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The use of hydrocolloids has greatly increased in recent years due to their pivotal role as functional ingredients. They can increase food consistency, control the microstructure that affects water absorption, alter texture and flavor, and improve the shelf life of baked products. In this work, we analyzed the effect of plant hemicelluloses such as Caesalpinia pulcherrima galactomannans and Tamarindus indica xyloglucans on the stability of frozen French bread dough. The amount of these additives used was optimized from an obtained central composite design- (CCD-) response surface methodology (RSM) of the alveograph parameters. Batches were characterized for moisture, water activity (aw), and texture using SEM, DSC, and TGA analyses when frozen for up to 60 days. Batches containing hemicelluloses showed better stability for aw over time. There was no difference between the texture parameters of the samples studied for 60 days. Both added hemicelluloses presented fewer fractures at 30 days and less wear at 60 days, in addition to better performance in the TGA analysis after less than 30 days of frozen storage. Batches containing only xyloglucans or galactomannans had a higher solidification peak temperature after 60 days. Both plant hemicelluloses reduced the damage caused by cold storage and improved stability for water activity. Also, dough preparations containing these additives showed better moisture retention, as well as less wear and tear and fewer fractures over their shelf life. Our evaluation suggests C. pulcherrima and T. indica are nonconventional sources of hydrocolloids that could be utilized in the bakery industry.

Encapsulating matrices are important to maintain the properties, promote the low and controlled release, and avoid these bioactive compounds' undesirable reactions. This study's objective was to evaluate cashew gum (CG) use combined with galactomannan (GAL) in the rosemary essential oil encapsulation by atomization. During the emulsification, the polysaccharides were crosslinked with sodium trimetaphosphate. The particles obtained after encapsulation were evaluated for moisture, solubility, particle size, encapsulation efficiency, morphology, antimicrobial activity, and chemical composition of the essential oil volatiles. GAL/CG blend showed higher encapsulation efficiency and lower oil release than the cashew gum matrix. Undoubtedly, galactomannan combined with cashew gum improved the microparticles' characteristics due to the galactomannan's high emulsifying property even in low concentration. Essential oil maintained its antimicrobial and chemical properties practically after the encapsulation procedure.

Background: Plant lectins have shown promising biological activities in the central nervous system (CNS). Objective: This study evaluated the effect of DAL, a lectin isolated from the seeds of the Dioclea altissima species, having binding affinity to D-glucose or D-mannose residues, on mice behavior. Methods: Mice (n=6/group) were treated (i.p.) with DAL (0.25, 0.5 or 1 mg/kg) or vehicle and subjected to several tests (open field/OFT, marble-burying/MBT, hole-board/HBT, elevated plus maze/PMT, tail suspension/ TST, forced swimming/FST or rotarod/RRT). Pizotifen, cyproheptadine, flumazenil, L-NAME, 7-NI, Larginine or yohimbine were administered 15 min before DAL (0.5 mg/kg) and the animals were evaluated on PMT. It was also verified whether the DAL effect depended on its structural integrity and ability to interact with carbohydrates. Results: The results showed there were no neurobehavioral changes in the mice at the RRT, FST and locomotion in the OFT. DAL (0.25, 0.5 or 1 mg/kg) increased the behavior of grooming and rearing in the OFT, head dips in the HBT, pedalling in the TST and decreased the number of marbles hidden in the MBT. In the PMT, DAL (0.25, 0.5 and 1 mg/kg) and Diazepam increased the frequency of entries in the open arms and the time of permanence in the open arms without affecting the locomotor activity. The effect of DAL was dependent on carbohydrate interaction and protein structure integrity and it prevented by pizotifen, cyproheptadine, flumazenil, L-NAME and 7-NI, but not by L-arginine or yohimbine. Conclusion: DAL was found to have an anxiolytic-like effect mediated by the 5-HT and GABAergic receptors and NO pathway.

Galactomannans are polysaccharides commercially used in foods as gelling agents, thickeners, and stabilizers. In this study, a galactomannan partially hydrolyzed from Adenanthera pavonina seeds was used as stabilizer, fat substitute, and source of dietary fibers in a mousse milk dessert. First, characterizations of intact (GM) and partially hydrolyzed (GMPH) galactomannan from A. pavonina were made. Through Rotational Central Compound Design (RCCD) and the response surface methodology, we evaluated creaming index, lipid content, moisture content, and texture of the mousses. The formulation F7 (1.75% of GMPH and 4.92% of fat) was chosen among the developed formulations, and it was characterized by low moisture content (44.13%), low lipid content (6.67%), low firmness (2.10 N), and high percentage of dietary fiber (4.21%). This formulation was compared with a traditional formulation of lemon mousse (FT) and with another formulation with the same fat percentage as the chosen formulation without GMPH (F7L). The three formulation were characterized physicochemically and microbiologically, and sensorial acceptance tests were performed. The results indicated that no structural modifications were found in the galactomannan after its partial hydrolysis. and good sensory acceptance when compared to FT. The lemon mousses produced met sanitary specifications, remaining stable for 21 days of storage. We concluded that the use of GMPH from Adenanthera pavonina seeds in lemon mousses proved to be a stable, light dessert alternative and a potential source of dietary fiber.

O consumo mundial crescente de fertilizantes e a consequente preocupação com as questões ambientais estimulam o desenvolvimento de tecnologias que, simultaneamente, promovam o aumento da produtividade agrícola e minimizem os danos ao ambiente. Uma alternativa que atende a esses requisitos seria o uso combinado de estimulantes vegetais de origem natural com os fertilizantes habitualmente utilizados e, nesse contexto, os oligossacarídeos despontam como potenciais bioestimulantes vegetais. Com este trabalho, objetivou-se realizar uma prospecção tecnológica através da busca por patentes relacionadas a preparações agrícolas que contêm oligossacarídeos na sua composição. A pesquisa foi feita nas bases de dados do Escritório Europeu de Patentes (EPO), da plataforma de busca de patentes da América Latina e Espanha (Latipat) e do Instituto Nacional de Propriedade Industrial (INPI). Os resultados mostram que houve um grande aumento no número de depósitos de patentes relacionadas a oligossacarídeos na agricultura nos últimos cinco anos, assim como uma forte tendência na utilização de oligossacarídeos de quitosana. A China é o principal detentor de patentes na área e o Brasil, embora não seja gerador desta tecnologia, encontra-se numa situação favorável para investir nesse ramo.

Nanoencapsulation is a valid alternative for the oral administration of peptide drugs and proteins, as nanoparticles protect them from proteolytic degradation in the gastrointestinal tract and promote the absorption of these macromolecules. The orofacial antinociceptive effect of frutalin (FTL), through the intraperitoneal route, has already been proven. This study aimed to develop, characterize, and evaluate the orofacial antinociceptive activity of an oral formulation containing FTL in acute and neuropathic preclinical tests. Nanoencapsulated FTL was administered by oral route. The acute nociceptive behavior was induced by administering capsaicin to the upper lip and NaCl to the right cornea. The nociceptive behavior was also induced by formalin injected into the temporomandibular joint. The neuropathic pain model involved infraorbital nerve transection (IONX), which induced mechanical hypersensitivity and was assessed by von Frey stimulation. Trpv1 gene expression was analyzed in the trigeminal ganglion. The analyzed sample did not show any cytotoxicity; 52.2% of the FTL was encapsulated, and the size of the nanocapsule was less than 200 nm, the polydispersion was 0.361, and the zeta potential was - 5.87 and - 12.8 mV, with and without FTL, respectively. Nanoencapsulated FTL administered by oral route had an orofacial antinociceptive effect in acute and neuropathic rodent models. The antinociceptive effect of FTL was prevented by ruthenium red, but not by camphor. FTL reduced Trpv1 gene expression. FTL promotes orofacial antinociception, probably due to the antagonism of TRPV1 channels, and the nanoformulation represents an effective method for the oral administration of this protein. HIGHLIGHTS: • Nanoformulation for oral protein administration. • Nanocapsule containing FTL prevents orofacial nociceptive acute and neuropathic pain. • Frutalin promotes orofacial antinociception behavior antagonism of TRPV1 channels.

Os compostos com atividade antiviral eficazes para o novo coronavírus ainda são pouco conhecidos, e os terpenos são um grupo interessante de agentes naturais com atividades antivirais de longo alcance. Esta investigação visa identificar a atividade antiviral in silico do ?-bisabolol, citral, cis-jasmona e eucaliptol contra nova glicoproteína Spike do SARS-COV-2. A interação entre os terpenos e a glicoproteína Spike foi analisada in silico por meio de simulação de docking molecular, buscando formar uma complexação estável. Todas as evidências químicas, espaciais e energéticas sugerem que essas 4 substâncias são capazes de complexar e estabilizar esta proteína chave na infecção viral e sustentam a hipótese de que complexações de terpenos dificultam a entrada do vírus nas células hospedeiras.

Iron nanoparticles (FeNP) present excellent magnetic properties and chemical stability, and for this reason, they are often configured into materials for a variety of potential uses in medical, biotechnological, and other applications. In this work, iron oxide nanoparticles functionalized with galactomannan (FeNP/Gal) from Caesalpinia pulcherrima were synthesized and submitted to characterization and evaluation of the cytotoxic activity. The functionalized nanoparticles were synthesized by co-precipitation and subjected to a process of surface modification with galactomannan and epichlorohydrin. These nanomaterials were characterized using infrared spectroscopy, X-ray diffraction (XRD), thermogravimetric analysis (TGA), differential thermogravimetric analysis (DTA), and scanning electron microscopy (SEM). 1D and 2D nuclear magnetic resonance (NMR) spectroscopy were also used in the structural analysis of the galactomannan. In addition, in vitro study was carried out to evaluate the cytotoxic activity of the FeNP/Gal nanoparticles on human cells of the HEK-293 strain (ATCC® CRL-1573). The FeNP/Gal nanoparticles had an average diameter of 13 ± 2 nm as opposed to 11 ± 2 nm for unreacted FeNP. The infrared spectrum of the FeNP/Gal nanoparticles presents characteristic absorbance bands of their chemical constituents, confirming that the iron oxide nanoparticles were functionalized with galactomannan. The cytotoxicity assay for the FeNP/Gal nanoparticles did not show significant cytotoxicity against HEK 293-Human embryonic kidney cell lines below 800 µg/mL However, this study points out the possibility of using hemicellulose and other plant-based polysaccharides to produce nanostructured materials for ​​tissue engineering and other biomedical applications.

Objective l-Asparaginase (ASNase) is an enzyme used in the treatment of acute lymphoblastic leukemia (ALL). As the therapeutic ASNases has bacterial origin, severe side effects are associated with its use, among them hypersensitivity and inactivation of the enzyme. In this context, the objective of this work was to produce a recombinant ASNase of bacterial origin in human cells in order to determine the presence and consequences of potential post-translational modifications on the enzyme. Results Recombinant ASNase was expressed in human cells with a molecular weight of 60 kDa, larger than in Escherichia coli, which is 35 kDa. N-glycosylation analysis demonstrated that the increased molecular weight resulted from the addition of glycans to the protein by mammalian cells. The glycosylated ASNase presented in vitro activity at physiological pH and temperature. Given that glycosylation can act to reduce antigenicity by masking protein epitopes, our data may contribute to the development of an alternative ASNase in the treatment of ALL in patients who demonstrate side effects to currently marketed enzymes.

A diet based on sunflower cake for lambs was assayed in order to reuse biodiesel industrial by‐products with the aim of reducing livestock costs and evaluating their influence on meat quality. To achieve these goals, sixteen male lambs were fed diets containing different levels of sunflower cake (control, 5%, 10% and 15%). Afterwards, their semimembranosus muscles were analysed by two‐dimensional electrophoresis coupled to mass spectrometry and their functional protein association was examined using STRING. Structural and metabolic proteins in the lambs’ proteomes changed significantly according to their diet. Fifteen proteins showed significant changes caused by the inclusion of sunflower cake, and the most differentially abundant structural proteins were detected in 2‐DE gels from the lambs. Differentially abundant metabolic proteins such as ENO3 (enolase 3), MDH1 (malate dehydrogenase) and ALDH1A1 (retinal dehydrogenase) have been proposed as biomarkers of quality parameters in other species.

Brazilian Somalis is a locally-adapted breed of rams raised in tropical climate and native pastures. The present study was conducted to evaluate gene expression and proteome of the reproductive tract of such rams. Samples were collected from testes, epididymides, seminal vesicles and bulbourethral glands of four rams. Expression of clusterin (CLU), osteopontin (OPN) and prostaglandin D2 synthase (PGDS) genes were evaluated in all samples by real-time PCR. Shotgun proteomic analysis was performed using samples from the head, corpus and cauda epididymides and from all other structures as well. Gene ontology terms and protein interactions were obtained from UniProtKB databases and MetaCore v.6.8 platform. CLU trasncripts were detected in the testes, epididymides, seminal vesicles and bulbourethral glands of the Somalis rams. The initial region and body of the epididymis had the greatest CLU expression. OPN mRNA was localized in all tissues of the ram reproductive tract. PGDS mRNA was detected in the testes and epididymides. Lable-free mass spectrometry allowed the identification of 137 proteins in all samples. Proteins of the epididymis head mainly participate in cellular processes and response to stimulus, participating in catalityc activity and binding. Proteins of epididymis body acted as regulatory proteins and in cellular processes, with binding and catalytic activity. Cauda epididymis molecules were associated with cellular processes and regulation, with binding function and catalytic activity as well. Testis proteins were mainly linked to cell processes and response to stimuli, and had catalytic function. Seminal vesicle proteins were involved in regulation and mainly with binding functions. Most bulbourethral gland proteins participated in cellular processes. The present study is the first to evaluate the proteome and gene expressions in the reproductive tract of Brazilian Somalis rams. Such pieces of information bring significant cointribution for the understanding of the reproductive physiology of locally-adapted livestock.

Physiologically mature ‘Paluma’ guava were treated with galactomannan (0.75%) and carnauba wax (0.9%) coating, stored at ambient (25 °C) and refrigerated (11 °C), and evaluated every 3rd day for postharvest quality and physiology. Ambient uncoated fruit were apt for consumption for only nine days; meanwhile, coated guavas were acceptable for 15 d with climacteric respiratory peak of 168.6 mg kg−1 h-1 at day 12, and firmness of 14.3 N attributed to lower lipid peroxidation and cell wall hydrolysis. Coating increased antioxidant enzymes SOD and CAT activities that led to 35% lower hydrogen peroxide content, when compared to uncoated control. Coating prevented guava degreening by inhibiting chlorophyll degradation with content of 45.05 mg kg-1, lower carotenoid content with 27.8 mg kg-1 and PPO activity, under refrigeration for 15 d. Refrigerated uncoated fruit exhibited as symptoms of chilling injury inhibition of softening and respiratory peak. Principal component analysis showed that coating and refrigeration treatment maintains guava quality for 15 d of storage similar to observed for control, at day zero. In conclusion, galactomannan-carnauba wax coating improved guava postharvest quality through maintenance of firmness and color, besides preventing chilling symptoms under refrigeration.

BACKGROUND Essential oils (EOs) and their volatile components (VCs) have varied biological and pharmacological activities, but low solubility and bioavailability hamper their applications, so that inclusion in cyclodextrins (CDs) is likely to improve their physicochemical properties and pharmacological effects. OBJECTIVE The authors conducted a systematic review to evaluate the biological activities and pharmacological applications of essential oils and their volatile components complexed with cyclodextrins. METHODS The search terms 'Cyxlodextrin', 'Inclusion Complex', 'Volatile oils', 'Essential oil' and 'Volatile components' were used to retrieve articles from the PUBMED, MEDLINE and SCOPUS databases. RESULTS A total of 38 articles were identified. A greater efficacy of EOs and their VCs complexed with different CDs types was found in in vitro and preclinical studies when compared to free forms in the various biological activities and animal models of the evaluated pharmacological tests. CONCLUSION This review of selected studies showed that the use of CDs promotes greater solubility, bioavailability and efficacy of EOs and their VCs, thus indicating an interesting alternative for the biotechnological development of new therapeutic formulations.

Frutalin (FTL) is a multiple-binding lectin belonging to the jacalin-related lectin (JRL) family and derived from Artocarpus incisa (breadfruit) seeds. This lectin specifically recognizes and binds α-d-galactose. FTL has been successfully used in immunobiological research for the recognition of cancer-associated oligosaccharides. However, the molecular bases by which FTL promotes these specific activities remain poorly understood. Here, we report the whole 3D structure of FTL for the first time, as determined by X-ray crystallography. The obtained crystals diffracted to 1.81 Å (Apo-frutalin) and 1.65 Å (frutalin–d-Gal complex) of resolution. The lectin exhibits post-translational cleavage yielding an α- (133 amino acids) and β-chain (20 amino acids), presenting a homotetramer when in solution, with a typical JRL β-prism. The β-prism was composed of three 4-stranded β-sheets forming three antiparallel Greek key motifs. The carbohydrate-binding site (CBS) involved the N-terminus of the α-chain and was formed by four key residues: Gly25, Tyr146, Trp147 and Asp149. Together, these results were used in molecular dynamics simulations in aqueous solutions to shed light on the molecular basis of FTL-ligand binding. The simulations suggest that Thr-Ser-Ser-Asn (TSSN) peptide excision reduces the rigidity of the FTL CBS, increasing the number of interactions with ligands and resulting in multiple-binding sites and anomeric recognition of α-d-galactose sugar moieties. Our findings provide a new perspective to further elucidate the versatility of FTL in many biological activities.

Plant lectins are carbohydrate-binding proteins, which can interact with cell surfaces to initiate anti-inflammatory pathways, as well as immunomodulatory functions. Here, we have extracted, purified and part-characterized the bioactivity of Jacalin, Frutalin, DAL and PNA, before evaluating their potential for wound healing in cultured human skin fibroblasts. Only Frutalin stimulated fibroblast migration in vitro, prompting further studies which established its low cytotoxicity and interaction with TLR4 receptors. Frutalin also increased p-ERK expression and stimulated IL-6 secretion. The in vivo potential of Frutalin for wound healing was then assessed in hybrid combination with the polysaccharide galactomannan, purified from Caesalpinia pulcherrima seeds, using both hydrogel and membrane scaffolds formulations. Physical-chemical characterization of the hybrid showed that lectin-galactomannan interactions increased the pseudoplastic behaviour of solutions, reducing viscosity and increasing Frutalin's concentration. Furthermore, infrared spectroscopy revealed -OH band displacement, likely caused by interaction of Frutalin with galactose residues present on galactomannan chains, while average membrane porosity was 100 μm, sufficient to ensure water vapor permeability. Accelerated angiogenesis and increased fibroblast and keratinocyte proliferation were observed with the optimal hybrid recovering the lesioned area after 11 days. Our findings indicate Frutalin as a biomolecule with potential for tissue repair, regeneration and chronic wound healing.

EnglishPequi oil is rich in bioactive compounds which can be encapsulated to increase protection against extrinsic environmental factors. A delayed degradation of pequi oil may occur by using microencapsulation technology, in addition to masking unpleasant flavors and aromas. Complex coacervation is a technique based on the electrostatic interaction between two oppositely charged biopolymers which form a matrix complexed around an agent of interest. However, cross-linking the particles is often necessary in order to make them more rigid. The objective of this research was to produce and characterize pequi oil microparticles in a cashew gum (CG) and gelatin (GE) matrix cross-linked with tannic acid. Cross-linked pequi oil microparticles were produced by varying the concentrations of biopolymers (0.5% to 1.5%) and tannic acid (0.3% to 8.1%) using a rotational central compound design. Ratio of cashew gum, gelatin and oil was 2:1:1 (m/m/m);respectively, at pH 4.5. The cross-linking process was performed with tannic acid for 30 minutes at 40 °C. The optimized formulation by means of the rotational central compound design for microparticle formation was 0.65% biopolymers (CG and GE) and 6.9% tannic acid. Increasing the tannic acid percentage in the cross-linking of the pequi oil particles had a higher yield and encapsulation efficiency. Cross-linking provided an increase in the degradation temperature of material; and consequently, improved the thermal stability of the particles. The cross-linking process was advantageous in producing the microparticles. EnglishO oleo de pequi e rico em compostos bioativos, os quais podem ser encapsulados para aumentar a protecao a fatores extrinsecos. A tecnologia de microencapsulamento, alem de retardar a degradacao do composto ativo, possibilita mascarar aromas e sabores indesejaveis. A coacervacao complexa e uma tecnica baseada na interacao eletrostatica entre dois biopolimeros com cargas opostas, que formam uma matriz complexada ao redor do agente de interesse. Entretanto, muitas vezes, se faz necessario o uso da reticulacao para tornar as particulas mais rigidas. O objetivo deste trabalho foi produzir e caracterizar microparticulas de oleo de pequi em matriz de goma de cajueiro (GC) e gelatina (GE) reticulada com acido tânico. As microparticulas de oleo de pequi reticuladas foram produzidas variando as concentracoes de biopolimeros (0,5% a 1,5% m/v) e do acido tânico, em relacao a massa de biopolimeros (0,3% a 8,1% m/m), a partir de um delineamento de composto central rotacional. A proporcao de GC, GE e oleo foi de 2:1:1 (m/m/m), respectivamente, em pH 4,5. O processo de reticulacao foi realizado com acido tânico por 30 minutos a 40 oC. A formulacao otimizada foi de 0,65% (m/v) de biopolimeros (GC e GE) e 6,9% (m/m) de acido tânico. O aumento do percentual de acido tânico na reticulacao das particulas de oleo de pequi conferiu maior rendimento e eficiencia de encapsulamento. A reticulacao proporcionou aumento na temperatura de degradacao do material, e consequente estabilidade termica das particulas. O processo de reticulacao foi vantajoso para a producao das microparticulas.

Background: Plant lectins have shown promising neuropharmacological activities in animal models. Objective: This study evaluated the effect of Dioclea altissima seed lectin (DAL) on adult zebrafish behavior. Method: Zebrafish (n=6/group) were treated (i.p.; 20 μL) with DAL (0.025; 0.05 or 0.1 mg/mL), vehicle or diazepam (DZP) and submitted to several tests (open field, light/dark preference or novel tank). Flumazenil, pizotifen or granisetron were administered 15 min before DAL (0.05 mg/mL), and the animals were evaluated on light/dark preference test. It was also verified whether the DAL effect depended on its structural integrity and ability to interact with carbohydrates. Results: DAL decreased the locomotor activity of adult zebrafish (0.025; 0.05 or 0.1 mg/mL), increased the time spent in the upper region of the aquarium (0.025 mg/mL), and decreased the latency time of adult zebrafish to enter the upper region on the novel tank test. DAL (0.05 mg/mL) also increased their permanence in the light zone of the light/dark preference test. The effect of DAL was dependent on carbohydrate interaction and protein structure integrity and was prevented by pizotifen, granizetron and flumazenil. Conclusion: DAL was found to have an anxiolytic-like effect mediated by the 5-HT and GABAergic receptors.

Dairy desserts containing Caesalpinia pulcherrima seed galactomannan were evaluated to determine their static and dynamic rheological behaviors. Variations in consistency index (k), flow behavior (n), yield stress and thixotropy of the desserts indicated that the galactomannan caused an increase in the shear stress and apparent viscosity of the system. All samples exhibited shear-thinning behavior with flow behavior index values (n) between 0.06 and 0.37. Dynamic rheological behavior was evaluated for MD (high solid levels) and MD/2 (half the amount of solids) groups, and both G’ and G’’ moduli were depended on the frequency. The MD and MD/2 groups showed variations in the elastic modulus (G’) throughout the temperature range (mainly at 50 °C), showing greater sensitivity at high temperatures. C. pulcherrima galactomannan was able to promote synergism with starch, milk protein and sucrose and to improve the development of stronger and more resistant gels. RESUMO: Sobremesas lácteas contendo galactomanana de semente de Caesalpinia pulcherrima tiveram suas propriedades reológicas estáticas e dinâmicas avaliadas. As variações nos índices de consistência (K) e comportamento de fluxo (n), assim como na tensão inicial de fluxo e na tixotropia das sobremesas mostraram o efeito da galactomanana sobre a tensão de cisalhamento e viscosidade aparente dos sistemas lácteos. Todas as sobremesas exibiram comportamento pseudoplástico, com índices de comportamento de fluxo (n) variando entre 0,06 e 0,37. A reologia dinâmica dos grupos MD (alto teor de sólidos) e MD/2 (metade do teor de sólidos), mostrou G’ > G’’ e módulos dependentes da frequência e da temperatura. Alterações químicas nos componentes das sobremesas foram observadas a 50° C em virtude da maior sensibilidade dos valores de G’ e G” a partir dessa temperatura. A galactomanana de C. pulcherrima contribuiu para o desenvolvimento de géis mais fortes e resistentes nas sobremesas lácteas, bem como mostrou sinergismo com amido, proteína do leite e sacarose.

The aim of this study was to determine the plasma protein profile of patients with primary antiphospholipid syndrome (PAPS) compared to healthy controls and identify proteins that might be used in the evaluation, diagnosis, and prognosis of this condition. The sample consisted of 14 patients with PAPS and 17 sex- and age-matched controls. Plasma samples were submitted to proteomic analysis (albumin and immunoglobulin G depletion, concentration, digestion, and label-free data-independent mass spectrometry). The software ExpressionEwas used to quantify intergroup differences in protein expression. The analysis yielded 65 plasma proteins of which 11 were differentially expressed (9 upregulated and 2 downregulated) in relation to controls. Four of these are known to play a role in pathophysiological mechanisms of thrombosis: fibrinogen α chain, fibrinogen α chain, apolipoprotein C-III, and α-1-glycoprotein-1. Our analysis revealed autoimmune response and the presence of proteins believed to be functionally involved in the induction of procoagulant activity in patients with PAPS. Further studies are necessary to confirm our findings and may eventually lead to the development of significantly more accurate diagnostic tools.

Neem fruit (Azadirachta indica A. Juss.) are popularly used to treat infections, diarrhea, fever, bronchitis, skin diseases, infected burns and hypertension. Although the antinociceptive and anti-inflammatory potential of A. indica has already been investigated in experimental models of pain and inflammation in mice, the current research is the first to report the evaluation of the capacity of A. indica fruit ethanolic extract (EtFrNeem) in acute pain attenuation using the adult zebrafish (Danio rerio) as an alternative model to the use in rodents. EtFrNeem was submitted to antioxidant action, preliminary chemical prospecting, FT-IR and determination of phenol and flavonoid content tests. Subsequently, EtFrNeem was tested for acute nociception and abdominal inflammation, locomotor activity, and acute toxicity in adult zebrafish. Possible neuromodulation mechanisms were also evaluated. EtFrNeem showed low antioxidant activity, but was shown to be rich in flavonoids. EtFrNeem showed no anti-inflammatory action, did not alter the locomotor system, and it was not toxic. However, EtFrNeem significantly reduced the nociceptive behavior induced by formalin, glutamate and acidic saline, when compared to the control group. These effects of EtFrNeem were significantly similar to those of morphine, used as a positive control. The antinociceptive effect of EtFrNeem was inhibited by naloxone, ketamine and amiloride. EtFrNeem has the pharmacological potential for acute pain treatment and this effect is modulated by the opioid system, NMDA receptors and ASICs channels. These results lead us to studies of isolation and characterization of EtFrNeem bioactive principles, using adult zebrafish as an experimental model.

The increased incidence of candidemia in terciary hospitals worldwide and the cross-resistance frequency require the new therapeutic strategies development. Recently, our research group demonstrated three semi-synthetic naphthofuranquinones (NFQs) with a significant antifungal activity in a fluconazole-resistant (FLC) C. tropicalis strain. The current study aimed to investigate the action's preliminary mechanisms of NFQs by several standardized methods such as proteomic and flow cytometry analyzes, comet assay, immunohistochemistry and confocal microscopy evaluation. Our data showed C. tropicalis 24 h treated with all NFQs induced an expression's increase of proteins involved in the metabolic response to stress, energy metabolism, glycolysis, nucleosome assembly and translation process. Some aspects of proteomic analysis are in consonance with our flow cytometry analysis which indicated an augmentation of intracellular ROS, mitochondrial dysfunction and DNA strand breaks (neutral comet assay and γ-H2AX detection). In conclusion, our data highlights the great contribution of ROS as a key event, probably not the one, associated to anti-candida properties of studied NFQs.

The healing performance of a hydrogel composed of hemicelluloses extracted from seeds of Caesalpinia pulcherrima (Fabaceae) and mixed with phytomodulatory proteins obtained from the latex of Calotropis procera was characterized on excisional wounds. The hydrogel did not induce dermal irritability. When topically used on excisional wounds, the hydrogel enhanced healing by wound contraction. Histology and the measurement of inflammatory mediators (myeloperoxidase, interleukin-1β, and interleukin-6) suggested that the inflammatory phase of the healing process was intensified, stimulating fibroplasia and neovascularization (proliferative phase) and tissue remodeling by increasing new collagen fiber deposition. In addition, reduction on levels of malondialdehyde in the groups that the hydrogel was applied suggested that the oxidative stress was reduced. The hydrogel performed better than the reference drug used, as revealed by the extended thickness of the remodeled epithelium.

The latex of Calotropis procera has previously been reported to contain osmotin. This protein (CpOsm) inhibited phytopathogens and this was mechanistically characterized. Here, the time-course profile of CpOsm transcripts was examined in the salt-stressed cultivated callus of C. procera in order to better understand its role in the physiology of the plant. Stressed callus (80 mM NaCl) showed an unbalanced content of organic compounds (proline and total soluble sugar) and inorganic ions (Na+, Cl−, and K+). Under salt treatment, the transcripts of CpOsm were detected after 12 h and slightly increased to a maximum at day seven, followed by reduction. Interestingly, CpOsm was not detected in the soluble protein fraction recovered from the salt-stressed callus as probed by electrophoresis, dot/Western blotting and mass spectrometry. The results suggested that (1) CpOsm is not constitutive in cultivated cells (laticifer-free tissues); (2) CpOsm transcripts appear under salt-stressed conditions; (3) the absence of CpOsm in the protein fractions of stressed cultivated cells indicated that salt-induced transcripts were not used for protein synthesis and this accounts to the belief that CpOsm may be a true laticifer protein in C. procera. More effort will be needed to unveil this process. In this study we show evidences that CpOsm gene is responsive to salt stress. However the corresponding protein is not produced in cultivated cells. Therefore, presently the hypothesis that CpOsm is involved in abiotic stress is not fully supported.

In this work, galactomannans and xyloglucans were isolated from the seed endosperm and cotyledon of Brazilian non-conventional sources, respectively. Extraction yields, monosaccharide ratios, macromolecular parameters and molar mass distributions were determined and compared to commercial guar gum and Locust Bean Gum (LBG). The extraction yield in relation to seed mass ranged from 7.0% to 40.63%, with xyloglucan yields being higher than galactomannan yields. Schizolobium parahyba and Caesalpinia pulcherrima galactomannans exhibited the lowest protein contents, 0.05% and 0.08%, respectively. Flow curves of 1% hemicellulose solutions (w:v) were measured by varying the shear rate from 0.1 to 100 s﹣1. The resulting data were fitted to a Power Law model, and all the hemicelluloses presented shear-thinning behavior. Galactomannans and xyloglucans with different monosaccharide ratios showed similar consistency indices. Rheological properties were also compared, and the results suggest new hemicellulose sources, which can be studied for additional applications in areas such as materials science, medicine and biology.

Visceral leishmaniasis is caused by parasites of the genus Leishmania. Prevalent in developing countries, the disease is on the list of the world’s most neglected diseases. Most cases (90%) in Latin America occur in Brazil, especially in the Northeast. The condition is treated mainly with pentavalent antimonials, an expensive and relatively inefficient drug known to induce resistance. In search of new antileishmanial drugs, much attention has been given to the medicinal properties of coumarin. In this study, coumarin was isolated from seeds of Amburana cearensis, purified and evaluated with regard to its ability to inhibit Leishmania chagasi promastigotes using the MTT test. The cell viability of macrophages treated with coumarin was also evaluated. The findings were submitted to one-way ANOVA for paired data, followed by the Bonferroni correction. The level of statistical significance was set at 5% (p 0.05), but was toxic to Leishmania chagasi promastigotes (p < 0.05). Our results represent a contribution to the development of new drugs for the control or prophylaxis of visceral leishmaniasis.

Background Diarrheal diseases are an important cause of morbidity and mortality among children in developing countries. We aimed to study the etiology and severity of diarrhea in children living in the low-income semiarid region of Brazil. Methodology This is a cross-sectional, age-matched case-control study of diarrhea in children aged 2–36 months from six cities in Brazil’s semiarid region. Clinical, epidemiological, and anthropometric data were matched with fecal samples collected for the identification of enteropathogens. Results We enrolled 1,200 children, 596 cases and 604 controls. By univariate analysis, eight enteropathogens were associated with diarrhea: Norovirus GII (OR 5.08, 95% CI 2.10, 12.30), Adenovirus (OR 3.79, 95% CI 1.41, 10.23), typical enteropathogenic Escherichia coli (tEPEC), (OR 3.28, 95% CI 1.39, 7.73), enterotoxigenic E. coli (ETEC LT and ST producing toxins), (OR 2.58, 95% CI 0.99, 6.69), rotavirus (OR 1.91, 95% CI 1.20, 3.02), shiga toxin-producing E. coli (STEC; OR 1.77, 95% CI 1.16, 2.69), enteroaggregative E. coli (EAEC), (OR 1.45, 95% CI 1.16, 1.83) and Giardia spp. (OR 1.39, 95% CI 1.05, 1.84). By logistic regression of all enteropathogens, the best predictors of diarrhea were norovirus, adenovirus, rotavirus, STEC, Giardia spp. and EAEC. A high diarrhea severity score was associated with EAEC. Conclusions Six enteropathogens: Norovirus, Adenovirus, Rotavirus, STEC, Giardia spp., and EAEC were associated with diarrhea in children from Brazil’s semiarid region. EAEC was associated with increased diarrhea severity., Author summary Most childhood diarrheal diseases studies focus on children from health centers or emergency hospitals that contribute to a lack of understanding of the etiology of community diarrhea. We aimed to investigate the etiology and severity of diarrheal diseases in children living in Brazil’s low-income, semiarid region communities. We used a case-control study of diarrhea (1,200 children, 596 cases and 604 controls) and showed that six enteropathogens (norovirus, adenovirus, rotavirus, STEC, Giardia and EAEC) are associated with diarrhea in these communities. Furthermore, enteroaggregative E. coli (EAEC) was associated with a high diarrhea clinical severity score. These findings help further the understanding of diarrheal disease etiology of previously unevaluated children from Brazil’s low-income semiarid region. This study contributes new information that expands our knowledge of diarrheal etiology in non-hospitalized children from developing countries. Moreover, we believe this work will support further research on specific enteropathogens, with special attention to the role of EAEC in severe cases of diarrhea and guide public health policies and physicians in the management of diarrheal diseases in Brazil’s low-income semiarid region.

The objective of the present study was to describe the relationship of seminal plasma and total sperm cell proteins with the semen freezability parameters of Guzerat bulls. Thirteen bulls were subjected to breeding soundness evaluation. Semen samples were collected, cryopreserved, and then post-thawing sperm kinetics were assessed, where high ( = 7) and low ( = 6) freezability groups were defined. Seminal plasma and total sperm proteins from the 2 groups were separated by 2-dimensional SDS-PAGE, and spots were identified by mass spectrometry. Semen parameters post-cryopreservation were as follows in the high and low freezability groups, respectively: mean total motility, 52.4 ± 20.5 and 13.7 ± 3.9; percentage of normal sperm, 89.0 ± 2.6 and 64.7 ± 14.0; and reactivity of hypo-osmotic swelling test, 38.9 ± 4.7 and 13.6 ± 3.7. Three seminal plasma proteins (osteopontin-K, DNase γ precursor, and DNASE1L3) and 6 proteins from sperm cells (acrosome formation-associated factor isoform 2, annexin A1, disintegrin and metalloproteinase domain-containing protein 2, dihydrolipoyl dehydrogenase, and glyceraldehyde-3-phosphate dehydrogenase) were highly expressed ( < 0.05) in the high freezability group. Another 6 seminal plasma proteins (acrosin inhibitor 1, glutathione peroxidase 3, metalloproteinase inhibitor 2, ephrin-A1, annexin A1, and platelet-activating factor acetylhydrolase) were significantly higher ( < 0.05) in the low freezability group. We described the associations of seminal plasma and sperm cell proteins with post-thawing sperm viability of Guzerat bulls raised in a typical semiarid environment. Such associations indicate that specific seminal plasma proteins more abundant in bulls of low semen freezability may be a response to an early oxidative stress that is not detected by conventional prefreezing semen evaluation. Moreover, specific sperm proteins were more associated with good freezability. The results presented here can serve as guidelines for future research aiming to develop better extenders and/or to improve bull semen selection for cryopreservation.

Chitinases (EC 3.2.1.14) found in the latex of Calotropis procera (Ait) R. Br. were studied. The proteins were homogeneously obtained after two ion exchange chromatography steps. Most proteins were identified individually in 15 spots on 2-D gel electrophoresis with isoelectric points ranging from 4.6 to 6.0 and molecular masses extending from 27 to 30 kDa. Additionally, 66 kDa proteins were identified as chitinases in SDS-PAGE. Their identities were further confirmed by mass spectrometry (MS) analysis of the tryptic digests of each spot and MS analysis of the non-digested proteins. Positive reaction for Schiff's reagent suggested the proteins are glycosylated. The chitinases exhibited high catalytic activity toward to colloidal chitin at pH 5.0, and this activity underwent decay in the presence of increasing amounts of reducing agent dithiothreitol. Spore germination and hyphae growth of two phytopathogenic fungi were inhibited only marginally by the chitinases but were affected differently. This suggested a complex relationship might exist between the specificity of the proteins toward the fungal species. The chitinases showed potent insecticidal activity against the Bruchidae Callosobruchus maculatus, drastically reducing survival, larval weight and adult emergence. It is concluded that closely related chitinases are present in the latex of C. procera, and the first experimental evidence suggests these proteins are involved more efficiently in defence strategies against insects rather than fungi.

Orofacial pain is a highly prevalent clinical condition, yet difficult to control effectively with available drugs. Much attention is currently focused on the anti-inflammatory and antinociceptive properties of lectins. The purpose of this study was to evaluate the antinociceptive effect of frutalin (FTL) using rodent models of inflammatory and neuropathic orofacial pain. Acute pain was induced by formalin, glutamate or capsaicin (orofacial model) and hypertonic saline (corneal model). In one experiment, animals were pretreated with l-NAME and naloxone to investigate the mechanism of antinociception. The involvement of the lectin domain in the antinociceptive effect of FTL was verified by allowing the lectin to bind to its specific ligand. In another experiment, animals pretreated with FTL or saline were submitted to the temporomandibular joint formalin test. In yet another, animals were submitted to infraorbital nerve transection to induce chronic pain, followed by induction of thermal hypersensitivity using acetone. Motor activity was evaluated with the rotarod test. A molecular docking was performed using the TRPV1 channel. Pretreatment with FTL significantly reduced nociceptive behaviour associated with acute and neuropathic pain, especially at 0.5 mg/kg. Antinociception was effectively inhibited by l-NAME and d-galactose. In line with in vivo experiments, docking studies indicated that FTL may interact with TRPV1. Our results confirm the potential pharmacological relevance of FTL as an inhibitor of orofacial nociception in acute and chronic pain mediated by TRPA1, TRPV1 and TRPM8 receptor.

The latex from Thevetia peruviana is rich in plant defense proteins, including a 120 kDa cysteine peptidase with structural characteristics similar to germin-like proteins. More than 20,000 plant species produce latex, including Apocynaceae, Sapotaceae, Papaveraceae and Euphorbiaceae. To better understand the physiological role played by latex fluids, a proteomic analysis of Thevetia peruviana (Pers.) Schum latex was performed using two-dimensional gel electrophoresis and mass spectrometry. A total of 33 proteins (86 %) were identified, including storage proteins, a peptidase inhibitor, cysteine peptidases, peroxidases and osmotins. An unusual cysteine peptidase, termed peruvianin-I, was purified from the latex by a single chromatographic step involving gel filtration. The enzyme (glycoprotein) was inhibited by E-64 and iodoacetamide and exhibited high specific activity towards azocasein (K m 17.6 µM), with an optimal pH and temperature of 5.0–6.0 and 25–37 °C, respectively. Gel filtration chromatography, two-dimensional gel electrophoresis, and mass spectrometry revealed that peruvianin-I possesses 120 kDa, pI 4.0, and six subunits (20 kDa). A unique N-terminal amino acid sequence was obtained to oligomer and monomers of peruvianin-I (1ADPGPLQDFCLADLNSPLFINGYPCRNPALAISDDF36). High-resolution images from atomic force microscopy showed the homohexameric structure of peruvianin-I may be organized as a trimer of dimers that form a central channel similar to germin-like proteins. Peruvianin-I exhibited no oxalate oxidase and superoxide dismutase activity or antifungal effects. Peruvianin-I represents the first germin-like protein (GLP) with cysteine peptidase activity, an activity unknown in the GLP family so far.

This study optimizes the pyrodextrinization of yam (Dioscorea sp.) starch isolated from tubers grown in Brazil to produce a yellow pyrodextrin with the lowest enzymatically available starch (AS) content and color difference (ΔE) index. At 140 °C (fixed heating temperature), the effects of acid concentration (0.65 - 2.99 g of HCl/kg of starch) and incubation time (53 - 307 min) on the response variables were evaluated using a response surface methodology. Some physicochemical characteristics were also determined on pyrodextrins. Both factors negatively affected the AS content, although positively influenced the ΔE (P 0.05). The yellow pyrodextrin produced with 1.82 g/kg and heating for 307 min, presented physicochemical properties similar to the commercial pyrodextrins from potato starch, with 46.6 % of AS, 24.5 of ΔE, high solubility and very low viscosity. The pyrodextrinization caused a decrease of 30 - 54 % in AS content (P 0.05), making these yam pyrodextrins a promising material for water-soluble and very low viscous dietary fiber.