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This clinical study investigates various metabolic and physiological parameters in dairy cows during puerperium. Retained fetal membranes (RFM) is a significant postpartum complication that can affect the overall health, fertility and productivity of dairy cattle. The research focuses on changes in total proteins, albumin, glucose, triglycerides, total cholesterol, aspartate aminotransferase (AST) and alanine aminotransferase (ALT), cortisol, insulin, and insulin-like growth factor 1 (IGF-1) levels among cows experiencing normal post-partum period (NP) and those with RFM. A significant increase in protein levels was noted during the post-partum period in the RFM group, indicating physiological impacts of RFM at this stage. Albumin levels showed significant differences, highlighting a significant biological effect of RFM in the post-partum period. Glucose levels varied significantly in the weeks leading to parturition, suggesting altered metabolic states in cows that suffered RFM. Triglyceride and cholesterol levels were significantly higher during the antepartum period in the group that experienced reproductive failure, indicating substantial alterations in lipid metabolism which could herald the apparition of RFM. AST and ALT levels provided insights into cellular stress and liver function, with significant increases noted around parturition which could be attributed to the substantial physiological strain of parturition itself. Cortisol levels were higher in RFM cows 2 weeks before parturition, which could indicate an increasing stress response or a physiological preparation for the upcoming labor, and may be more pronounced in cows predisposed to RFM. Insulin levels decreased significantly before and at parturition in RFM cows, indicating a strong energy deficit. IGF-1 levels decreased significantly in RFM cows after parturition. Significant changes in metabolic parameters, such as glucose, triglycerides, and cholesterol levels, delineate the pronounced metabolic challenges faced by cows with RFM. The study elucidates that while some variations are noted as parturition approaches, the most substantial impacts attributable to RFM on metabolic and physiological parameters occur after parturition. These changes may have implications for the health, recovery, and productivity of cows postpartum, suggesting the need for targeted management strategies to mitigate the effects of RFM.
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Urinary tract infections (UTIs) are a widespread condition in pets, with many antibiotics being prescribed, contributing to the rise in antimicrobial resistance, which is a worldwide threat. This study's main objective was to analyze the in vitro antimicrobial activity of Cornus mas and Sorbus aucuparia fruit hydro-ethanolic extracts towards bacteria identified in the urine of companion animals experiencing UTIs. Urine samples were collected from dogs and cats (n = 83; 47 negative, 36 positive); several bacterial strains were identified (n = 49) belonging to the Escherichia, Enterococcus, Staphylococcus, Proteus, Klebsiella, Enterobacter, Pseudomonas, Acinetobacter, Leclercia, and Kocuria genera. Bacterial susceptibility was tested using the disk diffusion method, with the majority being resistant to several beta-lactams, quinolones, trimethoprim/sulfamethoxazole, and nitrofurantoin. Subsequently, 13 resistant isolates were selected to evaluate the fruits extracts' antimicrobial potential using the agar well diffusion and broth microdilution methods. Cornus mas exhibited the greatest activity against Gram-negatives (primarily Pseudomonas luteola), while Sorbus aucuparia showed maximum effects towards Gram-positives (particularly Enterococcus faecalis). The MIC was 0.01 µg/µL for both extracts; the MBC was 0.08 µg/µL for Cornus mas and 0.05 µg/µL for Sorbus aucuparia. However, Cornus mas showed a stronger bactericidal effect. This is the first study to investigate these fruit extracts in UTI isolates of companion animals, and these extracts might be used as substitutes or adjuvants for antibiotics, thus contributing to a reduction in antimicrobial resistance.
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In dogs, periodontal disease (PD) is a highly common condition whose prevalence rises significantly with age. Studies on rats with experimentally induced PD and human clinical trials have found an association between oxidative stress (OS) and PD, as has been observed in many other inflammatory disorders. The objective of this research was to assess the main indicators of oxidative stress detected in the saliva of dogs and to compare them to the degree of inflammation and tartar amount on their teeth and gums. By identifying these correlations, we intended to identify early-stage PD markers that would allow us to diagnose the condition in dogs without requiring for invasive or traumatic procedures. The antioxidant enzyme superoxide dismutase (SOD), total antioxidant capacity (TAC), malondialdehyde (MDA) and 8-hydroxyguanosine (8OHG), matrix metalloproteinase-8 (MMP8), and the quantity of total proteins are the markers that have been investigated in order to accomplish these objectives. The appearance of specific forms of periodontal disease has been confirmed by an increase in the saliva concentration of MMP8, a marker commonly used in the diagnosis of these pathologies in humans. The study was carried out on 22 dogs divided in a control group (C) and study groups (S), the second one being divided into three subgroups according to the severity of the gum inflammation and the presence or absence of tartar: S1-small accumulation of tartar, lack of infection-related signs; S2-moderate accumulation of tartar, gums swollen, red, and bled easily; S3-significant accumulation of tartar; gums bright red and bleeding; marked pain and halitosis. A correlation between the concentration of the tested parameters and the severity of the local appearance of the gum and teeth was noted. In comparison to the results of individuals from the control group, MMP8 reported increases of 1.2, 2, and 3.7 times, respectively, in the case of individuals from subgroups S1, S2, and S3. Oxidative stress is caused by inflammatory processes in the oral cavity. The presence of oxidative stress was demonstrated by the evaluation of different indicators in the saliva: an increase in the general antioxidant status, which was associated with an increase in SOD activity; intensification of the lipid peroxidation processes, as demonstrated by the accumulation of the MDA peroxidation product; and an increase in the DNA oxidation processes, as demonstrated by the accumulation of the oxidation product 8OHdG. In consequence, it was observed that there were up to 2-fold increases in protein levels, total antioxidant activity, SOD, and MDA, and up to 8.6-fold increases in the case of 8-OHdG when comparing dogs from the S3 group (significant tartar accumulation; bright red and bleeding gums; marked pain and halitosis) to those in the control group.
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The aim of this research was to validate the use of a gentamicin (GEN) and nano-hydroxiapatite (nHAP)-loaded polycaprolactone nanostructured membrane (NM) as an innovative, highly efficient, low-cost treatment for periodontitis. We conducted an in vivo study on Wistar rats, in which we induced periodontitis by placing silk ligatures around the first right and left upper molars. The subjects were divided into three groups; the first group received no periodontal treatment, the second group received open flap debridement, and the third group received open flap debridement, together with the positioning of the GEN and nHAP-loaded nanostructured membrane as a treatment. The extent of periodontal regeneration was assessed by the periodontal pocket depth, bleeding on probing, tooth mobility, dental plaque, microbiological analysis, concentration of MMP-8 in saliva, plasma levels of CRP, and histological analysis. The results showed that using open flap debridement with the NM is more efficient, and it significantly reduces the probing depth, extent of bleeding on probing, dental mobility, bacterial plaque, and pathogenic flora. The concentrations of MMP-8 and CRP decrease. The histological analysis demonstrated that NM leads to bone regeneration. Our study indicates that gentamicin and nano-hydroxyapatite embedded in the fiber of the biodegradable membranes might be a promising therapeutic option for periodontitis treatment.
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Considering that Sorbus aucuparia fruits have been underutilized despite their tremendous potential, this study aimed to correlate the in vitro antioxidant, antibacterial and cell-protective abilities of fruit extracts derived from Sorbus aucuparia Romanian cultivars with their phytochemical composition. Therefore, following the preparation of ethanolic and carotenoid extracts, phytochemical screening was performed using UV-Vis and HPLC-DAD-ESI-MS methods. The antioxidant activity was analyzed using DPPH and FRAP tests. As the results revealed high contents of bioactive compounds (polyphenols 1.11 mg GAE/g DM, flavonoids 430.06 µg QE/g DM and carotenoids 95.68 µg/g DM) and an important antiradical action (DPPH 24.51 mg/mL and FRAP 0.016 µM TE/mL), we chose to further examine the fruits' biological properties. The antibacterial capacity was assessed employing agar well diffusion and broth microdilution techniques, with fruits displaying an intense activity against MSSA, MRSA and Enterococcus faecalis, but also E. coli and Pseudomonas aeruginosa. The cell-protective activity was analyzed on gentamicin-stressed renal cells, through MTT and Annexin V-FITC assays. Importantly, a significant increase in viability was registered on stressed cells following extract administration in low doses; nevertheless, viability was noticed to decline when exposed to elevated concentrations, potentially due to the cumulative actions of the extract and gentamicin. These findings offer novel light on the antibacterial activity of Sorbus aucuparia Romanian cultivars, as well as their cell-protective ability in renal cell injury.
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In spite of its well-known nephrotoxicity, gentamicin is nonetheless routinely used in humans and animals. However, no adjuvant treatments have been implemented to mitigate this harmful effect. Given this concern, medicinal plants represent a significant reservoir of natural antioxidants that could potentially reduce the renal oxidative stress induced by gentamicin. Therefore, the main objective of this research was to investigate the nephroprotective properties of Cornus mas and Sorbus aucuparia fruits in an experimental model of nephrotoxicity. The 3-week study was performed on male Wistar rats, which were randomly divided into six experimental groups, being subcutaneously treated with 50 mg/kg gentamicin and orally given Cornus mas and Sorbus aucuparia extracts, in doses of 40 mg/kg and 10 mg/kg, respectively. Antioxidant therapy significantly improved the nitro-oxidative stress parameters as well as the specific renal biomarkers KIM-1 and iNAG, demonstrating a considerable renal tubular protective impact. These outcomes were reinforced by biochemical and histopathological enhancements. Nevertheless, neither of the tested extracts succeeded in substantially diminishing BUN levels. Additionally, CysC did not significantly decline following extracts treatment, suggesting that the remedies did not effectively protect renal glomeruli against gentamicin stress. Future studies are required in order to determine the underlying mechanisms of these berries.
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Cornus , Insuficiencia Renal , Sorbus , Ratas , Humanos , Animales , Antioxidantes/farmacología , Antioxidantes/química , Ratas Wistar , Cornus/química , Gentamicinas/toxicidad , Sorbus/química , Estrés Oxidativo , Extractos Vegetales/farmacología , Extractos Vegetales/química , BiomarcadoresRESUMEN
Cornus mas L. is characterized by an increased quantity of bioactive compounds, namely polyphenols, monoterpenes, organic acids, vitamin C and lipophilic compounds such as carotenoids, being anciently used in the treatment of various diseases. This paper's objectives were to characterize the phytochemical profile of Cornus mas L. fruits and to evaluate the in vitro antioxidant, antimicrobial and cytoprotective effects on renal cells exposed to gentamicin. As such, two ethanolic extracts were obtained. The resulting extracts were used to assess the total polyphenols, flavonoids and carotenoids through spectral and chromatographic methods. The antioxidant capacity was assessed using DPPH and FRAP assays. Due to the high content of phenolic compounds analyzed in fruits and the results obtained regarding antioxidant capacity, we decided to further use the ethanolic extract to investigate the in vitro antimicrobial and cytoprotective effects on renal cells stressed with gentamicin. The antimicrobial activity was assessed using agar well diffusion and broth microdilution methods, with great results regarding Pseudomonas aeruginosa. The cytotoxic activity was assessed using MTT and Annexin-V assays. According to the findings, extract-treated cells had a higher cell viability. However, at high concentrations, viability was shown to decline, most likely due to the extract and gentamicin's additive effects.
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Mucoadhesive films loaded with doxycycline hyclate (Doxy Hyc), consisting of mixtures of hydroxypropylmethyl cellulose (HPMC) E3, K4 and polyacrylic acid (Carbopol 940), were prepared by casting method, aiming to design a formulation intended for application in the oral cavity. The obtained film formulations exhibited a Doxy Hyc content between 7.52 ± 0.42 and 7.83 ± 0.41%, which had adequate mechanical properties for application in the oral cavity and pH values in the tolerance range. The x-ray diffraction studies highlighted the amorphisation of Doxy Hyc in the preparation process and the antibiotic particles present on the surface of the films, identified in the TEM images, which ensured a burst release effect in the first 15 min of the in vitro dissolution studies, after which Doxy Hyc was released by diffusion, the data presenting a good correlation with the Peppas model, n < 0.5. The formulation F1, consisting of HPMC K4 combined with C940 in a ratio of 5:3, the most performing in vitro, was tested in vivo in experimentally-induced periodontitis and demonstrated its effectiveness in improving the clinical parameters and reducing the salivary levels of matrix metalloproteinase-8 (MMP-8). The prepared Doxy Hyc loaded mucoadhesive buccal film could be used as an adjuvant for the local treatment of periodontitis, ensuring prolonged release of the antibiotic after topical application.
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The immunomodulatory effect of a novel biomaterial obtained through electrospinning, based on polylactic acid (PLA) and nano-hydroxyapatite (nano-HAP), loaded with doxycycline (doxy) was evaluated in an animal model. The treatment capabilities as a local non-surgical treatment of periodontitis was investigated on the lower incisors of Wistar rats, after the induction of localized periodontitis using the ligature technique. Following the induction of the disease, the non-surgical treatment of scaling and root planing was applied, in conjunction with the application of the new material. The results of the treatment were evaluated clinically, using the tooth mobility and gingival index scores, as well as histologically. The salivary concentrations of matrix metalloproteinase 8 (MMP-8) and plasmatic concentrations of interleukin 1 (IL-1), interleukin 6 (IL-6) and tumor necrosis factor alpha (TNF-α) were also monitored. Two weeks after the ligature application, the periodontal disease was successfully induced in rats. The application of the novel biomaterial obtained through electrospinning was proven to be more effective in improving the clinical parameters, while decreasing the salivary MMP-8 and plasmatic IL-1 and TNF-α concentrations, compared to the simple scaling and root planing. Thus, the novel electrospun biomaterial could be a strong candidate as an adjuvant to the non-surgical periodontal therapy.
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Goat mastitis is still frequently diagnosed in dairy farms, with serious consequences on milk quality and composition. The aim of this study was to establish correlations between milk microorganisms and biochemical parameters in goats with no signs of clinical mastitis. Thus, 76 milk samples were collected from a dairy goat farm, Carpathian breed, followed by microbiological, molecular (16S rRNA sequencing) and somatic cells analysis, determination of lactate dehydrogenase (LDH), ß-glucuronidase, catalase (CAT), glutathione peroxidase (GPx) activity, total antioxidant capacity (TAC), nitric oxide (NO) and lipid peroxides (LPO) using spectrophotometry and the ELISA method for 8-hydroxy-deoxyguanosine (8-OHdG) as the oxidative DNA damage indicator. Samples positive for bacterial growth showed a significant (p < 0.05) increase in the number of somatic cells, LDH and ß-glucuronidase activity, as well as higher levels of CAT, GPx, NO, LPO and 8-OHdG compared with pathogen-free milk whereas TAC was lower in milk from an infected udder. These findings suggest that subclinical mastitis is associated with increased enzymatic activity and induction of oxidative stress. Nevertheless, changes in biochemical parameters tended to vary depending on the pathogen, the most notable mean values being observed overall in milk positive for Staphylococcus aureus.
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Metamizole (dipyrone) is a widely used non-opioid analgesic in both human and animal medicine. Metamizole's safety has been the topic of numerous opposing debates, given the fact that in certain countries metamizole is frequently used as an over-the-counter (OTC) medicine, while in others it is banned due to the risk of agranulocytosis. Further, small mammals such as rabbits, ferrets, rodents, and hedgehogs have become some of the most common pets present in veterinary practice, and each of these species has specific analgesic needs due to their anatomy and physiology. The key to providing appropriate medical care is in finding a substance that has minimal negative effects. In small mammals, analgesia is an important factor and, it happens frequently that, pain in these patients is not well managed. Post-operative pain management is an important topic in the welfare of animals. The objectives of this review, thus, were to provide a concise overview of analgesics that are used in the treatment of postoperative pain in small mammals (e.g., rabbits and rodents) and to highlight the importance of this product, metamizole, in veterinary medicine, as well as the potential of this substance as an alternative analgesic for the treatment of postoperative pain in small mammals.
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Dipirona , Manejo del Dolor , Animales , Humanos , Conejos , Dipirona/uso terapéutico , Hurones , Analgésicos/uso terapéutico , Dolor Postoperatorio/tratamiento farmacológico , Medicamentos sin Prescripción , Antiinflamatorios no Esteroideos/uso terapéuticoRESUMEN
Malignant melanoma is the most aggressive type of skin cancer, and due to the numerous limitations of current treatment methods, there is an urgent need to develop novel approaches for both the prevention and treatment of malignant melanoma, with research-oriented bioactive substances representing a notable first step. The current study decided to expand on previous rhodoxanthin research by investigating the possible anti-tumor effect as well as the effect on the antioxidant status in the case of murine melanoma in an experimental model. The 21-day study was carried out on female C57BL/6J mice. On the first day of the experiment, they were subcutaneously inoculated with 106 B16F10 cells and were given rhodoxanthin orally until the end of the study. Rhodoxanthin supplementation significantly reduced tumor growth (42.18%) and weight (15.74%). Furthermore, the epidermal growth factor (EGF) activity was reduced and the concentration of 8-OHdG dropped in the treated melanoma-bearing mice compared to the untreated ones, demonstrating the role of rhodoxanthin in slowing tumor growth, one of the mechanisms being the reduction of EGF level and the decrease of DNA oxidation. The administration of rhodoxanthin determined variations in antioxidant enzymes, both at the plasma level and at the tissue level.
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Obesity is a common feature in horses suffering from metabolic syndrome. While adipokines involvement as biomarkers is better established in human pathology, little data are available on horses. This study aimed to investigate the possible association and relationship between selected metabolic parameters and morphological traits in equine metabolic syndrome. Adiposity was evaluated using body condition score (BCS) and cresty neck score (CNS). Plasma levels of total cholesterol, insulin, NEFA, and adipokines (omentin and chemerin) were determined using enzyme-linked immunosorbent assays. Spearman correlation, univariate linear regression analysis and hierarchical clustering were performed. Significant positive correlations were observed between NEFA and bodyweight (r = 0.322; p = 0.006), BCS (r = 0.295; p = 0.013), and CNS (r = 0.267; p = 0.024), total cholesterol and bodyweight (r = 0.262; p = 0.027), and omentin and CNS (r = 0.234; p = 0.049). Cluster analysis supported these results and provided more details on the relationships between studied variables within and between the four resulting groups. These findings highlight NEFA, chemerin, and omentin as valuable biomarkers that could be further analyzed in other horse breeds for a better understanding of equine metabolic pathology.
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Taxus baccata L., an evergreen tree, was known until recently due to its high concentration of toxic compounds. The purpose of the present study was to focus on the only non-poisonous part, the red arils, which have recently been described as an important source of various bioactive constituents. To establish total phenolic, flavonoid, and carotenoid content, antioxidant capacity, and cytotoxic properties, two types of extracts were obtained. The chemical profile of the ethanolic extract was evaluated using chromatographic (HPLC-DAD-ESI+) and spectral (UV-Vis) methods, and the antioxidant activity of the ethanolic extract was assessed using DPPH and FRAP assays, yielding moderate results. In the second type of extract (methanol: ethyl acetate: petroleum ether (1:1:1, v/v/v)) we identified three carotenoids using open column chromatography and RP-PAD-HPLC, with rhodoxanthin being the most abundant. Considering the above and mainly because of the lack of information in the literature about this pigment and its biological effects, we decided to further investigate the cytotoxic activity of rhodoxanthin, the main carotenoid presented in aril, and its protective effect against H2O2-induced oxidative stress using two cell lines: human HaCaT keratinocytes and B16F10 murine malignant melanoma. The MTT and Annexin-V Apoptosis assays showed a substantial cytotoxic potential expressed in a dose-dependent manner towards the melanoma cell line, however, no obvious cytotoxic effects on human keratinocytes were noticed.
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Although conventional medicine, chemical drug synthesis and pharmaceutical research are advancing at a rapid pace, nature remains a major supplier of biological molecules. Natural bioactive compounds are studied closely especially as an alternative to the limitations of conventional therapy in many diseases, melanoma being one of them. Malignant melanoma is a highly aggressive type of cancer, and the current methods of treatment used are cryotherapy, external surgery, radiation therapy, chemotherapy, photodynamic therapy, biological therapy, and targeted drug therapy. Unfortunately, these treatment methods are often inefficient, extremely expensive and cause many side effects, which is why focusing on melanoma chemoprevention and adjuvant therapy with natural herbal phytoconstituents is an emerging strategy to prevent, cure or treat melanoma. This review aims to examine the latest discoveries in terms of potential natural bioactive compounds that possess important activity against the development and spread of murine melanoma cancer. In particular, the use of different phytochemicals such as phenolic acids, flavonoids, anthocyanins, terpenoids, essential oils and carotenoids in vitro and in vivo models will be discussed. These data are helpful in guiding researchers in the direction of studying phytonutrients with important effects in the prevention and treatment of melanoma.
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Melanoma , Neoplasias Cutáneas , Animales , Antocianinas/uso terapéutico , Melanoma/tratamiento farmacológico , Melanoma/patología , Ratones , Fitoquímicos/química , Fitoquímicos/farmacología , Fitoquímicos/uso terapéutico , Fitoterapia , Extractos Vegetales/química , Neoplasias Cutáneas/tratamiento farmacológico , Melanoma Cutáneo MalignoRESUMEN
Spore-based Bacillus spp. products are considered to have a higher probiotic potential compared to products containing only lactic acid bacteria because their viability in the gastrointestinal (GI) tract is higher, even when GI environmental conditions are unfavorable. The aim of this study was to assess the effect of a Bacillus subtilis, Bacillus licheniformis and Pediococcus acidilactici spore-based potential probiotic on the natural levels of postprandial endotoxemia. A total of 11 dogs completed the study: group 1-healthy dogs: n = 5; group 2-dogs with apparent dysbiosis: n = 6. For 30 days, the dogs were fed the probiotic product; clinical examinations and blood sampling were done before and after completion of the probiotic treatment. Endotoxin levels were assessed pre-meal, 6 h and 12 h post-meal, before initiation and after completion of the treatment. The results showed a decrease in endotoxin levels after treatment, especially 12 h post-meal (group 1: 20.60%; group 2: 44.93%). This study reports new information with regard to natural endotoxemia levels in dogs and suggests that a multi-strain formula (spore-based) consisting of B. subtilis, B. licheniformis and P. acidilactici is able to diminish endotoxin values.
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Syringa vulgaris L. (common lilac) is one of the most popular ornamental species, but also a promising not comprehensively studied source of bioactive compounds with important therapeutic potential. Our study was designed to characterize the chemical composition and to assess the antioxidant and cytotoxic properties of ethanolic extracts obtained from S. vulgaris L. flowers, leaves, bark, and fruit. The chemical profile of the ethanolic extracts was investigated using chromatographic (HPLC-DAD-ESI+, GC-MS) and spectral (UV-Vis, FT-IR) methods, while the protective effect against free radicals was evaluated in vitro by different chemical assays (DPPH, FRAP, CUPRAC). The cytotoxic activity was tested on two tumoral cell lines, HeLa, B16F10, using the MTT assay. Significant amounts of free or glycosylated chemical components belonging to various therapeutically important structural classes, such as phenyl-propanoids (syringin, acteoside, echinacoside), flavonoids (quercetin, kaempferol derivatives) and secoiridoids (secologanoside, oleuropein, 10-hydroxy oleuropein, demethyloleuropein, syringalactone A, nuzhenide, lingstroside) were obtained for the flowers, leaves and bark extracts, respectively. Furthermore, MTT tests pointed out a significant cytotoxic potential expressed in a non-dose-dependent manner toward the tumoral lines. The performed methods underlined that S. vulgaris extracts, in particular belonging to flowers and leaves, represent valuable sources of compounds with antioxidant and antitumoral potential.
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Antioxidantes/química , Flores/química , Estrés Oxidativo , Extractos Vegetales/química , Hojas de la Planta/química , Syringa/química , Animales , Línea Celular Tumoral , Cromatografía Líquida de Alta Presión , Flavonoides/análisis , Cromatografía de Gases y Espectrometría de Masas , Glucósidos , Glicósidos , Células HeLa , Humanos , Melanoma Experimental , Ratones , Fenoles , Fenilpropionatos , Hojas de la Planta/metabolismo , Quercetina/análisis , Espectroscopía Infrarroja por Transformada de FourierRESUMEN
Goat mastitis has become one of the most frequently diagnosed conditions in goat farms, with significant economic impact on the dairy industry. Inflammation of the mammary gland poses serious consequences on milk composition, with changes regarding biochemical parameters and oxidative stress markers. The aim of this paper is to present the most recent knowledge on the main biochemical changes that occur in the mastitic milk, as well as the overall effect of the oxidative and nitrosative stress on milk components, focusing on both enzymatic and nonenzymatic antioxidant markers. Mastitis in goats is responsible for a decrease in milk production, change in protein content with pronounced casein hydrolysis, and reduction in lactose concentration and milk fat. Milk enzymatic activity also undergoes changes, regarding indigenous enzymes and those involved in milk synthesis. Furthermore, during mastitis, both the electrical conductivity and the milk somatic cell count are increased. Intramammary infections are associated with a reduced milk antioxidant capacity and changes in catalase, lactoperoxidase, glutathione peroxidase or superoxide dismutase activity, as well as reduced antioxidant vitamin content. Mastitis is also correlated with an increase in the concentration of nitric oxide, nitrite, nitrate and other oxidation compounds, leading to the occurrence of nitrosative stress.
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ETHNOPHARMACOLOGICAL RELEVANCE: Combretum micranthum G. Don (CM) is extensively used in traditional medicine throughout West Africa and commonly known as "long-life herbal tea" or "plant to heal". Further, traditional healers frequently use the title plant to mitigate of renal disorders. AIM OF THE STUDY: To explore the nephroprotective property of standardised hydroalcoholic extract of Combretum micranthum in nicotinamide-streptozotocin induced diabetic nephropathy in rats. In addition, in-silico computational experiments were performed with bioactive compounds of the title plant against PPARα and PPARγ. MATERIAL AND METHODS: Male rats were made diabetic by a single intraperitoneal (ip) injection of STZ (50 mg/kg), 15 min after ip administration of NA (100 mg/kg) dissolved in normal saline. The diabetic rats received CM extract (200 and 400 mg/kg p.o.) daily, for eight weeks. Body weights and blood glucose (non-fasting and fasting) of rats were measured weekly. Daily food and water consumption were also measured. After 8 weeks of treatment, urine biochemical parameters such as N-Acetyl-ß-D-Glucosaminidase (NAG), urea (UR), uric acid (UA), creatinine (CRE), and serum markers of diabetes, kidney damage and liver damage such as insulin, lipid parameters), alanine aminotransferase (ALT), aspartate aminotransferase (AST) and gamma-glutamyl transferase (γGT), albumin (Alb), magnesium (Mg2+), calcium (Ca2+), phosphorus (P), were estimated. Blood glycosylated hemoglobin (HbA1C) were also estimated. kidney and liver were used for biochemical estimation of oxidative stress markers such as lipid peroxidation, superoxide dismutase (SOD) activity and glutathione peroxidase (GPx) activity. The kidney and pancreas were used for histopathological study. Further, HPLC chemoprofiling of CM extract and in-silico molecular simulation experiments were performed. RESULTS: At the end of eight weeks, renal damage induced by the consequence of prolong diabetic condition was confirmed by altered levels of serum and urine kidney and liver function markers, oxidative stress markers and histopathological variations in kidney. Treatment with CM extract ameliorated the diabetes mellitus-induced renal biochemical parameters and histopathological changes. Further, HPLC-UV & MS experiments revealed that CM extract contains several bioactive compounds including hyperozide (62.35 µg/mg of extract) and quercitrin (19.07 µg/mg of extract). In-silico experiment exhibited cianidanol (-17.133), epicatechin (-15.107) exhibited higher docking score against PPARα and luteoforol (-11.038), epigallocatechin (-10.736) against PPARγ. Based on docking and drug likeness score, four bioactive compounds were selected for molecular dynamic experiments. Cianidanol and epigallocatechin out of the 30 compounds are concluded as a potential candidate for the treatment of DN through activating PPARα and PPARγ target protein. CONCLUSIONS: Taken together, the present study provided the scientific footage for the traditional use of Combretum micranthum.
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Glucemia/efectos de los fármacos , Combretum , Diabetes Mellitus Experimental/tratamiento farmacológico , Nefropatías Diabéticas/prevención & control , Hipoglucemiantes/farmacología , Riñón/efectos de los fármacos , Extractos Vegetales/farmacología , Animales , Biomarcadores/sangre , Glucemia/metabolismo , Catequina/análogos & derivados , Catequina/aislamiento & purificación , Catequina/farmacología , Combretum/química , Diabetes Mellitus Experimental/inducido químicamente , Nefropatías Diabéticas/inducido químicamente , Nefropatías Diabéticas/metabolismo , Nefropatías Diabéticas/patología , Hipoglucemiantes/aislamiento & purificación , Riñón/metabolismo , Riñón/patología , Masculino , Simulación del Acoplamiento Molecular , Simulación de Dinámica Molecular , Niacinamida , Estrés Oxidativo/efectos de los fármacos , PPAR alfa/agonistas , PPAR alfa/metabolismo , PPAR gamma/agonistas , PPAR gamma/metabolismo , Extractos Vegetales/aislamiento & purificación , Ratas Wistar , Transducción de Señal , EstreptozocinaRESUMEN
Early detection of apoptotic cells on histological slides is of major importance for both diagnostic and research areas. In the current study, the aim was to propose a convenient method to stain the mitochondria and establish whether hepatocytes undergoing apoptosis can be identified in tissue sections using the proposed method. Liver tissue from five adult chinchillas was fixed with 10% neutral buffered formalin for Goldner's trichrome (GT) and Groat's iron hematoxylin and eosin (HE) stains and with Kolster's fixative for the Heidenhain's iron hematoxylin procedure. The HE and GT-stained sections showed the morphological features consistent with apoptosis i.e., homogenous intensely acidophilic cytoplasm, cell shrinkage with an irregular outline, nuclear shrinkage with cloudy karyoplasm, and karyopyknosis in the late stage. Sections stained with Heidenhain's iron hematoxylin method was used to pinpoint mitochondria and revealed cells which were undergoing the first stages of the apoptosis process i.e., disappearance of mitochondria from the cell, chromatin condensation and margination, paracentral localization of nucleoli, and vacuolated nuclei. In more advanced stages of apoptosis, cells presented significant nuclear and cytoplasmic changes. It was concluded that this is the first report targeting the mitochondria, by performing inexpensive histological staining techniques, in order to assess dead cells in situ.