Altered Enzyme Research Articles

Abstract P3041: DWORF Overexpression In The Heart Results In Enhanced Mitochondrial Function

Background: Calcium (Ca 2+ ) plays a pivotal role in regulating physiological processes in the heart including excitation-contraction coupling, Ca 2+ signaling, and mitochondrial function. It is well established that both Ca 2+ dysregulation and altered cardiac energy metabolism are universal features of heart failure (HF). Dwarf open reading frame (DWORF), a newly discovered microprotein, increases SERCA2a activity in part by relieving SERCA2a inhibition by phospholamban (PLN). DWORF overexpression via cardiac specific transgene or gene therapy is cardioprotective in genetic and experimental HF mouse models. In addition to activation of SERCA2a by PLN displacement, preliminary data indicates that enhanced mitochondrial function contributes to the cardioprotective effects of DWORF. Here we tested the effects of DWORF overexpression on cardiac mitochondrial function and Ca 2+ dynamics. Methods and Results: We isolated mitochondria from wildtype (WT) and DWORF transgenic (Tg) mouse hearts and measured O 2 consumption rates using a Seahorse mito stress test. We found that DWORF Tg mice have increased respiratory capacity compared to WT (DWORF Tg 1150.0 pmol/min, WT 799.4 pmol/min +/-146.2) and enhanced spare respiratory capacity (DWORF Tg 489.1 pmol/min, WT 229.4 pmol/min +/- 102.1). To measure mitochondrial Ca 2+ dynamics, we performed Ca 2+ retention capacity assays and found that DWORF Tg mice have increased Ca 2+ uptake rates compared to WT (DWORF Tg 32.32 s, WT 46.46 s +/- 4.39). Western blot analysis of the mitochondrial Ca 2+ uniporter (MCU) in mito extracts from WT and DWORF Tg hearts indicated elevated levels of MCU. However, electron transport chain (ETC) protein expression indicated no significant differences, indicating the enhanced mitochondrial activity observed in DWORF Tg mice was not due to increased ETC levels. Conclusions: DWORF Tg mouse heart mitochondria exhibit increased maximal respiration rates and enhanced spare respiratory capacity. However, ETC protein levels are unchanged, indicating altered enzyme activity rather than expression. Additionally, mitochondria from DWORF Tg hearts uptake Ca 2+ faster into the mitochondria through the increased levels of MCU, suggesting enhanced mitochondrial Ca 2+ dynamics.

Protective effect of Euphorbia thymifolia and Euphorbia hirta against carbon tetrachloride-induced hepatotoxicity in Wistar rats

Objectives The present study aims to investigate the protective effect of Euphorbia thymifolia and Euphorbia hirta extracts on in vitro antioxidant activity and in vivo analysis on hepatic marker enzyme levels and histopathological changes in the liver of carbon tetrachloride (CCl4) induced hepatotoxicity rats. Materials and methods This study includes 42 adult male Albino Wistar rats randomly divided into seven treatment groups, including control (basal diet, G1), CCl4-induced single dose (1.5 ml/kg, i.p.) as the negative control (G2), G1 supplemented with 300 mg/kg of ethanol extract of E. thymifolia (G3) and E. hirta (G4), G2 supplemented with 300 mg/kg of ethanol extract of E. thymifolia (G5), E. hirta (G6), and silymarin (25 mg/kg b.w.) used as a standard drug (G7) for 21-days experimental period. Results The ethanolic extracts of E. thymifolia and E. hirta exhibited potential in vitro antioxidant activity in a dose-dependent manner (25 μg/ml, 50 μg/ml, 100 μg/ml, 200 μg/ml and 250 μg/ml). Oxidative stress caused by CCl4-induced the liver damage, including changes in liver marker enzymes (aspartate aminotransferase, alanine aminotransferase, and alkaline phosphatase), enzymatic (superoxide dismutase and catalase), non-enzymatic antioxidants (lipid peroxides and glutathione) and hepatocellular alterations such as hydropic degeneration, irregular hepatocytes, and distention of the vein. Administration of E. thymifolia and E. hirta significantly (p < 0.05) restored the enzyme activity along with the histology of the liver. Conclusion The results from the current study demonstrate that E. thymifolia and E. hirta have the property of restoring hepatic redox capacity and antioxidant activities against CCl4-induced acute liver damage.

Serum liver enzyme alterations after laparoscopic cholecystectomy – a study.

The purpose of this study was to evaluate the effect of adoption of laparoscopic approach for cholecystectomy on liver function tests. This was a retrospective analysis of the data of 242 patients who had undergone laparoscopic cholecystectomy. Liver function tests were not ordered after laparoscopic cholecystectomy in routine surgical practice unless there was some specific reason in the postoperative period to initiate them. Only 14 patients had postoperative liver function tests and 2 had shown a transient rise in levels of enzymes. The rise in hepatic enzymes after laparoscopic cholecystectomy is transient and carries no clinical implications in patients with normal preoperative liver functions.

ISONIAZID-INDUCED LIVER INJURY: PHARMACOGENETIC ASPECTS

Inter-individual differences in anti-tuberculosis chemotherapy outcomes have been demonstrated in clinical practice, such as, achievement of intended therapeutic effect in some patients, and insufficiency or absence of response to treatment in others, with development of adverse drug reactions, especially, isoniazid-induced liver injury. Isoniazid-induced liver injuries are the main reason for isoniazid discontinuation, which further leads to considerably reduced effectiveness of anti-tuberculosis chemotherapy, increased relapse risk, and secondary drug-resistance of M. tuberculosis. Hepatotoxic reactions to chemotherapy are predicated by mutations in genes encoding enzymes participating in isoniazid biotransformation: N-acetyltransferase 2 (NAT2), cytochrome P450 2Е1, and glutathione-S-transferase. NAT2 gene polymorphism has been identified as risk factor for isoniazid hepatotoxicity. Nucleotide substitutions in NAT2 gene cause modification of enzyme protein structure, reduction of enzyme synthesis and alteration of its activity. Based on genetically-determined isoniazid acetylation rate, patients are referred to three acetylator types: rapid, intermediate, and slow. Correlations have been established in meta-reviews and systematic reviews between slow acetylator type and frequency of hepatotoxic reactions to isoniazid. Based on study findings, interrelation was shown between cytochrome P450 CYP2E1 gene polymorphism and increased risk of liver injury during chemotherapy with isoniazid. Contribution of GSTM1 and GSTT1 genotypes to isoniazid toxicity requires further exploration, as the obtained results were ambiguous and controversial.

Arsenic exposure diminishes ovarian follicular reserve and induces abnormal steroidogenesis by DNA methylation

Arsenic contamination is a worldwide public health problem, and the effect of arsenic on male reproduction has been extensively studied; however, data on the biotoxicity of arsenic in terms of female reproduction are more scarce. In this study, a human-cell-animal translational strategy was applied to explore the effect of arsenic exposure on ovarian steroidogenesis and its potential mechanism. We conducted a 1:1 propensity score matched case–control study involving 127 diminished ovarian reserve (DOR) cases and 127 healthy controls. The ovarian follicular fluid levels of 21 metal elements, including arsenic, were measured. The results showed that there were significant differences in follicular fluid metal profiles between DOR patients and controls and that arsenic, molybdenum, and strontium played important roles in DOR progression [OR (95 % CI): 2.203 (1.385, 3.503), 2.308 (1.490, 3.575) and 2.922 (1.864, 4.580), respectively]. In the primary ovarian granulosa cell culture model, we found that treatment with 8 μM arsenic for 24 and 48 h induced a decrease in human granulosa cell viability. The estradiol (E2) level was significantly decreased after arsenic exposure (P < 0.05), which was dependent on significant alterations (P < 0.05) in key enzymes in steroidogenesis. In addition, a model for sodium arsenite exposure through water in rats from weaning to sexual maturity was established. We evaluated ovarian development by monitoring the estrous cycle, observing ovarian pathology, and calculating the follicular proportion. RT–qPCR, Western blotting, and bisulfite-sequencing PCR were used to investigate the effect of arsenic exposure on ovarian steroidogenesis and its possible mechanism. The results indicated that steroidogenic factor-1 (SF-1) was an important target of the steroidogenesis disorder induced by arsenic exposure. Arsenic significantly increased the DNA methylation level (P < 0.05) in the promoter region of SF-1 to reduce its expression, subsequently decreasing the levels of steroidogenic acute regulatory protein (StAR), P450 cholesterol side-chain cleavage enzyme (CYP11A1), and aromatase (CYP19A1) (P < 0.05), leading to premature depletion of ovarian follicles.

Application of gamma irradiation on morphological, biochemical, and molecular aspects of wheat (Triticum aestivum L.) under different seed moisture contents

Nuclear technology is currently used as a tool in mutation breeding to improve crops by increasing genetic variation. The ionization of gamma rays produces large amounts of free radicals, simulating stressors in the natural environment. To avoid gamma-ray-induced oxidative stress, plants use antioxidant defense systems. Exposure of plants to irradiation can affect the germination, growth, and production of metabolites. Plants' sensitivity to irradiation depends on genetic and environmental factors such as moisture content. For this purpose, the effects of different gamma irradiation doses [0, 100, 200, 300, and 400 Gray (Gy)] and different seed moisture contents (7, 13, and 19%) on traits such as seed germination, seedling growth, molecular and biochemical alterations in antioxidant enzymes were examined in the current study. Based on the results, the highest seed germination percentage was observed in the interaction effect of seed moisture at 13% with an irradiation dose of 400 Gy (98.89%). Seedling survival percent and seedling length decreased with increasing doses of gamma irradiation at different seed moisture contents. Increasing gamma irradiation doses were reduced root and stem fresh and dry weight, and root and stem length. The highest level of catalase enzyme activity and expression was observed at 200 and 300 Gy irradiation doses at different moisture contents. The peroxidase and polyphenol oxidase gene expression were reduced at all contents of gamma irradiation doses and seed moisture compared to the control. It can be concluded that the dose of 200–300 Gy of gamma irradiation reduced plant growth by 30% in terms of fresh and dry weight and length of plants, as well as enhanced the expression of antioxidant enzymes. The results of this study could help plant breeders select an appropriate dose rate in wheat for further research.

Loss-of-function variant in chymotrypsin like elastase 3B (CELA3B) is associated with non-alcoholic chronic pancreatitis

BackgroundGenetic alterations in digestive enzymes have been associated with chronic pancreatitis (CP). Recently, chymotrypsin like elastase 3B (CELA3B) emerged as a novel risk gene. Thus, we evaluated CELA3B in two European cohorts with CP. MethodsWe analyzed all 8 CELA3B exons in 550 German non-alcoholic CP (NACP) patients and in 241 German controls by targeted DNA sequencing. In addition, we analyzed exons 6 and 7 by Sanger sequencing and the c.129+1G>A variant by melting curve analysis in 1078 further German controls. As replication cohort, we investigated up to 243 non-German European NACP patients and up to 1665 controls originating from Poland, Hungary, and Sweden. We assessed the cellular secretion and the elastase activity of recombinant CELA3B variants. ResultsIn the German discovery cohort, we detected a splice-site variant in intron 2, c.129+1G>A, in 9/550 (1.64%) CP patients and in 5/1319 (0.38%) controls (P=0.007, OR=4.4, 95% CI=1.5–13.0). In the European replication cohort, this variant was also enriched in patients (9/178 [5.06%]) versus controls (13/1247 [1.04%]) (P=0.001, OR=5.1, 95% CI=2.1–12.0). We did not find the two previously reported codon 90 variants, p.R90C and p.R90L. ConclusionsOur data indicate that CELA3B is a susceptibility gene for CP. In contrast to previous reports suggesting that increased CELA3B activity is associated with CP risk, the splice-site variant identified here is predicted to cause diminished CELA3B expression. How reduced CELA3B function predisposes to pancreatitis remains to be elucidated.

Abstract 300: Matrix metalloproteinase-9 responsive active targeted silica nanoparticles for pancreatic cancer detection by multispectral optoacoustic tomography

Abstract Purpose: Pancreatic ductal adenocarcinoma (PDAC) is the most lethal disease and the leading cause of cancer death worldwide. The survival rate of patients with this form of cancer is about 8%. The physiological barrier of the tumor microenvironment composed of a dense stroma and disorganized blood vessels creates a barrier for early identification and treatment of this deadly disease. In recent years, nanoparticle-based controlled delivery systems were developed to exploit the pathophysiology of biological systems such as acidic tumor microenvironment or the altered tumor-specific enzymes to improve the diagnosis and treatment efficacy. Here, we demonstrate the collagenase IV-mediated tumor site-selective release of the IR-780 imaging probe from the M-Ge-SDC1 nanoparticles, revealing the feasibility of the collagenase IV (MMP-9) responsive target specificity for diagnosing pancreatic cancer by multispectral optoacoustic tomography (MSOT) imaging. Methods: Mesoporous silica nanoparticles (MSN) with wormhole pore topology were synthesized and were characterized by dynamic light scattering (DLS) and transmission electron microscopy (TEM). The surface of MSN was conjugated with Gelatin-A to obtain M-Ge. The M-Ge particles were loaded with propidium Iodide (PI) or IR780 infrared imaging dye. The M-Ge surface was further conjugated with Syndecan-1 (SDC1) to improve the target specificity to release imaging cargo from the nanoparticles. Female athymic mice were orthotopically implanted with S2VP10 tumor cells. After a week of tumor implantation, mice were intravenously injected with M-Ge-SDC1 nanoparticles containing IR780 dye and were imaged with MSOT and AMI. Results: In the current study, Mesoporous silica nanoparticles with 27 nm diameter were synthesized. The Gelatin-A crosslinking on the surface of MSN particles as a gatekeeper was developed that could degrade upon contact with collagenase IV in the tumor microenvironment. The conjugation of SDC1 further improved the tumor specificity. The athymic mice orthotopically implanted with S2VP10 cells closely resemble human PDAC. Our results demonstrated that intravenous delivery of M-Ge-SDC1 nanoparticles could enzymatically degrade (MMP-9) and release IR780 at the tumor site and conjugation of SDC1 further improved the tumor specificity to detect the orthotopically implanted pancreatic tumors (p&amp;lt;0.0001,n=5). Conclusion: Due to the lack of effective screening tools, PDAC has the lowest survival rate and limited therapeutic efficacy for current FDA-approved drugs compared to other malignancies. Innovative technologies to develop engineered nanoparticles with active targeting moiety and dynamic imaging technology can overcome these limitations. Implementing such systems can enhance PDAC detection that can be translated into the clinic to improve health care. Citation Format: Abhilash Samykutty, Molly McNally, William M. MacCuaig, Jordan Hagood, Girish Mishra, Barish H. Edil, William E. Grizzle, Lacey R. McNally. Matrix metalloproteinase-9 responsive active targeted silica nanoparticles for pancreatic cancer detection by multispectral optoacoustic tomography [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 300.

The Effects of Bisphenol A on Oxidative Stress, Antioxidant Defence, Histopathological Alterations and Lysozyme Activity in Narrow-Clawed Crayfish (Pontastacus leptodactylus)

This study aimed to investigate of bisphenol A (BPA) on histopathological tissue (hepatopancreas and gills), immune ability (lysozyme activity), oxidative stress and antioxidant defence in crayfish (Pontastacus leptodactylus). Crayfish were exposed to BPA to various concentrations (0 (control), and 10, 50, 100 μg/L) for 5 and 20 days in triplicates. Histopathological findings showed moderate to severe hyperemia and inflammatory cell infiltrations, irregular epithelial cell arrangements, degeneration, necrosis and sloughing at the cells, collapse of the hepatopancreas tubules and hyperemia, oedema and sloughing at the epithelial cells of gills. In addition to swelling of gills, accumulation of hemocytes in the hemocoelic space of the gill lamellae relating to dosage were noticed. Antioxidant-related enzyme activities which superoxide dismutase (SOD), Glutathione-S-transferase GST, and glutathione reductase (GR) activity in 10, 50, 100 μg/L BPA group were significantly lower than control at 5 days (p&lt;0.05). Lysozyme activity (LSZ) was no significantly enhanced according to control (p&gt;0.05). These results suggest that the alterations in the antioxidant enzymes and the histological structure of the hepatopancreas, and gill tissue of crayfish can be used as potential biomarkers for risk assessment in aquatic ecosystem.

Correlation Between Sialidase NEU1 mRNA Expression Changes in Autism Spectrum Disorder.

Abnormal alterations in enzymes functioned in sialic acid modifications may be associated with ASD. In order to study the differences in peripheral blood sialidase (neuraminidase 1; NEU1) mRNA expression between autism spectrum disorder (ASD) children and healthy control, and to examine the correlation between NEU1 mRNA expression and the main behavioral phenotypes in children with ASD, we performed RT-qPCR to measure NEU1 mRNA expression in peripheral blood of 42 children with ASD and 42 healthy controls. In addition, we used the Autism Diagnostic Observation Schedule, Second Edition (ADOS-2) to measure and evaluate the behavioral phenotypes of children with ASD. Our results showed that NEU1 mRNA in the ASD group was significantly higher than in the control group (P < 0.0001). In addition, the ADOS-2 diagnostic scores of 42 children with ASD were correlated with their NEU1 mRNA expression results (R = 0.344, P = 0.0257). Moreover, in general, NEU1 mRNA expression was also positively correlated with the Social Affect (SA) of ADOS-2 (R = 0.3598, P = 0.0193) but not with the Restricted and Repetitive Behavior (RRB) (R = 0.15, P = 0.3432). Our results indicated that sialidase NEU1 mRNA was significantly increased in children with ASD, and its expression was correlated with the SA of children with ASD, which suggested that sialidase NEU1 may affect the SA of ASD. Our data highlighted the potential of NEU1 expression change may play an important role in ASD disease and lay the foundation for further studies on the relationship between NEU1 and ASD.

Antioxidant potential and protective effects of bee pollen extract against Salmonella induced hepatic and renal toxicity in BALB/c mice

Apicultural products comprise honey, bee pollen, propolis, bee wax and royal jelly which are known for their medicinal and health promoting properties. Among these, bee collected pollen allure much attention for its high nutritional properties. Here, we have investigated the protective role of bee pollen against Salmonella typhimurium induced biochemical alteration in BALB/c mice. Experimental animals (BALB/c mice) were divided equally into 10 different groups including normal and treated. Oxidative stress was induced by injecting Salmonella typhimurium (0.2 mL of 2×104 CFU/mL) intraperitoneally in mice. Bacteria induced sufficient alterations in serum enzymes within 5 days. Aqueous extracts of bee pollen of different crops (250 mg/kg) were administrated orally to control and experimental mice for 21 days. Then, hepatic and renal enzymes were measured with the help of standardized kits. Results of this study have revealed that bacterial infection increases the levels of the hepatic and renal enzymes levels (P <0.001) but after treatment with bee pollen extracts, altered levels of enzymes were normalized up to the normal levels. This normalization was highest with bee pollen of Helianthus annus. Administration of bee pollen alone did not produce any negative effects in mice.

Epigenetics: Through the pediatric urology looking glass

Epigenetics: Through the pediatric urology looking glass

Plantain peels restore sexual performance, hormonal imbalance, and modulate nitric oxide production and key enzymes of penile function in paroxetine-sexually impaired male rats.

This study focused on the effect of unripe (UPP) and ripe (RPP) plantain peels' extracts (200 and 400 mg/kg) on sexual behavior, hormonal profiles [testosterone, follicle-stimulating hormone (FSH), and luteinizing hormone (LH)], and enzymes [acetylcholinesterase (AChE), phosphodiesterase-5' (PDE-5), arginase, angiotensin-I converting enzyme (ACE), ecto-5'neucleotidase, and adenosine deaminase (ADA)] in paroxetine (PAR)-induced penile dysfunction rats. From the result, arginase, PDE-5', ACE, ecto-5'-nucleotidase ADA, and AChE activities, sexual activities, hormonal profile, and NO level were reduced, while thiobarbituric acid reactive species (TBARS) level increased (p < .05) relative to normal control rats. However, treatment with UPP and URP reduced the activities of these enzymes, decreased TBARS levels, and increased hormones, and penile NO levels in PAR-induced rats. Thus, the use of UPP and RPP could be channeled towards the improvement of sexual performance in erectile dysfunction (ED) disorder. PRACTICAL APPLICATIONS: Plantain fruits are a tropical staple food crop commonly consumed at various stages of ripeness and cooking methods. However, its peels are regarded as a waste product with reported cases of environmental menace. Interestingly, plantain peel is being used as a major raw material for industrial applications in the agro-based industries and in folklore for the treatment of many human ailments due to its rich phytochemicals such as polyphenols, carotenoids, alkaloids, etc., which have been reported. A prelude study has also indicated its usefulness in ED management, but further pharmacological investigations are needed to proffer information on its effect in ED management and its anti-androgenic activity in male Wistar rats. The information from this study could be of pharmaceutical importance in designing natural remedies capable of improving penile rigidity, hormone profiles, and alteration of enzymes linked with ED.

Molecular and Biochemical Detection of Insecticide Resistance in the Leishmania Vector, Phlebotomus papatasi (Diptera: Psychodidae) to Dichlorodiphenyltrichloroethane and Pyrethroids, in Central Iran.

The aim of the present study was to explore resistance markers and possible biochemical resistance mechanisms in the Phlebotomine sand fly Phlebotomus papatasi in Esfahan Province, central Iran. Homogenous resistant strains of sand flies were obtained by exposing P. papatasi collected from Esfahan to a single diagnostic dose of DDT. The adults from the colony were tested with papers impregnated with four pyrethroid insecticides: Permethrin 0.75%, Deltamethrin 0.05%, Cyfluthrin 0.15%, and Lambdacyhalothrin 0.05% to determine levels of cross-resistance. To discover the presence of mutations, a 440 base pair fragment of the voltage gated sodium channel (VGSC) gene was amplified and sequenced in both directions for the susceptible and resistant colonies. We also assayed the amount of four enzymes that play a key role in insecticide detoxification in the resistant colonies. A resistance ratio (RR) of 2.52 folds was achieved during the selection of resistant strains. Sequence analysis revealed no knockdown resistance (kdr) mutations in the VGSC gene. Enzyme activity ratio of the resistant candidate and susceptible colonies were calculated for α-esterases (3.78), β-esterases (3.72), mixed function oxidases (MFO) (3.21), and glutathione-S-transferases (GST) (1.59). No cross-resistance to the four pyrethroids insecticides was observed in the DDT resistant colony. The absence of kdr mutations in the VGSC gene suggests that alterations in esterase and MFO enzymes are responsible for the resistant of P. papatasi to DDT in central Iran. This information could have significant predictive utility in managing insecticide resistant in this Leishmania vector.

Assessment of Liver Enzyme Abnormality among Antiretroviral Therapy Experienced HIV- Seropositive Patients in Asmara, Eritrea

Liver disease are predominant in HIV/AIDS patients and a wide range of the population have been affected. However, studies that have assessed the burden and risks of liver enzyme alterations among antiretroviral therapy experienced patients are hardly available in Eritrea. Thus, the present study targets to determine the prevalence and the risks related with abnormal liver enzymes among HIVinfected individuals.

Pomegranate juice intake enhances the effects of aerobic training on insulin resistance and liver enzymes in type 2 diabetic men: a single-blind controlled trial

BackgroundLifestyle interventions are the first-line treatment for Type 2 diabetes mellitus (T2DM), highly prevalent in the community. This study aimed to examine the 8-week separate and combined effects of aerobic training (AT) and pomegranate juice intake (PJI) on insulin resistance and serum levels of liver enzymes, liver enzymes, and insulin resistance in men with T2DM.MethodsThis study evaluated the alterations of anthropometric indices, insulin resistance, and liver enzymes in 40 middle-aged men (40–50) with T2DM. Participants were randomly assigned into four groups: AT+PJI (n = 10); AT (n = 10); PJI (n = 10), and control (C) (n = 10). The AT program consisted of 60–75% of maximum heart rate (HRmax), 40–60 min/day, and 3 days/wk. Participants in the PJI group consumed 240 ml of pomegranate juice (sugar or additive-free) daily for 8 weeks.ResultsAT+PJI, PJI, and AT groups decreased anthropometric indices, HOMA-IR, and liver enzymes after 8 weeks. In contrast, the C group significantly increased the mentioned variables after the intervention. The result showed that AT+PJI significantly lowered liver enzymes, anthropometric indices, and HOMA-IR than AT or PJI alone. Also, the results of this study showed no significant difference between AT and PJI groups. However, in these groups, significant improvements in these variables were observed compared to the control group.ConclusionDue to the effect of combined AT+PJI in improving T2DM risk factors, it could be recommended for T2DM patients to prevent increased liver enzymes and insulin resistance.

Effect of Dietary Polyherbal Mixture on Growth Performance, Haemato-Immunological Indices, Antioxidant Responses, and Intestinal Morphometry of African Catfish, Clarias gariepinus

A 56-day feeding trial was conducted to evaluate the dietary effect of Nigeria polyherbal mixture (PHB) on growth, haemato-immune parameters, antioxidant enzyme activities, and intestinal morphometry of African catfish, Clarias gariepinus. Four diets were formulated with PHB powder at inclusion of 0 g/kg (control), 0.5 g/kg (PHB 0.5), 1.0 g/kg (PHB 1.0), and 2.0 g/kg (PHB 2.0) and fed to African catfish ( 6.32 ± 0.02 g /fish, 15 fish/50 L tank) in triplicates. Fish fed PHB 2.0 recorded higher final weight, weight gain, and specific growth rate ( p &lt; 0.05 ) compared to the control. There were no statistical differences ( p &gt; 0.05 ) in feed conversion ratio and protein efficiency ratio among the dietary groups. The haematological profile indicates that the fish fed PHB-1.0 had higher Hb, RBCs count, and Hct, and blood performance ( p &lt; 0.05 ) compared to the control. ALT levels were found higher in the control and the lowest values recorded in PHB groups ( p &lt; 0.05 ); however, AST did not differ significantly among the groups ( p &gt; 0.05 ). The highest heterophil counts and respiratory burst activity were recorded in PHB-1.0 and PHB-0.5 and PHB-2.0, respectively, while a numerically improved total immunoglobulin value was recorded in PHB-0.5 compared to the control ( p &gt; 0.05 ). Antioxidant enzymes such as superoxide dismutase and catalase had a significant improvement in fish fed PHB-based diet compared to the control ( p &lt; 0.05 ), while glutathione peroxidase showed no statistical differences between the groups ( p &gt; 0.05 ). Intestinal morphometric measurements showed that the fish fed PHB-1.0 had the highest villus height, area of absorption, and villus height/cryptal depth ratio compared to the control. Muscle thickness increased ( p &lt; 0.05 ) with increasing PHB level with the highest value recorded PHB-2.0. In conclusion, dietary PHB could improve growth, blood profile, immunity, antioxidant enzyme capacity, and intestinal morphometry of African catfish without any significant alteration in the liver function enzymes.

Astragalin attenuates AlCl3/D-galactose-induced aging-like disorders by inhibiting oxidative stress and neuroinflammation

Astragalin (AST) is a natural flavonoid with excellent antioxidant and anti-inflammatory activities. However, whether AST is an effective chemical for neuronal protection and its underlying mechanisms remain to be elucidated. In this study, we established a mouse model of cognitive impairment and aging-like phenotype induced by sequential administration of AlCl3 and D-galactose (Gal). We found that AST effectively ameliorated cognitive impairment in the model mice and improved their learning and memory performance in the Morris water maze (MWM) test. AlCl3/Gal-induced activation of astrocytes and microglia and inflammation were observed by immunohistochemistry and immunofluorescence, but could be attenuated by AST. In addition, alterations in oxidative stress-regulating enzymes or markers, including T-SOD, T-AOC, CAT, GSH-Px, and MDA, as well as the pro-inflammatory factors TNF-α, IL-1β, and IL-6, were restored. At the mechanistic level, AlCl3/Gal-intoxicated mice showed a significant elevation of Notch/HES-1 and NF-κB signaling axis corresponding to microglia activation and inflammation. AST attenuated the activation of Notch/HES-1 and NF-κB signaling axis, thus reducing the inflammation. In summary, AST is a promising natural product to protect neurons from toxin-induced injury, indicating its therapeutic potential for neurological disorders.

Alterations of Oxidative Stress Indicators, Antioxidant Enzymes, Soluble Sugars, and Amino Acids in Mustard [Brassica juncea (L.) Czern and Coss.] in Response to Varying Sowing Time, and Field Temperature.

The impact of elevated temperature at the reproductive stage of a crop is one of the critical limitations that influence crop growth and productivity globally. This study was aimed to reveal how sowing time and changing field temperature influence on the regulation of oxidative stress indicators, antioxidant enzymes activity, soluble sugars (SS), and amino acids (AA) in Indian Mustard. The current study was carried out during the rabi 2017–2018 and 2018–2019 where, five varieties of mustard viz. Pusa Mustard 25 (PM-25) (V1), PM-26 (V2), BPR-541-4 (V3), RH-406 (V4), and Urvashi (V5) were grown under the field conditions on October 30 (normal sowing; S1), November 18 (late sowing; S2) and November 30 (very late sowing; S3) situations. The S1 and S3 plants, at mid-flowering stage, showed a significant variation in accumulation of SS (8.5 and 17.3%), free AA (235.4 and 224.6%), and proline content (118.1 and 133%), respectively, and played a crucial role in the osmotic adjustment under stress. The results showed that S3 sowing, exhibited a significant induction of the hydrogen peroxide (H2O2) (110.2 and 86.6%) and malondialdehyde (23.5 and 47.5%) concentrations, respectively, which indicated the sign of oxidative stress in plants. Interestingly, the polyphenol oxidase, peroxidase, superoxide dismutase, and catalase enzyme activities were also significantly increased in S3 plants compared to S1 plants, indicating their significant roles in ameliorating the oxidative stress. Furthermore, the concentration of fatty acid levels such as palmitic, stearic, oleic, and linoleic acids level also significantly increased in S3 plants, which influenced the seed and oil quality. The study suggests that the late sowing significantly impaired the biochemical mechanisms in Indian mustard. Further, the mustard variety V4 (RH-406) was found to be effective for cultivation as well as environmental stress adoption in Indian soils, and it could be highly useful in breeding for developing heat-tolerant genotypes for ensuring the food security.

No Back-up Plan: Loss of Isozyme Diversity as a Promising Therapeutic Strategy for Cancer

Metabolic rewiring in cancer cells supports many aspects of tumor growth. Understanding the mechanisms that result in metabolic rewiring, such as altered enzyme expression, is key to identifying therapeutic vulnerabilities that selectively target cancer cells. In this issue of Cancer Research, Marczyk and colleagues analyze matched tumor-normal enzyme expression across 14 different cancer types and report that cancer cells exhibit a general loss of isozyme diversity (LID) relative to corresponding normal tissue. The authors hypothesized that the presence of a cancer dominant isozyme may reduce metabolic plasticity and uniquely sensitize cancer cells to isozyme-specific inhibitors. Several LID targets were identified, including acetyl-CoA carboxylase 1 (ACC1), which the authors validated using a clinically available inhibitor of ACC1/2. This study is the first to systematically evaluate isozymes affected by LID, which represents a promising strategy to target the unique metabolic demands of cancer. See related article by Marczyk et al., p. 1698.