RESUMO
The study investigates the efficacy of an enzymatic preparation primarily with α-galactosidase activity for improving the quality of white sugar from poor-quality sugar beets. Focused on overcoming raffinose accumulation challenges in sugar beets, especially those harvested prematurely or stored for extended periods, an innovative exploration of enzymatic application in an industrial setting for the first time was conducted. By integrating theoretical calculations and experimental data, the findings reveal that α-galactosidase preparation notably diminishes raffinose content in beet juice, thus enhancing the sucrose yield and overall sugar quality. A reliable method to process lower-quality beets, promising enhanced efficiency in sugar production, was presented. The study also highlights the economic benefits of incorporating enzyme preparation into the production process, demonstrating a notable return on investment and underscoring the potential of enzymatic treatments to address industry challenges.
Assuntos
Beta vulgaris , Rafinose , alfa-Galactosidase , Rafinose/química , Rafinose/metabolismo , Beta vulgaris/química , alfa-Galactosidase/metabolismo , alfa-Galactosidase/química , Açúcares/química , Açúcares/metabolismo , CatáliseRESUMO
The presence of antinutrients and undesirable flavours in kidney bean flour poses challenges to consumer acceptance. Although extrusion can mitigate antinutrients to some extent, its impact on reducing beany flavour in bean flour remains underexplored. This study investigated the effects of injecting acetic acid or sodium carbonate solutions at three concentration levels (0.05, 0.1, 0.15 mol/L), in conjunction with three temperature profiles (40/60/80/80/90, 40/60/80/90/110, 50/70/90/110/130 °C) and two feed moisture levels (25, 30 %), on the removal of antinutrients (condensed tannins, trypsin inhibitor activity, phytic acid, raffinose family oligosaccharides) and reduction of volatile compounds that contribute to beany flavour in whole kidney bean flour. The results showed that all concentrations of acetic acid and sodium carbonate solutions effectively reduced condensed tannins compared to water, especially at 130 °C extrusion temperature. Introducing acetic acid and sodium carbonate solutions at a concentration of 0.15 mol/L led to 72 and 90 % reduction of total raffinose family oligosaccharide content, respectively, in contrast to the 17 % reduction observed with water alone. The incorporation of sodium carbonate solution reduced the total volatile compounds by 45-58 % as compared with water (23-33 %) and acetic acid (11-27 %). This reduction was primarily due to the reduction of aldehydes, alcohols, and aromatic hydrocarbons. These results indicate that injecting sodium carbonate solution during extrusion can effectively reduce antinutrients and beany flavour compounds in kidney bean flour.
Assuntos
Ácido Acético , Carbonatos , Farinha , Manipulação de Alimentos , Phaseolus , Ácido Fítico , Paladar , Farinha/análise , Carbonatos/química , Manipulação de Alimentos/métodos , Ácido Fítico/análise , Phaseolus/química , Proantocianidinas/análise , Rafinose/química , Temperatura , Concentração de Íons de Hidrogênio , Compostos Orgânicos Voláteis/análiseRESUMO
Segmental grafts from living donors have advantages over grafts from deceased donors when used for small intestine transplantation. However, storage time for small intestine grafts can be extremely short and optimal graft preservation conditions for short-term storage remain undetermined. Secreted factors from mesenchymal stem cells (MSCs) that allow direct activation of preserved small intestine grafts. Freshly excised Luc-Tg LEW rat tissues were incubated in preservation solutions containing MSC-conditioned medium (MSC-CM). Preserved Luc-Tg rat-derived grafts were then transplanted to wild-type recipients, after which survival, injury score, and tight junction protein expression were examined. Luminance for each graft was determined using in vivo imaging. The findings indicated that 30-100 and 3-10 kDa fractions of MSC-CM have superior activating effects for small intestine preservation. Expression of the tight-junction proteins claudin-3, and zonula occludens-1 preserved for 24 h in University of Wisconsin (UW) solution containing MSC-CM with 50-100 kDa, as shown by immunostaining, also indicated effectiveness. Reflecting the improved graft preservation, MSC-CM preloading of grafts increased survival rate from 0% to 87%. This is the first report of successful transplantation of small intestine grafts preserved for more than 24 h using a rodent model to evaluate graft preservation conditions that mimic clinical conditions.
Assuntos
Intestino Delgado , Células-Tronco Mesenquimais , Preservação de Órgãos , Ratos Endogâmicos Lew , Animais , Intestino Delgado/transplante , Ratos , Preservação de Órgãos/métodos , Masculino , Soluções para Preservação de Órgãos , Sobrevivência de Enxerto , Meios de Cultivo Condicionados , Proteína da Zônula de Oclusão-1/metabolismo , Claudina-3/metabolismo , Ratos Transgênicos , Glutationa , Rafinose , Alopurinol , Insulina , AdenosinaRESUMO
Soaking pulses in water is a traditional practice widely used both by many households and by the food industry, and depending on the specific conditions used, can effectively reduce α-galactosides. Monitoring changes in α-galactoside content in pulses under different steeping conditions can provide insights into the degradation mechanisms and help overcome the barrier to consumption caused by digestive problems. In this study, we analyzed the impact of steeping at different temperatures (30, 45, 60, 75, and 90 °C) and at different pH (4.0, 5.0, and 6.0) on α-galactosides content in chickpeas, lentils, and beans. Our results showed that the lower the pH, the faster the α-galactosides were reduced. Moreover, steeping at lower temperatures (30 °C and 45 °C) favored hydrolysis of α-galactosides, whereas steeping at higher temperatures (60, 75, and 90 °C) favored diffusion. Soaking at 45 °C at a pH of 4.0 for 3 h resulted in acceptable levels of α-galactosides (less than 1 g/100 g), i.e. a reduction of up to 65 % in chickpeas, 85 % in lentils, and 52 % in beans.
Assuntos
Cicer , Lens (Planta) , Oligossacarídeos , Rafinose , Temperatura , Concentração de Íons de Hidrogênio , Hidrólise , Rafinose/química , Rafinose/análise , Oligossacarídeos/química , Oligossacarídeos/análise , Lens (Planta)/química , Cicer/química , Manipulação de Alimentos/métodos , Galactosídeos/química , DifusãoRESUMO
Raffinose family oligosaccharides (RFOs) are synthesized from sucrose and subsequent addition of galactose moieties which was provided by galactinol. Galactinol is synthesized from UDP-galactose and myo-inositol. RFOs accumulate at late stage of seed development and play important roles in seed longevity. RFOs are major components in seeds of many plant species. Here, we document a methodology for extraction and quantitative analysis of raffinose metabolism-related soluble sugars or the derivative alcohols in plant seeds. This protocol, based on high-performance liquid chromatography (HPLC), achieves the efficient separation and accurate quantification of sucrose, myo-inositol, galactinol, and raffinose within 25 min of retention time.
Assuntos
Rafinose , Sementes , Sacarose , Rafinose/metabolismo , Sementes/metabolismo , Sementes/crescimento & desenvolvimento , Cromatografia Líquida de Alta Pressão/métodos , Sacarose/metabolismo , Inositol/metabolismo , Inositol/análogos & derivadosRESUMO
Cassava is a crucial staple crop for smallholder farmers in tropical Asia and Sub-Saharan Africa. Although high yield remains the top priority for farmers, the significance of nutritional values has increased in cassava breeding programs. A notable negative correlation between provitamin A and starch accumulation poses a significant challenge for breeding efforts. The negative correlation between starch and carotenoid levels in conventional and genetically modified cassava plants implies the absence of a direct genomic connection between the two traits. The competition among various carbon pathways seems to account for this relationship. In this study, we conducted a thorough analysis of 49 African cassava genotypes with varying levels of starch and provitamin A. Our goal was to identify factors contributing to differential starch accumulation. Considering carotenoid levels as a confounding factor in starch production, we found that yellow- and white-fleshed storage roots did not differ significantly in most measured components of starch or de novo fatty acid biosynthesis. However, genes and metabolites associated with myo-inositol synthesis and cell wall polymer production were substantially enriched in high provitamin A genotypes. These results indicate that yellow-fleshed cultivars, in comparison to their white-fleshed counterparts, direct more carbon toward the synthesis of raffinose and cell wall components. This finding is underlined by a significant rise in cell wall components measured within the 20 most contrasting genotypes for carotenoid levels. Our findings enhance the comprehension of the biosynthesis of starch and carotenoids in the storage roots of cassava.
Assuntos
Carbono , Parede Celular , Inositol , Manihot , Raízes de Plantas , Rafinose , Amido , Amido/metabolismo , Manihot/genética , Manihot/metabolismo , Carbono/metabolismo , Raízes de Plantas/metabolismo , Raízes de Plantas/genética , Parede Celular/metabolismo , Inositol/metabolismo , Rafinose/metabolismo , Genótipo , Carotenoides/metabolismoRESUMO
A series of intricate and dynamic physiological healing processes are involved in the healing of skin wounds. Herein, a multifunctional hydrogel is firstly designed and constructed by L-arginine-grafted O-carboxymethyl chitosan (CMCA), catechol-modified oxidized hyaluronic acid (DOHA), and dopamine nanoparticles (pDA-NPs). pDA-NPs were loaded in hydrogel for inherently powerful antimicrobial properties and could be as a cross-linking agent to construct hydrogels. Raffinose (Raf) was further incorporated to obtain CMCA-DOHA-pDA2@Raf hydrogel for its function of modulating epidermal differentiation. The hydrogel has good physicochemical properties and could promote cell proliferation and migration, which shows superior hemostatic capabilities in animal models of hemorrhage. The hydrogel significantly promoted wound healing on rat skin defect models by upregulating VEGF and CD31 and decreasing IL-6 and TNF-α, stimulating neovascularization and collagen deposition in epithelial structures. This multifunctional hydrogel implies the potential to be a dynamic wound dressing.
Assuntos
Quitosana , Dopamina , Hidrogéis , Nanopartículas , Rafinose , Cicatrização , Cicatrização/efeitos dos fármacos , Animais , Hidrogéis/química , Hidrogéis/farmacologia , Nanopartículas/química , Dopamina/química , Dopamina/farmacologia , Ratos , Quitosana/química , Quitosana/análogos & derivados , Quitosana/farmacologia , Rafinose/química , Rafinose/farmacologia , Proliferação de Células/efeitos dos fármacos , Ácido Hialurônico/química , Ácido Hialurônico/farmacologia , Humanos , Masculino , Reagentes de Ligações Cruzadas/química , Ratos Sprague-Dawley , Pele/efeitos dos fármacos , Movimento Celular/efeitos dos fármacosRESUMO
Drought is the most severe form of stress experienced by plants worldwide. Cucumber is a vegetable crop that requires a large amount of water throughout the growth period. In our previous study, we identified that overexpression of CsHSFA1d could improve cold tolerance and the content of endogenous jasmonic acid in cucumber seedlings. To explore the functional diversities of CsHSFA1d, we treat the transgenic plants under drought conditions. In this study, we found that the heat shock transcription factor HSFA1d (CsHSFA1d) could improve drought stress tolerance in cucumber. CsHSFA1d overexpression increased the expression levels of galactinol synthase (CsGolS3) and raffinose synthase (CsRS) genes, encoding the key enzymes for raffinose family oligosaccharide (RFO) biosynthesis. Furthermore, the lines overexpressing CsHSFA1d showed higher enzymatic activity of GolS and raffinose synthase to increase the content of RFO. Moreover, the CsHSFA1d-overexpression lines showed lower reactive oxygen species (ROS) accumulation and higher ROS-scavenging enzyme activity after drought treatment. The expressions of antioxidant genes CsPOD2, CsAPX1 and CsSOD1 were also upregulated in CsHSFA1d-overexpression lines. The expression levels of stress-responsive genes such as CsRD29A, CsLEA3 and CsP5CS1 were increased in CsHSFA1d-overexpression lines after drought treatment. We conclude that CsHSFA1d directly targets and regulates the expression of CsGolS3 and CsRS to promote the enzymatic activity and accumulation of RFO to increase the tolerance to drought stress. CsHSFA1d also improves ROS-scavenging enzyme activity and gene expression indirectly to reduce drought-induced ROS overaccumulation. This study therefore offers a new gene target to improve drought stress tolerance in cucumber and revealed the underlying mechanism by which CsHSFA1d functions in the drought stress by increasing the content of RFOs and scavenging the excessive accumulation of ROS.
Assuntos
Cucumis sativus , Galactosiltransferases , Regulação da Expressão Gênica de Plantas , Oligossacarídeos , Proteínas de Plantas , Plantas Geneticamente Modificadas , Rafinose , Espécies Reativas de Oxigênio , Cucumis sativus/genética , Cucumis sativus/fisiologia , Cucumis sativus/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Rafinose/metabolismo , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genética , Oligossacarídeos/metabolismo , Galactosiltransferases/metabolismo , Galactosiltransferases/genética , Secas , Fatores de Transcrição de Choque Térmico/metabolismo , Fatores de Transcrição de Choque Térmico/genética , Estresse Fisiológico/genéticaRESUMO
Gluten-free foods (GF) availability on supermarket shelves is growing and it is expected to continue expanding in the years ahead. These foods have been linked to a lower content of fermentable oligosaccharides, disaccharides, monosaccharides and polyols (FODMAPs), molecules that trigger gastrointestinal symptoms in sensitive persons. In this study, the FODMAP content of 25 cereal-based GF foods in Spain (breakfast cereals, pasta, bread, biscuits, bakery products, and dough and puff pastry) and 25 gluten-containing equivalents (GC) available in the same supermarket were analysed and compared. Lactose, fructose, glucose, sorbitol, mannitol, raffinose, stachyose and fructans were quantified. In a like-by-like analysis, GF foods were found to generally contain fewer FODMAPs than their GC counterparts. The ingredients used in the manufacture of GF cereal-based foods may contribute to this fact. When the individually wrapped size was considered, the proportion of samples classified as high-FODMAPs in GC and GF foods showed a trend towards fewer samples in the GF. However, not all the GF samples were low-FODMAP. Altogether, our findings provide essential information for FODMAP content databases of GF products in Spain.
Assuntos
Dieta Livre de Glúten , Dissacarídeos , Grão Comestível , Glutens , Monossacarídeos , Oligossacarídeos , Polímeros , Grão Comestível/química , Espanha , Monossacarídeos/análise , Glutens/análise , Oligossacarídeos/análise , Dissacarídeos/análise , Polímeros/análise , Fermentação , Frutanos/análise , Lactose/análise , Pão/análise , Humanos , Rafinose/análise , Frutose/análiseRESUMO
Mitogen-activated protein kinase (MAPK) cascades and raffinose have been observed to increase in plants exposed to cold. However, it remains elusive whether and how MAPK regulates raffinose synthesis under cold stress. Here, overexpression of SlMAPK3 promoted the accumulation of galactinol and raffinose under cold stress, while CRISPR/Cas9-mediated mutants showed the opposite results. Moreover, SlMAPK3 promoted the expression of SlWRKY46 at low temperatures and interacted with SlWRKY46 protein. Overexpression of SlWRKY46 enhanced cold resistance. Furthermore, SlWRKY46 directly bound to the promoter of SlGols1 to enhance its expression and promoted the accumulation of raffinose. Virus-induced gene-silencing (VIGS)-mediated knockdown of SlGols1 remarkably elevated cold sensitivity and reduced raffinose content. Meanwhile, exogenous supplementation of raffinose could improve the cold tolerance of tomato plants. Thus, our data indicates that SlMAPK3 modulates cold resistance by regulating raffinose content and SlWRKY46 expression. SlWRKY46 also promotes the accumulation of raffinose by inducing the expression of SlGols1.
Assuntos
Solanum lycopersicum , Rafinose/metabolismo , Proteínas de Plantas/metabolismo , Temperatura Baixa , Proteínas Quinases Ativadas por Mitógeno/genética , Regulação da Expressão Gênica de Plantas , Plantas Geneticamente Modificadas/metabolismoRESUMO
To investigate the role of sugar metabolism in desiccation-sensitive seeds, we performed a natural desiccation treatment on Phoebe chekiangensis seeds in a room and systematically analyzed the changes in seed germination, sugar compounds, malondialdehyde, and relative electrical conductivity during the seed desiccation. The results revealed that the initial moisture content of P. chekiangensis seed was very high (37.06%) and the seed was sensitive to desiccation, the germination percentage of the seed decreased to 5.33% when the seed was desiccated to 22.04% of moisture content, therefore, the seeds were considered recalcitrant. Based on the logistic model, we know that the moisture content of the seeds is 29.05% when the germination percentage drops to 50% and that it is desirable to keep the seed moisture content above 31.74% during ambient transportation. During seed desiccation, sucrose and trehalose contents exhibited increasing trends, and raffinose also increased during the late stage of desiccation, however, low levels of the non-reducing sugar accumulations may not prevent the loss of seed viability caused by desiccation. Glucose and fructose predominated among sugar compounds, and they showed a slight increase followed by a significant decrease. Their depletion may have contributed to the accumulation of sucrose and raffinose family oligosaccharides. Correlation analysis revealed a significant relationship between the accumulation of sucrose, trehalose, and soluble sugars, and the reduction in seed viability. Sucrose showed a significant negative correlation with glucose and fructose. Trehalose also exhibited the same pattern of correlation. These results provided additional data and theoretical support for understanding the mechanism of sugar metabolism in seed desiccation sensitivity.
Assuntos
Dessecação , Açúcares , Açúcares/metabolismo , Dessecação/métodos , Rafinose/metabolismo , Trealose/metabolismo , Sementes/metabolismo , Germinação , Sacarose/metabolismo , Glucose/metabolismo , Frutose/metabolismoRESUMO
BACKGROUND: Primary dysfunction and rejection are more common in donor liver tissues with steatosis. AMP-activated protein kinase (AMPK) assumes organ-protective functions during ischemia. Metformin was used for the activation of AMPK in hepatocytes. The aim of this study is to investigate the effectiveness of metformin administration for the reversal of cold-ischemia-induced damage in hepatosteatosis. MATERIAL AND METHODS: Seven-week-old C7BL56 male-mice (n = 109) were separated into four groups depending on diet type and metformin use. A specific diet model was followed for 10 weeks to induce hepatosteatosis. A group of the animals was administered with metformin for the last four weeks via oral gavage. After resection, the liver tissues were perfused and kept for 0-6-12-24 h in the UW solution. Histopathological examinations were performed, and Western blot was utilized to analyze p-AMPK and AMPK expression levels. RESULTS: Hepatosteatosis decreased significantly with metformin. The steatotic liver group had more prominent pericentral inflammation, necrosis as well as showing a decreased and more delayed AMPK response than the non-fat group. All these alterations could be corrected using metformin. CONCLUSION: Metformin can increase the resistance of livers with hepatosteatosis to cold-ischemia-induced damage, which in turn may pave the way for successful transplantation of fatty living-donor livers.
Assuntos
Fígado Gorduroso , Transplante de Fígado , Metformina , Soluções para Preservação de Órgãos , Traumatismo por Reperfusão , Masculino , Camundongos , Animais , Humanos , Metformina/farmacologia , Metformina/uso terapêutico , Proteínas Quinases Ativadas por AMP/metabolismo , Doadores Vivos , Fígado/patologia , Fígado Gorduroso/tratamento farmacológico , Fígado Gorduroso/etiologia , Glutationa , Rafinose , Alopurinol , Insulina , AdenosinaRESUMO
Vascularized composite allotransplantation (VCA) represents a promising reconstructive solution primarily conducted to improve quality of life. However, tissue damage caused by cold-ischemia (CI) storage prior to transplant represents a major factor limiting widespread application. This study investigates the addition of the novel free radical scavenger PrC-210 to UW Organ Preservation Solution (UW Solution) to suppress CI-induced skeletal muscle injury in a rat hind limb amputation model. Lewis rats received systemic perfusion of UW solution +/- PrC-210 (0 mM control, 10 mM, 20 mM, 30 mM, or 40 mM), followed by bilateral transfemoral amputation. Limbs were stored in 40 mL of the same perfusate at 4 °C for 48 h. Muscle punch biopsies were taken at set times over the 48 h cold-storage period and analyzed for caspase-3,7 activity, cytochrome C levels, and qualitative histology. A single 15 s perfusion of PrC-210-containing UW Solution conferred a dose-dependent reduction in CI-induced muscle cell death over 48 h. In the presence of PrC-210, muscle cell mitochondrial cytochrome C release was equivalent to 0 h controls, with profound reductions in the caspase-3,7 apoptotic marker that correlated with limb histology. PrC-210 conferred complete prevention of ROS-induced mitochondrial lysis in vitro, as measured by cytochrome C release. We conclude that the addition of 30 mM PrC210 to UW Solution conferred the most consistent reduction in CI limb damage, and it warrants further investigation for clinical application in the VCA setting.
Assuntos
Aloenxertos Compostos , Diaminas , Soluções para Preservação de Órgãos , Traumatismo por Reperfusão , Compostos de Sulfidrila , Ratos , Animais , Sequestradores de Radicais Livres , Caspase 3 , Aloenxertos Compostos/patologia , Citocromos c , Qualidade de Vida , Ratos Endogâmicos Lew , Glutationa/farmacologia , Alopurinol/farmacologia , Insulina/farmacologia , Isquemia , Preservação de Órgãos , Temperatura Baixa , Traumatismo por Reperfusão/patologia , Rafinose , AdenosinaRESUMO
BACKGROUND: The composition of organ preservation solutions is crucial for maintaining graft integrity and early graft function after transplantation. The aim of this study is to compare new organ preservation solution PERLA® with the gold standard preservation solution University of Wisconsin (UW) regarding oxidative stress and early graft injury. METHODS: In order to assess oxidative stress after cold storage, kidney grafts have been preserved for 18 h at 4° C in either UW solution or PERLA® solution and then assessed for oxidative stress injury (protocol 1). To assess kidney injuries and oxidative stress after reperfusion, rat kidneys were harvested, stored in cold UW or in PERLA® solutions for 18 h at 4 °C and then transplanted heterotopically for 6 h (protocol 2). PERLA® is a high Na+/low K+ solution including PEG-35 (1 g/L), trimetazidine (1 µM), carvedilol (10 µM) and tacrolimus (5 µM). RESULTS: Our results showed that preservation of kidneys in PERLA® solution significantly attenuates oxidative stress parameters after cold storage and reperfusion. We found a significant decrease in oxidative damage indicators (MDA, CD and CP) and a significant increase in antioxidant indicators (GPx, GSH, CAT, SOD and PSH). Moreover, PERLA® solution decreased kidney injury after reperfusion (creatinine, LDH and uric acid). CONCLUSION: PERLA® solution was more effective than UW storage solution in preserving rat's kidney grafts.
Assuntos
Transplante de Rim , Soluções para Preservação de Órgãos , Traumatismo por Reperfusão , Humanos , Ratos , Animais , Transplante de Rim/efeitos adversos , Traumatismo por Reperfusão/prevenção & controle , Traumatismo por Reperfusão/metabolismo , Soluções para Preservação de Órgãos/farmacologia , Rim/metabolismo , Alopurinol/farmacologia , Estresse Oxidativo , Adenosina , Glutationa , Insulina , RafinoseRESUMO
Plasmodium vivax malaria is increasingly recognized as a major global health problem and the socio-economic impact of P.vivax-induced burden is huge. Vaccine development against P. vivax malaria has been hampered by the lack of an in vitro culture system and poor access to P. vivax sporozoites. The recent generation of Plasmodium falciparum parasites that express a functional P. vivax AMA1 molecule has provided a platform for in vitro evaluation of PvAMA1 as a potential blood stage vaccine. Three so-called PvAMA1 Diversity Covering (DiCo) proteins were designed to assess their potential to induce a functional and broad humoral immune response to the polymorphic PvAMA1 molecule. Rabbits were immunized with the mixture of three, Pichia-produced, PvAMA1 DiCo proteins, as well as with 2 naturally occurring PvAMA1 alleles. For these three groups, the experimental adjuvant raffinose fatty acid sulfate ester (RFASE) was used, while in a fourth group the purified main mono-esterified constituent (RSL10) of this adjuvant was used. Animals immunized with the mixture of the three PvAMA1 DiCo proteins in RFASE showed high anti-PvAMA1 antibody titers against three naturally occurring PvAMA1variants while also high growth-inhibitory capacity was observed against P. falciparum parasites expressing PvAMA1. This supports further clinical development of the PvAMA1 DiCo mixture as a potential malaria vaccine. However, as the single allele PvAMA1 SalI-group showed similar characteristics in antibody titer and inhibition levels as the PvAMA1 DiCo mixture-group, this raises the question whether a mixture is really necessary to overcome the polymorphism in the vaccine candidate. RFASE induced strong humoral responses, as did the animals immunized with the purified component, RSL10. This suggests that RSL10 is the active ingredient. However, one of the RSL10-immunized animal showed a delayed response, necessitating further research into the clinical development of RSL10.
Assuntos
Vacinas Antimaláricas , Malária Falciparum , Malária Vivax , Parasitos , Animais , Coelhos , Proteínas de Protozoários/genética , Plasmodium vivax , Rafinose , Sulfatos , Proteínas de Membrana/genética , Antígenos de Protozoários/genética , Adjuvantes Imunológicos , Malária Falciparum/prevenção & controle , Plasmodium falciparum , Malária Vivax/prevenção & controle , Anticorpos AntiprotozoáriosRESUMO
For domestic cats ovaries, recommended cold-storage limit is 24â¯h in Phosphate Buffered Saline (PBS) or Dulbecco`s PBS (DPBS). Here, we attempted to verify wheatear cat ovaries may benefit from more complex solutions during prolonged cold-storage (>24â¯h). First, the preservation capabilities of extracellular (SP+), intracellular (UW) solutions and DPBS supplemented with glutathione (DPBS+GSH) were compared using ovary fragments from the same ovary (n=10). Intact ovary stored in DPBS served as a control. Ovaries were kept at 4⯰C for 48â¯h, and 72â¯h. In the second experiment, first ovary was stored in DPBS, second in SP+ or UW solution for 48â¯h (nâ¯=â¯12). Ovaries pairs stored in DPBS for 24â¯h served as a control (n=8). Tissue samples were evaluated directly after cold-storage and after following 24â¯h in vitro culture. Ovarian follicle morphology, apoptosis rates (cleaved caspase-3, TUNEL), and follicular growth activation (Ki-67) were assessed. Ovary fragmentation impaired follicular morphology preservation upon cold-storage comparing to intact ovary. However, ovarian fragments stored in UW for 48â¯h and in SP+ for 72â¯h presented better morphology than DPBS+GSH group. Comparison of intact ovaries cold-storage for 48â¯h showed that SP+ provided superior follicular morphology over DPBS, and it was comparable to the outcome of 24-hour storage. No follicular activation after in vitro culture was observed. Nevertheless, tissue culture increased considerably caspase-3 cleavage and TUNEL detection. The ovary fragmentation prior to cold-storage is not recommended in domestic cats. Replacement of DPBS with SP+ solution for whole ovary and UW solution for ovarian tissue fragments improves follicular structure preservation during 48-hour cold-storage.
Assuntos
Soluções para Preservação de Órgãos , Ovário , Feminino , Animais , Gatos , Ovário/fisiologia , Caspase 3 , Folículo Ovariano/fisiologia , Glutationa , Rafinose , Alopurinol , Insulina , AdenosinaRESUMO
The University of Wisconsin (UW) solution is the most effective preservation solution currently used; however, to safely use expanded-criteria donor grafts, a new cold storage solution that alleviates graft injury more effectively is required. We prepared a heavy water (D2O)-containing buffer, Dsol, and observed strong protective effects during extended cold storage of rat hearts and livers. In the current study, we modified Dsol (mDsol) and tested its efficacy. The aim of the present study was to determine whether mDsol could protect the rat liver more effectively than the UW solution and to clarify the roles of D2O and deferoxamine (DFX). Rat livers were subjected to cold storage for 48 hours in test solutions: UW, mDsol, mDsol without D2O or DFX (mDsol-D2O[-], mDsol-DFX[-]), and subsequently reperfused on an isolated perfused rat liver for 90 minutes at 37°C. In the UW group, the liver was dehydrated during cold storage and rapidly expanded during reperfusion. Accordingly, the cumulative weight change was the highest in the UW group, together with augmented portal veinous resistance and ALT leakage and decreased oxygen consumption rate and bile production. These changes were significantly suppressed in the mDsol-treated group. In the mDsol-D2O(-) and mDsol-DFX(-) groups offered partial protection. In conclusion, mDsol appeared to be superior to the UW solution for simple cold storage of the rat liver, presumably due to improved microcirculation in the early phase of reperfusion. Both heavy water and deferoxamine are essential for alleviating seamless organ swelling that occurs during cold storage and subsequent reperfusion.
Assuntos
Transplante de Fígado , Soluções para Preservação de Órgãos , Humanos , Ratos , Animais , Óxido de Deutério/farmacologia , Desferroxamina/farmacologia , Fígado , Soluções para Preservação de Órgãos/farmacologia , Reperfusão , Glutationa/farmacologia , Alopurinol/farmacologia , Insulina/farmacologia , Rafinose/farmacologia , Preservação de Órgãos , AdenosinaRESUMO
The milk fat globule membrane (MFGM) can protect probiotic bacteria from bile stress. However, its potential mechanism has not been reported. In this study, the viability, morphology and gene transcriptional response of Bifidobacterium longum ssp. infantis ATCC 15697 (BI_15697) stressed by bile salts with or without MFGM were investigated. It was shown that MFGM alleviated the reduction in BI_15697 population induced by 0.2% porcine bile stress and restored the population to the control levels. MFGM ameliorated the shrunken, fragmented appearance and irregular morphology of BI_15697 and maintained cell integrity disrupted by bile stress. RNA-sequencing results showed that MFGM increased transport of glucose and raffinose and decreased that of branched-chain amino acids (BCAA) in the presence of bile salts. MFGM stimulated the expression of genes involved in the synthesis of raffinose in galactose metabolism and the metabolism of BCAA, suggesting that MFGM stimulated the accumulation of raffinose and BCAA in the presence of bile. In addition, MFGM stimulated the expression of 2 bile efflux transporters under bile stress. Together, the multifactorial response helps BI_15697 excrete bile salts and maintain cellular integrity in response to bile stress. This study proposes a mechanism for the protection of BI_15697 against bile salt stress by MFGM, thereby providing a molecular basis for its application in incorporation of probiotics.
Assuntos
Bifidobacterium longum subspecies infantis , Bile , Glicoproteínas , Animais , Suínos , Rafinose , Glicolipídeos , Gotículas Lipídicas , Ácidos e Sais BiliaresRESUMO
The Raffinose Family of Oligosaccharides (RFOs), including Galactinol, Raffinose, and Stachyose, are pivotal carbohydrates with significant roles in abiotic stress tolerance and growth within dynamic environments. Plant development is profoundly influenced by light, a major environmental signal. Despite this, the interconnections between the biosynthesis of secondary sugars and light signaling have remained unexplored. This study reveals that exposure to light induces the expression of Galactinol synthase (AtGolS1), a key enzyme in the RFO biosynthesis pathway. The light-inducible response of AtGolS1 operates downstream of ELONGATED HYPOCOTYL 5 (HY5), a central regulator in light signaling. Mutant seedlings with disrupted HY5 function (hy5-215) exhibit reduced AtGolS1 transcript accumulation compared to wild-type (WT) and HY5 overexpression seedlings. DNA-protein interaction studies demonstrate that HY5 directly binds to light-responsive cis-elements in the promoter region of AtGolS1, thereby mediating its light responsiveness. Quantification of galactinol revealed a diminished accumulation in the hy5-215 mutant compared to wild-type (WT) and HY5 overexpression seedlings. Consequently, these findings shed light on the intricate crosstalk between RFO biosynthesis and light signaling in Arabidopsis.
Assuntos
Proteínas de Arabidopsis , Arabidopsis , Dissacarídeos , Galactosiltransferases , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Fatores de Transcrição de Zíper de Leucina Básica/genética , Fatores de Transcrição de Zíper de Leucina Básica/metabolismo , Regulação da Expressão Gênica de Plantas , Hipocótilo/metabolismo , Rafinose/metabolismo , Plântula/genética , Plântula/metabolismoRESUMO
Pulmonary infections caused by multidrug-resistant bacteria have become a significant threat to human health. Bacterial biofilms exacerbate the persistence and recurrence of pulmonary infections, hindering the accessibility and effectiveness of antibiotics. In this study, a dry powder inhalation (DPI) consisting of polymyxin B sulfate (PMBS) inhalable microparticles and high-lectin-affinity (HLA) sugar (i.e., raffinose) carriers was developed for treating pulmonary infections and targeting bacterial lectins essential for biofilm growth. The formulated PMBS-HLA DPIs exhibited particle sizes of approximately 3 µm, and surface roughness varied according to the drug-to-carrier ratio. Formulation F5 (PMBS: raffinose = 10:90) demonstrated the highest fine particle fraction (FPF) value (64.86%), signifying its substantially enhanced aerosol performance, potentially attributable to moderate roughness and smallest mass median aerodynamic particle size. The efficacy of PMBS-HLA DPIs in inhibiting biofilm formation and eradicating mature biofilms was significantly improved with the addition of raffinose, suggesting the effectiveness of lectin-binding strategy for combating bacterial biofilm-associated infections. In rat models with acute and chronic pulmonary infections, F5 demonstrated superior bacterial killing and amelioration of inflammatory responses compared to spray-dried PMBS (F0). In conclusion, our HLA carrier-based formulation presents considerable potential for the efficient treatment of multidrug-resistant bacterial biofilm-associated pulmonary infections.