classes of available antifungal drugs. Moreover, the mature biofilm formed by these fungi presents higher antifungal resistance when compared to planktonic cells. Nevertheless, the resistance mechanisms developed by the biofilm lifestyle are not completely elucidated. In the current study, we have investigated the mainly known resistance mechanisms to azoles (voriconazole and fluconazole) and polyenes (amphotericin B [AMB]) in S. apiospermum, S. minutisporum, S. aurantiacum, and L. prolificans (formerly S. prolificans) biofilms. Both classes of antifungals can physically bind to the extracellular matrix of mature biofilms, preventing the drugs from reaching their targets on biofilm-forming cells, which precludes their activity and toxicity. In addition, the activity of efflux pumps, measured by Rhodamine 6 G, was increased along with the maturation of the biofilm. The efflux pump's inhibition by L-Phe-L-Arg-ß-naphthylamide culminated in a 2- to 16-fold increase in azole susceptibility in conidial cells, but not in mature biofilms. Finally, we demonstrated by using specific inhibitors that in conidia, but not in biofilms, AMB induced the production of reactive oxygen species through the activity of the oxidative phosphorylation system (complex I-IV and alternative oxidases). However, the cellular redox imbalance caused by AMB was well-coped with the high activity of antioxidative enzymes, such as superoxide dismutase and catalase. Altogether, our results revealed that Scedosporium/Lomentospora biofilm resistance occurs through various mechanisms that operate concomitantly, which could explain the huge challenge in the clinical treatment of scedosporiosis/lomentosporiosis. LAY SUMMARY: Scedosporium/Lomentospora spp. are multidrug-resistant pathogens able to cause diverse types of infections with typical biofilm characteristics, which makes the treatment a hard issue. We deciphered the resistance mechanisms to classical antifungals developed in the biofilm formed by these fungi.

DHA/EPA supplementation decreases anxiety-like behaviour, but it does not ameliorate metabolic profile in obese male rats.

Obesity is a major public health problem that predisposes to several diseases and higher mortality in patients with COVID-19. Obesity also generates neuroinflammation, which predisposes to the development of neuropsychiatric diseases. Since there is a lack of effective treatments for obesity, the search for new strategies to reverse its consequences is urgent. In this perspective, the anti-inflammatory properties of omega-3 polyunsaturated fatty acids such as DHA/EPA might reduce the harmful effects of obesity. Here, we used the cafeteria diet (CAF) model to induce obesity in Wistar rats. Animals received ultra-processed food for 20 weeks, and DHA/EPA supplementation (500 mg/kg per d) was performed between the 16th and the 20th week. At the end of the experiment, it was evaluated: body weight, visceral fat deposition, plasma glucose, insulin and triglycerides, and it was also measured the levels of inflammatory cytokines TNF-α and IL-6 in plasma and liver, and TNF-α in the prefrontal cortex. The elevated plus maze test was performed to analyse anxiety-like behaviour. Our results demonstrated that DHA/EPA could not reverse weight and fat gain and did not modify plasma dosages. However, there was a decrease in IL-6 in the liver (DHA/EPA effect: P = 0.023) and TNF-α in the brain (CAF compared with CAF + DHA/EPA, P < 0.05). Also, there was a decrease in the anxiety index in CAF + DHA/EPA compared with the CAF group (P < 0.01). Thus, DHA/EPA supplementation is helpful to reverse the consequences of obesity in the brain.

Hemispheric lateralization, endothelial function, and arterial compliance in chronic post-stroke individuals: A cross-sectional exploratory study.

Purpose/Aim: Cardiovascular function is controlled and regulated by a functional brain-heart axis. Although the exact mechanism is not fully understood, several studies suggest a hemispheric asymmetry in the neural control of cardiovascular function. Thus, the purpose of this study was to examine whether endothelial function and arterial compliance differ between individuals with left- and right-sided strokes.Materials and Methods: This was a cross-sectional exploratory study. Thirty individuals more than 6 months after stroke participated in the study. The endothelial function was assessed by ultrasound-measured flow-mediated dilation of the nonparetic arm brachial artery (baFMD). The arterial stiffness was assessed by measuring carotid-femoral pulse wave velocity (cfPWV) and central aortic pulse wave analysis [augmentation index (AIx), augmentation index normalized to a heart rate of 75 bpm (AIx@75) and reflection magnitude (RM)] using applanation tonometry. Results: Participants with right-sided stroke had worse endothelial function than those with left-sided stroke. This difference (baFMD = 2.51%) was significant (p = 0.037), and it represented a medium effect size (r = 0.38). Likewise, they had higher arterial stiffness than those with left-sided stroke. This difference (AIx = 10%; RM = 7%) was significant (p = 0.011; p = 0.012), and it represented a medium effect size (r = 0.48; r = 0.47).Conclusions: Our findings suggest that individuals with right-sided stroke have reduced endothelial function and arterial compliance compared to those with left-sided stroke. These data may indicate that those with right-sided strokes are more susceptible to cardiovascular events.

Differences in Charge Distribution in Leishmania tarentolae Leishmanolysin Result in a Reduced Enzymatic Activity.

Leishmania tarentolae is a non-pathogenic trypanosomatid isolated from lizards widely used for heterologous protein expression and extensively studied to understand the pathogenic mechanisms of leishmaniasis. The repertoire of leishmanolysin genes was reported to be expanded in L. tarentolae genome, but no proteolytic activity was detected. Here, we analyzed L. tarentolae leishmanolysin proteins from the genome to the structural levels and evaluated the enzymatic activity of the wild-type and overexpressing mutants of leishmanolysin. A total of 61 leishmanolysin sequences were retrieved from the L. tarentolae genome. Five of them were selected for phylogenetic analysis, and for three of them, we built 3D models based on the crystallographic structure of L. major ortholog. Molecular dynamics simulations of these models disclosed a less negative electrostatic potential compared to the template. Subsequently, L. major LmjF.10.0460 and L. tarentolae LtaP10.0650 leishmanolysins were cloned in a pLEXSY expression system into L. tarentolae. Proteins from the wild-type and the overexpressing parasites were submitted to enzymatic analysis. Our results revealed that L. tarentolae leishmanolysins harbor a weak enzymatic activity about three times less abundant than L. major leishmanolysin. Our findings strongly suggest that the less negative electrostatic potential of L. tarentolae leishmanolysin can be the reason for the reduced proteolytic activity detected in this parasite.

The industrial versatility of Gluconobacter oxydans: current applications and future perspectives.

Gluconobacter oxydans is a well-known acetic acid bacterium that has long been applied in the biotechnological industry. Its extraordinary capacity to oxidize a variety of sugars, polyols, and alcohols into acids, aldehydes, and ketones is advantageous for the production of valuable compounds. Relevant G. oxydans industrial applications are in the manufacture of L-ascorbic acid (vitamin C), miglitol, gluconic acid and its derivatives, and dihydroxyacetone. Increasing efforts on improving these processes have been made in the last few years, especially by applying metabolic engineering. Thereby, a series of genes have been targeted to construct powerful recombinant strains to be used in optimized fermentation. Furthermore, low-cost feedstocks, mostly agro-industrial wastes or byproducts, have been investigated, to reduce processing costs and improve the sustainability of G. oxydans bioprocess. Nonetheless, further research is required mainly to make these raw materials feasible at the industrial scale. The current shortage of suitable genetic tools for metabolic engineering modifications in G. oxydans is another challenge to be overcome. This paper aims to give an overview of the most relevant industrial G. oxydans processes and the current strategies developed for their improvement.

Anti-Leishmania braziliensis activity of 1,10-phenanthroline-5,6-dione and its Cu(II) and Ag(I) complexes.

Leishmaniasis, included in the priority list of the WHO, remains as a neglected disease caused by parasites of the Leishmania genus. There is no vaccine available for human leishmaniasis, and the current treatment is based on old drugs that cause serious side effects. Herein, we initially studied the cellular distribution of the virulence factor gp63, the major metallopeptidase, in a virulent strain of Leishmania braziliensis, and then we measured the inhibitory effects of 1,10-phenanthroline-5,6-dione (phendione), and its metal complexes, [Cu(phendione)3](ClO4)2.4H2O and [Ag(phendione)2]ClO4, on both cellular and extracellular metallopeptidases produced by promastigotes. The action of the three compounds on parasite viability and on parasite-macrophage interaction was also determined. Gp63 molecules were detected in several parasite compartments, including the cytoplasm, the membrane lining the cell body and flagellum, and in the flagellar pocket, which explains the presence of gp63 in the culture medium. The test compounds inhibited parasite metallopeptidases in a typical dose-dependent manner, and they also caused a significant and irreversible inhibition of parasite motility. Moreover, the pre-treatment of promastigotes with the test compounds induced a decrease in the association index with macrophages. Collectively, phendione and its Cu(II) and Ag(I) complexes are excellent prototypes for the development of new anti-L. braziliensis drugs.

Photobiomodulation on trigeminal neuralgia: systematic review.

Trigeminal neuralgia is a recurrent episode of facial pain, that may be associated with other conditions such as multiple sclerosis, neoplasms, and nerve compromises or may occur due to an unknown cause. The available treatments are pharmacotherapy or surgery; however, both are susceptible to develop side effects. Photobiomodulation could be a promising alternative therapy for trigeminal neuralgia. A systematic review of literature was carried out using the PRISMA protocol, in the PubMed/MEDLINE, Embase, and Web of Science databases. Risk of bias by ROB 2.0 protocol was performed in included studies. Initially, 20 identified articles were collected varying between the years of 1983-2018, from which 6 were included. A total of 193 patients were evaluated; photobiomodulation was compared to conventional therapies, TENS, and therapy combinations with pharmacotherapy. The overall risk of bias was low, with some concerns in the randomization and double-blinding process; moreover, there are few reports in the literature. Photobiomodulation appears to be as effective as conventional therapies, being a coadjutant therapeutic opportunity for the treatment of trigeminal neuralgia.

Exploring Innovative Leishmaniasis Treatment: Drug Targets from Pre-Clinical to Clinical Findings.

Leishmaniasis is a group of tropical diseases caused by parasitic protozoa belonging to the genus Leishmania. The disease is categorized in cutaneous leishmaniasis (CL), mucocutaneous leishmaniasis (MCL), and visceral leishmaniasis (VL). The conventional treatment is complex and can present high toxicity and therapeutic failures. Thus, there is a continuing need to develop new treatments. In this review, we focus on the novel molecules described in the literature with potential leishmanicidal activity, categorizing them in pre-clinical (in vitro, in vivo), drug repurposing and clinical research.

Innovation in the teaching of human physiology at university and school: pedagogical process based on interdisciplinarity and learning station rotation.

This paper presents the description and analysis of a didactic experience involving the participation of a university and a community school, developed as part of the National Science and Technology Week, at a public university in northeastern Brazil. For this purpose, the use of learning station rotation enabled innovation in the teaching of physiology integrated with biochemistry and health education contents. The didactic approach consisted of creating a learning circuit comprising seven stations. The central theme of the stations emphasized physiology, with specific foci on biochemistry and cardiorespiratory and endocrine health. Each station provided unique activities related to the central theme, including a station concerning digital technology in physiology. The school students were divided into small groups (6 or 7 people) that rotated through the stations, with a total of 81 students visiting each station. A qualitative assessment was performed using a Likert-scale questionnaire to measure the level of satisfaction of the students. It was found that this didactic approach increased the receptivity of the students to the contents, facilitated student-teacher dialogue, and provided an excellent tool for establishing an interface between the university and the community school. Overall, 76.5% of the students rated the activity as excellent.

Effect of dietary inclusion of dried apple pomace on faecal butyrate concentration and modulation of gut microbiota in dogs.

This research aimed to evaluate the apparent total tract digestibility (ATTD) of nutrients, metabolisable energy (ME) and palatability of the diet, as well as products of intestinal fermentation and faecal microbiota of dogs fed with dried apple. For this purpose, three experiments were performed. In Experiment I, digestibility and ME of four diets containing 0%, 3%, 6% and 9% dried apple were evaluated, in addition to the faecal characteristics of the dogs. The diets were offered to eight adult dogs, distributed in double Latin square (4 × 4), totalling eight repetitions per treatment. In Experiment II, products of intestinal fermentation and faecal microbiota from 16 adult dogs fed diets containing 0% and 9% dried apple for 30 d (n = 8) were evaluated. Finally, Experiment III compared the dietary preference of 0 vs. 9% dried apple using 15 adult dogs. The inclusion of dried apple in the diet (p < 0.05) showed a linear reduction in the ATTD of dry matter (DM), crude protein (CP), and acid hydrolysed ether extract (EEA), and a linear increase in the ATTD of total dietary fibre (TDF). Consumption of 9% of dried apple increased faecal butyrate and reduced propionate and ammonia (p < 0.05). With this diet, there was also an increase (p < 0.05) in the faecal concentration of Faecalibacterium, Erysipelatoclostridium, Blautia, and Bacteroides. No differences were found in the palatability of the diets. The inclusion of up to 9% of dried apple in the diet reduces the digestibility of nutrients and does not influence the dogs' food preference; however, it improves some indicators of dogs' intestinal functionality.

Soybean oil supplementation for sows in the first three days after farrowing.

One of the main causes of death in newborn piglets is the low level of energy reserves to maintain their body temperature, which can lead to hypothermia and, subsequently, death. Thus, the objective of this study was to evaluate the effect of soybean oil supplementation to sows in the first three days after farrowing to save piglet fat reserves through the higher nutritional intake of sow milk. In total, 604.5, 750, 1000, 1250 and 1500 g of soybean oil were provided for each sow during the three days of supplementation. A total of 60 sows were evaluated per treatment, distributed in a random block design, supplemented in the first three days after farrowing with soybean oil added on top of the feed at the time of feeding. Performance and reproductive data and milk samples were collected from the sows to determine fat levels. Piglets were evaluated for fall-back rate and survival. There was no significant effect of soybean oil supplementation on any of the parameters evaluated for both sows and their milk. Therefore, soybean oil supplementation for sows in the first three days after farrowing does not influence performance parameters, reproduction and milk fat of the sows and mortality and fall-back rate of the piglets.

Core of the saliva microbiome: an analysis of the MG-RAST data.

BACKGROUND: Oral microbiota is considered as the second most complex in the human body and its dysbiosis can be responsible for oral diseases. Interactions between the microorganism communities and the host allow establishing the microbiological proles. Identifying the core microbiome is essential to predicting diseases and changes in environmental behavior from microorganisms. METHODS: Projects containing the term "SALIVA", deposited between 2014 and 2019 were recovered on the MG-RAST portal. Quality (Failed), taxonomic prediction (Unknown and Predicted), species richness (Rarefaction), and species diversity (Alpha) were analyzed according to sequencing approaches (Amplicon sequencing and Shotgun metagenomics). All data were checked for normality and homoscedasticity. Metagenomic projects were compared using the Mann-Whitney U test and Spearman's correlation. Microbiome cores were inferred by Principal Component Analysis. For all statistical tests, p < 0.05 was used. RESULTS: The study was performed with 3 projects, involving 245 Amplicon and 164 Shotgun metagenome datasets. All comparisons of variables, according to the type of sequencing, showed significant differences, except for the Predicted. In Shotgun metagenomics datasets the highest correlation was between Rarefaction and Failed (r = - 0.78) and the lowest between Alpha and Unknown (r = - 0.12). In Amplicon sequencing datasets, the variables Rarefaction and Unknown (r = 0.63) had the highest correlation and the lowest was between Alpha and Predicted (r = - 0.03). Shotgun metagenomics datasets showed a greater number of genera than Amplicon. Propionibacterium, Lactobacillus, and Prevotella were the most representative genera in Amplicon sequencing. In Shotgun metagenomics, the most representative genera were Escherichia, Chitinophaga, and Acinetobacter. CONCLUSIONS: Core of the salivary microbiome and genera diversity are dependent on the sequencing approaches. Available data suggest that Shotgun metagenomics and Amplicon sequencing have similar sensitivities to detect the taxonomic level investigated, although Shotgun metagenomics allows a deeper analysis of the microorganism diversity. Microbiome studies must consider characteristics and limitations of the sequencing approaches. Were identified 20 genera in the core of saliva microbiome, regardless of the health condition of the host. Some bacteria of the core need further study to better understand their role in the oral cavity.

Design and Validation of a Simulator for Feline Cephalic Vein Cannulation-A Pilot Study.

In recent years there has been an increased use of alternative methods for teaching veterinary clinical skills, since ethical considerations preclude the use of live animals for demonstration or practice of many procedures. Skills training on cats (i.e., feline venipuncture) is a particularly challenging area. This study aimed to develop a simulator for cephalic venipuncture in cats and to validate this simulator using questionnaires answered by undergraduate students and experienced veterinarians. The simulator was developed to provide an experience that was close to reality, including an artificial blood system that flows through the catheter when venipuncture is correctly performed, while at the same time using simple methodology and accessible materials so that it could be reproduced in other universities. The experienced vets agreed (44.4%) or strongly agreed (55.6%) that the simulator was good for venipuncture training, and the most useful feature was the experience of catheter manipulation and fixation on the cat's limb. All the students agreed that the practical class with the simulator was important for learning this skill. Both groups (students and experienced veterinarians) unanimously agreed that it is important to train using a simulator before trying the procedure on a live cat. This simulator offers undergraduate students an alternative way to learn and practice venipuncture in cats helping to reduce the use of live animals in practical classes.

Surface, adhesiveness and virulence aspects of Candida haemulonii species complex.

The emerging opportunistic pathogens comprising the Candida haemulonii complex (C. haemulonii [Ch], C. duobushaemulonii [Cd] and C. haemulonii var. vulnera[Chv]) are notable for their intrinsic antifungal resistance. Different clinical manifestations are associated with these fungal infections; however, little is known about their biology and potential virulence attributes. Herein, we evaluated some surface properties of 12 clinical isolates of Ch (n = 5), Cd (n = 4) and Chv (n = 3) as well as their virulence on murine macrophages and Galleria mellonella larvae. Scanning electron microscopy demonstrated the presence of homogeneous populations among the species of the C. haemulonii complex, represented by oval yeasts with surface irregularities able to form aggregates. Cell surface hydrophobicity was isolate-specific, exhibiting high (16.7%), moderate (25.0%) and low (58.3%) hydrophobicity. The isolates had negative surface charge, except for one. Mannose/glucose- and N-acetylglucosamine-containing glycoconjugates were evidenced in considerable amounts in all isolates; however, the surface expression of sialic acid was poorly detected. Cd isolates presented significantly higher amounts of chitin than Ch and Chv. Membrane sterol and lipid bodies, containing neutral lipids, were quite similar among all fungi studied. All isolates adhered to inert surfaces in the order: polystyrene > poly-L-lysine-coated glass > glass. Likewise, they interacted with murine macrophages in a quite similar way. Regarding in vivo virulence, the C. haemulonii species complex were able to kill at least 80% of the larvae after 120 hours. Our results evidenced the ability of C. haemulonii complex to produce potential surface-related virulence attributes, key components that actively participate in the infection process described in Candida spp.