Manganese Superoxide Dismutase Research Articles

Deoxynivalenol induces spleen damage, apoptosis, and inflammation in mice by increasing mitochondrial reactive oxygen species: Protective effects of curcumin.

Deoxynivalenol (DON), a Fusarium mycotoxin, causes spleen apoptosis and inflammation, which damage the organ. Curcumin (Cur) is a member of the ginger family. It has anti-apoptotic and anti-inflammatory effects that maintain the health of the organism's immune system. Here, the protective effects of Cur against DON-induced spleen damage were explored. First, we found DON (2.4mg/kg body weight) decreased the expression of manganese superoxide dismutase, mitochondrial membrane potential, adenosine triphosphate, and disturbed hematoxylin and eosin staining in mice spleen. The results confirmed that DON causes mitochondrial reactive oxygen species (mtROS) overproduction leading to spleen damage. Second, we found DON decreased the expression of mitochondrial apoptosis-inducing factor (AIF) and B-cell lymphoma-2 (Bcl-2), and increased the expression of nuclear AIF, Bcl2-associated X (Bax), cysteine-aspartate protease-3 (caspase-3), caspase-9. Mitoquinone is a mitochondria-targeted antioxidant that can prevent of mitochondrial oxidative damage. These expression increases were not observed in the mitoquinone-treated group, confirming that mtROS was an upstream regulatory target of apoptosis and inflammation in DON-exposed mice spleens. Finally, we confirmed that Cur (50 or 100mg/kg body weight) attenuated DON-induced apoptosis and inflammation by inactivating mtROS. Collectively, these results confirm that DON causes spleen damage by increasing mtROS, and the protective effects of curcumin.

Effects of Enterocytozoon hepatopenaei infection on intestinal immune defense and physiological processes of Penaeus vannamei

As the main pathogen causing growth retardation, EHP is considered to be mainly parasitic in the hepatopancreas of shrimp. However, the intestines of shrimp infected with EHP frequently exhibit syndromes such as jejunum and white midgut. Therefore, the challenge experiment was carried out in this study to compare the differences in intestinal histology, digestion and absorption, immune defense and oxidative stress of P. vannamei between the control group and EHP infection group. Histological analysis showed that EHP infection significantly damaged the intestine of the shrimp, including intestinal villus rupture and outer membrane impairment. Concurrently, EHP infection can trigger intestinal immune response, and the expression of key immune genes like Toll, myeloid differentiation factor, anti-lipopolysaccharide factor, and Relish was significantly enhanced, while the expression of IMD and alkaline phosphatase was suppressed. Additionally, antioxidant genes manganese superoxide dismutase, catalase, glutathione peroxidase, glutathione-S-transferase, and nuclear factor E2-related factor 2 were up-regulated to varying extents in EHP infection group, and the contents of lipid peroxides and malondialdehyde were heavily accumulated. Moreover, the expression levels of key genes involved in nutrient absorption, transport and synthesis, such as glucose transporter 1, Na+-K+ATPase, fatty acid synthase, acetyl-CoA carboxylase, rapamycin kinase, mTOR regulation-related protein, eukaryotic translation initiation factor 4E binding protein, ribosomal protein S6 kinase, were significantly up-regulated. However, the activities of amylase, lipase, and trypsin were inhibited in EHP infection group throughout the experiment. In summary, EHP infection damaged the intestine of P. vannamei, accompanied by immune response and oxidative stress. At the same time, nutrient transport and synthesis pathways were activated, while digestive enzyme activities were inhibited, indicating that in order to maintain survival, shrimps must accelerate material transport. Unfortunately, it remains in a state of nutrient deficiency that ultimately affects growth.

Integrating Forensic Autopsies with Proteomic Profiling for Suicide Risk Assessment: A Comprehensive Review of Literature.

Suicide is a major global public health concern that affects people of all ages, with over 700000 individuals intentionally ending their lives every year. Suicide is a multifactorial event related to multiple risk factors interlocking with each other, among which neurobiological factors are considered to be an objective measure of the incidence of this phenomenon and can be used as a measurable tool for evaluating suicidal tendencies. The aim of this study is to thoroughly examine available data and assess candidate proteins as prospective biomarkers for predicting suicides and ascertaining the manner of death in forensic cases. An electronic search was conducted on PubMed, Science Direct Scopus, and the Excerpta Medica Database. The systematic review adhered to PRISMA guidelines and encompassed case series, prospective and retrospective studies, and short communications published in English. The focus was on proteomics and suicide, specifically, those studies where researchers conducted human proteomic analyses on specimens obtained from individuals who completed or attempted suicide. A total of 14 studies met the inclusion criteria, resulting in a dataset of numerous candidate protein biomarkers. These include tenascin-C, potassium voltage-gated channel subfamily Q member 3, vimentin-immunoreactive astrocytes, glutathione S-transferase theta 1, iron transport proteins, Acrystallin chain B, manganese superoxide dismutase, glial fibrillary acidic protein, various glycolytic pathway proteins, 14-3-3 eta and 14-3-3 theta proteins, specific cytoskeleton proteins, C-reactive protein, serum amyloid A protein 1, extrinsic coagulation pathway proteins, the vacuolar-type proton pump ATPase subunit, plasma apolipoprotein A-IV, and ER stress proteins. These proteins are proposed as a panel of biomarkers to be evaluated in conjunction with other clinical predictors of suicide. This review aims to provide a comprehensive summary of all proteomic studies conducted on cases of attempted or completed suicide. By doing so, it seeks to bridge existing gaps in knowledge and pave the way for future investigations. The ultimate goal is to potentially identify a suicide biomarker.

Effects of vitamin D and common nettle (Urtica dioica) extract administration on Mn-SOD and Catalase (CAT) secretion in the colon tissues of rats with experimentally induced Crohn's disease.

We have examined some effects of administering vitamin D and extract of common nettle (Urtica dioica) to rats with experimentally induced Crohn's disease (CR). Body weight and colon length were lower in the CR group than in normal controls, whereas scores for histopathologic changes seen in sections stained by the H&E and PAS methods were lower in rats with CR than in those that also received either vitamin D (CRD) or nettle extract (CRI). Strong manganese-superoxide dismutase (Mn-SOD) immunoreactivity was detected in the crypt epithelium of the CR and CRI groups and in the lymphoid tissue of the CRD group. Weak catalase (CAT) immunoreactivity in the crypt epithelium in the CR, CRI, and CRD groups and strong CAT immunoreactivity in the lymphoid tissue in the CR group were also observed. Our results reveal that administering either vitamin D and common nettle extract can have augment Mn-SOD and CAT expression in colon tissues and contribute to alleviation of some complications of experimental Crohn's disease.

White Tea Aqueous Extract: A Potential Anti-Aging Agent Against High-Fat Diet-Induced Senescence in Drosophila melanogaster.

White tea has been scientifically proven to exhibit positive biological effects in combating chronic diseases, including cancer, metabolic syndrome, etc. Nevertheless, the anti-aging activity and mechanism of white tea on organisms exposed to a high-fat diet remain unexplored. Herein, we prepared a white tea aqueous extract (WTAE) from white peony in Fuding and assessed its in vivo antioxidant and anti-aging effects by employing a Drosophila melanogaster senescence model induced by lard, delving into the underlying molecular mechanisms through which the WTAE contributes to lifespan improvement. Notably, the WTAE significantly extended the lifespan of Drosophila fed a high-fat diet and partially restored the climbing ability of Drosophila on a high-fat diet, accompanied by increased activities of copper-zinc superoxide dismutase, manganese-superoxide dismutase, and catalase and decreased lipid hydroperoxide levels in Drosophila. Furthermore, transcriptomic analysis indicated that the WTAE countered aging triggered by a high-fat diet via activating oxidative phosphorylation, neuroactive ligand-receptor interactions, and more pathways, as well as inhibiting circadian rhythm-fly, protein processing in the endoplasmic reticulum, and more pathways. Our findings suggest that WTAE exhibits excellent inhibitory activity against high-fat diet-induced senescence and holds promising potential as an anti-aging agent that can be further developed.

6-Gingerol Induced Apoptosis and Cell Cycle Arrest in Glioma Cells via MnSOD and ERK Phosphorylation Modulation.

6-gingerol, a bioactive compound from ginger, has demonstrated promising anticancer properties across various cancer models by inducing apoptosis and inhibiting cell proliferation and invasion. In this study, we explore its mechanisms against glioblastoma multiforme (GBM), a notably aggressive and treatment-resistant brain tumor. We found that 6-gingerol crosses the blood-brain barrier more effectively than curcumin, enhancing its potential as a therapeutic agent for brain tumors. Our experiments show that 6-gingerol reduces cell proliferation and triggers apoptosis in GBM cell lines by disrupting cellular energy homeostasis. This process involves an increase in mitochondrial reactive oxygen species (mtROS) and a decrease in mitochondrial membrane potential, primarily due to the downregulation of manganese superoxide dismutase (MnSOD). Additionally, 6-gingerol reduces ERK phosphorylation by inhibiting EGFR and RAF, leading to G1 phase cell cycle arrest. These findings indicate that 6-gingerol promotes cell death in GBM cells by modulating MnSOD and ROS levels and arresting the cell cycle through the ERFR-RAF-1/MEK/ ERK signaling pathway, highlighting its potential as a therapeutic agent for GBM and setting the stage for future clinical research.

7-Hydroxy-3-(4'-methoxyphenyl) coumarin (C12) attenuates bleomycin-induced acute lung injury and fibrosis through activation of SIRT3.

Silent mating-type information regulation 2 homology 3 (SIRT3) is a member of the sirtuins family expressed in mitochondria performs deacetylation of metabolic enzymes and promotes longevity. 7-hydroxy-3-(4'-methoxyphenyl) coumarin (C12) is a small molecule first ever known for its direct activation of SIRT3. SIRT3 performs its function by balancing the redox system by activating manganese superoxide dismutase (MnSOD) and 8-Oxoguanine glycosylase (OGG1). For the first time, we reported that activation of SIRT3 by C12 attenuated bleomycin (BLM)-)-induced acute lung injury and pulmonary fibrosis. C12 prevented the oxidative stress and injury caused by BLM in alveolar epithelial cells (BEAS-2B) in in vitro and inhibited the fibrosis in transforming growth factor-beta (TGF-β) induced fibrosis in fibroblasts (MRC-5). Additionally, activation of SIRT3 by C12 in vivo mice model alleviated BLM-induced inflammation, collagen accumulation, cellular infiltration, and restoration of alveolar architecture by inhibiting TGF-β, smooth muscle actin (α-SMA), collagen-1A, collagen-3A, and mesenchymal markers. The protective effect of C12 was through activation of MnSOD and OGG1 in both in vitro and in vivo models suggesting C12 can be a potent SIRT3 activator and helps to treat fibrotic-related diseases.

Ginkgo biloba for Tardive Dyskinesia and Plasma MnSOD Activity: Association with MnSOD Ala-9Val Variant: A Randomized, Double-blind Trial.

Excessive free radicals are implicated in the pathophysiology of tardive dyskinesia (TD), and Ginkgo biloba extract (EGb761) scavenges free radicals, thereby enhancing antioxidant enzymes such as mitochondrial manganese superoxide dismutase (MnSOD). This study examined whether EGb761 treatment would improve TD symptoms and increase MnSOD activity, particularly in TD patients with specific MnSOD Val-9Ala genotype. An EGb761 (240 mg/day) 12-week double-blind clinical trial with 157 TD patients was randomized. The severity of TD was measured by the Abnormal Involuntary Movement Scale (AIMS) and plasma MnSOD activity was assayed before and after 12 weeks of treatment. Further, in an expanded sample, we compared MnSOD activity in 159 TD, 227 non-TD and 280 healthy controls, as well as the allele frequencies and genotypes for the MnSOD Ala-9Val polymorphism in 352 TD, 486 non-TD and 1150 healthy controls. EGb761 significantly reduced TD symptoms and increased MnSOD activity in TD patients compared to placebo (both p < 0.01). Moreover, we found an interaction between genotype and treatment response (p < 0.001). Furthermore, in the EGb761 group, patients carrying the Ala allele displayed a significantly lower AIMS total score than patients with the Val/Val genotype. In addition, MnSOD activity was significantly lower at baseline in TD patients compared with healthy controls or non-TD patients. EGb761 treatment enhanced low MnSOD activity in TD patients and produced greater improvement in TD symptoms in patients with the Ala allele of the MnSOD Ala-9Val polymorphism.

Exogenous MnSO4 Improves Productivity of Degenerated Volvariella volvacea by Regulating Antioxidant Activity.

Manganese is one of the trace elements necessary for organisms to maintain normal biological activities and is also a cofactor for manganese superoxide dismutase (Mn-SOD) and manganese peroxidase (MnP). In order to find a simple and effective method to rejuvenate the degenerated V. volvacea strains, we explored the effect of the exogenous addition of MnSO4 on the antioxidant vigour and productivity of degenerated strains of V. volvacea. The results showed that the exogenous MnSO4 had no significant effect on the non-degenerated strain T0, but it could effectively increase the mycelial growth rate, mycelial biomass, and LBL decolouring ability of the degenerated strains T10 and T19, and reduce the production cycle and increased the biological efficiency of T10; it helped the severely degenerated T19 to regrow its fruiting body; and it also significantly increased the viability of the matrix-degrading enzymes such as EG, Lac, Xyl, etc. of T10 and T19. Meanwhile, exogenous MnSO4 significantly increased the activity of GPX, GR, CAT, SOD, and the content of GSH, polyphenols, minerals, and polysaccharides in T10 and T19 strains, which resulted in a significant decrease in the accumulation of ROS, such as O2- and H2O2 in T10 and T19. The correlation analysis showed that there was a significant correlation between antioxidant activity and the production ability of V. volvacea. This study can provide theoretical reference and technical support for the rejuvenation research of degenerated strains of V. volvacea and other edible fungi.

Resynthesis of Damaged Fe-S Cluster Proteins Protects Aspergillus fumigatus Against Oxidative Stress in the Absence of Mn-Superoxide Dismutase.

The importance of manganese superoxide dismutase (Mn-SOD), an evolutionarily ancient metalloenzyme that maintains the integrity and function of mitochondria, was studied in oxidative stress-treated Aspergillus fumigatus cultures. Deletion of the Mn-SOD gene (sodB) increased both the menadione sodium bisulfite (MSB)-elicited oxidative stress and the deferiprone (DFP)-induced iron limitation stress sensitivity of the strain. Moreover, DFP treatment enhanced the MSB sensitivity of both the gene deletion mutant and the reference strain. The lack of SodB also increased the susceptibility of conidia to killing by human macrophages. Concurring with the stress sensitivity data, RNS sequencing data also demonstrated that the deletion of sodB largely altered the MSB-induced oxidative stress response. The difference between the oxidative stress responses of the two strains manifested mainly in the intensity of the response. Importantly, upregulation of "Ribosome protein", "Iron uptake", and "Fe-S cluster assembly" genes, alterations in the transcription of "Fe-S cluster protein" genes, and downregulation of "Heme binding protein" genes under MSB stress were characteristic only for the ΔsodB gene deletion mutant. We assume that the elevated superoxide level generated by MSB treatment may have destroyed Fe-S cluster proteins of mitochondria in the absence of SodB. This intensified the resynthesis of Fe-S cluster proteins, which was accompanied with enhanced translation and iron acquisition, leading to increased DFP sensitivity.

MnSOD Mimetics in Therapy: Exploring Their Role in Combating Oxidative Stress-Related Diseases.

Reactive oxygen species (ROS) are double-edged swords in biological systems-they are essential for normal cellular functions but can cause damage when accumulated due to oxidative stress. Manganese superoxide dismutase (MnSOD), located in the mitochondrial matrix, is a key enzyme that neutralizes superoxide radicals (O2•-), maintaining cellular redox balance and integrity. This review examines the development and therapeutic potential of MnSOD mimetics-synthetic compounds designed to replicate MnSOD's antioxidant activity. We focus on five main types: Mn porphyrins, Mn salens, MitoQ10, nitroxides, and mangafodipir. These mimetics have shown promise in treating a range of oxidative stress-related conditions, including cardiovascular diseases, neurodegenerative disorders, cancer, and metabolic syndromes. By emulating natural antioxidant defenses, MnSOD mimetics offer innovative strategies to combat diseases linked to mitochondrial dysfunction and ROS accumulation. Future research should aim to optimize these compounds for better stability, bioavailability, and safety, paving the way for their translation into effective clinical therapies.

Post-sepsis chronic muscle weakness can be prevented by pharmacological protection of mitochondria.

Sepsis, mainly caused by bacterial infections, is the leading cause of in-patient hospitalizations. After discharge, most sepsis survivors suffer from long-term medical complications, particularly chronic skeletal muscle weakness. To investigate this medical condition in detail, we previously developed a murine severe sepsis-survival model that exhibits long-term post-sepsis skeletal muscle weakness. While mitochondrial abnormalities were present in the skeletal muscle of the sepsis surviving mice, the relationship between abnormal mitochondria and muscle weakness remained unclear. Herein, we aimed to investigate whether mitochondrial abnormalities have a causal role in chronic post-sepsis muscle weakness and could thereby serve as a therapeutic target. Experimental polymicrobial abdominal sepsis was induced in 16-18months old male and female mice using cecal slurry injection with subsequent antibiotic and fluid resuscitation. To evaluate the pathological roles of mitochondrial abnormalities in post-sepsis skeletal muscle weakness, we utilized a transgenic mouse strain overexpressing the mitochondria-specific antioxidant enzyme manganese superoxide dismutase (MnSOD). Following sepsis development in C57BL/6 mice, we evaluated the effect of the mitochondria-targeting synthetic tetrapeptide SS-31 in protecting mitochondria from sepsis-induced damage and preventing skeletal muscle weakness development. In vivo and in vitro techniques were leveraged to assess muscle function at multiple timepoints throughout sepsis development and resolution. Histological and biochemical analyses including bulk mRNA sequencing were used to detect molecular changes in the muscle during and after sepsis RESULTS: Our time course study revealed that post sepsis skeletal muscle weakness develops progressively after the resolution of acute sepsis and in parallel with the accumulation of mitochondrial abnormalities and changes in the mitochondria-related gene expression profile. Transgenic mice overexpressing MnSOD were protected from mitochondrial abnormalities and muscle weakness following sepsis. Further, pharmacological protection of mitochondria utilizing SS-31 during sepsis effectively prevented the later development of muscle weakness. Our study revealed that the accumulation of mitochondrial abnormalities is the major cause of post-sepsis skeletal muscle weakness. Pharmacological protection of mitochondria during acute sepsis is a potential clinical treatment strategy to prevent post-sepsis muscle weakness.

Molecular identification and characterization of the superoxide dismutase (SOD) gene family in Tetranychus urticae (Acari: Tetranychidae) and the role of TuSOD2 gene under short-term heat stress

Superoxide dismutases (SODs) are key enzymes for scavenging insects' excess reactive oxygen species (ROS) and play a crucial role in resisting a variety of stresses. The deletion of SOD genes can alter the response of insects to abiotic stresses. In this study, four genes of the superoxide dismutase family in Tetranychus urticae were identified and analyzed, among which three copper/zinc SODs and a manganese SOD. The result of quantitative real-time PCR showed that TuSOD2 expression level was up-regulated significantly under short-term heat stress, but TuSOD1, TuSOD2, and TuSOD4 were down-regulated or no significant difference, which indicated that T. urticae responded to oxidative stress induced by heat stress by raising the expression level of superoxide dismutase gene. Among them, TuSOD2 gene plays a key role in response to heat stress in T. urticae, and other SOD genes may be responsible for other oxidative stresses. RNA interference experiments showed that the knocking down of TuSOD2 reduced the activities of various antioxidant enzymes, resulting in a decline in the tolerance to high temperature and an increase in the mortality of T. urticae under short-term heat stress. To summarize, SOD, especially TuSOD2 plays an important role in treating short-term heat stress in T. urticae.

Low protein diet influences mineral absorption and utilization in medium-growing yellow-feathered broilers from 1 to 30 days of age

Reduced-protein diet can save protein ingredients and reduce nitrogen (N) losses. However, the effect of low protein diet on the mineral uptake and utilization in broilers needs to be explored. The aim of this study was to investigate the effect of low-protein diet on the growth performance, N deposition, mineral contents in serum, tissues and excreta, and the activities and gene expression of related enzymes in tissues of medium-growing yellow-feathered broilers, so as to elucidate the relationship between dietary protein level and the absorption and utilization of minerals in broilers. A total of 72 1-d-old Spotted-Brown male broilers were randomly allotted to 1 of 2 treatments with 6 replicate cages of 6 birds per cage for each treatment. The dietary crude protein (CP) levels for the two treatments were 21 % (the control treatment) and 19 % (low protein treatment), respectively. The experimental period was 30 d. The results showed that no differences (P > 0.05) were detected in average daily gain, average daily feed intake and feed: gain ratio of broilers during 1 to 30 d between the two treatments. However, low protein intake increased (P < 0.05) N retention rate, serum P, Cu and Mn, and excreta Cu, Mn and Zn, and decreased (P < 0.05) liver P and excreta P. In addition, birds fed low protein diet had higher (P < 0.05) manganese superoxide dismutase, and total superoxide dismutase activities in liver, and total antioxidant capacity and malondialdehyde content in heart, and lower (P < 0.05) copper-zinc superoxide dismutase (CuZnSOD) and succinate dehydrogenase activities in liver and CuZnSOD mRNA level in heart. In conclusion, the reduction of dietary CP content from 21 % to 19 % improved N retention, the absorption of P, Cu and Mn, as well as the antioxidant ability of liver and heart, and influenced metabolic utilization of P, Cu, Zn, Fe and Mn in medium-growing yellow-feathered broilers from 1 to 30 d of age.

TP53 Codon 72 Polymorphism Impacts Macrophage Activation through Reactive Oxygen Species-Dependent Cell Signaling Alterations.

The role of the most common TP53 single-nucleotide polymorphism (SNP) at codon 72, which encodes for proline (P72) or arginine (R72), in the regulation of the immune system has not yet been thoroughly explored. We found that this SNP contributes to aggravated inflammatory response in COVID-19 patients resulting from biased macrophage activation. R72-P53 inhibits mitochondrial manganese superoxide dismutase, leading to impaired reactive oxygen species scavenging, oxidation of phosphatase and tensin homolog deleted on chromosome 10 (PTEN), and, consequently, its inhibition. Reduced PTEN activity causes constitutive activation of the PI3K/Akt pathway, which restricts proinflammatory (M1) and promotes anti-inflammatory (M2) phenotypes through NF-κB and p53 inhibition. In contrast, PTEN-reduced PI3K/Akt activity, in P72 carrying cells, favors M1 phenotypes. LPS-stimulated R72 macrophages fail to reduce tumor growth in a mouse model of cancer, in contrast with P72 macrophages, which preserve M1 phenotype invivo and reduce tumor growth by enhancing antitumor T cell responses, consistent with antitumor functions of M1 macrophages. In addition, P72 macrophages contributed to increased mortality in a mouse model of LPS-induced endotoxemia. Therefore, given the high frequency of P72 in African Americans, cell signaling alterations driven by codon 72 of TP53 SNP may potentially contribute to differences in clinical outcomes and health disparities in common diseases associated with dysregulated macrophage activation.

MATE transporter OsMATE2 mediates root growth, grain size and weight by interacting with Mn-SOD and PABP in rice

Multidrug and toxic compound extrusion proteins (MATE) can transport small organic molecules in and out of cells and participate in detoxification, nutrient absorption, disease resistance and plant development processes. These compounds are widely distributed in plants. However, the mechanism by which MATE affects grain development remains elusive. In this study, we studied a MATE transporter, OsMATE2, which localized on the membrane. The CRISPR-Cas9 (CR) knockout line of OsMATE2 presented obvious decreases in grain weight. In addition, root development was also affected. Two proteins that interact with OsMATE2, namely, manganese-superoxide dismutase (Mn-SOD) and poly(A)-binding protein (PABP), were identified from a screening of yeast library. The results were validated through yeast two-hybrid and bimolecular fluorescence complementation experiments. The CRISPR-Cas9 (CR) knockout lines of Mn-SOD and PABP presented increased grain size and weight. Our findings demonstrated that OsMATE2 interacts with Mn-SOD and PABP to regulate grain development in rice.

Effect of increasing levels of Hermetia illucens in a fishmeal-free diet at sea bream (Sparus aurata, L.) gastrointestinal level

The impact of a fishmeal-free, high plant-protein-based diet, in which vegetable proteins (VPs) were partially substituted with the proteins derived from Hermetia illucens pupae meal (Hi) was examined in the gilthead sea bream (Sparus aurata). The physiological response at the gastrointestinal level was investigated by considering the gene expression responses and histology at the liver, stomach, pyloric caeca, foregut, midgut, and hindgut. To this end, three iso-proteic (crude protein, Nx6.25, 45 %) and iso-lipidic (crude fat, 20 %) extruded diets were formulated. A control diet (CV), comprising a substantial proportion of vegetable plant-protein (VPs) derivatives, was formulated alongside two test diets. These were created by replacing graded levels of the VPs (20 and 40 %, respectively) with protein derived from Hi (Hi20 and Hi40, respectively). In all dietary treatments, the foregut, midgut, and hindgut exhibited general structural integrity, displaying intact intestinal folds, enterocytes, and no signs of inflammation. However, occasional mild lymphocytic infiltrations of the epithelium and lamina propria were observed in the pyloric caeca of the CV and Hi20 diets. The histological examination of the stomach revealed that the CV diet had induced moderately-severe lymphocytic gastritis, accompanied by a down-regulation of heat shock protein 70 (hsp70). Moderate lymphocytic infiltrations were observed in liver of fish fed with the CV diet, which correlated to a down-regulation of copper-zinc superoxide dismutase (Sod1). In the hindgut, an increase in the expression of Sod1, manganese superoxide dismutase (Sod2), and intestinal alkaline phosphatase (iap) and a decrease in the expression of hsp70 were observed in insect meal-containing diet relative to the CV. Overall, the present study showed that in sea bream a plant-protein based diet lacking in fishmeal negatively affects stomach mucosa, but HI included in a plant-protein-based diet helps in ameliorate histological distortion of tissue through specific correlated gene activations.

Quantum refinement in real and reciprocal space using the Phenix and ORCA software.

X-ray and neutron crystallography, as well as cryogenic electron microscopy (cryo-EM), are the most common methods to obtain atomic structures of biological macromolecules. A feature they all have in common is that, at typical resolutions, the experimental data need to be supplemented by empirical restraints, ensuring that the final structure is chemically reasonable. The restraints are accurate for amino acids and nucleic acids, but often less accurate for substrates, inhibitors, small-molecule ligands and metal sites, for which experimental data are scarce or empirical potentials are harder to formulate. This can be solved using quantum mechanical calculations for a small but interesting part of the structure. Such an approach, called quantum refinement, has been shown to improve structures locally, allow the determination of the protonation and oxidation states of ligands and metals, and discriminate between different interpretations of the structure. Here, we present a new implementation of quantum refinement interfacing the widely used structure-refinement software Phenix and the freely available quantum mechanical software ORCA. Through application to manganese superoxide dismutase and V- and Fe-nitrogenase, we show that the approach works effectively for X-ray and neutron crystal structures, that old results can be reproduced and structural discrimination can be performed. We discuss how the weight factor between the experimental data and the empirical restraints should be selected and how quantum mechanical quality measures such as strain energies should be calculated. We also present an application of quantum refinement to cryo-EM data for particulate methane monooxygenase and show that this may be the method of choice for metal sites in such structures because no accurate empirical restraints are currently available for metals.

Surface-enhanced Raman scatting microfluidic chip based on the identification competition strategy were used for rapid and simultaneous detection of liver cancer related proteins

Biomarker testing plays a crucial role in the early detection of liver cancer. Herein, we developed a dual-signal amplification approach utilizing magnetic aggregation and a recognition competition strategy for the simultaneous detection of alpha-fetoprotein (AFP) and manganese superoxide dismutase (MnSOD) in serum. 4-MBA@AuNPs@H1 and DTNB@AuNPs@H2 were synthesized by functionalizing Raman signaling molecules and aptamer complementary chains onto the surface of gold nanoparticles (AuNPs). The detection complex Raman signal molecule@AuNPs@H-Fe3O4@cDNA was assembled by conjugating 4-MBA@AuNPs@H1 and DTNB@AuNPs@H2 with two nucleic acid aptamers (cDNA1 and cDNA2) modified with Fe3O4 through partial base complementary pairing. The target protein exhibited specific binding with the aptamer, leading to the competitive displacement of 4-MBA@AuNPs@H1 and DTNB@AuNPs@H2 from the Fe3O4 array surface, consequently resulting in a reduction of the Surface-Enhanced Raman Spectroscopy (SERS) signal. Through this approach, the limit of detection (LOD) for AFP and MnSOD in serum was achieved at remarkably low levels of 5.89 pg/mL and 6.23 pg/mL, respectively, with a reaction incubation period of only 5 min. Finally, the platform was utilized for the quantification of AFP and MnSOD in the serum of a nude mouse hepatocellular carcinoma model. The results obtained through SERS were consistent with those from enzyme-linked immunosorbent assay (ELISA), validating its accuracy. This methodology presents a novel approach for the swift and concurrent detection of proteins, holding significant clinical promise for the early diagnosis of hepatocellular carcinoma.

Unlocking Selective Anticancer Mechanisms: Dinuclear Manganese Superoxide Dismutase Mimetics Combined with Pt(II) Complexes.

We conducted an in-depth exploration of the in vitro activities of the dinuclear Mn2L2Ac and Mn2L2 complexes (where HL=2-{[di(2-pyridyl)methylamino]-methyl}phenol), possessing dual superoxide dismutase (SOD) and catalase (CAT) activity. We investigated these complexes both individually and in conjunction with various Pt(II)-complexes, either as mixtures or as the Mn2-Pt adducts. Our findings revealed a notable up to 50 % enhancement in the viability of healthy human breast cells, contrasted with a viability decrease as low as 50 % in breast cancer cells upon combined treatments with Mn2 SOD mimics and Pt(II) complexes. Specifically, we synthesized and characterized the self-assembled Mn2-Pt adducts (isolated Mn2L2Pt and in situ Mn2L2Pt'), linking Mn2L2-core with the carboxylate group of PtDAPCl2 (dichloro(2,3-diaminopropionic acid) platinum(II)). The SOD activity of the isolated Mn2L2Pt adduct (kSOD=1.7×107 M-1 s-1) remained intact. Through in vitro cell viability assessments, ROS levels, cellular Mn uptake and proteomics measurements, we elucidated key mechanisms underlying the observed biological effects. We demonstrated that Mn2-containing formulations predominantly target mitochondrial processes, differently affecting the proteome of cancerous and healthy cells. They induced downregulation of H2S signaling and expression of mitochondrial complex I and III, as well as increased oxidative phosphorylation pathways and upregulation of EGFR in cancer cells. In contrast, healthy cells showed a decrease in EGFR expression and a moderate enrichment in oxidative phosphorylation pathways.