Alternative to antimicrobial growth promoters in the diets of broilers challenged with subclinical necrotic enteritis.

Necrotic enteritis (NE) is a disease of worldwide distribution, which affects young broilers and causes economic losses on a scale of 6 billion dollars per year. For decades, NE was controlled in poultry flocks by dietary administration of low doses of antimicrobial growth promoters (AGPs). However, an increase in NE incidence was noted after the AGP ban. This study aimed to compare the effect of an antibiotic (Enramycin) diet to a combination of sodium butyrate, hydrolyzed yeast, and zinc proteinate (ViligenTM) on broiler diets regarding performance, blood parameters, intestinal permeability, morphology and lesions, and carcass yield of broilers challenged with Eimeria spp. and Clostridium perfringens to simulate subclinical necrotic enteritis. A total of 1,150 one-day-old male broiler chickens with an initial average weight of 43.9 ± 0.65 g were allocated to 50 experimental pens. Animals were divided into 5 groups: Negative control (NC) without additives; Positive control (PC) with 0.12 g/ton of Enramycin (8%); V500, V1000, and V1500 with the addition of 500, 1.000, and 1.500 g/ton of Viligen, respectively. All animals were challenged by Eimeria spp. at 7 d of age and by C. perfringens at 17, 18, and 19 d for induction of subclinical NE. The broilers fed with all concentrations of Viligen showed similar performance, blood parameters, intestinal permeability, and carcass yield compared to PC broilers. However, NC broilers showed higher FCR compared to PC broilers from 1 to 33 d (1.42 vs. 1.39) (P = 0.048) and from 1 to 42 d (1.51 vs. 1.49) (P < 0.001). V1500 broilers had fewer intestinal lesions at 28 d when compared to the PC treatment (P < 0.05) and showed that higher Viligen inclusion resulted in lower intestinal damage. At 21 d, the V500 group showed higher intestinal morphology characteristics (VH:VD 4.9 vs. 3.5) compared to the PC treatment (P < 0.001). Thus, in this study, the dietary addition of Viligen to broilers challenged by an experimental model of subclinical NE resulted in lower intestinal damage and similar performance to that obtained by the addition of Enramycin.

Syagrus coronata fixed oil prevents rotenone-induced movement disorders and oxidative stress in Drosophila melanogaster.

One prominent aspect of Parkinson's disease (PD) is the presence of elevated levels of free radicals, including reactive oxygen species (ROS). Syagrus coronata (S. coronata), a palm tree, exhibits antioxidant activity attributed to its phytochemical composition, containing fatty acids, polyphenols, and flavonoids. The aim of this investigation was to examine the potential neuroprotective effects of S. coronata fixed oil against rotenone-induced toxicity using Drosophila melanogaster. Young Drosophila specimens (3-4 d old) were exposed to a diet supplemented with rotenone (50 µM) for 7 d with and without the inclusion of S. coronata fixed oil (0.2 mg/g diet). Data demonstrated that rotenone exposure resulted in significant locomotor impairment and increased mortality rates in flies. Further, rotenone administration reduced total thiol levels but elevated lipid peroxidation, iron (Fe) levels, and nitric oxide (NO) levels while decreasing the reduced capacity of mitochondria. Concomitant administration of S. coronata exhibited a protective effect against rotenone, as evidenced by a return to control levels of Fe, NO, and total thiols, lowered lipid peroxidation levels, reversed locomotor impairment, and enhanced % cell viability. Molecular docking of the oil lipidic components with antioxidant enzymes showed strong binding affinity to superoxide dismutase (SOD) and glutathione peroxidase (GPX1) enzymes. Overall, treatment with S. coronata fixed oil was found to prevent rotenone-induced movement disorders and oxidative stress in Drosophila melanogaster.

Dry residue of cassava on slow-growing broiler diets, with or without the addition of carbohydrases.

Dry residue of cassava was studied on the digestibility, performance, intestinal measurements, with or without inclusion of carbohydrases, of slow-growing broilers. 160 Label Rouge broiler chickens, 21-d-old, were distributed in a randomized, 2x5 factorial arrangement (male and female x 0, 10, 20, 30 and 40% residue) (metabolism trial). 1,100 male chicks were distributed in a 2x5 factorial arrangement (with/without carbohydrases x 0; 2.5; 5.0; 7.5; and 10.0% residue), with five replicates (performance trial). Increasing residue levels led to increases in energetic values. Feed intake from 1-21-d-old and 1-63-d-old decreased linearly. At 42 d-old, feed intake and weight gain levels exhibited a quadratic response, which predicted a highest value at 3.32% and 4.77%, respectively, for diets without carbohydrases. For 21- and 42-d-old chickens, the inclusion of carbohydrases reduced the weight and length of the small intestine. The energetic values of the diets were positively influenced by the residue and had similar digestibility values for both sexes. Inclusion of up to 10% of residue in slow-growing broiler diets does not impaired performance and intestinal morphology. The addition of carbohydrases reduced the viscosity of the digesta but it was not enough to improve the performance of the birds.

Translation and validation into Brazilian Portuguese of the English version of the International Consultation on Incontinence Modular Questionnaire for Male Lower Urinary Tract Symptoms (ICIQ-MLUTS).

OBJECTIVES: To translate and cross-culturally adapt the English version of the International Consultation on Incontinence Modular Questionnaire for Male Lower Urinary Tract Symptoms (ICIQ-MLUTS) into Brazilian Portuguese and evaluate its psychometric properties. INTRODUCTION: Male lower urinary tract symptoms (LUTS) are frequent and commonly assessed with questionnaires. The ICIQ-MLUTS is a robust instrument that investigates the main aspects of LUTS in men and their impact on quality of life. Although highly recommended, Grade A is not as popular as the International Prostate Symptom Score (IPSS) and remained untranslated and unvalidated for Brazilian Portuguese. METHODS: After authorization by the Advisory Board of the International Consultation on Incontinence (ICIQ) the translation process was conducted according to the standard guidelines and the ICIQ validation protocol. Internal consistency was assessed using Cronbach's ⍺ coefficient and values > 0.7 were considered satisfactory. To assess test-retest reliability and reproducibility, Spearman's correlation coefficient and intraclass correlation coefficient were used. For group data, a Spearman correlation coefficient or an intraclass correlation coefficient of at least 0.70 demonstrates good test-retest reliability. A p < 0.05 was considered significant. RESULTS: One hundred and eighty-six, aged 61.41 ± 11.01 years, suffering from LUTS participated in the study between January 2021 and October 2022. Cronbach's ⍺, 0.875, demonstrated the internal consistency of the Portuguese version of ICIQ-MLUTS. The intraclass correlation coefficient of 0.912 (0.882; 0.935 - 95% CI) for the test-retest evidenced the stability and validity of the instrument. Likewise, Spearman's correlation coefficient highlighted the agreement between IPSS and ICIQ-MLUTS, 0.906, <0.001. DISCUSSION: The Portuguese version of the ICIQ-MLUTS demonstrated internal consistency, stability, and validity, in addition to agreement with the IPSS. CONCLUSION: The ICIQ-MLUTS, translated and validated into Brazilian Portuguese, is a robust and reliable instrument to assess LUTS in Brazilian men and can be used in the evaluation of treatment and research.

Performance and bone characteristics of broilers fed diets supplemented with vitamin A at different concentrations.

This work evaluated the influence of vitamin A on performance, organ weight, and bone and skin characteristics in broilers (Cobb 500) at 21 and 42 days of age. A total of 1920 chickens were distributed in a randomised design, considering six vitamin A supplementation levels (0, 6000, 16,000, 26,000, 36,000, and 46,000 IU kg-1 ), with 16 replicates and 20 chickens per experimental unit, established due to rising the range of vitamin levels observed in the literature to evaluate the effect of vitamin A on broilers. At 22 days, half of the replicates from each treatment continued receiving the initial diet, and the other eight repetitions received diets without vitamin A (0 IU kg-1 ) until 42 days. The level of vitamin A influenced feed intake (FI) and body weight gain (BWG) until 21 days for all treatments. Broilers at 21 days of age had a more significant BWG at a vitamin A supplementation level of 28,209 IU kg-1 . At 42 days, vitamin A influenced the BWG and FI of broilers at treatments that were not supplemented after 21 days. Treatments supplemented up to 42 days showed quadratic responses to vitamin A for BWG, FI, and feed conversion. The vitamin A levels influenced the relative weights of the small intestine, pancreas, gizzard, abdominal fat, Seedor index, and breaking strength at 42 days, where the adequate supplementation of vitamin A improved these characteristics in broilers. Vitamin A supplementation from 22 to 42 days old did not affect broiler performance. An increased BWG was obtained when vitamin A supplementation occurred until 42 days, with supplementation of 29,375 IU kg-1 and a lower response of feed conversion with the addition of 27,775 IU kg-1 .

Effects of feeding diets with zinc-l-selenomethionine on growth performance of broilers subjected to cyclic heat stress.

Limited information exists on the use of zinc-l-selenomethionine (Zn-L-SeMet) in broiler diets and its effects on the growth performance, body temperature, mortality rates, blood profile, and gene expression, especially when animals are reared under cyclic heat stress conditions. This study aimed to investigate the impact of Zn-L-SeMet in broiler diets from 1 to 42 days of age reared under cyclic heat stress and its effects on growth performance, cloacal temperatures, mortality rate, blood parameters, and insulin-like growth factor 1 (IGF-1) and growth hormone receptor (GHR) gene expression in the breast muscle. A total of 1000 male Cobb 500® broiler chicks were randomly assigned to five treatments: 0, 0.15, 0.23, 0.47, and 1.30 mg/kg of Zn-L-SeMet. Each treatment consisted of 10 replicates with 20 birds each. No statistically significant differences in growth performance were observed from 1 to 21 days of age (P > 0.05). However, from 1 to 42 days, feed intake (FI) and feed conversion ratio (FCR) decreased linearly (P < 0.05). Cloacal temperatures showed no significant effects (P > 0.05), while overall mortality rate exhibited a quadratic response (P < 0.05), with the optimal inclusion level predicted to reduce broiler mortality at 0.71 mg/kg. Triglyceride (TRG) levels increased with 0.97 mg/kg (P < 0.05), and gama-glutamil transferase (GGT) levels decreased with the inclusion of 1.19 mg/kg (P < 0.05). No significant effects on IGF-1 and GHR gene expression were found (P > 0.05). In conclusion, the inclusion of 1.30 mg/kg of Zn-L-SeMet in diets of heat-stressed broilers improved growth performance from 1 to 42 days of age. An inclusion of 0.71 mg/kg reduced mortality rate, while 0.97 mg and 1.19 mg increased and reduced TRG and GGT levels, respectively.

Therapeutic hypothermia is limited in preventing developmental impairments after neonatal hypoxia-ischemia.

The only current treatment for neonatal hypoxia-ischemia (HI) is therapeutic hypothermia (TH), which still shows some limitations. Specific effects of TH in the several processes involved in brain injury progression remain unclear. In this study, the effects of TH treatment on developmental parameters, behavioral outcomes, and peripheral leukocytes were evaluated in neonatal male and female rats. In P7, animals were submitted to right common carotid artery occlusion followed by hypoxia (8% oxygen). TH was performed by reducing the animal scalp temperature to 32°C for 5 h. Behavioral parameters and developmental landmarks were evaluated. Animals were euthanized at P9 or P21, and cerebral hemispheres, spleen, and thymus were weighed. White blood cells (WBC) were counted in blood smears. There was a reduction in the weight of the brain hemisphere ipsilateral to the carotid occlusion in HI and TH groups, as well as a reduction in body weight gain and a delay in the opening of the ipsilateral eye. Latency in negative geotaxis was increased by HI at P12. TH did not prevent brain weight loss, developmental impairments, or WBC number changes but prevented negative geotaxis impairment and spleen weight reduction. These data reinforce that a better understanding of the events that occur after HI and TH in both males and females is necessary and would allow the development of more adequate and sex-specific therapeutic approaches.

Structural and thermodynamic properties of quasi-2D Mo(1-x)Wx(S, Se, Te)2monolayer alloys: a statistical first principle study.

In this work, we report anab initiostudy of the structural and thermodynamic properties of two-dimensional transition-metal dichalcogenides (2D-TMDC) alloys, Mo(1-x)Wx(S, Se, Te)2, using the cluster expansion framework to compute the Helmholtz free energy of alloys as a function of alloy composition and temperature, in the framework of the generalized quasi-chemical approximation. We consider alloying only on the metal sublayer. Our results indicate a weak dependence of the structural properties (lattice constants, nearest-neighbor bond lengths, and layer width) on the alloy composition (i.e. concentrations of W and Mo atoms), in line with the very similar values of the atomic radii of Mo and W atoms. A stronger dependence on the chalcogen is obtained, a trend that reflects the larger variations in atomic radii among the three chalcogen species. As a function of composition, the structural parameters we examined show similar trends, with negligible bowing (i.e. deviations from a Vegard's law interpolation between end compounds), for the three alloys. Moreover, already at 300 K the behavior of these structural features as a function of composition is very similar to that of the standard-regular-solution (SRS) high-temperature limit. In contrast, the electronic band gaps of the the three alloys as a function of composition show small but significant bowing, as high as -1% to -2% near thex= 0.5 alloy composition. Similarly to the structural features, the band gaps attain the high-temperature SRS limit already at 300 K. Regarding thermodynamic properties, we obtain negative values of the internal energy of mixing for the three alloys over the full range of compositions. Therefore, the theoretical alloying phase diagram for the three alloys is featureless, with stability of a fully-mixed alloy at all temperatures and compositions, with no miscibility gap (hence no bimodal nor spinodal decomposition lines). The thermodynamic potentials (mixing internal energy, mixing entropy, and mixing free energy) reach the high-temperature limit at ∼1000 K, the temperature range of synthesis of 2D-TMDC alloys. These trends of structural and electronic properties of the 2D-TMDC alloys are due to the very similar atomic radii and the nearly identical coordination chemistry of Mo and W. Our results are in agreement with experimental work on the alloying of Mo and W atoms, for samples of Mo(1-x)WxS2monolayer alloys, that found that the random mixed alloy is the thermodynamically stable state for this alloy, with no segregation or phase separation.

Zeolite and corn with different compositions in broiler chickens feeding.

The objective of this work was to evaluate the behavior of zeolite against different types of corn in broiler chickens' diets. 1,200 male broiler chickens were assigned to 6 diets in a completely randomized design and a 2 × 3 factorial arrangement, consisting of 2 types of corn (higher or lower mycotoxin contamination) and the inclusion of zeolite (0; 5,000 and 10,000 g ton-1). In the period from d 1 to 21, there was an interaction (P = 0.0040) between types of corn and the inclusion of zeolite for feed conversion ratio (FCR). In the phase from 1 to 42 d, there was an interaction (P = 0.0322) on the serum levels of creatinine (CREA) and digestible gross energy (dGE); corn with lower mycotoxin level contamination (LMLC), caused a reduction in body weight gain (BWG) (P = 0.0046) and increase in the relative weight of abdominal fat (P = 0.0256). Inclusion rates of zeolite promoted an increase in the digestible CP (P = 0.0477) and digestible ash (P < 0.0001), as well as an increase (P < 0.0001) in hot carcass yield (P = 0.0433). The results indicate that the inclusion of zeolite, in the amounts used, did not alter the performance, serum levels, intestinal development and litter quality of the birds at 42 d of age. However, it was responsible for the improvement in the percentage of digestible nutrients (CP, GE, and MM).

Short-term effects of therapeutic hypothermia following hypoxia-ischemia in neonatal male and female rats.

Nowadays, the only treatment for human babies suffering from hypoxia-ischemia (HI) is therapeutic hypothermia (TH). However, a better understanding of the specific effects of TH in males and females is important to improve its clinical application. The present study evaluated the short-term effects of TH on the brain injury and behavioral outcomes in male and female neonatal rats submitted to neonatal HI. Seven-day-old Wistar rats underwent a surgery for unilateral occlusion of the right common carotid artery and were exposed to a hypoxic atmosphere (8% oxygen) for 75 min. Then, the animals in the TH group were submitted to TH (scalp temperature of 32°C) for 5 h. In the behavioral tests, no remarkable differences triggered by HI or TH were observed relative to SHAM animals. Only females of the HI group presented lower latency to complete the righting reflex test. TH reduced the volume of brain injury in males, but not in females. The animals of the HI group showed a reduction in the number of neurons in the CA1 and dentate gyrus (DG) regions of the hippocampus and TH partially prevented neuronal death. In the CA1 region of the hippocampus, animals from the HI group showed more degenerating cells relative to the SHAM, which was reversed by TH. In the DG, animals from the HI group showed an increase in the number of degenerating neurons, which was partially reversed by TH only in males. Our data show that HI leads to a brain injury, which was attenuated by TH in a sex-dependent way and clarify the importance of the assessment of males and females in order to outline specific strategies for the treatment of each sex in newborns suffering from HI.

AbobotulinumtoxinA is effective in patients with urinary incontinence due to neurogenic detrusor overactivity regardless of spinal cord injury or multiple sclerosis etiology: Pooled analysis of two phase III randomized studies (CONTENT1 and CONTENT2).

BACKGROUND: Neurogenic detrusor overactivity incontinence (NDOI) is often inadequately managed with oral therapy. OBJECTIVE: To assess efficacy and safety of abobotulinumtoxinA (aboBoNT-A; Dysport®; Ipsen Ltd.) according to etiology of NDOI. DESIGN, SETTING, AND PARTICIPANTS: Two phase III, randomized, double-blind studies (CONTENT1 [NCT02660138] conducted in Asia, Europe and North America; CONTENT2 [NCT02660359] conducted in the Americas, Asia, Europe and Oceania) both included patients with spinal cord injury (SCI) or multiple sclerosis (MS), with inadequately managed NDOI, regularly performing clean intermittent catheterization (CIC). INTERVENTION: Patients in CONTENT1 and CONTENT2 received aboBoNT-A injections 600 U (n = 162)/800 U (n = 161), or placebo (n = 162) into the detrusor muscle. OUTCOME MEASUREMENTS AND STATISTICAL ANALYSIS: Primary endpoint: mean change from baseline in number of NDOI episodes/week at Week 6. Secondary endpoints: proportion of patients with no NDOI episodes; incontinence-related quality of life (I-QoL); urodynamic parameters; and time-to-retreatment. Safety was also assessed. Statistical analyses were conducted for pooled populations by etiology (aboBoNT-A doses vs. placebo). RESULTS AND LIMITATIONS: Of 485 randomized patients, 341 (70%) and 144 (30%) had SCI and MS etiologies, respectively. A significant reduction was observed in mean NDOI episodes/week at Week 6 with both aboBoNT-A doses versus placebo in the SCI (all p < 0.001) and MS (all p < 0.01) groups, as well as significant improvements in I-QoL and urodynamic parameters. Median time-to-retreatment was longer in patients with MS (48-62 weeks across doses) than those with SCI (39-44 weeks). Safety data were similar between etiologies. Urinary tract infection was the most frequent adverse event; similar numbers were reported across treatment groups. CONCLUSIONS: AboBoNT-A was well tolerated and significantly improved continence and bladder function, and QoL, in patients with SCI or MS with NDOI performing regular CIC. PATIENT SUMMARY: AboBoNT-A injections improved QoL, symptoms, and bladder function in patients with SCI or MS with bladder muscle overactivity that causes incontinence.

Liquid chromatography-mass spectrometry for simultaneous determination of spironolactone and canrenone in plasma samples

Abstract n our study, we aimed to validate a method based on liquid chromatography-mass spectrometry (LC-MS) to quantify spironolactone (SPI) and its active metabolite canrenone (CAN) simultaneously in plasma samples to support in vivo experiments. Compounds were separated by using a C18 column with the isocratic elution of a mobile phase composed of 0.1% (v/v) formic acid in methanol-water (60:40 v/v) at a flow rate of 0.4 mL min−1. SPI and CAN were detected in na electrospray interface operating in a positive ionization mode and quantified using the selective ion mode monitoring of mass-charge ratios (m/z) of 439.0 for SPI and 363.1 for CAN. After calculating the matrix effect using theoretical equations, we observed the strong interference of plasma in the equipment-generated signal, which required creating analytical curves using the matrix as a solvent. The method was nevertheless linear (r 2 > 0.999) in a concentration range of 0.4-5.0 µg mL−1, as well as precise, with a coefficient of variation less than 5%. SPI's and CAN's recovery rates from the plasma ranged from 87.4% to 112.1%, while their limits of detection (i.e., 0.07 µg mL−1 and 0.03 µg mL−1, respectively) and quantification (i.e., 0.20 µg mL−1 and 0.08 µg mL−1, respectively) in the presence of plasma contaminants were low. Therefore, the bioanalytical method seems to be feasible for quantifying SPI and CAN in plasma

Pyrazolines as potential anti-Alzheimer's agents: DFT, molecular docking, enzyme inhibition and pharmacokinetic studies.

Alzheimer's disease (AD) is a neurodegenerative disease that is characterized as the main dementia in the elderly. Eighteen pyrazolines were synthesized and evaluated for their inhibitory effects against acetylcholinesterase (AChE) in vitro. Possible interactions between pyrazolines and the enzyme were explored by in silico experiments. Compound 2B of the series was the most active pyrazoline with an IC50 value of 58 nM. Molecular docking studies revealed two important π-π interactions with residues Trp 286 and Tyr 341. A correlation between the HOMO-1 surface and AChE inhibition was observed. ADMET assays demonstrated a good profile for compound 2B. From the abovementioned findings, a new avenue of compound 2B analogues could be explored to develop anti-AD agents.

Influence of dietary corn distillers' dried grains with solubles on nutrient digestibility, growth performance, and carcass traits in rabbits.

Two experiments were performed to evaluate the digestibility and growth performance of New Zealand White rabbits fed corn distillers' dried grains with solubles (DDGS). For the digestibility trial, 20 rabbits were housed in metabolic cages. The animals were distributed in two treatments, T1 (control diet without inclusion of DDGS) and T2 (control diet + 300 g kg-1 DDGS). For the growth performance trial, 100 rabbits (50 males and 50 females) were assigned in a factorial design 2 × 5 (2 gender × 5 inclusions of DDGS) and five replicates. The treatments were composed of diets with inclusions of 0, 60, 120, 180, and 240 g kg-1 of DDGS. The rabbits were housed at 35 days old, and the experiment lasted 35 days. The DDGS showed high digestibility for protein (74.10%) and lipids (81.51%) and a high content of digestible energy (2979 kcal kg-1). In the second trial, growth performance and carcass yield and organ relative weights were evaluated. There were no interactions between gender and DDGS inclusions (P > 0.05). A linear decrease was observed for feed intake (FI) for the period from days 35 to 50 (FI, P = 0.001) and FI and FCR from days 35 to 70 (FI, P = 0.004; FCR, P = 0.001) with the increasing levels of DDGS. Rabbits supplemented with 240 g kg-1 had lower (P < 0.05) whole carcass yield (WCY) and carcass without head yield compared with the control rabbits. DDGS is highly digestible in rabbits, and when supplemented up to 240 g kg-1 in diets, it improved FCR but reduced FI and WCY.

Performance, metabolism, and meat quality of broilers fed dry brewery residue.

A study was conducted to assess the chemical composition and apparent metabolizable energy (AME), nitrogen-corrected AME (AMEn), and their respective coefficients of dry brewery residue (DBR) for broilers. A second study was conducted to evaluate the increasing inclusions of DBR in broiler diets and its effects on growth performance, carcass traits, blood metabolites, and meat quality. In the first trial (metabolism assay), a total of 250 male broilers were assigned to 5 diets (0, 100, 200, 300, and 400 g DBR/kg) composed by 5 replications with 10 animals per cage in a completely randomized design in the period from days 11 to 21. There was an increasing linear effect for AME, AMEn, metabolizable coefficient of gross energy (MCGE), and MCGE corrected for N (MCGEn). In the second experiment, a total of 840 one-day-old male broiler chickens were assigned to 6 treatments (0, 20, 40, 60, 80, and 100 g DBR/kg) with 7 replications and 20 animals per pen, from days 0 to 42. Growth performance was determined at days 21 and 42. There was an increasing linear effect for weight gain in the period from days 0 to 21 and for feed intake from days 0 to 42. A linear decreasing effect was observed at day 21 for cholesterol and quadratic effect for uric acid (UA), aspartate aminotransferase, and albumin. At day 42, there was a quadratic effect for UA and creatinine. At day 21, an increasing linear effect was observed for protein deposition rate. In conclusion, the inclusion of DBR up to 100 g/kg improved WG in the starter phase and did not negatively affect the productive parameters of broilers from days 0 to 42.

Pouteria macrophylla Fruit Extract Microemulsion for Cutaneous Depigmentation: Evaluation Using a 3D Pigmented Skin Model.

Here, we verify the depigmenting action of Pouteria macrophylla fruit extract (EXT), incorporate it into a safe topical microemulsion and assess its effectiveness in a 3D pigmented skin model. Melanocytes-B16F10- were used to assess the EXT effects on cell viability, melanin synthesis, and melanin synthesis-related gene transcription factor expression, which demonstrated a 32% and 50% reduction of intra and extracellular melanin content, respectively. The developed microemulsion was composed of Cremophor EL®/Span 80 4:1 (w/w), ethyl oleate, and pH 4.5 HEPES buffer and had an average droplet size of 40 nm (PdI 0.40 ± 0.07). Skin irritation test with reconstituted epidermis (Skin Ethic RHETM) showed that the formulation is non-irritating. Tyrosinase inhibition was maintained after skin permeation in vitro, in which microemulsion showed twice the inhibition of the conventional emulsion (20.7 ± 2.2% and 10.7 ± 2.4%, respectively). The depigmenting effect of the microemulsion was finally confirmed in a 3D culture model of pigmented skin, in which histological analysis showed a more pronounced effect than a commercial depigmenting formulation. In conclusion, the developed microemulsion is a promising safe formulation for the administration of cutite fruit extract, which showed remarkable depigmenting potential compared to a commercial formulation.

Therapeutic hypothermia for the treatment of neonatal hypoxia-ischemia: sex-dependent modulation of reactive astrogliosis.

Therapeutic hypothermia (TH) is the standard treatment for neonatal hypoxia-ischemia (HI) with a time window limited up to 6 h post injury. However, influence of sexual dimorphism in the therapeutic window for TH has not yet been elucidated in animal models of HI. Therefore, the aim of this study was to investigate the most effective time window to start TH in male and female rats submitted to neonatal HI. Wistar rats (P7) were divided into the following groups: NAÏVE and SHAM (control groups), HI (submitted to HI) and TH (submitted to HI and TH; 32ºC for 5 h). TH was started at 2 h (TH-2 h group), 4 h (TH-4 h group), or 6 h (TH-6 h group) after HI. At P14, animals were subjected to behavioural tests, volume of lesion and reactive astrogliosis assessments. Male and female rats from the TH-2 h group showed reduction in the latency of behavioral tests, and decrease in volume of lesion and intensity of GFAP immunofluorescence. TH-2 h females also showed reduction of degenerative cells and morphological changes in astrocytes. Interestingly, females from the TH-6 h group showed an increase in volume of lesion and in number of degenerative hippocampal cells, associated with worse behavioral performance. Together, these results indicate that TH neuroprotection is time- and sex-dependent. Moreover, TH started later (6 h) can worsen volume of brain lesion in females. These data indicate the need to develop specific therapeutic protocols for each sex and reinforce the importance of early onset of the hypothermic treatment.

Sulfated ß-glucan from Agaricus subrufescens inhibits flavivirus infection and nonstructural protein 1-mediated pathogenesis.

Despite substantial morbidity and mortality, no therapeutic agents exist for treatment of dengue or Zika, and the currently available dengue vaccine is only recommended for dengue virus (DENV)-immune individuals. Thus, development of therapeutic and/or preventive drugs is urgently needed. DENV and Zika virus (ZIKV) nonstructural protein 1 (NS1) can directly trigger endothelial barrier dysfunction and induce inflammatory responses, contributing to vascular leak in vivo. Here we evaluated the efficacy of the (1-6,1-3)-ß-D-glucan isolated from Agaricus subrufescens fruiting bodies (FR) and its sulfated derivative (FR-S) against DENV-2 and ZIKV infection and NS1-mediated pathogenesis. FR-S, but not FR, significantly inhibited DENV-2 and ZIKV replication in human monocytic cells (EC50 = 36.5 and 188.7 µg/mL, respectively) when added simultaneously with viral infection. No inhibitory effect was observed when FR or FR-S were added post-infection, suggesting inhibition of viral entry as a mechanism of action. In an in vitro model of endothelial permeability using human pulmonary microvascular endothelial cells (HPMECs), FR and FR-S (0.12 µg/mL) inhibited DENV-2 NS1- and ZIKV NS1-induced hyperpermeability by 50% and 100%, respectively, as measured by Trans-Endothelial Electrical Resistance. Treatment with 0.25 µg/mL of FR and FR-S inhibited DENV-2 NS1 binding to HPMECs. Further, FR-S significantly reduced intradermal hyperpermeability induced by DENV-2 NS1 in C57BL/6 mice and protected against DENV-induced morbidity and mortality in a murine model of dengue vascular leak syndrome. Thus, we demonstrate efficacy of FR-S against DENV and ZIKV infection and NS1-induced endothelial permeability in vitro and in vivo. These findings encourage further exploration of FR-S and other glycan candidates for flavivirus treatment alone or in combination with compounds with different mechanisms of action.

Novel trypanocidal thiophen-chalcone cruzain inhibitors: structure- and ligand-based studies.

Background: Chagas disease is a neglected tropical disease that affects millions of people worldwide and for which no effective treatment is available. Materials & methods: 17 chalcones were synthesized, for which the inhibition of cruzain and trypanocidal activity were investigated. Results: Chalcone C8 showed the highest cruzain inhibitory (IC50 = 0.536 µm) and trypanocidal activity (IC50 = 0.990 µm). Molecular docking studies showed interactions involving Asp161 and the thiophen group interacting with the S2 subsite. Furthermore, quantitative structure-activity relationship (q2 = 0.786; r2 = 0.953) and density functional theory studies were carried out, and a correlation between the lowest unoccupied molecular orbital surface and trypanocidal activity was observed. Conclusion: These results demonstrate that these chalcones are worthwhile hits to be further optimized in Chagas disease drug discovery programs.

Validation of a simple chromatographic method for naringenin quantification in skin permeation experiments.

Naringenin is a flavonoid that can be found in citrus fruits (e.g., Citrus sp). This natural compound is known for its antioxidant activity, antitumor, and neuroprotective potential, also acting directly in controlling the inflammatory response. Topical and transdermal routes are attractive alternatives that could circumvent the low oral bioavailability. However, a simple analytical method capable of determining naringenin in skin layers is still demanded. Thus, this work aimed to validate a selective and straightforward chromatographic method for naringenin determination in skin permeation studies. The developed method uses a reversed-phase C18 column as stationary phase and a mobile phase composed of methanol/phosphoric acid 0.01 M (65:35, v/v), eluted at a flow rate of 0.6 mL/min with detection at 290 nm. The method was linear (r2 > 0.99) in a broad concentration range of 0.5-10.0 µg/mL, precise with an overall variation coefficient lower than 2%, and accurate with naringenin recovery from the skin layers higher than 85%. Additionally, the method was sensitive (LD = 0.10 µg/mL, LQ = 0.20 µg/mL), selective against skin matrices as well as naringenin degradation products, and robust regarding methodology parameters. Therefore, the method was suitable to be used in skin permeation studies employing naringenin.