STING orchestrates the neuronal inflammatory stress response in multiple sclerosis.

Inflammation-induced neurodegeneration is a defining feature of multiple sclerosis (MS), yet the underlying mechanisms remain unclear. By dissecting the neuronal inflammatory stress response, we discovered that neurons in MS and its mouse model induce the stimulator of interferon genes (STING). However, activation of neuronal STING requires its detachment from the stromal interaction molecule 1 (STIM1), a process triggered by glutamate excitotoxicity. This detachment initiates non-canonical STING signaling, which leads to autophagic degradation of glutathione peroxidase 4 (GPX4), essential for neuronal redox homeostasis and thereby inducing ferroptosis. Both genetic and pharmacological interventions that target STING in neurons protect against inflammation-induced neurodegeneration. Our findings position STING as a central regulator of the detrimental neuronal inflammatory stress response, integrating inflammation with glutamate signaling to cause neuronal cell death, and present it as a tractable target for treating neurodegeneration in MS.

Detoxified castor seed meal replaces soybean meal in the supplement for Holstein-Zebu crossbred steers finished on tropical pasture during the rainy season.

The objective of this study was to examine the effect of replacing soybean meal (SBM) with detoxified castor seed meal (DCM) on the intake, digestibility, feeding behavior, and performance of pasture-finished (rainy season) steers supplemented with concentrate at 0.4% of their body weight. Forty ½ Holstein + ½ Zebu steers (initial weight: 283.3 ± 36.3 kg) were allocated to four treatments in a completely randomized experimental design. Treatments consisted of diets in which DCM replaced 0, 30, 60, and 90% of SBM in the supplement dry matter (DM). The steers were finished on an Urochloa brizantha pasture and the experiment lasted 112 days. Replacing SBM with DCM did not influence (P > 0.05) the intake or apparent digestibility of DM, crude protein, or neutral detergent insoluble fiber of the animals. Grazing time increased (P < 005), whereas the intake and rumination efficiencies of the steers did not change (P > 0.05) with the substitution. The replacement of SBM with DCM in the supplement fed to the steers also did not influence (P > 0.05) their final weight, average daily gain, or feed conversion (P > 0.05). We recommend replacing up to 90% (DM basis) of SBM with DCM in the concentrate supplement of steers grazing on Urochloa brizantha pasture during rainy season while supplemented with concentrate at 0.4% of their body weight.

Obtaining Aromatic Extracts from Portuguese Thymus mastichina L. by Hydrodistillation and Supercritical Fluid Extraction with CO2 as Potential Flavouring Additives for Food Applications.

Humans often respond to sensory impulses provided by aromas, and current trends have generated interest in natural sources of fragrances rather than the commonly used synthetic additives. For the first time, the resulting aroma of a selected culture of Thymus mastichina L. was studied as a potential food ingredient. In this context, dried (DR) and fresh (FR) samples were submitted to carbon dioxide (CO2) supercritical extraction (SFE) and hydrodistillation (HD) methods. The extracts were characterised according to their volatile composition by GC-MS, cytotoxicity against a non-tumour cell culture, and sensory attributes (odour threshold and olfactive descriptors). The most abundant aromas were quantified, and the analysis performed by GC-MS revealed an abundance of terpenoids such as thymol chemotype, followed by the precursors α-terpinene and p-cymene. DR and FR extracts (EX) obtained from SFE-CO2 show the highest content of thymol, achieving 52.7% and 72.5% of the isolated volatile fraction. The DR essential oil (EO) contained the highest amount of terpenoids, but it was also the most cytotoxic extract. In contrast, SFE-CO2 products showed the lowest cytotoxic potential. Regarding FR-OE, it had the lowest extraction yield and composition in aroma volatiles. Additionally, all samples were described as having green, fresh and floral sensory notes, with no significant statistical differences regarding the odour detection threshold (ODT) values. Finally, FR-EX of T. mastichina obtained by SFE-CO2 presented the most promising results regarding food application.

Inclusion of palm kernel cake in the supplement reduces nutrient digestibility but does not interfere with the performance of steers finished on tropical pasture.

The aim of this study was to evaluate the effect of increasing inclusion levels of palm kernel cake (PKC) in the supplement of steers finished on a Urochloa brizantha cv. Marandu pasture on intake, digestibility, feeding behavior, weight gain, and carcass traits. Forty 1/2 Holstein × 1/2 Zebu crossbred steers with an average body weight of 352.80 ± 3.32 kg, at 36 months of age, were supplemented with concentrate at 0.4% of their body weight. Four treatments were formulated (supplement containing 0, 15, 30, or 45% PKC, dry matter [DM] basis) and the animals were monitored for 240 days, weighed, and slaughtered. The inclusion of PKC in the supplement did not influence the intakes of total DM, crude protein, or neutral detergent fiber, but increased (P < 0.05) ether extract intake and reduced (P < 0.05) non-fibrous carbohydrate intake. The apparent digestibility of DM, crude protein, and neutral detergent fiber decreased (P < 0.05), whereas intake and rumination efficiencies by the dietary treatments. Average daily gain, carcass weight, and backfat thickness did not change in response to the inclusion of PKC in the supplement. We recommend the inclusion of 45% palm kernel cake in the concentrate of steers supplemented at 0.4% of their body weight and finished on Urochloa brizantha cv. Marandu pasture.

Paracoccin Overexpression in Paracoccidioides brasiliensis Enhances Fungal Virulence by Remodeling Chitin Properties of the Cell Wall.

BACKGROUND: The thermodimorphic fungi Paracoccidioides spp. are the etiological agents of paracoccidioidomycosis. Although poorly studied, paracoccin (PCN) from Paracoccidioides brasiliensis has been shown to harbor lectinic, enzymatic, and immunomodulatory properties that affect disease development. METHODS: Mutants of P. brasiliensis overexpressing PCN (ov-PCN) were constructed by Agrobacterium tumefaciens-mediated transformation. ov-PCN strains were analyzed and inoculated intranasally or intravenously to mice. Fungal burden, lung pathology, and survival were monitored to evaluate virulence. Electron microscopy was used to evaluate the size of chito-oligomer particles released by ov-PCN or wild-type strains to growth media. RESULTS: ov-PCN strains revealed no differences in cell growth and viability, although PCN overexpression favored cell separation, chitin processing that results in the release of smaller chito-oligomer particles, and enhanced virulence. Our data show that PCN triggers a critical effect in the cell wall biogenesis through the chitinase activity resulting from overexpression of PCN. As such, PCN overexpression aggravates the disease caused by P. brasiliensis. CONCLUSIONS: Our data are consistent with a model in which PCN modulates the cell wall architecture via its chitinase activity. These findings highlight the potential for exploiting PCN function in future therapeutic approaches.

p24 Family Proteins Are Involved in Transport to the Plasma Membrane of GPI-Anchored Proteins in Plants.

p24 proteins are a family of type-I membrane proteins that cycle between the endoplasmic reticulum (ER) and the Golgi apparatus via Coat Protein I (COPI)- and COPII-coated vesicles. These proteins have been proposed to function as cargo receptors, but the identity of putative cargos in plants is still elusive. We previously generated an Arabidopsis (Arabidopsis thaliana) quadruple loss-of-function mutant affecting p24 genes from the δ-1 subclass of the p24 delta subfamily (p24δ3δ4δ5δ6 mutant). This mutant also had reduced protein levels of other p24 family proteins and was found to be sensitive to salt stress. Here, we used this mutant to test the possible involvement of p24 proteins in the transport to the plasma membrane of glycosylphosphatidylinositol (GPI)-anchored proteins. We found that GPI-anchored proteins mostly localized to the ER in p24δ3δ4δ5δ6 mutant cells, in contrast to plasma membrane proteins with other types of membrane attachment. The plasma membrane localization of GPI-anchored proteins was restored in the p24δ3δ4δ5δ6 mutant upon transient expression of a single member of the p24 δ-1 subclass, RFP-p24δ5, which was dependent on the coiled-coil domain in p24δ5. The coiled-coil domain was also important for a direct interaction between p24δ5 and the GPI-anchored protein arabinogalactan protein4 (AGP4). These results suggest that Arabidopsis p24 proteins are involved in ER export and transport to the plasma membrane of GPI-anchored proteins.

Insights on the Extraction Performance of Alkanediols and Glycerol: Using Juglans regia L. Leaves as a Source of Bioactive Compounds.

Glycerol and alkanediols are being studied as alternative solvents to extract phytochemicals from plant material, often as hydrogen bond donors in deep eutectic solvents (DESs). Many of those alcohols are liquid at room temperature, yet studies of their use as extraction solvents are scarce. In this work, glycerol and a series of alkanediols (1,2-ethanediol, 1,2-propanediol, 1,3-propanediol, 1,3-butanediol, 1,2-pentanediol, 1,5-pentanediol, and 1,2-hexanediol) were studied for the extraction of phenolic compounds from Juglans regia L. leaves, a rich source of this class of bioactive compounds. The extraction yield was quantified, and the bioactivity of both extracts and pure solvents was evaluated by measuring the anti-inflammatory and cytotoxic activities. The solvents showing the best combined results were 1,2 and 1,3-propanediol, as their extracts presented a high amount of phenolic compounds, close to the results of ethanol, and similar cytotoxicity against cervical carcinoma cells, with no impact on non-tumor porcine liver cells in the studied concentration range. On the other hand, none of the extracts (and solvents) presented anti-inflammatory activity. Overall, the results obtained in this work contribute to the study of alternative solvents that could potentially be used also as formulation media, highlighting the importance of walnut leaves as a source of bioactive compounds.

An integrated pipeline for sample preparation and characterization at the EMBL@PETRA3 synchrotron facilities.

The characterization of macromolecular samples at synchrotrons has traditionally been restricted to direct exposure to X-rays, but beamline automation and diversification of the user community has led to the establishment of complementary characterization facilities off-line. The Sample Preparation and Characterization (SPC) facility at the EMBL@PETRA3 synchrotron provides synchrotron users access to a range of biophysical techniques for preliminary or parallel sample characterization, to optimize sample usage at the beamlines. Here we describe a sample pipeline from bench to beamline, to assist successful structural characterization using small angle X-ray scattering (SAXS) or macromolecular X-ray crystallography (MX). The SPC has developed a range of quality control protocols to assess incoming samples and to suggest optimization protocols. A high-throughput crystallization platform has been adapted to reach a broader user community, to include chemists and biologists that are not experts in structural biology. The SPC in combination with the beamline and computational facilities at EMBL Hamburg provide a full package of integrated facilities for structural biology and can serve as model for implementation of such resources for other infrastructures.

Nutritional and Biochemical Profiling of Leucopaxillus candidus (Bres.) Singer Wild Mushroom.

The wild mushroom Leucopaxillus candidus (Bres.) Singer was studied for the first time to obtain information about its chemical composition, nutritional value and bioactivity. Free sugars, fatty acids, tocopherols, organic and phenolic acids were analysed by chromatographic techniques coupled to different detectors. L. candidus methanolic extract was tested regarding antioxidant potential (reducing power, radical scavenging activity and lipid peroxidation inhibition). L. candidus was shown to be an interesting species in terms of nutritional value, with high content in proteins and carbohydrates, but low fat levels, with the prevalence of polyunsaturated fatty acids. Mannitol was the most abundant free sugar and ß-tocopherol was the main tocopherol isoform. Other compounds detected were oxalic and fumaric acids, p-hydroxybenzoic and cinnamic acids. The methanolic extract revealed antioxidant activity and did not show hepatoxicity in porcine liver primary cells. The present study provides new information about L. candidus.

Wild Morchella conica Pers. from different origins: a comparative study of nutritional and bioactive properties.

BACKGROUND: Morchella conica Pers. is a species of fungus that belongs to the Morchellaceae family and was studied in order to obtain more information about this species, by comparing Portuguese and Serbian wild samples. Free sugars, fatty acids, tocopherols, organic and phenolic acids were analysed by chromatographic techniques. M. conica methanolic extracts were tested regarding antioxidant and antimicrobial properties. The absence of hepatotoxicity was confirmed in porcine liver primary cells. RESULTS: The nutrition value varied between the two samples and the Serbian one gave the highest energy contribution and content in sugars, polyunsaturated fatty acids and phenolic compounds, while the Portuguese sample was richer in saturated and monounsaturated fatty acids, tocopherols and organic acids. In general, the Portuguese sample revealed the highest antioxidant potential (except for lipid peroxidation inhibition), while the Serbian one displayed higher antimicrobial activity. None of the samples revealed toxicity towards liver cells. CONCLUSION: The present study provides new data concerning chemical characterization and bioactivity of Morchella conica Pers.

Expanding current knowledge on the chemical composition and antioxidant activity of the genus Lactarius.

Despite the presence of toxic compounds in inedible mushrooms, the question whether the chemical nutrients and non-nutrients compositions in edible and inedible Lactarius species are similar remains unanswered. To answer this question, Lactarius citriolens Pouzar and Lactarius turpis (Weinm.) Fr., two inedible species, were studied in order to obtain information about their chemical composition and bioactivity. Free sugars, fatty acids, tocopherols, organic and phenolic acids were analysed by chromatographic techniques coupled to different detectors. L. citriolens and L. turpis methanolic extracts were tested regarding antioxidant potential (reducing power, radical scavenging activity and lipid peroxidation inhibition). The composition of macronutrients varied among the two species, but the profiles were similar between them and among other Lactarius species; L. citriolens gave the highest energy contribution, saturated fatty acids and organic acids, while the L. turpis sample was richer in free sugars, mono- and polyunsaturated fatty acids, tocopherols and phenolic compounds. L. turpis methanolic extract showed the highest antioxidant activity. The absence of hepatoxicity of the methanolic extracts was confirmed in porcine liver primary cells (in vitro conditions). The present study provided new information about wild L. citriolens and L. turpis, comparing their chemical composition and antioxidant properties with other Lactarius species, and expanding the knowledge about this genus.

Comparative anatomy of the pelvic vessels in the bearded capuchin (Sapajus libidinosus) and baboons, apes and modern humans.

Cebus/Sapajus has shown high cognitive and manipulatory behaviour as well as intermittent bipedalism. Although the function of the muscles and bones of this genus has been widely investigated, the arterial system that supports these tissues has not been studied in much detail, and a full description of the blood vessels of the pelvis is still missing. Therefore, we studied the vessels of the pelvis of Sapajus libidinosus in terms of their origin, distribution and muscle irrigation and compared them with those of other primates available in the literature. In general, the distribution pattern and origin of arteries in the pelvis of the bearded capuchin are more similar to those of baboons compared to other primates. This similarity may be because both have a tail, a similar body shape and use, preferentially, quadrupedal movement.

Prevalence and risk factors associated with goat gastrointestinal helminthiasis in the Sertão region of Paraíba State, Brazil.

Gastrointestinal helminthiasis represents an obstacle to goat raising, causing severe damage to herds such as growth retardation, weight loss, and even death. In this study, we aimed to determine the prevalence and risk factors associated to goat gastrointestinal helminthiasis in the Sertão region of Paraíba State, Brazil. A total of 256 goats from 54 farms were systematically sampled. Blood and fecal samples were collected from each animal for egg per gram (EPG), larval culture, and packed cell volume (PCV) analyses. We found that 79.3% of the goats investigated were parasitized with gastrointestinal helminths. Significant correlation (p = 0.004) was observed between the EPG and PCV of the animals studied, and it was observed that the EPG increases as the PCV decreases. In the larval culture, the most prevalent helminth was Haemonchus sp. (83.2%). Age and sex were significant variables (p ≤ 0.20) for the development of gastrointestinal helminths: 86.8% of animals over 36 months of age and 81.7% of females were infected. The variable type of animal exploitation was also significant, with 90.3% (p ≤ 0.20) of the animals presenting double suitability (milk and meat). The Sertão region of Paraíba State presents high prevalence of gastrointestinal helminthiasis in goats, and age and type of animal exploitation are the most relevant risk factors to the development of these parasites.

Dimethyl 2-nitro-biphenyl-4,4'-di-carboxyl-ate.

The title compound, C16H13NO6, exhibits a biphenyl unit with a dihedral angle between the two aryl rings of 56.01 (5)°. The two ester groups vary slightly from planarity, with ar-yl-ester dihedral angles of 4.57 (5) and 16.73 (5)°. The nitro group is turned from the aromatic unit with an ar-yl-nitro dihedral angle of 48.66 (4)°. In the crystal, mol-ecules are connected by weak C-H⋯O inter-actions, forming a three-dimensional network.

Anthelmintic efficacy of pumpkin seed (Cucurbita pepo Linnaeus, 1753) on ostrich gastrointestinal nematodes in a semiarid region of Paraíba State, Brazil.

The aim of this study was to verify the in vivo effectiveness of pumpkin seed (Curcubita pepo Linnaeus, 1753) in naturally infected ostriches in the Cariri zone, semiarid region of Paraíba State, Brazil. Forty-eight ostriches were used, African Black breed, of 14 to 36 months old, naturally infected by gastrointestinal nematodes. These animals were divided into four groups of 12 ostriches. Group 1 consists of animals treated with 0.5 g/kg live weight (l. w.) of pumpkin seed meal; group 2 received 1 g/kg l. w. of pumpkin seed meal; group 3 was treated with Albendazole 5 %, at the dosage of 1 mL/10 kg l. w.; and Group 4 was the control group and do not received treatment. Groups 1 and 2 received the treatment for three consecutive days, orally, at intervals of 7 days, totaling nine administrations. The Albendazole 5 % was administered one time, at the beginning of the experiment, according to the manufacturer's recommendations. The groups treated with pumpkin seed showed a significant decrease in egg counts per gram of feces (EPG), wherein group 2 (1 g/kg l. w.) was the most effective. The control and drug groups showed no reduction in EPG. The results of the present study demonstrate that the administration of pumpkin seed was effective in controlling gastrointestinal helminths in naturally infected ostriches.

The journey of cardosin A in young Arabidopsis seedlings leads to evidence of a Golgi-independent pathway to the protein storage vacuole.

The aspartic proteinase cardosin A is a vacuolar enzyme found to accumulate in protein storage and lytic vacuoles in the flowers and protein bodies in the seeds of the native plant cardoon. Cardosin A was first isolated several decades ago and has since been extensively characterized, both in terms of tissue distribution and enzyme biochemistry. In the native system, several roles have been attributed to cardosin A, such as reproduction, reserve mobilization, and membrane remodeling. To participate in such diverse events, cardosin A must accumulate and travel to different compartments within the cell: protein storage vacuoles, lytic vacuoles, and the cytoplasmic membrane (and eventually outside the cell). Several studies have approached the expression of cardosin A in Arabidopsis thaliana and Nicotiana tabacum with promising results for the use of these systems to study of cardosin A trafficking. A poly-sorting mechanism has been uncovered for this protein, as two different vacuolar sorting determinants, mediating different vacuolar routes, have been described. The first is a conventional C-terminal domain, which delivers the protein to the vacuole via the Golgi, and the second is a more unconventional signal-the plant-specific insert (PSI)-that mediates a Golgi-independent route. The hypothesis that these two signals are activated according to cell needs and in organs with high metabolic activity is investigated here. An Arabidopsis line expressing cardosin A under an inducible promoter was used to understand the dynamics of cardosin A regarding vacuolar accumulation during seed germination events. Using antibodies against different regions of the protein and combining them with immunofluorescence and immunocytochemistry assays in different young seedling tissues, cardosin A was detected along the secretory pathway to the protein storage vacuole, often associated with the endoplasmic reticulum. More interestingly, upon treatment with the drug Brefeldin A, cardosin A was still detected in protein storage vacuoles, indicating that the intact protein can bypass the Golgi in this system, contrary to what was observed in N. tabacum. This study is a good starting point for further research involving the use of fluorescent fusions and exploring in more detail the relationship between cardosin A trafficking and plant development.

Sand fly population dynamics in areas of American cutaneous leishmaniasis, Municipality of Paraty, Rio de Janeiro, Brazil.

The present study is the second and last part of the study that investigated the fauna and behavior of sand flies in areas prone to cutaneous leishmaniasis outbreaks, in the State of Rio de Janeiro, in the municipality of Paraty. To collect the sand flies, CDC and Shannon light traps were used, installed in the peridomiciliary and forest areas, and manual suction tubes on the walls of the home and in the animal shelters. A total of 102,937 sand flies, belonging to nine genera and 23 species were captured from October 2009 to September 2012. Regarding the monthly frequency of sand flies, the period of highest density was from November to March, with a maximum peak in January. The lowest density was observed in June and July. In the studied area, the species of epidemiological importance, Nyssomyia intermedia, Pintomyia fischeri, Migonemyia migonei and Nyssomyia whitmani, were found in all months of the year, a period in which residents may be in contact with these vectors of the etiological agent of cutaneous leishmaniasis.

The potential of almonds, hazelnuts, and walnuts SFE-CO2 extracts as sources of bread flavouring ingredients.

Nuts have been part of the human diet since our early ancestors, and their use goes beyond nutritional purposes, for example, as aromatic sources for dairy products. This work explores the potential of almond (Prunus dulcis (Mill.) DA Webb), hazelnut (Corylus avellana L.), and walnut (Juglans regia L.) extracts as sources of food flavouring agents, suggesting a new added-value application for lower quality or excess production fruits. The extracts were obtained by supercritical fluid extraction with carbon dioxide and characterized by: quantification of the volatile fraction by HS-SPME GC-MS; sensory perception and description; and cytotoxicity against Vero cells. All extracts revealed potential as flavouring ingredients due to terpene abundance. No significant differences were observed for the minimal sensory perception, in which the odour threshold values ranged from 8.3 × 10-4 to 6.9 × 10-3 µg·mL-1 for walnuts and almonds extracts, respectively. In contrast, the cytotoxic potential differed significantly among the extracts, and P. dulcis extract presented lower cytotoxicity. Notes as woody, fresh, and green were identified in the volatile intensifiers obtained from the P. dulcis extract. Thus, almond extract was identified as the most promising ingredient to increase the sensory value of food products, namely bread. This potential was verified by an increase in the odour perception of bread after adding 4 µL of extract to each 100 g of bread dough. The quantified eucalyptol and d-limonene terpenes - found in the P. dulcis extract - have improved the release of the pleasant and natural volatile compounds from bread crust and crumb compared to the control bread chemical and sensory profiles.

Calcium channel ß3 subunit regulates ATP-dependent migration of dendritic cells.

Migratory dendritic cells (migDCs) continuously patrol tissues and are activated by injury and inflammation. Extracellular adenosine triphosphate (ATP) is released by damaged cells or actively secreted during inflammation and increases migDC motility. However, the underlying molecular mechanisms by which ATP accelerates migDC migration is not understood. Here, we show that migDCs can be distinguished from other DC subsets and immune cells by their expression of the voltage-gated calcium channel subunit ß3 (Cavß3; CACNB3), which exclusively facilitates ATP-dependent migration in vitro and during tissue damage in vivo. By contrast, CACNB3 does not regulate lipopolysaccharide-dependent migration. Mechanistically, CACNB3 regulates ATP-dependent inositol 1,4,5-trisphophate receptor-controlled calcium release from the endoplasmic reticulum. This, in turn, is required for ATP-mediated suppression of adhesion molecules, their detachment, and initiation of migDC migration. Thus, Cacnb3-deficient migDCs have an impaired migration after ATP exposure. In summary, we identified CACNB3 as a master regulator of ATP-dependent migDC migration that controls tissue-specific immunological responses during injury and inflammation.

MicroRNA-92a-CPEB3 axis protects neurons against inflammatory neurodegeneration.

Neuroinflammation causes neuronal injury in multiple sclerosis (MS) and other neurological diseases. MicroRNAs (miRNAs) are important modulators of neuronal stress responses, but knowledge about their contribution to neuronal protection or damage during inflammation is limited. Here, we constructed a regulatory miRNA-mRNA network of inflamed motor neurons by leveraging cell type-specific miRNA and mRNA sequencing of mice undergoing experimental autoimmune encephalomyelitis (EAE). We found robust induction of miR-92a in inflamed spinal cord neurons and identified cytoplasmic polyadenylation element-binding protein 3 (Cpeb3) as a key target of miR-92a-mediated posttranscriptional silencing. We detected CPEB3 repression in inflamed neurons in murine EAE and human MS. Moreover, both miR-92a delivery and Cpeb3 deletion protected neuronal cultures against excitotoxicity. Supporting a detrimental effect of Cpeb3 in vivo, neuron-specific deletion in conditional Cpeb3 knockout animals led to reduced inflammation-induced clinical disability in EAE. Together, we identified a neuroprotective miR-92a-Cpeb3 axis in neuroinflammation that might serve as potential treatment target to limit inflammation-induced neuronal damage.