Positive Control Research Articles

Herbs Used for the Management of Hypertension: A Systematic Review

Background: Hypertension is characterized by persistent high blood pressure and has emerged as a critical risk factor for severe cardiovascular diseases. Although several drugs have been designated to reduce blood pressure, these drugs can potentially cause side effects for patients. Therefore, medicinal plants are used to complement conventional drugs to treat various ailments. Methods: This study reviews the anti-hypertensive potential of herbs and plants and their mechanism of action in reducing blood pressure at their evaluated effective doses. Databases such as PubMed, Scopus, Science Direct, and Google Scholar were used to search articles from January 2016 to Sept 2022. The key search terms included “hypertension”, “lowers blood pressure”, “anti-hypertensive”, and “plants”. These generated 807 documents and using the PRISMA guidelines, thirty plants were identified for this review. Research studies with similar plant parts extracted from similar solvents, more than one dose, and animal models with a positive control were considered in this review. Studies that did not show significant blood pressure reduction were excluded. Results: Based on this criterion, the mechanism of actions of these plants was thematically grouped into three categories, namely 1) antioxidant, 2) angiotensin-converting enzyme (ACE) inhibition, and 3) calcium influx in vascular smooth muscle. The effective doses, plant parts used, and traditional medicine usage are presented in this review. Conclusion: Further research is highly recommended to identify the active compounds and to potentially develop them into anti-hypertensive drugs as well as to establish the safe doses and standardization of these plant extracts. This review is partially funded by grants from the Ministry of Higher Education.

Hederagenin Improves Adriamycin-induced Nephropathy by Inhibiting the JAK/STAT Signaling Pathway

ABSTRACT Background: Glomerulonephritis, a common kidney disease and major cause of end-stage renal disease, lacks effective treatment options. Hederagenin (HDG) exerts potent anti-inflammatory and protective effects on the kidneys and exhibits promise for the treatment of glomerulonephritis. This study aimed to investigate the therapeutic effects and mechanism of action of hederagenin in the context of adriamycin-induced nephropathy (ADN). Methods: C57BL/6 mice were randomly divided into 5 groups that included the control, model, low-dose HDG (20 mg/kg), high-dose HDG (40 mg/kg), and positive control (10 mg/kg irbesartan) groups. ADN was established in mice by administering a single injection of 10 mg/kg adriamycin. Renal pathology and fibrosis were assessed using haematoxylin and eosin (H & E) and Masson’s trichrome staining, whereas in vitro studies were conducted using cultured mouse podocytes (MPC5). Immunofluorescence staining and western blotting were performed to detect inflammation and the protein levels of signaling pathways. Results: The results revealed that HDG significantly improved adriamycin-induced abnormal serum creatinine, albumin, and urea nitrogen levels. HDG treatment reduced glomerular injury and fibrosis, particularly at high doses. Additionally, HDG effectively reduced adriamycin-induced activation of Janus kinase-signal transducer and activator of transcription (JAK/STAT) signaling and renal fibrosis while suppressing CD4+/CD8+ cell ratios in the kidneys and enhancing the immune response. Interestingly, when the JAK/STAT signaling pathway was activated by an agonist, the ameliorative effects of HDG on ADN were inhibited, thus suggesting that JAK/STAT signaling is a key target of HDG. Conclusion: HDG may represent a promising treatment option for glomerulonephritis by inhibiting JAK/STAT-mediated immune-inflammatory responses.

Probiotic potential of a novel endophytic Streptomyces griseorubens CIBA-NS1 isolated from Salicornia sp. against Vibrio campbellii infection in shrimp

A novel endophytic Streptomyces griseorubens CIBA-NS1 was isolated from a salt marsh plant Salicornia sp. The antagonistic effect of S. griseorubens against Vibrio campbellii, was studied both in vitro and in vivo. The strain was validated for its endophytic nature and characterized through scanning electron microscopy, morphological and biochemical studies and 16SrDNA sequencing. The salinity tolerance experiment has shown that highest antibacterial activity was at 40‰ (16 ± 1.4 mm) and lowest was at 10 ‰ salinity (6.94 ± 0.51 mm). In vivo exclusion of Vibrio by S. griseorubens CIBA-NS1 was studied in Penaeus indicus post larvae and evaluated for its ability to improve growth and survival of P. indicus. After 20 days administration of S. griseorubens CIBA-NS1, shrimps were challenged with V. campbellii. The S. griseorubens CIBA-NS1 reduced Vibrio population in test group when compared to control, improved survival (60.5 ± 6.4%) and growth, as indicated by weight gain (1.8 ± 0.05g). In control group survival and growth were 48.4 ± 3.5% and 1.4 ± 0.03 g respectively. On challenge with V. campbellii, the S. griseorubens CIBA-NS1 administered group showed better survival (85.6 ± 10%) than positive control (64.3 ± 10%). The results suggested that S. griseorubens CIBA-NS1 is antagonistic to V. campbellii, reduce Vibrio population in the culture system and improve growth and survival. This is the first report on antagonistic activity of S. griseorubens isolated from salt marsh plant Salicornia sp, as a probiotic candidate to prevent V. campbellii infection in shrimps.

Isolation and characterization of secondary metabolites from Bryophylum pinnatum (Lam.) Oken and assessment of wound healing efficacy using animal model

Wound healing is a complex and intrinsic process that implicates various stages in skin tissue regeneration, may further improvise using secondary metabolites. During a screening of plants traditionally utilized for wound healing, especially by tribals, we encountered Bryophyllum pinnatum (Lam.) Oken Plant. Further, we considered the leaf extract of Bryophyllum pinnatum (Lam.) Oken in various solvents for In vivo characterization of wound healing. The leaf extracts were prepared in petroleum ether, ethyl acetate, and ethanol and further identified the phytochemicals via advanced analytical techniques. The solvent extracts in the ointment base (1 % and 2 % w/w) were prepared and tested against the acute and burn wound models along with control and positive control (Cipladine 1 %). The ethanolic extract had higher phytochemical constituents was tested against the rat model, was found that 2 % w/w ethanolic ointment base stimulated the wound healing area, hydroxyproline content, and wound index and simultaneously reduced the epithelization period compared to the control. The present study concluded that the Ethanolic extract of Bryophyllum pinnatum leaves loaded ointment further promote the % of wound contraction rate, epithelization, and promotion of angiogenesis, exhibited the potential to be utilized to treat wounds.

Two Lactones Non-toxic to Humans Are Effective Repellents for the German Cockroach.

Lactones are cyclic esters of hydroxy carboxylic acids, present in several fruits and animals consumed by humans. There is evidence that some lactones modify insect behavior. The aim of the present study was to evaluate the repellent effect of four lactones (γ- and δ-nonalactone, and γ- and δ-dodecalactone) in first instar nymphs of the German cockroach, Blattella germanica (Linnaeus). To assess repellency, a nymph was placed on a circle of filter paper, half of which had been treated with lactone dissolved in acetone and the other half with acetone alone. The behavior of the nymph was recorded and the time the nymph spent in each half of the paper was quantified using Ethovision XT 10.1 software. Values of Distribution Coefficient (DC) were calculated: DC = (Tt - TA) / Tt, where Tt is the experimental time and TA is the time the nymph spent in the area treated with the repellent agent. DC can vary between 0 and 1. Values significantly higher than 0.5 indicate repellency. N,N-diethyl-meta-toluamide (DEET) was used as a positive control. DEET, δ- and γ-nonalactone caused repellency as from 77.9µg/cm2, whereas γ- and δ-dodecalactone had a repellent effect starting at 779.0µg/cm2. The values of DC for these concentrations were 0.89 (DEET), 0.86 (γ-nonalactone), 0.87 (δ-nonalactone), 0.83 (γ-dodecalactone), and 0.72 (δ-dodecalactone). To our knowledge, this is the first report of repellency produced by lactones in the German cockroach. This work allowed to identify two lactones that have a repellent effect similar to DEET.

Synthesis of New Glucose-containing 5-Arylisoxazoles and their Enzyme Inhibitory Activity

Abstract: Carbohydrates are an important group of biomolecules that have received special attention due to their significant role in the design and synthesis of new bioactive compounds. In this study, a new class of 5-arylisoxazole-glucose hybrids was designed and synthesized for evaluation of their inhibitory effects on α-glucosidase, α-amylase, and tyrosinase. The target compounds depicted selective α-glucosidase inhibitory activity over α-amylase, which is an important factor in reducing probable gastrointestinal problems in the treatment of type 2 diabetes. In this respect, compound 9a, possessing the phenylisoxazole group, was found to be the most potent α-glucosidase inhibitor (IC50 = 292.2 ± 0.1 μM) compared to acarbose (IC50 = 750.2 ± 0.1 μM) as the positive control. All compounds were also evaluated for their anti-tyrosinase effect, and among them, compound 9j, containing a fluoroaryl moiety, showed potent activity (IC50 = 50.1 ± 6.3 μM) in comparison to kojic acid (IC50 = 23.6 ± 2.6 μM). Also, docking studies were performed to investigate the probable mode of action, which indicated the construction of important H-bonding interactions between the sugar moiety and the enzyme’s active sites. According to the results, hybrids containing heterocycles attached to glucose can be used to inhibit α-glucosidase.

Pterostilbene-Loaded Polydopamine Nanoparticles Down-Regulate Tumor Necrosis Factor-α and Improve Myocardial Function in Mice with Acute Myocardial Infarction

TNF-α expression is related to myocardial function damage and recovery in patients with acute myocardial infarction, but its mechanism is not clear. 50 SD mice, 10 in each group, were in this study divided into TNF-α group, Notch1/eIF3a agonist group, model group, positive control group, and control group. The cardiac function score, myocardial infarction volume, myocardial cell apoptosis index, TNF-α expression, and Notch1/eIF3a pathway factor expression were observed. The size of polydopamine nanoparticles carrying pterostilbene was about 15.5 nm, and cardiac function score, myocardial infarction volume, myocardial cell apoptosis index, and myocardial cell apoptosis number in the model group and Notch1/eIF3a agonist group were higher than model group and Notch1/eIF3a agonist group (P < 0.05). Compared with model group, the Notch1/eIF3a agonist group, TNF-α group, and positive control group showed no differences (P > 0.05). The model group and Notch1/eIF3a agonist group had highest inflammatory response and lowest oxidative stress, which were significantly different from other groups (P < 0.05). The expression of TNF-α in Notch1/eIF3a agonist group, model group, and positive control group all decreased, which was significantly different from other groups (P < 0.05). The expressions of p-IL-6 and p-eIF3a in model group, Notch1/eIF3a agonist group and positive control group were all highest (P < 0.05). Carrying Pterostilbene-loaded polydopamine nanoparticles (PPNs) therefore inhibits apoptosis of cardiomyocytes, Notch1/eIF3a signaling pathway and inflammatory response and oxidative stress of myocardial system, and protects cardiomyocytes of model mice.

Sesquiterpene coumarins with anti-vitiligo and cytotoxic activities from Ferula samarkandica

Two undescribed sesquiterpene coumarins (1–2) and eight known analogues (3–10) were isolated from the roots of Ferula samarkandica Korovin, growing in Uzbekistan. Their structures were established by comprehensive spectroscopic analysis including NMR, HRESIMS, UV and IR data. The absolute configurations were determined based on ECD calculations and confirmed by single-crystal X-ray diffraction analysis. Six compounds carrying 7′-carbonyl, 3′-carbonyl and 3′-acetoxy (4, 5, 6, 7, 9, 10) demonstrated anti-vitiligo potentials, in addition to five (4, 7, 8, 9, 10) with cytotoxic effects. Evaluation of anti-vitiligo activity in B16 cells revealed that compounds 4–7 and 9 exhibited promotion effects on melanin synthesis in B16 cells more potent than the positive control 8-methoxypsoralen (8-MOP), while 5, 7, 9 and 10 could also stimulate the tyrosinase activity superior to 8-MOP. Besides, compounds 4 and 7–10 also presented certain cytotoxic activities against HeLa, HT-29 and A549 cell lines, wherein 8 displayed a significant cytotoxicity against HT-29 cells with an IC50 value of 8.33 ± 0.34 μM.

Sulforaphane decreases oxidative stress and inhibits NLRP3 inflammasome activation in a mouse model of ulcerative colitis

Excessive oxidative stress and NLRP3 inflammasome activation are considered the main drivers of inflammatory bowel disease (IBD), and inhibition of inflammasomes ameliorates clinical symptoms and morphological manifestations of IBD. Herein, we examined the roles of NLRP3 activation in IBD and modulation of NLRP3 by sulforaphane (SFN), a compound with multiple pharmacological activities that is extracted from cruciferous plants. To simulate human IBD, we established a mouse colitis model by administering dextran sodium sulfate in the drinking water. SFN (25, 50 mg·kg−1·d−1, ig) or the positive control sulfasalazine (500 mg/kg, ig) was administered to colitis-affected mice for 7 days. Model mice displayed pathological alterations in colon tissue as well as classic symptoms of colitis beyond substantial tissue inflammation. Expression of NLRP3, ASC, and caspase-1 was significantly elevated in the colonic epithelium. The expression of NLRP3 inflammasomes led to activation of downstream proteins and increases in the cytokines IL-18 and IL-1β. SFN administration either fully or partially reversed these changes, thus restoring IL-18 and IL-1β, substantially inhibiting NLRP3 activation, and decreasing inflammation. SFN alleviated the inflammation induced by LPS and NLRP3 agonists in RAW264.7 cells by decreasing the levels of reactive oxygen species. In summary, our results revealed the pathological roles of oxidative stress and NLRP3 in colitis, and indicated that SFN might serve as a natural NLRP3 inhibitor, thereby providing a new strategy for alternative colitis treatment.

The effect of ethanolic Acalypha Wilkesiana extract on blood sugar, sodium, potassium levels in alloxan-induced diabetic and salt-induced hypertensive Wistar rats

Diabetes Mellitus and hypertension often occur simultaneously. Hypertension affects approximately 70% of patients with diabetes, and the risk of cardiovascular disease in diabetic patients is three times higher than that in healthy individuals. The study was aimed at investigating the problem of diabetes and hypertension, and a possible remedy for it. Twenty-five (25) male wistar rats were assigned into five (5) groups of five (5) rats each. Group A served as negative control, and received water and feed only. Group B served as the positive control, and received alloxan160 mg/kg BW+ salt-loaded diet. Group C-E received alloxan 160 mg/kg BW+ salt-loaded diet + 100 mg/kg BW, 200 mg/kg BW, and 300 mg/kg BW of ethanolic acalypha wilkesiana (EAW) respectively. All administrations were given orally twice daily for 21 days. At the end of the treatment, blood samples were collected to estimate blood glucose, insulin, sodium, and potassium levels. Results are presented as mean±SEM, and were analysed using One-way ANOVA, statistical significance was considered at (P≤0.05). The results showed decrease in blood glucose levels and increase in insulin levels in groups C, D, and E compared to group B. There was also a decrease in sodium ion levels, and a concomitant increase in potassium ion levels in all wistar rat treatment groups when compared to group B. In summary, the results showed improved blood sugar, insulin, sodium, and potassium ion levels following EAW administration. It is therefore concluded that EAW administration was effective in improving diabetic and hypertensive complications in wistar rats.

The effect of garlic formulation on some hematological parameters and organ weight of male and female Wister albino rats

The study was conducted to investigate the impact of garlic formulation on certain haematological parameters and organ weights in both male and female rats. Haematological data, including the count of red blood corpuscles (RBC), the volume of packed cells (PCV), the count of white blood corpuscles (WBC), and the count of platelets, were evaluated using a fully automated haematology analyser. The liver, right kidney, left kidney, heart, and spleen of the rats were weighed to determine their respective organ weights. A total of thirty (30) albino Wister rats, consisting of both males and females, were randomly assigned to five groups, with six animals in each group. Group 1 acted as the positive control and was given 100% standard feed. Group II rats were given a mixture of 10% garlic and 90% feed. Group III animals were provided with a diet consisting of 20% garlic and 80% feed. Group IV received a diet containing 40% garlic and 60% feed. The final group, Group V, was fed a diet consisting of 80% garlic and 20% feed daily for a period of 21 days. The feed formulation was assessed at concentrations of 10, 20, 40, and 80 g/dl, based on the weight of the animal. The findings indicated a noteworthy (P<0.05) reduction in white blood cell count (WBC) and lymphocyte count (LYM). There were no statistically significant changes (P>0.05) observed in the levels of haemoglobin (HGB), haematocrit (HCT), mean corpuscular haemoglobin concentration (MCHC), mean corpuscular volume (MCV), and mean corpuscular haemoglobin (MCH) when compared to the control group. Furthermore, there was a noteworthy (P<0.05) reduction in HGB, MCV, and MCH when the rats reached a body weight of 80 g/100dl. The results indicated a statistically significant (P<0.05) reduction in the weight of the left kidney, heart, and spleen, as well as in the liver and right kidney weights at 20, 40, and 80 g/dl body weight. There were no statistically significant differences (P>0.05) in the body weight of the liver and right kidney, which were both 10g/dl. Ultimately, the use of garlic formulation in appropriate dosages can impact certain haematological markers and the weights of organs.

Effect of ethanolic stem bark extract of Annona muricata (soursop) on reproductive hormones and ovary in female Wistar rats

The effect of ethanol extract of stem bark of Annona muricata (soursop) on reproductive hormones and ovaries of Wistar rats was studied. The phytochemical analysis revealed the presence of alkaloids, flavonoids, flavones, flavonols, xanthones, flavanonols, flavonones, and condensed tannins. Twenty (20) experimental animals were randomly grouped into five groups of 4 rats per group and were allowed to acclimatize for 2 weeks. Rats in group A served as the control and were fed with rat feed and water, rats in group B severed as positive control and received low dose clomiphene citrate (20 mg/kg) daily for 21 days, rat feed and water, rats in groups C to E were fed with normal rat feed, water and received different doses of fresh stem bark of Annona muricata extract at 250, 500 and 1000 mg/kg respectively for 21 days. The animals were sacrificed under anesthesia after the experiment and the ovaries were surgically extracted, weighed and sent for histology. Blood samples were collected for analysis (hormonal assay of luteinizing hormone (LH), estrogen, follicle stimulating hormone (FSH), progesterone and prolactin. This was carried out using enzyme linked immunosorbent assay (ELISA) technique. The results showed that there were significant changes in the hormone level of the test groups when compared with the control rats. Significant increase was recorded in progesterone, estrogen, prolactin, follicle stimulating hormone and luteinizing hormone p< 0.05. Hence Annona muricata improves fertility in female Wistar rats. There were no histological changes in the ovaries and hence, it was concluded that Annona Muricata does not cause organ damage with increasing dose.

A Meta-Analysis of the Effects of Dietary Yeast Mannan-Rich Fraction on Broiler Performance and the Implication for Greenhouse Gas Emissions from Chicken Production

Dietary supplementation of yeast-derived mannan-rich fraction (MRF) could improve the gastrointestinal health and production efficiency of broilers, and, consequently, lower the environmental impacts of chicken production. The objective of this meta-analysis was to quantify the retrospective effects of feeding MRF (Actigen®, Alltech Inc., Nicholasville, KY) on the production performance of broilers. The meta-analysis database included 27 studies and consisted of 66 comparisons of MRF-supplemented diets vs. basal (i.e., negative control) and antibiotic-supplemented (i.e., positive control) diets. A total of 34,596 broilers were involved in the comparisons and the average final age of the birds was 35 days. Additionally, the impact of feeding MRF on the carbon footprint (feed and total emission intensities) of chicken production was evaluated using the meta-analysis results of broiler performance (MRF vs. basal diets) to develop a scenario simulation that was analyzed by a life cycle assessment (LCA) model. A database of all trials (MRF vs. basal and antibiotic diets) indicated that feeding MRF increased (p < 0.01) average daily feed intake (ADFI; +3.7%), final body weight (FBW; +3.5%), and average daily gain (ADG; 4.1%) and improved (p < 0.01) feed conversion ratio (FCR; −1.7%) without affecting (p > 0.05) mortality. A subdatabase of MRF vs. basal diets indicated that dietary MRF increased ADFI (+4.5%), FBW (+4.7%), and ADG (+6.3%) and improved FCR (−2.2%) and mortality (−21.1%). For the subdatabase of MRF vs. antibiotic diets, both treatments exhibited equivalent effects (p > 0.05) on broiler performance parameters, suggesting that MRF could be an effective alternative to in-feed antibiotics. Subgroup analysis revealed that different study factors (year of study, breed/strain, production challenges, and MRF feeding duration) influenced the effect of dietary MRF on broiler performance. Simulated life cycle analysis (LCA) indicated that feeding MRF decreased feed and total emission intensities, on average, by −2.4% and −2.1%, respectively. In conclusion, these results demonstrate that dietary MRF is an effective nutritional solution for improving broiler performance, an effective alternative to in-feed antibiotic growth promoters, and reduces the environmental impact of poultry meat production.

In vivo anthelmintic effect of Artemisia annua L. on oxyurid nematodes of laboratory mice

Oxyurids are common intestinal parasites of mammals, and they are the cause of recurrent helminthic infections, especially in children Hence natural herbal compounds are necessary for a repeatable cure. This study aimed to investigate the antinematodal effect of Artemisia annua L. n-hexane extract on oxyurids in vivo. In this study, A. annua L. n-hexane extract was applied to mice naturally infected with Syphacia obvelata and Aspiculuris tetraptera species orally at 300, 600, and 1200 mg/kg doses for seven days and the change occurring in the number of oxyurid eggs was investigated by the fecal flotation method. The nematodes in the necropsy were compared on the 8th day in all groups in terms of number, gender, and species. Albendazole (ABZ) was applied at a 5 mg/kg dose for three days as positive drug control, and corn oil was used as solvent control. The arithmetic mean of oxyurid eggs counted in fecal flotation 43.51% decrease in the drug control ABZ group and showed a 21.12% increase in the A. annua 1200 mg/kg group on the last day of application. When compared with the corn oil group, a 34.08%, 34.76% and 36.91% decrease in ABZ 5 mg/kg, A. annua 300 mg/kg and A. annua 1200 mg/kg group respectively were found in their necropsies in terms of nematode number. Non-polar compounds of A. annua L., very low concentration of ABZ and as a dilution solvent corn oil are probably responsible for this result.

Design, Synthesis, and Antifungal Evaluation of Diverse Heterocyclic Hydrazide Derivatives as Potential Succinate Dehydrogenase Inhibitors.

Plant pathogenic fungi pose a significant threat to agricultural production, necessitating the development of new and more effective fungicides. The ring replacement strategy has emerged as a highly successful approach in molecular design. In this study, we employed the ring replacement strategy to successfully design and synthesize 32 novel hydrazide derivatives containing diverse heterocycles, such as thiazole, isoxazole, pyrazole, thiadiazole, 1,3,4-oxadiazole, 1,2,4-oxadiazole, thiophene, pyridine, and pyrazine. Their antifungal activities were evaluated in vitro and in vivo. Bioassay results revealed that most of the title compounds displayed remarkable antifungal activities in vitro against four tested phytopathogenic fungi, including Fusarium graminearum, Botrytis cinerea, Sclerotinia sclerotiorum, and Rhizoctonia solani. Especially, compound 5aa displayed a broad spectrum of antifungal activity against F. graminearum, B. cinerea, S. sclerotiorum, and R. solani, with the corresponding EC50 values of 0.12, 4.48, 0.33, and 0.15 μg/mL, respectively. In the antifungal growth assay, compound 5aa displayed a protection efficacy of 75.5% against Fusarium head blight (FHB) at a concentration of 200 μg/mL. In another in vivo antifungal activity evaluation, compound 5aa exhibited a noteworthy protective efficacy of 92.0% against rape Sclerotinia rot (RSR) at a concentration of 100 μg/mL, which was comparable to the positive control tebuconazole (97.5%). The existing results suggest that compound 5aa has a broad-spectrum antifungal activity. Electron microscopy observations showed that compound 5aa might cause mycelial abnormalities and organelle damage in F. graminearum. Moreover, in the in vitro enzyme assay, we found that the target compounds 5aa, 5ab, and 5ca displayed significant inhibitory effects toward succinate dehydrogenase, with the corresponding IC50 values of 1.62, 1.74, and 1.96 μM, respectively, which were superior to that of boscalid (IC50 = 2.38 μM). Additionally, molecular docking and molecular dynamics simulation results revealed that compounds 5aa, 5ab, and 5ca have the capacity to bind in the active pocket of succinate dehydrogenase (SDH), establishing hydrogen-bonding interactions with neighboring amino acid residues.

Ultra-diluted complex additive in the diet reveals benefits in the intestinal tract of nursery-phase piglets.

This study aimed to assess an ultra-diluted (UD) complex, as a replacement for an antimicrobial growth promoter in diets, on growth performance, intestinal health, and inflammatory response of nursery piglets. The experiment lasted 37 days and involved 126 animals weaned at 21 ± 1.3 d, with an initial body weight of 5.62 ± 1.16kg. Piglets were assigned to six dietary treatments in a randomized block design with seven replicates and three piglets per pen as experimental unit. The treatments were: positive control (PC)- basal diet + 120mg/kg of chlorohydroxyquinoline; negative control (NC)- basal diet without additives; and NC containing 4.5; 6.0; 7.5 or 9.0kg of UD additive/ton diet. Performance data were calculated, and daily diarrhea was observed. Blood samples were collected for hematological analysis. At the end of the experiment, one animal per pen was slaughtered for organ weighing, pH, and the collection of intestinal samples for histopathology. Feces and cecal contents were collected for microbiological and antibiogram analyses. There was no difference in the performance between the treatments. Throughout the study, UD levels were equal to those of PC for diarrhea occurrence. Higher levels of UD complex led to higher total leukocyte counts. The 4.5 treatment showed a reduction in total and thermotolerant Enterobacteriaceae populations in piglet feces and an increase in lactic acid bacteria compared to PC. All treatments resulted in fewer duodenal histopathological alterations than those in the NC group. The use of UD additives, especially at 4.5kg/ton, is a good alternative to chlorohydroxyquinoline in piglet diets.

Antidepressant-like Effects of Chinese Quince (Chaenomeles sinensis) Fruit Based on In Vivo and Molecular Docking Studies

In this study, we examined the potential antidepressant-like effects of Chinese quince fruit extract (Chaenomeles sinensis fruit extract, CSFE) in an in vivo model induced by repeated injection of corticosterone (CORT)-induced depression. HPLC analysis determined that chlorogenic acid (CGA), neo-chlorogenic acid (neo-CGA), and rutin (RT) compounds were major constituents in CSFE. Male ICR mice (5 weeks old) were orally administered various doses (30, 100, and 300 mg/kg) of CSFE and selegiline (10 mg/kg), a monoamine oxidase B (MAO-B) inhibitor, as a positive control following daily intraperitoneal injections of CORT (40 mg/kg) for 21 days. In our results, mice treated with CSFE exhibited significant improvements in depressive-like behaviors induced by CORT. This was evidenced by reduced immobility times in the tail suspension test and forced swim test, as well as increased step-through latency times in the passive avoidance test. Indeed, mice treated with CSFE also exhibited a significant decrease in anxiety-like behaviors as measured by the elevated plus maze test. Moreover, molecular docking analysis indicated that CGA and neo-CGA from CSFE had stronger binding to the active site of MAO-B. Our results indicate that CSFE has potential antidepressant effects in a mouse model of repeated injections of CORT-induced depression.

In vitro digestion and fermentation of animal-derived fermentable substrates using canine and feline faecal inoculum

Abstract There is increasing interest in understanding the fermentative benefits of animal-derived fermentable substrates (ADFS) in pet foods. While previous research has assessed various ADFS using faecal inoculum derived from the cat, there is no published literature available for the dog. Additionally, very little is understood of the fermentation profiles of ADFS, such as skin and bone. Therefore, faeces were collected from a cohort of cats and dogs fed a complete and balanced high-protein diet and a selection of substrates were analysed in this study. Individual ADFS (tendon, bone cake, mechanically deboned meat (MDM), corium and hydrolysed collagen (a positive control)) were digested in vitro, followed by fermentation using either canine or feline faecal inoculum. Concentrations of butyrate, indole and ammonia were determined after 24 h of fermentation. Regardless of whether cat or dog faecal inoculum was used, fermentation of hydrolysed collagen produced the highest (P < 0.01) concentrations of butyrate and ammonia and the lowest concentrations of indole. For the other substrates, there were differences in the fermentation profiles between the canine and feline inocula. During the feline faecal fermentations, in comparison to the other substrates bone cake produced high (P < 0.05) butyrate concentrations, whereas in the dog faecal fermentations, MDM resulted in high (P < 0.05) butyrate concentrations. In conclusion, ADFS from different alternative co-products lead to different fermentation products, providing valuable information which may be considered in canine and feline dietary formulations.

Effect of high plant protein diet supplemented with Lysophospholipids-butyrate on the growth performance, liver health and intestinal morphology of Largemouth bass (Micropterus salmoides)

This study was conducted to evaluate the effects of dietary supplementation with lysophospholipids-butyrate (LPB) in a high plant protein diet on the growth performance, liver health, and intestinal morphology of Largemouth bass (Micropterus salmoides). Three experimental diets were formulated: a positive control diet (PC) containing 15.00 % soybean meal (SBM) and 13 % chicken meal；a high-plant-protein diet (HPP) with 28.00 % SBM; a diet (LPB) supplemented LPB at 0.1 % in HPP. Largemouth bass with an average initial body weight of 9.3 g were fed for 60 days, with 4 replicates per group and 20 fish per replicate. The results demonstrated that the HPP group exhibited significantly lower final body weight (FBW), weight gain rate (WGR) and specific growth rate (SGR) compared to the PC group. Moreover, the HPP diet induced a notable increase in liver collagen fiber and liver lipid content, which was alleviated by LPB supplementation. In comparison to the HPP group, the LPB group showed significant increases in serum albumin and albumin/globulin, along with a significant decrease in alanine aminotransferase activity (P < 0.05). Compared with the PC group, the mRNA levels of acetyl-CoA carboxylase-1 (ACC-1), diacylglycerol acyltransferase (DGAT), carnitine palmitoyl transferase-1 (CPT-1) and tumor necrosis factor-α (TNF-α) in HPP group were increased significantly (P < 0.05), while adipose triglyceride lipase (ATGL), hormone-sensitive lipase (HSL) as well as apoptotic gene B-cell lymphoma-2 (Bcl-2) were significantly down-regulated. The mRNA levels of fatty acid synthase (FAS), ACC-1, peroxisome proliferator activated receptor-α (PPAR-α), ATGL, interleukin-1β (IL-1β), transforming growth factor-β (TGF-β) and Bcl-2 were up-regulated (P < 0.05), while DGAT and TNF-α were down-regulated in the LPB supplemented groups than that in the HPP group (P < 0.05). In conclusion, the supplementation of 0.1 % LPB can alleviate liver lipid accumulation and fibrosis caused by high level of soybean meal.

Paeoniflorin alleviates TGF-β2-mediated extracellular matrix remodeling and oxidative stress in human trabecular meshwork cells.

The multifunctional profibrotic cytokine transforming growth factor-beta2 (TGF-β2) is implicated in the pathophysiology of primary open angle glaucoma. Paeoniflorin (PAE) is a monoterpene glycoside with multiple pharmacological efficacies, such as antioxidant, anti-fibrotic, and anti-inflammatory properties. Studies have demonstrated that paeoniflorin protects human corneal epithelial cells, retinal pigment epithelial cells, and retinal microglia from damage. Here, the biological role of PAE in TGF-β2-dependent remodeling of the extracellular matrix (ECM) within the trabecular meshwork (TM) microenvironment. Primary or transformed (GTM3) human TM (HTM) cells conditioned in serum-free media were incubated with TGF-β2 (5ng/mL). PAE (300μM) was added to serum-starved confluent cultures of HTM cells for 2h, followed by incubation with TGF-β2 for 22h. SB-431542, a TGF-β receptor inhibitor (10μM), was used as a positive control. The levels of intracellular ROS were evaluated by CellROX green dye. Western blotting was used to measure the levels of TGF-β2/Smad2/3 signaling-related molecules. Collagen 1α1, collagen 4α1, and connective tissue growth factor (CTGF) expression was evaluated by RT-qPCR. Immunofluorescence assay was conducted to measure collagen I/IV expression in HTM cells. Phalloidin staining assay was conducted for evaluating F-actin stress fiber formation in the cells. PAE attenuated TGF-β2-induced oxidative stress and suppressed TGF-β2-induced Smad2/3 signaling in primary or transformed HTM cells. Additionally, PAE repressed TGF-β2-induced upregulation of collagen 1α1, collagen 4α1, and CTGF expression and reduced TGF-β2-mediated collagen I/IV expression and of F-actin stress fiber formation in primary or transformed HTM cells. PAE alleviates TGF-β2-induced ECM deposition and oxidative stress in HTM cells through inactivation of Smad2/3 signaling.