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Parasitology Research Dec 2023Fish trypanosomiasis is a common blood parasitic disease transmitted by aquatic invertebrates, such as leeches. This study aims to shed light on the cytotoxicity of...
Fish trypanosomiasis is a common blood parasitic disease transmitted by aquatic invertebrates, such as leeches. This study aims to shed light on the cytotoxicity of Trypanosoma sp. on erythrocytes and its impacts on the innate immune response (serum lysozyme activity, nitric oxide production, phagocytic activity, serum total protein, and globulin) in wild African catfish, Clarias gariepinus. One hundred catfish were examined using blood smears stained with Giemsa and confirmed with PCR. The prevalence of infection was found to be 10% by microscope detection and 15% by PCR. The morphological identification of Trypanosoma as Trypanosoma mukasai was determined. Additionally, this study included previously undescribed features of Trypanosoma, such as the width of the anterior and posterior body, the length of the posterior pale region, and the number of folds. Various alterations in erythrocytes were observed, totaling 54.57%. Nuclear abnormalities, including fragmented nuclei, eccentric nuclei, and micronuclei, were also reported. Infected fish showed a reduction in serum total protein and globulin levels, while nitric oxide production, lysozyme activity, and phagocytic activity exhibited a significant increase compared to non-infected fish. We believe that our findings will contribute valuable data to the morphological and molecular identification of Trypanosoma sp. in African catfish, as well as their cytotoxic impact.
Topics: Animals; Catfishes; Muramidase; Nitric Oxide; Trypanosoma; Globulins
PubMed: 38057596
DOI: 10.1007/s00436-023-08026-8 -
International Journal of Biological... Feb 2024Glycation is a spontaneous chemical reaction, which affects the structure and function of proteins under normal physiological conditions. Therefore, organisms have...
Glycation is a spontaneous chemical reaction, which affects the structure and function of proteins under normal physiological conditions. Therefore, organisms have evolved diverse mechanisms to combat glycation. In this study, we show that the Escherichia coli glycolytic enzyme phosphoglucose isomerase (Pgi) exhibits deglycation activity. We found that E. coli Pgi catalyzes the breakdown of glucose 6-phosphate (G6P)-derived Amadori products (APs) in chicken lysozyme. The affinity of Pgi to the glycated lysozyme (K, 1.1 mM) was ten times lower than the affinity to its native substrate, fructose 6-phosphate (K, 0.1 mM). However, the high kinetic constants of the enzyme with the glycated lysozyme (k, 396 s and kK, 3.6 × 10 M s) indicated that the Pgi amadoriase activity may have physiological implications. Indeed, when using total E. coli protein (20 mg/mL) as a substrate in the deglycation reaction, we observed a release of G6P from the bacterial protein at a Pgi specific activity of 33 μmol/min/mg. Further, we detected 11.4 % lower APs concentration in protein extracts from Pgi-proficient vs. deficient cells (p = 0.0006) under conditions where the G6P concentration in Pgi-proficient cells was four times higher than in Pgi-deficient cells (p = 0.0001). Altogether, these data point to physiological relevance of the Pgi deglycation activity.
Topics: Glucose-6-Phosphate Isomerase; Escherichia coli; Muramidase; Escherichia coli Proteins; Phosphates
PubMed: 38056730
DOI: 10.1016/j.ijbiomac.2023.128541 -
European Journal of Pharmaceutics and... Jan 2024Molecular mobility in form of alpha and beta relaxations is considered crucial for characterization of amorphous lyophilizates and reflected in the transition...
Molecular mobility in form of alpha and beta relaxations is considered crucial for characterization of amorphous lyophilizates and reflected in the transition temperatures Tg and Tg. Based on an overview of applied methods to study beta relaxations, Dynamic Mechanical analysis was used to measure Tg and Tg in amorphous freeze-dried samples. Lysozyme and trehalose as well as their mixtures in varying ratios were investigated. Three different residual moisture levels, ranging from roughly 0.5-7 % (w/w), were prepared via equilibration of the freeze-dried samples. Known plasticising effects of water on Tg were confirmed, also via differential scanning calorimetry. In addition and contrary to expectations, an influence of water on the Tg also was observed. On the other hand, an increasing amount of trehalose lowered Tg but increased Tg showing that Tg and Tg are not paired. The findings were interpreted with regard to their underlying molecular mechanisms and a correlation with the known influences of water and trehalose on stability. The results provide encouraging hints for future stability studies of freeze-dried protein formulations, which are urgently needed, not least for reasons of sustainability.
Topics: Muramidase; Trehalose; Water; Freeze Drying; Transition Temperature; Calorimetry, Differential Scanning
PubMed: 38029940
DOI: 10.1016/j.ejpb.2023.11.019 -
Journal of Pharmaceutical Analysis Oct 2023Muramidase-released protein (MRP) is now being recognized as a critical indicator of the virulence and pathogenicity of (). However, the identification of viable...
Muramidase-released protein (MRP) is now being recognized as a critical indicator of the virulence and pathogenicity of (). However, the identification of viable therapeutics for infection was hindered by the absence of an explicit mechanism for MRP-actuated inflammation. Dihydroartemisinin (DhA) is an artemisinin derivative with potential anti-inflammatory activity. The modulatory effect of DhA on the inflammatory response mediated by the virulence factor MRP remains obscure. This research aimed to identify the signaling mechanism by which MRP triggers the innate immune response in mouse spleen and cultured macrophages. With the candidate mechanism in mind, we investigated DhA for its ability to dampen the pro-inflammatory response induced by MRP. The innate immune response in mice was drastically triggered by MRP, manifesting as splenic and systemic inflammation with splenomegaly, immune cell infiltration, and an elevation in pro-inflammatory cytokines. A crucial role for Toll-like receptor 4 (TLR4) in coordinating the MRP-mediated inflammatory response via nuclear factor-kappa B (NF-κB) activation was revealed by TLR4 blockade. In addition, NF-κB-dependent transducer and activator of transcription 3 (STAT3) and mitogen-activated protein kinases (MAPKs) activation was required for the inflammatory signal transduction engendered by MRP. Intriguingly, we observed an alleviation effect of DhA on the MRP-induced immune response, which referred to the suppression of TLR4-mediated actuation of NF-κB-STAT3/MAPK cascades. The inflammatory response elicited by MRP is relevant to TLR4-dependent NF-κB activation, followed by an increase in the activity of STAT3 or MAPKs. DhA mitigates the inflammation process induced by MRP via blocking the TLR4 cascade, highlighting the therapeutic potential of DhA in targeting infection diseases.
PubMed: 38024861
DOI: 10.1016/j.jpha.2023.05.013 -
Sheng Wu Gong Cheng Xue Bao = Chinese... Nov 2023Human-derived lysozyme is a general term for a group of naturally occurring alkaline proteins in the human body that are capable of lysing bacterial cell walls. Its... (Review)
Review
Human-derived lysozyme is a general term for a group of naturally occurring alkaline proteins in the human body that are capable of lysing bacterial cell walls. Its action is characterized by its ability to cleave the β-(1,4)-glycosidic bond between N-acetylglucosamine and N-acetylmuramic acid in peptidoglycan. Human-derived lysozyme has a variety of properties such as antibacterial, anti-inflammatory, antiviral and immune enhancing, and is therefore widely used in the domestic and international pharmaceutical markets. This review summarizes the structural features, expression sites, biological functions of human-derived lysozymes and its market applications.
Topics: Humans; Muramidase; Anti-Bacterial Agents
PubMed: 38013179
DOI: 10.13345/j.cjb.230241 -
The Journal of Pain May 2024Spinal cord injury (SCI) affects ∼500,000 people worldwide annually, with the majority developing chronic neuropathic pain. Following SCI, approximately 60% of these...
Spinal cord injury (SCI) affects ∼500,000 people worldwide annually, with the majority developing chronic neuropathic pain. Following SCI, approximately 60% of these individuals are diagnosed with comorbid mood disorders, while only ∼21% of the general population will experience a mood disorder in their lifetime. We hypothesize that nociceptive and depressive-like dysregulation occurs after SCI and is associated with aberrant macrophage infiltration in segmental pain centers. We completed moderate unilateral C5 spinal cord contusion on LysM-eGFP reporter mice to visualize infiltrating macrophages. At 6-weeks post-SCI, mice exhibit nociceptive and depressive-like dysfunction compared to naïve and sham groups. There were no differences between the sexes, indicating that sex is not a contributing factor driving nociceptive or depressive-like behaviors after SCI. Utilizing hierarchical cluster analysis, we classified mice based on endpoint nociceptive and depressive-like behavior scores. Approximately 59.3% of the SCI mice clustered based on increased paw withdrawal threshold to mechanical stimuli and immobility time in the forced swim test. SCI mice displayed increased myeloid cell presence in the lesion epicenter, ipsilateral C7-8 dorsal horn, and C7-8 DRGs as evidenced by eGFP, CD68, and Iba1 immunostaining when compared to naïve and sham mice. This was further confirmed by SCI-induced alterations in the expression of genes indicative of myeloid cell activation states and their associated secretome in the dorsal horn and dorsal root ganglia. In conclusion, moderate unilateral cervical SCI caused the development of pain-related and depressive-like behaviors in a subset of mice and these behavioral changes are consistent with immune system activation in the segmental pain pathway. PERSPECTIVE: These experiments characterized pain-related and depressive-like behaviors and correlated these changes with the immune response post-SCI. While humanizing the rodent is impossible, the results from this study inform clinical literature to closely examine sex differences reported in humans to better understand the underlying shared etiologies of pain and depressive-like behaviors following central nervous system trauma.
Topics: Animals; Spinal Cord Injuries; Neuralgia; Depression; Mice; Female; Male; Mice, Transgenic; Disease Models, Animal; Green Fluorescent Proteins; Myeloid Cells; Mice, Inbred C57BL; Behavior, Animal; Muramidase
PubMed: 38007034
DOI: 10.1016/j.jpain.2023.11.016 -
BMC Microbiology Nov 2023Dental caries is a chronic oral disease caused by microbial infections, which result in erosion of the dental enamel and cause irreversible damage. Therefore, proper...
BACKGROUND
Dental caries is a chronic oral disease caused by microbial infections, which result in erosion of the dental enamel and cause irreversible damage. Therefore, proper disease management techniques and the creation of an environment that prevents intraoral growth and biofilm formation of Streptococcus mutans in the early stages, are crucial to prevent the potential progression of dental plaque to disease. Here, we aimed to investigate antimicrobial and antibiofilm effects of the Bacillus velezensis ID-A01 supernatant (ID23029) against S. mutans, and its inhibitory effects on acidogenesis.
RESULTS
A killing kinetics assay showed a peak lethality percentage of 94.5% after 6 h of exposure to ID23029. In sucrose-exposed conditions, ID23029 inhibited lactic acid formation, preventing the pH from falling below the threshold for enamel demineralization, and inhibited up to 96.6% of biofilm formation. This effect was maintained in the presence of lysozyme. Furthermore, ID23029 retained up to 92% lethality, even at an intraoral concentration at which lysozyme is ineffective against S. mutans.
CONCLUSIONS
This study demonstrates the potential of the B. velezensis ID-A01 supernatant for the prevention and treatment of dental caries. Its eventual use in dental practice is encouraged, although further studies are required to confirm its beneficial effects.
Topics: Humans; Muramidase; Streptococcus mutans; Dental Caries; Anti-Infective Agents; Biofilms
PubMed: 37996837
DOI: 10.1186/s12866-023-03114-2 -
Poultry Science Jan 2024The objective of the present studies was to evaluate muramidase (MUR) supplementation in broilers under Eimeria and/or Clostridium perfringens challenge. For this, 2...
A microbial muramidase improves growth performance and reduces inflammatory cell infiltration in the intestine of broilers chickens under Eimeria and Clostridium perfringens challenge.
The objective of the present studies was to evaluate muramidase (MUR) supplementation in broilers under Eimeria and/or Clostridium perfringens challenge. For this, 2 experiments were conducted. Experiment 1. A total of 256 one-day old male Cobb 500 chicks were placed in battery cages in a completely randomized design, with 5 treatment groups, 7 replicate cages per treatment and 8 birds per cage. The treatments were: nonchallenged control (NC), challenged control (CC), CC + MUR at 25,000 or 35,000 LSU(F)/kg, and CC + Enramycin at 10 ppm (positive control-PC). Challenge consisted of 15× the recommended dose of coccidiosis vaccine at placement, and Clostridium perfringens (10 CFU/bird) inoculation at 10, 11, and 12 d. Macro and microscopic evaluation, immunohistochemistry, and gene expression were evaluated at 7, 14, 21, and 28 d of age. Experiment 2. A total of 1,120 one-day old male Cobb 500 chicks were placed in floor pens with fresh litter in a completely randomized design, with 4 treatment groups, 8 replicate pens per treatment, and 35 birds per pen. The treatments were: Control, supplementation of MUR at 25,000 or 45,000 LSU(F)/kg, and a positive control (basal diet plus Enramycin). At 10, 11, and 12 d of the experiment all the birds were inoculated by oral gavage with a fresh broth culture of a field isolate Clostridium perfringens (0.5 mL containing 10 CFU/bird). It was observed that in Experiment 1 MUR supplementation reduced the infiltration of macrophages and CD8+ lymphocytes in the liver and ileum of infected birds, downregulated IL-8 and upregulated IL-10 expression. In Experiment 2, MUR linearly improved the growth performance of the birds, increased breast meat yield, and improved absorption capacity. MUR supplementation elicited an anti-inflammatory response in birds undergoing a NE challenge model that may explain the improved growth performance of supplemented birds.
Topics: Animals; Male; Eimeria; Clostridium perfringens; Chickens; Muramidase; Coccidiosis; Clostridium Infections; Intestines; Diet; Poultry Diseases; Animal Feed
PubMed: 37995420
DOI: 10.1016/j.psj.2023.103226 -
Changes in physicochemical parameters of duck eggs and extra-embryonic structures during incubation.Animal : An International Journal of... Dec 2023Duckling embryogenesis should be deepened due to the hatching technology and its modification possibilities. Many changes occur in incubated eggs, which expose the...
Duckling embryogenesis should be deepened due to the hatching technology and its modification possibilities. Many changes occur in incubated eggs, which expose the embryo to hazards. The study aimed to analyse the physicochemical properties of eggshell, yolk, thick albumen (TA), and amniotic fluid (AF) of incubated hatching eggs from 52-week-old Cherry Valley ducks. The morphological features of 18 fresh eggs were analysed. Over 28 days, a total of 800 eggs underwent incubation. Eggshell surface temperature and egg weight loss were measured on days 1, 4, 7, 10, 14, 18, 21, and 25. Eggshell, TA, AF, and yolk were collected from eggs at incubation days 1-21 (every week). TA was collected on days 0, 1, and 7, while AF on days 7, 14, and 21. The analysis covered a range of physicochemical parameters. Eggshell thickness decreased with incubation, reaching its lowest point posthatch (P < 0.001). The highest pH for TA was recorded on day 1, while the lowest was on day 7 when comparing days 0, 1, and 7 (P < 0.001). TA pH was consistently higher than in AF (P < 0.001). However, the pH of TA was the highest on day 1 and the lowest on day 7 (P < 0.001). Yolk pH increased from days 1 to 21 (P < 0.001). There was also a noticeable in egg weight loss (0.34% daily) (P < 0.001). Vitelline membrane strength decreased from day 0 to day 1 (P < 0.001). Lysozyme activity in thick albumen on day 7 was higher than on days 0 and 1 (P < 0.001). Lysozyme activity in AF was higher on day 21 than days 7 and 14 (P < 0.001). TA viscosity was highest on day 0 and lowest on day 1, compared to other days (P < 0.001). AF viscosity and CP content exhibited an increase on day 21 as compared to days 7 and 14 (P < 0.001). The CP content in TA was notably higher on day 7 than on days 0 and 1 (P < 0.001). Polyunsaturated fatty acids declined, while monounsaturated and transfatty acids increased (P < 0.001). Viscosity and lysozyme activity increased on day 7 in TA and day 21 in AF. TA and the amniotic cavity appeared to facilitate the transfer of substances, particularly CP. Viscosity could be an indicator for optimising incubation conditions, as incorrect changes can affect embryo mortality. The results showed the different utilisation of nutrients, such as fatty acids. It could support research on the in-ovo administration of various substances.
Topics: Animals; Ducks; Muramidase; Ovum; Egg Shell; Weight Loss; Eggs; Chickens
PubMed: 37981451
DOI: 10.1016/j.animal.2023.101024 -
Analytica Chimica Acta Dec 2023Antifouling coatings are critically necessary for optical biosensors for various analytical application sectors, from medical diagnostics to foodborne pathogen...
Antifouling coatings are critically necessary for optical biosensors for various analytical application sectors, from medical diagnostics to foodborne pathogen detection. They help avoid non-specific protein/cell attachment on the active biosensor surface and catch the analytes directly in the complex media. Advances in antifouling plasmonic surfaces have been mainly focused on detecting clinical biomarkers in real biofluids, whereas developing antifouling coatings for direct analysis of analytes in complex media has been scarcely investigated for food quality control and safety. Herein, we propose a new low-fouling poly-l-lysine (PLL)-based surface layer for directly detecting an allergen protein, lysozyme, in the food matrix using surface plasmon resonance. The PLL-based polymer contains densely immobilized anionic oligopeptide side chains to create an electric charge-balanced layer able to repel the non-specific adsorption of undesired molecules on the biosensor surface. It also includes sparsely attached aptamer probes for capturing lysozyme directly in food sources with no pre-analytical sample treatment. We optimized the surface layer fabrication condition and tested the dual-functional surface to evaluate its ability to detect the target protein selectively. The developed analytical approach allowed for achieving a limit of detection of 0.04 μg mL (2.95 nM) and a limit of quantification of 0.13 μg mL (8.95 nM). Lysozyme was successfully quantified in milk samples using the plasmonic dual-functional aptasensor without sample pre-treatment or target isolation, illustrating the device's utility.
Topics: Biofouling; Muramidase; Surface Plasmon Resonance; Biosensing Techniques; Allergens
PubMed: 37977796
DOI: 10.1016/j.aca.2023.341979