Optimization of spermatozoa analysis in mice: A comprehensive protocol.

Sperm quality is critical to predict reproductive alterations caused by immunological factors or toxicant agents. Yet, no detailed protocol has been published focusing on analyses of sperm parameters in mice. Our aim was to evaluate the most efficient diluent for mice sperm analyses and to optimize the sperm morphology classification, through the comparison of different staining methods. The diluents assessed were PBS (baseline), HTF, DMEM, 1 % BSA in PBS and 9 % skimmed powdered milk diluted in PBS. Spermatozoa were evaluated for vitality, motility, and morphology, smears were stained with Papanicolaou, HE, Giemsa, and Rapid staining. Sperm vitality and total motility reached better scores in milk based and DMEM diluents. HE raised up as an effective option since its combination with any of the diluents we tested, resulted in a fair staining, which was appropriated to evaluate mice spermatozoa. Finally, based on WHO manual, we have updated the current morphological classification for mice sperm, since we have detailed the head defects as well as included midpiece and tail defects on it. Taken together, we presented a useful, low cost, and reliable method to assess sperm morphology that could be employed worldwide by laboratories dedicated to study reproductive biology on mice model.

Stereotactic radiotherapy for liver oligometastases: a pooled analysis following the estro/eortc consensus recommendations.

A large pooled analysis of liver oligometastases, classified accordingly to the ESTRO/EORTC recommendations, treated by stereotactic radiotherapy (SBRT) and Radiosurgery (SRS) was carried out. The clinical and dosimetric data of patients who underwent SBRT/SRS for liver metastases were analysed in terms of efficacy and toxicity profile. In particular, the Local Control (LC), the Distant Metastases Free Survival (DMFS), the Disease-Free Survival (DFS), the Overall Survival (OS), and the Next Systemic Therapy Free Survival (NEST-FS) rates were analysed. 113 patients (M/F: 49/64), accounting for a total of 150 hepatic lesions (March 2006-February 2023) in two Italian radiotherapy Institutions were evaluated. Median age was 67 years old (36-92) and 48 (42.5%) patients had at least one comorbidity. The majority of the lesions were induced (30.7%) or repeated oligoprogressive (12.7%) metastases. 98 lesions were treated with more than one daily fraction (mainly 50 Gy in 5 fractions), while 52 were radiosurgery treatments (mainly 32 Gy). The treatment response at 3-4 months was evaluable in 147 lesions: complete response was 32.0%, partial response 17.0%, and stable disease 32.0%. Actuarial LC, DMFS, DFS, OS, and NEST-FS at 1 year were 75.8%, 37.7%, 34.9%, 78.7%, and 59.4% respectively; while actuarial LC, DMFS, DFS, OS, and NEST-FS at 2 years were 52.1%, 24.9%, 21.9%, 51.3%, and 36.8%, respectively. The achievement of complete response, synchronous oligometastases, and no treatment interruptions correlated with a more favorable outcomes. As per the toxicity profile, we registered only two acute and one late toxicity cases higher than grade 2. Stereotactic treatment for liver metastases seems to be a safe and promising option in terms of local control. The best results in term of outcomes have been obtained in patients with complete response, synchronous oligometastases, favorable histology, and no treatment interruptions.

Systematic review of literature to evaluate global distribution of species of the Sporothrix genus stored in culture collections.

Introduction: Sporotrichosis is a subcutaneous mycosis caused by fungi of the genus Sporothrix sp. Phenotypic and genotypic differences have been associated with their geographic distribution, virulence, or clinical manifestation of sporotrichosis. In the past decade, the interest in identifying species of the Sporothrix sp. has been increasing, due to its epidemiological importance and, in consequence, is important to know how to preserve them for future studies, in culture collection. Aims: The purposes of this study were to analyze the global distribution of environmental isolates and/or causal agents of sporotrichosis identified by polyphasic taxonomy, with mandatory use of molecular identification, and to evaluate the percentages and distribution of isolates stored in culture collections. Methods: A systematic review of articles on animal and human sporotrichosis and/or environmental isolation of the fungus, from 2007 to 2023, was done. Results: Our results demonstrated that, S. globosa, S. schenckii, and S. brasiliensis were the most identified species. With respect to the deposit and maintenance of species, we observed that only 17% of the strains of Sporothrix sp. isolated in the world are preserved in a culture collection. Conclusions: This systematic review confirmed a difficulty in obtaining the frequency of Sporothrix species stored in culture collection and insufficient data on the molecular identification mainly of animal sporotrichosis and isolation of Sporothrix sp. in environmental samples.

Field Hip Stability Isometric Test (F-HipSIT): Reliability of Assessing the Hip Posterolateral Muscle Strength in Sports Settings.

CONTEXT: The Hip Stability Isometric Test (HipSIT) is commonly employed in clinical settings for evaluating the hip posterolateral muscle strength. In this study, we introduced the "Field Hip Stability Isometric Test" (F-HipSIT) and assessed the intrarater and interrater reliability of this strength assessment specifically designed for sports settings. DESIGN: Reliability study. METHODS: Two independent raters (A and B) went to athletes' training facilities to conduct 2 sessions of F-HipSIT spaced at least 1 week apart. The average peak force value from 3 valid attempts of each leg was recorded and normalized by the participant's body mass for statistical analysis. RESULTS: Thirty male and 30 female amateur athletes took part in this study. Rater A obtained similar values in the first (0.39 [0.05] and 0.44 [0.07] kg·f/kg) and second (0.39 [0.06] and 0.45 [0.07] kg·f/kg) testing days for men and women, respectively. Rater B also found similar values in the first (0.35 [0.06] and 0.42 [0.08] kg·f/kg) and second (0.36 [0.06] and 0.45 [0.08] kg·f/kg) testing days for men and women, respectively. Excellent intrarater intraclass correlation coefficient (ICC) values were found for men (ICC = .922) and women (ICC = .930), with coefficient of variation of 6% to 8% and minimal detectable change of 0.06 to 0.10 kg·f/kg. The F-HipSIT presented good interrater reliability for men (ICC = .857) and women (ICC = .868), with coefficient of variation of 5% and minimal detectable change of 0.05 to 0.06 kg·f/kg. CONCLUSION: The F-HipSIT intrarater and interrater reliability among male and female recreational athletes supports this field test as a quick and convenient screening tool to monitor hip posterolateral muscle strength in sports settings.

Metabolomic analysis of retinopathy stages and amputation in type 2 diabetes.

BACKGROUND: Diabetic retinopathy (DR) and limb amputation are frequent complications of diabetes that cannot always be explained by blood glucose control. Metabolomics is a science that is currently being explored in the search for biomarkers or profiles that identify clinical conditions of interest. OBJECTIVE: This study aimed to analyze, using a metabolomic approach, peripheral blood samples from type 2 diabetes mellitus (DM2) individuals, compared with those with diabetic retinopathy and limb amputation. METHODS: The sample consisted of 128 participants, divided into groups: control, DM2 without DR (DM2), non-proliferative DR (DRNP), proliferative DR (DRP), and DM2 amputated (AMP). Metabolites from blood plasma were classified by spectra using nuclear magnetic resonance (NMR), and the metabolic routes of each group using metaboanalyst. RESULTS: We identified that the metabolism of phenylalanine, tyrosine, and tryptophan was discriminant for the DRP group. Histidine biosynthesis, on the other hand, was statistically associated with the AMP group. The results of this work consolidate metabolites such as glutamine and citrulline as discriminating for DRP, and the branched-chain amino acids as important for DR. CONCLUSIONS: The results demonstrate the relationship between the metabolism of ketone bodies, with acetoacetate metabolite being discriminating for the DRP group and histidine being a significant metabolite in the AMP group, when compared to the DM2 group.

Do athletes with hamstring strain injury have shorter muscle fascicles in the injured limb?

INTRODUCTION: Previous studies have suggested that a reduced length of the biceps femoris long head (BFlh) fascicles may increase the risk of hamstring strain injury (HSI). However, it remains unclear whether the BFlh fascicles of the injured limb are shorter than those of the contralateral limb in athletes with an acute HSI. OBJECTIVE: To investigate the between-limb asymmetry of BFlh fascicle length in amateur athletes with an acute HSI. METHODS: Male amateur athletes were evaluated using ultrasound scans within five days following an HSI. The BFlh fascicle length was estimated using a validated equation. RESULTS: Eighteen injured athletes participated in this study. There was no significant difference (p = 0.27) in the length of BFlh fascicles between the injured limb (9.53 ± 2.55 cm; 95%CI 8.26 to 10.80 cm) and the uninjured limb (10.54 ± 2.87 cm; 95%CI 9.11 to 11.97 cm). Individual analysis revealed high heterogeneity, with between-limb asymmetries (percentage difference of the injured limb compared to the uninjured limb) ranging from -42% to 25%. Nine out of the 18 athletes had a fascicle length that was more than 10% shorter in the injured limb compared to the uninjured limb, five athletes had a difference of less than 10%, and four athletes had a fascicle length that was more than 10% longer in the injured limb compared to the uninjured limb. CONCLUSION: The architecture characteristics of injured and uninjured muscles is not consistent among athletes with HSI. Therefore, rehabilitation programs focused on fascicle lengthening should be evaluated on a case-by-case basis.

Nutritional and antioxidant potential of Pleurotus djamor (Rumph. ex Fr.) Boedijn produced on agronomic wastes banana leaves and sugarcane bagasse substrates.

Global food production faces challenges concerning access to nutritious and sustainably produced food. Pleurotus djamor, however, is an edible mushroom that can be cultivated on agricultural waste. Considering that nutritional and functional potential of mushrooms can change based on cultivation conditions, we examined the influence of substrates with different compositions of banana leaf and sugarcane bagasse on the nutritional, mycochemical, and antioxidant properties of P. djamor. The mushrooms were grown for 120 days and dried in a circulating air oven at 45 °C for three days. We conducted bromatological analyses and mycochemical characterization (1H-NMR, total phenolics, and flavonoids) of the mushrooms and assayed the antioxidant activity of extracts from the dried mushrooms using an ethanol/water solution (70:30 v/v). In general, the substrates produced mushrooms with high protein (18.77 ± 0.24% to 17.80 ± 0.34%) and dietary fiber content (18.02 ± 0.05% to 19.32 ± 0.39%), and with low lipid (0.28 + 0.08% to 0.4 + 0.6%), and caloric content (maximum value: 258.42 + 8.49), with no significant differences between the groups (p ≥ 0.05). The mushrooms also exhibited high levels of total phenolics and flavonoids. The mushrooms cultivated on sugarcane bagasse substrates presented the highest values (p < 0.05). Analysis of the 1H-NMR spectra indicates an abundant presence of heteropolysaccharides, ß-glucans, α-glucans, and oligosaccharides, and all the mushroom extracts exhibited high antioxidant activity. In conclusion, our study demonstrates that agricultural residues permit sustainable production of edible mushrooms while maintaining nutritional and functional properties.

Genotoxic and antiproliferative properties of Endopleura uchi bark aqueous extract.

The bark extract from Endopleura uchi has been widely used in traditional medicine to treat gynecological-related disorders, diabetes, and dyslipidemias albeit without scientific proof. In addition, E. uchi bark extract safety, especially regarding mutagenic activities, is not known. The aim of this study was to determine the chemical composition, antitumor, and toxicological parameters attributed to an E. uchi bark aqueous extract. The phytochemical constitution was assessed by colorimetric and chromatographic analyzes. The antiproliferative effect was determined using sulforhodamine B (SRB) assay using 4 cancer cell lines. Cytotoxic and genotoxic activities were assessed utilizing MTT and comet assays, respectively, while mutagenicity was determined through micronucleus and Salmonella/microsome assays. The chromatographic analysis detected predominantly the presence of gallic acid and isoquercitrin. The antiproliferative effect was more pronounced in human colon adenocarcinoma (HT-29) and human breast cancer (MCF-7) cell lines. In the MTT assay, the extract presented an IC50 = 39.1 µg/ml and exhibited genotoxic (comet assay) and mutagenic (micronucleus test) activities at 20 and 40 µg/ml in mouse fibroblast cell line (L929) and mutagenicity in the TA102 and TA97a strains in the absence of S9 mix. Data demonstrated that E. uchi bark possesses bioactive compounds which exert cytotoxic and genotoxic effects that might be associated with its antitumor potential. Therefore, E. uchi bark aqueous extract consumption needs to be approached with caution in therapeutic applications.

Ebselen: A Promising Repurposing Drug to Treat Infections Caused by Multidrug-Resistant Microorganisms.

Bacterial multiresistance to drugs is a rapidly growing global phenomenon. New resistance mechanisms have been described in different bacterial pathogens, threatening the effective treatment of even common infectious diseases. The problem worsens in infections associated with biofilms because, in addition to the pathogen's multiresistance, the biofilm provides a barrier that prevents antimicrobial access. Several "non-antibiotic" drugs have antimicrobial activity, even though it is not their primary therapeutic purpose. However, due to the urgent need to develop effective antimicrobials to treat diseases caused by multidrug-resistant pathogens, there has been an increase in research into "non-antibiotic" drugs to offer an alternative therapy through the so-called drug repositioning or repurposing. The prospect of new uses for existing drugs has the advantage of reducing the time and effort required to develop new compounds. Moreover, many drugs are already well characterized regarding toxicity and pharmacokinetic/pharmacodynamic properties. Ebselen has shown promise for use as a repurposing drug for antimicrobial purposes. It is a synthetic organoselenium with anti-inflammatory, antioxidant, and cytoprotective activity. A very attractive factor for using ebselen is that, in addition to potent antimicrobial activity, its minimum inhibitory concentration is very low for microbial pathogens.

Suprachoroidal gene transfer with nonviral nanoparticles in large animal eyes.

Suprachoroidal nonviral gene therapy with biodegradable poly(ß-amino ester) nanoparticles (NPs) provides widespread expression in photoreceptors and retinal pigmented epithelial (RPE) cells and therapeutic benefits in rodents. Here, we show in a human-sized minipig eye that suprachoroidal injection of 50 µl of NPs containing 19.2 µg of GFP expression plasmid caused GFP expression in photoreceptors and RPE throughout the entire eye with no toxicity. Two weeks after injection of 50, 100, or 200 µl, there was considerable within-eye and between-eye variability in expression that was reduced 3 months after injection of 200 µl and markedly reduced after three suprachoroidal injections at different locations around the eye. Reduction of bacterial CpG sequences in the expression plasmid resulted in a trend toward higher expression. These data indicate that nonviral suprachoroidal gene therapy with optimized polymer, expression plasmid, and injection approach has potential for treating photoreceptors throughout the entire retina of a human-sized eye.

Molecular Profile of Variants Potentially Associated with Severe Forms of COVID-19 in Amazonian Indigenous Populations.

Coronavirus disease 2019 (COVID-19) is an infection caused by SARS-CoV-2. Genome-wide association studies (GWASs) have suggested a strong association of genetic factors with the severity of the disease. However, many of these studies have been completed in European populations, and little is known about the genetic variability of indigenous peoples' underlying infection by SARS-CoV-2. The objective of the study is to investigate genetic variants present in the genes AQP3, ARHGAP27, ELF5L, IFNAR2, LIMD1, OAS1 and UPK1A, selected due to their association with the severity of COVID-19, in a sample of indigenous people from the Brazilian Amazon in order to describe potential new and already studied variants. We performed the complete sequencing of the exome of 64 healthy indigenous people from the Brazilian Amazon. The allele frequency data of the population were compared with data from other continental populations. A total of 66 variants present in the seven genes studied were identified, including a variant with a high impact on the ARHGAP27 gene (rs201721078) and three new variants located in the Amazon Indigenous populations (INDG) present in the AQP3, IFNAR2 and LIMD1 genes, with low, moderate and modifier impact, respectively.

Harnessing Brazilian biodiversity database: identification of flavonoids as potential inhibitors of SARS-CoV-2 main protease using computational approaches and all-atom molecular dynamics simulation.

SARS-CoV-2 (Severe Acute Respiratory Syndrome Coronavirus 2) is the etiological agent responsible for the global outbreak of COVID-19 (Coronavirus Disease 2019). The main protease of SARS-CoV-2, Mpro, is a key enzyme that plays a vital role in mediating viral replication and transcription. In this study, a comprehensive computational approach was employed to investigate the binding affinity, selectivity, and stability of natural product candidates as potential new antivirals acting on the viral polyprotein processing mediated by SARS-CoV-2 Mpro. A library of 288 flavonoids extracted from Brazilian biodiversity was screened to select potential Mpro inhibitors. An initial filter based on Lipinski's rule of five was applied, and 204 compounds that did not violate any of the Lipinski rules were selected. The compounds were then docked into the active site of Mpro using the GOLD program, and the poses were subsequently re-scored using MM-GBSA (Molecular Mechanics Generalized Born Surface Area) binding free energy calculations performed by AmberTools23. The top five flavonoids with the best MM-GBSA binding free energy values were selected for analysis of their interactions with the active site residues of the protein. Next, we conducted a toxicity and drug-likeness analysis, and non-toxic compounds were subjected to molecular dynamics simulation and free energy calculation using the MM-PBSA (Molecular Mechanics Poisson-Boltzmann Surface Area) method. It was observed that the five selected flavonoids had lower MM-GBSA binding free energy with Mpro than the co-crystal ligand. Furthermore, these compounds also formed hydrogen bonds with two important residues, Cys145 and Glu166, in the active site of Mpro. Two compounds that passed the drug-likeness filter showed stable conformations during the molecular dynamics simulations. Among these, NuBBE_867 exhibited the best MM-PBSA binding free energy value compared to the crystallographic inhibitor. Therefore, this study suggests that NuBBE_867 could be a potential inhibitor against the main protease of SARS-CoV-2 and may be further examined to confirm our results.

Frederic Lewy: how the two World Wars changed his life, work, and name.

In 1912, Friedrich Lewy described the inclusion bodies present in Parkinson disease and in Lewy body dementia. Throughout his life, Lewy fought in two wars - on opposite sides. He was born in Berlin in a Jewish family, and served in the German Army in World War I. In the following years, on many occasions he had to change his line of research due to Nazi persecution. Lewy became a naturalized American, changed his name to Frederic Henry Lewey, and served in the US Army as a lieutenant colonel. Lewy died in 1950 and never used the famous eponym in his papers.

Em 1912, Friedrich Lewy descreveu os corpos de inclusão presentes na doença de Parkinson e na demência com corpos de Lewy. Ao longo sua vida, Lewy lutou em duas guerras, em lados opostos. Ele nasceu em Berlin em uma família de origem judaica e serviu no Exército alemão na Primeira Guerra Mundial. Nos anos seguintes, passou muitas vezes teve de mudar sua linha de pesquisa devido à perseguição Nazista. Lewy naturalizou-se americano, mudou seu nome para Frederic Henry Lewey, e serviu no Exército americano como tenente-coronel. Lewy morreu em 1950 e nunca usou o famoso epônimo em seus artigos.

Quantification of the liver structure of zebrafish (Danio rerio) submitted to different diets and physical exercise.

The zebrafish has been used in research for over 80 years. In the last three decades, discoveries about the fundamental properties of development, regeneration, cancer, and other diseases have established the zebrafish as an important model organism in biomedical research. This study aimed to evaluate liver alterations in zebrafish by quantitatively assessing the areas occupied by hepatocytes, as well as connective and adipose tissues. Forty-eight adult Danio rerio (38 males and 10 females) of approximately 13 months of age were used. They were divided into four groups, with 12 animals each. The fish were randomly distributed to form the groups, which received a maintenance and/or hypercaloric diet, with or without the addition of physical exercise. The animals underwent six hours of forced exercise (5 cm/s) for thirteen weeks. The animals that practiced physical exercise had a higher volumetric density of the area occupied by hepatocytes (65.92%±1.81 - GMex and 50.75%±2.24 GHex) among the groups. The GH group had a higher volumetric density of the area occupied by connective tissue (15.12%±0.72), followed by the GHex group (13.53%±1.43). Regarding the volumetric density of the area occupied by adipose tissue, the GH group had a higher density (27.21%±1.36), followed by the GHex group (21.66%±1.11) with statistically significant differences. The GMex had a volumetric density of the area occupied by adipose tissue of 3.5%±0.76, while the GM had 5.7%±0.5, with statistical difference. In relation to the animals in the GHex group, they had 20.39% less fat than the animals in the GH group. The animals in the GMex group had 72.47% less fat than those in the GM group. It is concluded that the different dietary constitutions and the imposition of physical exercise were able to modify the structural architecture of the liver of Danio rerio. These are acceptable criteria for modulations, thus aiming at the control and possible interferences directly related to the metabolism of the species and therefore the control of diseases.

Exposure to progesterone before an ovulation synchronization protocol increases the follicular diameter and fertility of multiparous suckled Bos taurus cows.

The objective of this study was to evaluate the effect of administering injectable progesterone (P4i) before a timed artificial insemination (TAI) protocol on the follicular growth, ovulation, and pregnancy rate of Bos taurus suckled cows. The effect of P4i administration before the TAI on the pregnancy rate (P/AI) was evaluated in 576 suckled Bos taurus cows at 30-90 days postpartum. In addition, the effect of P4i administration before TAI on follicular dynamics was evaluated in subgroup of 401 suckled Bos taurus cows. On Day -10 (D-10), cows were divided into two experimental groups (Control and P4i). In this moment, P4i cows received i.m. 150 mg of injectable long-action progesterone. After that, both experimental groups received a synchronization protocol (Day 0; D0) that consisted of administration i.m. of 2 mg of estradiol benzoate and a progesterone intravaginal insert on D0. On Day 8 (D8), the progesterone insert was removed, and the cows received 500 µg of cloprostenol, 400 IU of eCG, and 1 mg of estradiol cypionate. TAI was performed 48 h after the removal of the progesterone insert. The ultrasound exams were performed in a subgroup of cows on Days 0, 8, 10 and 12 to evaluate the diameter of the largest follicle, rate of follicular growth and risks of single and double ovulation. The pregnancy diagnosis was performed 30 days after TAI in all cows to determine the pregnancy rate. The diameter of the largest follicle, on D10 (P = 0.84), rate of follicular growth (P = 0.14), ovulation rate (P = 0.40) and double ovulation rates (P = 0.23) did not differ between experimental groups. The pregnancy rate was greater in the P4i group [Control 46.2 % (133/288) vs. P4i 55.6 % (160/288); P = 0.03]. The diameter of the largest follicles (LF) on D0 (Control 11.6 ± 0.2 vs. P4i 13.3 ± 0.3) was greater (P = 0.01) in the P4i group. In conclusion, injectable progesterone before the ovulation synchronization protocol increased the diameter of the largest follicle on the D0 and the pregnancy rate in multiparous Bos taurus suckled beef cows.

Twenty-first-century demographic and social inequalities of heat-related deaths in Brazilian urban areas.

Population exposure to heat waves (HWs) is increasing worldwide due to climate change, significantly affecting society, including public health. Despite its significant vulnerabilities and limited adaptation resources to rising temperatures, South America, particularly Brazil, lacks research on the health impacts of temperature extremes, especially on the role played by socioeconomic factors in the risk of heat-related illness. Here, we present a comprehensive analysis of the effects of HWs on mortality rates in the 14 most populous urban areas, comprising approximately 35% of the country's population. Excess mortality during HWs was estimated through the observed-to-expected ratio (O/E) for total deaths during the events identified. Moreover, the interplay of intersectionality and vulnerability to heat considering demographics and socioeconomic heterogeneities, using gender, age, race, and educational level as proxies, as well as the leading causes of heat-related excess death, were assessed. A significant increase in the frequency was observed from the 1970s (0-3 HWs year-1) to the 2010s (3-11 HWs year-1), with higher tendencies in the northern, northeastern, and central-western regions. Over the 2000-2018 period, 48,075 (40,448-55,279) excessive deaths were attributed to the growing number of HWs (>20 times the number of landslides-related deaths for the same period). Nevertheless, our event-based surveillance analysis did not detect the HW-mortality nexus, reinforcing that extreme heat events are a neglected disaster in Brazil. Among the leading causes of death, diseases of the circulatory and respiratory systems and neoplasms were the most frequent. Critical regional differences were observed, which can be linked to the sharp North-South inequalities in terms of socioeconomic and health indicators, such as life expectancy. Higher heat-related excess mortality was observed for low-educational level people, blacks and browns, older adults, and females. Such findings highlight that the strengthening of primary health care combined with reducing socioeconomic, racial, and gender inequalities represents a crucial step to reducing heat-related deaths.

The upcoming European Soil Monitoring Law: An effective instrument for the protection of terrestrial ecosystems?

Soils are a precious resource consistently placed under several threats and urgently in need of protection within a regulatory framework at the European level. Soils are central to the provision of environmental services as well as human existence on earth. The need to protect soil has been identified by several recent European strategies and fortunately, a specific European regulation for soil protection is on the way-the European Soil Monitoring Law (formerly: Soil Health Law). However, efforts need to ensure that the upcoming Soil Monitoring Law closes gaps between existing regulations for chemicals and acknowledges current European strategies for environmental protection and sustainability. This brief communication started from a fruitful discussion among SETAC Global Soils Interest Group members on a recent public consultation on the newly proposed Soil Monitoring Law of the European Commission and highlights critical points focusing on the chemical pollution of soils. We emphasize urgent needs such as the essential definition of a "healthy state" of soils; the implementation of a suitable set of indicators and quality standards for the description of physical, chemical, and biological states of soils; the enforcement of the "polluter-pays" principle; and the establishment of a Europe-wide monitoring program. Results from monitoring need to be fed back into regulatory frameworks, including the regulation of chemicals. Guidance documents for the risk assessment of chemicals are outdated and need to be updated. Finally, actions need to be taken to foster healthy soils, stop biodiversity decline, and ensure the functioning of ecosystem services for future generations. Integr Environ Assess Manag 2024;20:316-321. © 2023 The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC).

Antifungal activity of 2-chloro-N-phenylacetamide: a new molecule with fungicidal and antibiofilm activity against fluconazole-resistant Candida spp / Atividade antifúngica de 2-cloro-N-fenilacetamida: uma nova molécula com atividade fungicida e anti-biofilme contra Candida spp. resistentes a fluconazol

In the current context of emerging drug-resistant fungal pathogens such as Candida albicans and Candida parapsilosis, discovery of new antifungal agents is an urgent matter. This research aimed to evaluate the antifungal potential of 2-chloro-N-phenylacetamide against fluconazole-resistant clinical strains of C. albicans and C. parapsilosis. The antifungal activity of 2-chloro-N-phenylacetamide was evaluated in vitro by the determination of the minimum inhibitory concentration (MIC), minimum fungicidal concentration (MFC), inhibition of biofilm formation and its rupture, sorbitol and ergosterol assays, and association between this molecule and common antifungal drugs, amphotericin B and fluconazole. The test product inhibited all strains of C. albicans and C. parapsilosis, with a MIC ranging from 128 to 256 µg.mL-1, and a MFC of 512-1,024 µg.mL-1. It also inhibited up to 92% of biofilm formation and rupture of up to 87% of preformed biofilm. 2-chloro-N-phenylacetamide did not promote antifungal activity through binding to cellular membrane ergosterol nor it damages the fungal cell wall. Antagonism was observed when combining this substance with amphotericin B and fluconazole. The substance exhibited significant antifungal activity by inhibiting both planktonic cells and biofilm of fluconazole-resistant strains. Its combination with other antifungals should be avoided and its mechanism of action remains to be established.

No atual contexto de patógenos fúngicos resistentes emergentes tais como Candida albicans e Candida parapsilosis, a descoberta de novos agentes antifúngicos é uma questão urgente. Esta pesquisa teve como objetivo avaliar o potencial antifúngico da 2-cloro-N-fenilacetamida contra cepas clínicas de C. albicans e C. parapsilosis resistentes a fluconazol. A atividade antifúngica da substância foi avaliada in vitro através da determinação da concentração inibitória mínima (CIM), concentração fungicida mínima (CFM), ruptura e inibição da formação de biofilme, ensaios de sorbitol e ergosterol, e associação entre esta molécula e antifúngicos comuns, anfotericina B e fluconazol. O produto teste inibiu todas as cepas de C. albicans e C. parapsilosis, com uma CIM variando de 128 a 256 µg.mL-1, e uma CFM de 512-1,024 µg.mL-1. Também inibiu até 92% da formação de biofilme e causou a ruptura de até 87% de biofilme pré-formado. A 2-cloro-N-fenilacetamida não promoveu atividade antifúngica pela ligação ao ergosterol da membrana celular fúngica, tampouco danificou a parede celular. Antagonismo foi observado ao combinar esta substância com anfotericina B e fluconazol. A substância exibiu atividade antifúngica significativa ao inibir tanto as células planctônicas quanto o biofilme das cepas resistentes ao fluconazol. Sua combinação com outros antifúngicos deve ser evitada e seu mecanismo de ação deve ser estabelecido.

Antifungal activity of 2-chloro-N-phenylacetamide: a new molecule with fungicidal and antibiofilm activity against fluconazole-resistant Candida spp

Abstract In the current context of emerging drug-resistant fungal pathogens such as Candida albicans and Candida parapsilosis, discovery of new antifungal agents is an urgent matter. This research aimed to evaluate the antifungal potential of 2-chloro-N-phenylacetamide against fluconazole-resistant clinical strains of C. albicans and C. parapsilosis. The antifungal activity of 2-chloro-N-phenylacetamide was evaluated in vitro by the determination of the minimum inhibitory concentration (MIC), minimum fungicidal concentration (MFC), inhibition of biofilm formation and its rupture, sorbitol and ergosterol assays, and association between this molecule and common antifungal drugs, amphotericin B and fluconazole. The test product inhibited all strains of C. albicans and C. parapsilosis, with a MIC ranging from 128 to 256 µg.mL-1, and a MFC of 512-1,024 µg.mL-1. It also inhibited up to 92% of biofilm formation and rupture of up to 87% of preformed biofilm. 2-chloro-N-phenylacetamide did not promote antifungal activity through binding to cellular membrane ergosterol nor it damages the fungal cell wall. Antagonism was observed when combining this substance with amphotericin B and fluconazole. The substance exhibited significant antifungal activity by inhibiting both planktonic cells and biofilm of fluconazole-resistant strains. Its combination with other antifungals should be avoided and its mechanism of action remains to be established.

Resumo No atual contexto de patógenos fúngicos resistentes emergentes tais como Candida albicans e Candida parapsilosis, a descoberta de novos agentes antifúngicos é uma questão urgente. Esta pesquisa teve como objetivo avaliar o potencial antifúngico da 2-cloro-N-fenilacetamida contra cepas clínicas de C. albicans e C. parapsilosis resistentes a fluconazol. A atividade antifúngica da substância foi avaliada in vitro através da determinação da concentração inibitória mínima (CIM), concentração fungicida mínima (CFM), ruptura e inibição da formação de biofilme, ensaios de sorbitol e ergosterol, e associação entre esta molécula e antifúngicos comuns, anfotericina B e fluconazol. O produto teste inibiu todas as cepas de C. albicans e C. parapsilosis, com uma CIM variando de 128 a 256 µg.mL-1, e uma CFM de 512-1,024 µg.mL-1. Também inibiu até 92% da formação de biofilme e causou a ruptura de até 87% de biofilme pré-formado. A 2-cloro-N-fenilacetamida não promoveu atividade antifúngica pela ligação ao ergosterol da membrana celular fúngica, tampouco danificou a parede celular. Antagonismo foi observado ao combinar esta substância com anfotericina B e fluconazol. A substância exibiu atividade antifúngica significativa ao inibir tanto as células planctônicas quanto o biofilme das cepas resistentes ao fluconazol. Sua combinação com outros antifúngicos deve ser evitada e seu mecanismo de ação deve ser estabelecido.

Hábitos de sono e autoavaliação miofuncional orofacial de crianças com risco para distúrbios respiratórios do sono / Sleep habits and orofacial myofunctional self-assessment of children at risk for sleep breathing disorders

RESUMO Objetivo Identificar e correlacionar as queixas de alterações miofuncionais orofaciais e de risco para distúrbios respiratórios do sono com hábitos de sono na infância. Método Participaram 71 pais ou responsáveis de crianças de 6 a 11 anos, matriculadas em uma escola pública. Foi aplicado um formulário com questões semiestruturadas/anamnese e os protocolos Nordic Orofacial Test-Screening - entrevista, Pediatric Obstructive Sleep Apnea Screening Tool Questionnaire, e Children's Sleep Habits Questionnaire - todos em suas versões em português/Brasil no formato online. Para a análise estatística, realizou-se o teste de correlação de Spearman, considerando o nível de significância de 5%. Resultados 29 crianças eram do sexo feminino (40,8%) e 42 do sexo masculino (59,2%), com média de idade de 8,52 anos. As queixas miofuncionais orofaciais encontradas foram relacionadas às funções de respiração (35,2%), mastigação e deglutição (32,4%) e hábitos deletérios (33,8%). Todas as crianças apresentaram um baixo risco para distúrbios respiratórios do sono e quanto aos hábitos de sono, 23 crianças (32,39%) apresentaram uma somatória menor, enquanto que 48 crianças (67,61%) ultrapassaram 41 pontos. Conclusão Houve correlação entre risco para distúrbios respiratórios do sono em crianças com as queixas de alterações miofuncionais orofaciais e com a baixa qualidade/maus hábitos de sono.

ABSTRACT Purpose To identify orofacial myofunctional complaints and sleep-disordered breathing and correlate them with sleep habits in childhood. Methods The study included 71 parents/guardians of public school children aged 6 to 11 years. They answered a form with semi-structured medical history questions and the Nordic Orofacial Test-Screening - interview, the Pediatric Obstructive Sleep Apnea Screening Tool Questionnaire, and the Children's Sleep Habits Questionnaire - all of them in their Portuguese/Brazilian versions in an online format. Statistical analyses used Spearman's correlation, setting the significance level at 5%. Results There were 29 female children (40.8%) and 42 male ones (59.2%), with a mean age of 8.52 years. The study found orofacial myofunctional complaints related to breathing functions (35.2%), chewing and swallowing (32.4%), and deleterious habits (33.8%). All children were at a low risk of sleep-disordered breathing. As for sleep habits, 23 children (32.39%) had a lower total score, whereas 48 children (67.61%) exceeded 41 points. Conclusion There was a correlation between the risk of sleep-disordered breathing in children with complaints of orofacial myofunctional disorders and poor sleep quality/habits.