Biochemical Level Research Articles

Evaluation of Iron Deficiency Anemia in Pregnancy

Anemia in pregnancy was the common nutritional deficiency and a frequent medical concern that leads to numerous known complications, impacting both maternal and fetal health. Objective: To evaluate the biochemical parameters such as serum Iron, Ferritin TIBS and TfS in anemic and non-anemic pregnant women. Methods: In this cross-sectional study, pregnant women age 18-45 years attending the gynecological department of Niazi welfare foundation teaching hospital, Sargodha from April 2023 to January 2024 were included. Serum iron, ferritin, total iron-binding capacity, and transferrin saturation were assessed from blood samples. Independent sample t-test in SPSS version 26.0 was applied to see significant difference in biochemical parameters at value of <0.05. Results: Findings revealed mean hemoglobin level of 10.26 ± 1.52 g/dl in all participants. Anemic results were found in majority, accounting for 268 (70.9%) cases. Among these anemic patients, 149 (55.5%) had mild anemia, 107 (40%) had moderate anemia, and 12 (4.5%) had severe anemia. Anemic patients had significantly lower serum ferritin level (p = 0.02), serum iron (p < 0.001), and transferrin saturation (p < 0.002) than non-anemic patients. Additionally, total iron-binding capacity, (p < 0.001) indicated the anemic status of the pregnant women. Conclusions: Biochemical parameters of pregnant women with anemia were reduced compared to those without anemia. Healthcare providers should closely monitor pregnant women with reduced biochemical levels to prevent complications for both the mother and the fetus.

One-Year Impact of Occupational Exposure to Polycyclic Aromatic Hydrocarbons on Sperm Quality.

Polycyclic aromatic hydrocarbons (PAHs) have toxic potential, especially as carcinogens, neurotoxins, and endocrine disruptors. The objective of this study is to know the impact of exposure to PAHs on the reproductive health of male workers who operate in solar thermal plants. Case-control study. A total of 61 men were included: 32 workers exposed to PAH at a solar thermal plant and 29 unexposed people. Seminal quality was studied both at the cellular level (quantity and quality of sperm) and at the biochemical level (magnitudes of oxidative stress in seminal plasma). In exposure to PAHs, a significantly higher seminal leukocyte infiltration was observed, as well as lower activity in seminal plasma of superoxide dismutase (SOD) and a reduced glutathione/oxidised glutathione (GSH/GSSG) ratio. The oxidative stress parameters of seminal plasma did not show a relationship with sperm cellularity, neither in those exposed nor in those not exposed to PAH. One year of exposure to PAH in a solar thermal plant does not have a negative impact on the sperm cellularity of the worker, either quantitatively (sperm count) or qualitatively (motility, vitality, morphology, or cellular DNA fragmentation). However, PAH exposure is associated with lower antioxidant capacity and higher leukocyte infiltration in seminal plasma.

Hepatoprotective potentials of Usnea longissima Ach. and Xanthoparmelia somloensis (Gyelnik) Hale extracts in ethanol-induced liver injury

In our study, the antioxidant and anti-inflammatory effects of different lichen applications were investigated in rats using an experimental ethanol toxicity model. 48 rats were used in the study and they were divided into 6 groups with 8 rats in each group. These groups were: control, ethanol (2 g/kg), ethanol + Usnea longissima Ach. (200 mg/kg), ethanol + Usnea longissima Ach. (400 mg/kg), ethanol + Xanthoparmelia somloensis (Gyelnik) Hale (100 mg/kg) and ethanol + Xanthoparmelia somloensis (Gyelnik) Hale (200 mg/kg). The experimental work continued for 21 days. Lichen extracts and ethanol were administered by gavage to rats divided into groups. According to the experimental protocol, the experimental animals were sacrificed and their liver tissues were isolated. Biochemical parameters in serum, histological examinations, oxidative stress and inflammation parameters both at biochemical and molecular level in liver tissues were performed. Oxidative stress and inflammatory response were increased in the liver tissue of rats treated with ethanol for 21 days, and liver functions were impaired. It was found that U. longissima and X. somloensis extracts showed good antioxidant activity and conferred protective effects against ethanol-induced oxidative stress and inflammation. This could be attributed to the presence of secondary metabolites in the extract, which act as natural antioxidants and could be responsible for increasing the defence mechanisms against free radical production induced by ethanol administration.

Virus-Induced galactinol-sucrose galactosyltransferase 2 Silencing Delays Tomato Fruit Ripening.

Tomato fruit ripening is an elaborate genetic trait correlating with significant changes at physiological and biochemical levels. Sugar metabolism plays an important role in this highly orchestrated process and ultimately determines the quality and nutritional value of fruit. However, the mode of molecular regulation is not well understood. Galactinoal-sucrose galactosyltransferase (GSGT), a key enzyme in the biosynthesis of raffinose family oligosaccharides (RFOs), can transfer the galactose unit from 1-α-D-galactosyl-myo-inositol to sucrose and yield raffinose, or catalyze the reverse reaction. In the present study, the expression of SlGSGT2 was decreased by Potato Virus X (PVX)-mediated gene silencing, which led to an unripe phenotype in tomato fruit. The physiological and biochemical changes induced by SlGSGT2 silencing suggested that the process of fruit ripening was delayed as well. SlGSGT2 silencing also led to significant changes in gene expression levels associated with ethylene production, pigment accumulation, and ripening-associated transcription factors (TFs). In addition, the interaction between SlGSGT2 and SlSPL-CNR indicated a possible regulatory mechanism via ripening-related TFs. These findings would contribute to illustrating the biological functions of GSGT2 in tomato fruit ripening and quality forming.

49 Crossing-talk between micronutrient and macronutrient metabolism: Implication in health and disease

Abstract Recent goals of swine ration formulation have evolved from preventing nutritional deficiency diseases and maximizing growth performance to optimizing immune function and disease resistance with least climate impact. Thus, certain micronutrients such as selenium (Se) are widely considered to be health beneficial at dietary concentrations above nutrient requirements. During the past decade, we have conducted a number of experiments to determine impacts and mechanisms of dietary Se concentrations (0.03 to 1 or 3 mg/kg) on metabolism of carbohydrate, lipid, and protein in pigs from growing to finishing. Strikingly, pigs fed high-Se diets developed hyperinsulinemia and insulin resistance. Those pigs also had increased lipid, protein, saturated fatty acids (myristic, palmitic, stearic, and behenic acids), and aspartic acid, glutamine, and alanine and decreased polyunsaturated fatty acids (dihomo-γ-linolenic, oleic, α-linolenic, and eicosatrienoic acids) in the liver and(or) adipose tissues. The high-Se diets increased and decreased amounts of 5 and 2 selenoproteins, respectively, in tissues of pigs. These effects were associated with down-regulated insulin signaling (INSR and AKT), glycolysis (glucokinase) and gluconeogenesis (PEPCK) and up-regulated lipogenesis (FOXO1, SREBP1, ACC and FAS), and protein synthesis (mTOR and RPS6/S6). In contrast, high dietary fat intakes up- or down-regulated expressions of all selenoproteins except for TXNRD2 and TXNRD3 and showed a strong effect on expressions of DIO2, GPX6, and SELENOK. These induced selenogenome expression changes were associated with increased serum malondialdehyde and plasma concentrations of tumor necrosis factor alpha and interleukin-6, along with risk of obesity and chronic inflammation. Altogether, metabolisms of micronutrients and macronutrients in pigs are coordinated through interdependent and mutually regulated crossing talks at tissue, cellular, biochemical, and signal levels.

β-glucans, SAM, and GSH fluctuations in barley anther tissue culture conditions affect regenerants’ DNA methylation and GPRE

BackgroundMicrospore embryogenesis is a process that produces doubled haploids in tissue culture environments and is widely used in cereal plants. The efficient production of green regenerants requires stresses that could be sensed at the level of glycolysis, followed by the Krebs cycle and electron transfer chain. The latter can be affected by Cu(II) ion concentration in the induction media acting as cofactors of biochemical reactions, indirectly influencing the production of glutathione (GSH) and S-adenosyl-L-methionine (SAM) and thereby affecting epigenetic mechanisms involving DNA methylation (demethylation—DM, de novo methylation—DNM). The conclusions mentioned were acquired from research on triticale regenerants, but there is no similar research on barley. In this way, the study looks at how DNM, DM, Cu(II), SAM, GSH, and β-glucan affect the ability of green plant regeneration efficiency (GPRE).ResultsThe experiment involved spring barley regenerants obtained through anther culture. Nine variants (trials) of induction media were created by adding copper (CuSO4: 0.1; 5; 10 µM) and silver salts (AgNO3: 0; 10; 60 µM), with varying incubation times for the anthers (21, 28, and 35 days). Changes in DNA methylation were estimated using the DArTseqMet molecular marker method, which also detects cytosine methylation.Phenotype variability in β-glucans, SAM and GSH induced by the nutrient treatments was assessed using tentative assignments based on the Attenuated Total Reflectance-Fourier Transform Infrared (ATR-FTIR) spectroscopy. The effectiveness of green plant regeneration ranged from 0.1 to 2.91 plants per 100 plated anthers. The level of demethylation ranged from 7.61 to 32.29, while de novo methylation reached values ranging from 6.83 to 32.27. The paper demonstrates that the samples from specific in vitro conditions (trials) formed tight groups linked to the factors contributing to the two main components responsible for 55.05% of the variance (to the first component DNM, DM, to the second component GSH, β-glucans, Cu(II), GPRE).ConclusionsWe can conclude that in vitro tissue culture conditions affect biochemical levels, DNA methylation changes, and GPRE. Increasing Cu(II) concentration in the IM impacts the metabolism and DNA methylation, elevating GPRE. Thus, changing Cu(II) concentration in the IM is fair to expect to boost GPRE.

Responses of survival, antioxidant system and intestinal microbiota of native snail Bellamya purificata to the invasive snail Pomacea canaliculata

Pomacea canaliculata is one of the most successful invader in worldwide, adversely affecting native ecosystem through direct predation or indirect competition, while the mechanism of indirect effects on native species remain poorly understood. To clarify the effects of P. canaliculata on the native near-niche species, Bellamya purificata, a widespread freshwater gastropod in China, was selected as the research subject. The changes of mortality, histology, antioxidant system as well as the intestinal flora diversity of B. purificata were explored in present study. The results showed that the median lethal dose of P. canaliculata culture solution for B. purificata was 23.76 ind/L and a concentration-dependent damage of both the gonad and hepatopancreas were observed, the gonadal villi were dissolved and the hepatopancreas cells were broken at 20 ind/L. Furthermore, different concentrations of P. canaliculata culture solution leading to the antioxidant damage on the enzyme or non-enzyme systems of B. purificata at various degrees. Additionally, a decrease in the diversity of the intestinal flora was observed, accompanied by an increase in the abundance of pathogenic bacteria such as Pseudomonas and Aeromonas after the exposure of the culture solution of P. canaliculata. Last, after being recovered in freshwater for 24 h, the antioxidant damage of B. purificata and the disturbance of intestinal flora diversity were still not recovered especially in the high concentration group. The indirect competitive mechanism of P. canaliculata culture solution on B. purificata were explored from the aspects of tissue, biochemical level and intestinal flora, which enriched the research of P. canaliculata invasion on native snails in China, and provided new insights for the study of the invasion strategy of P. canaliculata.

Nectopsyche sp (Trichoptera: Leptoceridae) sublethal effects caused by different concentrations of arsenic (As): a biochemical markers approach.

Environmental impacts related to arsenic (As) contamination are a persistent issue of particular interest in Latin American countries with increasing mining activities. In Ecuador, the redefinition of public policies to promote the increase in mining since 2008 has led to a significant rise in the presence of this heavy metal in rivers and effluents, sometimes exceeding the 0.1 mg L-1, limit recommended by Ecuadorian Environmental Regulations. This study aimed to evaluate the sublethal effects through the detection of biochemical biomarker changes (Catalase, Antioxidant capacity by FRAP, and Glutathione S-transferase) generated in larvae of Nectopsyche sp following prolonged exposure to different concentrations of As (C1 = 0.05 mg L-1, C2 = 0.1 mg L-1, C3 = 0.8 mg L-1) in a controlled environment, emulating the maximum limits allowed by current Ecuadorian legislation. While As concentration levels in water increased, so did levels in the tissue of Nectopsyche sp specimens. On the other hand, behavioral parameters (mortality and mobility) did not show differences in either time or As concentrations. However, both Catalase and Antioxidant capacity by FRAP levels tended to decrease with increasing As concentration, and in both cases, the differences were significant. Additionally, Glutathione S-transferase activity did not increase significantly. These results preliminarily demonstrate that biochemical responses change with varying As concentrations in Nectopsyche sp and are affected at behavioral and biochemical levels produced by the As at chronic levels.

The Dynamics of Allelochemicals and Phytotoxicity in Eisenia fetida during the Decomposition of Eucalyptus grandis Litter.

Allelopathy is an underlying and controversial mechanism for detrimental environmental effects in the management of Eucalyptus plantations. However, little attention has been paid to the dynamics of allelochemicals and phytotoxicity in soil fauna during litter decomposition. To explore the relationship between the dynamics of phytotoxicity and allelochemicals, a decomposition experiment was conducted using 4-year-old and 8-year-old Eucalyptus grandis litter (0, 10, 20, 30, and 45 days). The acute toxicity of Eisenia fetida was assessed, and a chemical analysis of the eucalyptus leaves was performed. Biochemical markers, including total protein, acetylcholinesterase (AChE) activity, and oxidative stress levels (SOD and MDA) were measured. A comet assay was used to determine DNA damage in E. fetida cells. The results showed that after 20-30 days of decomposition, E. grandis litter exhibited stronger phytotoxic effects on E. fetida in terms of growth and biochemical levels. After 20 days of decomposition, the weight and total protein content of E. fetida first decreased and then increased over time. SOD activity increased after 20 days but decreased after 30 days of decomposition before increasing again. MDA content increased after 20 days, then decreased or was stable. AChE activity was inhibited after 30 days of decomposition and then increased or stabilized with further decomposition. Soluble allelochemicals, such as betaine, chlorogenic acid, and isoquercitrin, significantly decreased or disappeared during the initial decomposition stage, but pipecolic acid significantly increased, along with newly emerging phenolic fractions that were present. More allelochemicals were released from 8-year-old litter than from 4-year-old E. grandis litter, resulting in consistently more severe phytotoxic responses and DNA damage in E. fetida. Scientific management measures, such as the appropriate removal of leaf litter in the early stages of decomposition, might help support greater biodiversity in E. grandis plantations.

Assessment of Antifungal Potential of Rhizospheric Bacterial Isolates of Daucus carota against Rhizoctonia solani

Rhizoctonia solani is an important plant pathogen which is responsible for causing a number of plant diseases in a variety of cropping plants. The current studies were carried out to explore antifungal potential of bacteria isolated from rhizospheric region of Daucus carota against Rhizoctonia. Of the total 21 bacterial isolates isolated, only 3 isolates namely CVO-3, CVO-4 and CRY-O-2 were shown antifungal potential against Rhizoctonia. Besides having antifungal potential these isolates also have some plant growth promoting attributes that will be an extra benefit of using these isolates as a biofungicides against R. solani. Thus, these isolates could be used as biofungicides against Rhizoctonia to prevent a variety of plant diseases like sheath blight in Oryza sativa, stem canker and black scurf in Solanum tuberosum and seedling diseases in Brassica napus. The isolates CVO-3, CVO-4 and CRY-O-2 were also characterized biochemical level by using methods based on Determinative Bacteriology given in Bergey’s manual and at molecular level using 16S rRNA sequencing and sequences of 16S rDNA segment were submitted to NCBI in GenBank database with the accession numbers OR775574 (Lysinibacillus boronitolerans ss3cvo3), OR775575 (Lysinibacillus boronitolerans ss4cvo4) and OR708523 (Shouchella rhizosphaerae ss1cryo2). The present investigation is an approach towards the use of these bacterial isolates from carrot rhizosphere to control the activities in the rhizosphere of other cropping plants.

Resveratrol alleviates thiram-induced tibial dyschondroplasia by regulating BMP-2, RUNX-2 and HIF-1α expressions in broiler chickens

Tetramethylthiuram disulfide (thiram) is a pesticide and fungicide, widely used in agriculture, and is a recognized cause of tibial dyschondroplasia (TD), a skeletal abnormality in fast-growing broiler chickens. This study aimed to investigate the effect of resveratrol (RSV) against TD induced by thiram in broilers. A total of 240 broilers were evenly divided into three groups (n = 80) i.e., Control, TD and RSV. All groups received feed and water ad libitum, whereas the TD and RSV groups received standard diet with an addition of thiram (50 mg/kg of feed) from 4–7 days to induce TD. After the cessation of thiram on day 8, the RSV group was administered RSV at 400 mg/kg body weight orally from 8–18 days. Histopathological changes in tibia were assessed using hematoxylin and eosin (H&E) staining. In addition, the mRNA and protein expressions of targeted genes were analyzed using RT-qPCR, immunohistochemistry, and western blotting. Thiram feeding resulted in an increased growth plate (GP) width, decreased vascularization, irregular shaped chondrocytes with pyknotic nuclei, lameness, poor production performance, reduced levels of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), total antioxidant capacity (T-AOC), alkaline phosphatase (ALP) and elevated levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and malondialdehyde (MDA). The mRNA and protein expressions of BMP-2 and RUNX-2 were downregulated, whereas HIF-1α was upregulated in TD affected broilers (p < 0.05). RSV administration resulted in a significant increase in mRNA and protein levels of BMP-2 and RUNX-2 (p < 0.05) and a decrease in the levels of HIF-1α. In addition, RSV improved vascularization, angiogenesis, alleviated lameness, and enhanced production efficiency. Furthermore, it restored the GP width, serum biochemical levels and liver antioxidant enzymes. In conclusion, this study indicated that resveratrol mitigated the adverse effects of thiram toxicity in broiler chickens.

Evaluation of Hormonal and Biochemical Profile of Ladakhi Cow during Estrous Cycle: The Future Perspective of Reproductive Management of Ladakhi Cow

The Ladakhi cow is an indigenous breed from Ladakh, India, and has a unique genetic makeup to adapt to high-altitude hypobaric stress. Furthermore, little study is available on examining the Ladakhi cow’s blood biochemicals and reproductive hormonal status during various stages of the estrus cycle. Hence, the current study was conducted on eight reproductively mature Ladakhi cows at the experimental dairy cattle unit in Leh-Ladakh, India, to determine the changes in serum biochemical and hormonal levels during the estrus cycle. The estrus behaviour was initially observed before the blood sampling on days 0 (the day the cow exhibited heat symptoms), 2nd, 8th, 17th, and 21st days. The serum was isolated for biochemical and hormonal profiling as per standard procedures using commercially available kits. The results revealed that serum biochemicals, viz. Ca, Fe, ALT, AST, ALP, TG, TP, Albumin, Glucose, UA, Creatinine, and Urea levels were within the normal range of cows at high altitudes. However, reproductive hormones, e.g. GnRH, FSH, LH, progesterone, estrogen, PGF2α, IGF-1, and Insulin varied from the reference level at different stages of the estrus cycle. This study’s findings have provided new insights into the serum biochemical and hormonal profiling of Ladakhi cows during the estrus cycle and this information may help to develop a reference range and baseline value for the serum biochemicals and hormonal levels of Ladakhi cows for other breeding programs and health monitoring.

AlphaFold2 Reveals Structural Patterns of Seasonal Haplotype Diversification in SARS-CoV-2 Nucleocapsid Protein Variants.

The COVID-19 pandemic saw the emergence of various Variants of Concern (VOCs) that took the world by storm, often replacing the ones that preceded them. The characteristic mutant constellations of these VOCs increased viral transmissibility and infectivity. Their origin and evolution remain puzzling. With the help of data mining efforts and the GISAID database, a chronology of 22 haplotypes described viral evolution up until 23 July 2023. Since the three-dimensional atomic structures of proteins corresponding to the identified haplotypes are not available, ab initio methods were here utilized. Regions of intrinsic disorder proved to be important for viral evolution, as evidenced by the targeted change to the nucleocapsid (N) protein at the sequence, structure, and biochemical levels. The linker region of the N-protein, which binds to the RNA genome and self-oligomerizes for efficient genome packaging, was greatly impacted by mutations throughout the pandemic, followed by changes in structure and intrinsic disorder. Remarkably, VOC constellations acted co-operatively to balance the more extreme effects of individual haplotypes. Our strategy of mapping the dynamic evolutionary landscape of genetically linked mutations to the N-protein structure demonstrates the utility of ab initio modeling and deep learning tools for therapeutic intervention.

Evaluation of muscular apoptotic changes and myogenin gene expression in experimental trichinosis after stem cells and atorvastatin added to ivermectin treatment

Trichinosis is a common parasitic disease that affects the striated skeletal muscles, causing apoptotic and degenerative changes associated with myogenin expression in the affected myocytes. Hence, this study aimed to assess the ameliorative effects of stem cells and atorvastatin added to ivermectin on the infected myocytes during the muscular phase of murine trichinosis. 120 laboratory Swiss albino male mice were divided into 10 groups, and each group was subdivided into intestinal and muscular phases (each n = 6); uninfected control; untreated infected control; infected received ivermectin monotherapy; infected received atorvastatin monotherapy; infected received stem cells monotherapy; infected received ivermectin and atorvastatin dual therapy; infected received ivermectin and stem cells dual therapy; infected received atorvastatin and stem cells dual therapy; infected received ivermectin 0.2, atorvastatin 40, and stem cells triple therapy; and infected received ivermectin 0.1, atorvastatin 20, and stem cells triple therapy. Intestinal phase mice were sacrificed on the 5th day post-infection, while those of the muscular phase were sacrificed on the 35th day post-infection. Parasitological, histopathological, ultrastructural, histochemical, biochemical, and myogenin gene expression assessments were performed. The results revealed that mice that received ivermectin, atorvastatin, and stem cell triple therapies showed the maximum reduction in the adult worm and larvae burden, marked improvement in the underlying muscular degenerative changes (as was noticed by histopathological, ultrastructural, and histochemical Feulgen stain assessment), lower biochemical levels of serum NK-κB and tissue NO, and lower myogenin expression. Accordingly, the combination of stem cells, atorvastatin, and ivermectin affords a potential synergistic activity against trichinosis with considerable healing of the underlying degenerative sequel.

Iron and Manganese concentrations in leaf tissues of Rhizophora mangle (Rhizophoraceae): implications for energetic metabolism

Introduction: Iron (Fe) and manganese (Mn) are bioessential micronutrients for plants but can impair the energetic metabolism when present at high levels. Objective: To assess the photosynthetic performance and oxidative damages in Rhizophora mangle L. leaf tissues at low and high concentrations of Fe (74 and 195 mg kg-1; Feleaf) and Mn (65 and 414 mg kg-1; Mnleaf). Methods: Photosynthetic pigments, chlorophyll a fluorescence, leaf CO2 assimilation and gas exchange and DPPH• radical scavenging capacity were sampled in R. mangle growing in estuarine forests in the North region of Espírito Santo State and the extreme South of Bahia State (Brazil) showing low and high Feleaf and Mnleaf. Results: Effects of high Fe and Mn were not identified on pigment levels. The increase in Feleaf and Mnleaf at the levels observed in this assessment had a positive effect on the number of reaction centers and on the efficiency of the oxygen-evolving complex, evaluated as K-band, while no changes were found in the parameters related to the excitation trapping efficiency at the active center of photosystem II. Distinct interference patterns of Fe and Mn on the functional processes of photosynthesis were identified, especially on CO2 assimilation and reactive oxygen species metabolism, with major effects on CO2 assimilation and carboxylation efficiency of Rubisco at high Mnleaf. Conclusion: These findings demonstrate the efficiency of R. mangle in positively regulating the electron transport chain in response to high Fe and Mn, at least in terms of the preservation of structure and functionality of the plant photosynthetic apparatus. Moreover, interference of high Mnleaf in R. mangle occurs at non-stomatal and biochemical levels. There is an antagonistic interference of these trace elements with the physiology of R. mangle, which is a dominant species in Brazilian mangroves.

Metabolic characterization and transcriptional profiling of polyphenols in Cannabis sativa L. inflorescences with different chemical phenotypes.

After the most comprehensive analysis of the phenolic composition in Cannabis reported to date, a total of 211 compounds were identified, phenolic profiles were able to discriminate cannabis varieties and a complex regulatory network for phenolics accumulation in Cannabis chemovars was highlighted. Female inflorescences of Cannabis sativa L. are plenty of secondary metabolites, of which flavonoids and phenolic acids have been investigated by far less than phytocannabinoids and terpenoids. Understanding the biochemical composition in phenylpropanoids of Cannabis inflorescences, the molecular basis of flavonoid synthesis and how their content can be modulated by specific transcription factors will shed light on the variability of this trait in the germplasm, allowing the identification of biologically active metabolites that can be of interest to diverse industries. In this work, an untargeted metabolomic approach via UHPLC-HRMS was adopted to investigate the composition and variability of phenylpropanoids in thirteen Cannabis genotypes differentiated for their profile in phytocannabinoids, highlighting that phenolic profiles can discriminate varieties, with characteristic, unique genotype-related patterns. Moreover, the transcription profile of candidate phenolics regulatory MYB and bHLH transcription factors, analyzed by RT-qPCR, appeared strongly genotype-related, and specific patterns were found to be correlated between biochemical and transcriptional levels. Results highlight a complex regulatory network for phenolic accumulation in Cannabis chemovars that will need further insights from the functional side.

New insights about the toxicity of 2,4-D: Gene expression analysis reveals modulation on several subcellular responses in Chironomus riparius

Herbicides are the main class of pesticides applied in crops and are capable of polluting the surrounding freshwater system; thus, understanding their impact on non-target species, whose mechanism of action is not described, helps to elucidate the real risks of these pollutants to the environment. 2,4-dichlorophenoxyacetic acid (2,4-D) is frequently detected in water and, due to its persistence, poses a risk to wildlife. In this way, the present work aimed to describe the implication of exposure to concentrations of 2,4-D already reported in aquatic environments in several physiological mechanisms of C. riparius at molecular and biochemical levels. To achieve this, bioassays were conducted with fourth instar larvae exposed to three concentrations of 2,4-D (0.1, 1.0, and 7.5 μg L−1). Larvae were collected after 24 and 96 h of exposure, and the expression of 42 genes, related to six subcellular mechanisms, was assessed by Real-Time PCR (RT-PCR). Besides, the activity of the enzymes catalase (CAT), glutathione S-transferase (GST), and acetylcholinesterase (AChE) was determined. The main metabolic route altered after exposure to 2,4-D was the endocrine system (mainly related to 20-hydroxyecdysone and juvenile hormone), confirming its endocrine disruptor potential. Four of the eleven stress response genes studied were down-regulated, and later exposure modulated DNA-repair genes suggesting genotoxic capacity. Moreover, only one gene from each detoxification phase was modulated at short exposure to 1.0 μg L−1. The molecular responses were not dose-dependent, and some early responses were not preserved after 96 h, indicating a transient response to the herbicide. Exposure to 2,4-D did not alter the activity of CAT, GST, and AChE enzymes. The responses described in this study reveal new mechanistic pathways of toxicity for 2,4-D in non-target organisms and highlight potential ecological consequences for chironomids in aquatic systems at the edges of agricultural fields.

Spinach flavonoid-rich extract: Unleashing plant defense mechanisms against cucumber powdery mildew

In response to the escalating demand for organic commodities, the exploration of non-chemical alternatives such as phytochemicals for plant disease management has gained momentum. This investigation focused on assessing the efficacy of a sustainable compound, Spinach Flavonoid-Rich Extract (SFRE), in mitigating cucumber powdery mildew (Podosphaera fusca) under controlled greenhouse conditions, and its potential in eliciting plant defense mechanisms at both biochemical and genetic levels. The application of SFRE resulted in a significant reduction in P. fusca disease severity, with a decrease of approximately 99%. High-Performance Liquid Chromatography (HPLC) analysis identified spinacetin as the dominant flavonoid in SFRE, with a concentration of 81.97 μg per 100 mg of the extract's dry weight. Furthermore, SFRE was found to induce systemic resistance in the infected seedlings, as evidenced by the upregulation of defense-related enzymes, including β-1,3-glucanase, chitinase, peroxidase, phenylalanine ammonia-lyase, and polyphenol oxidase, and the enhanced expression of β-1,3-glucanase, chitinase, and phenylalanine ammonia-lyase genes. In addition, SFRE application led to an accumulation of flavonoid/phenolic compounds and an enhancement in chlorophyll synthesis. A robust correlation was observed between phenolic compounds and the activities of defense enzymes, as well as the expression of the tested genes. This study substantiates that SFRE, as a reservoir of natural compounds, presents an effective and environmentally friendly alternative to chemical fungicides for P. fusca control, and could be incorporated into a comprehensive approach to managing cucumber powdery mildew through integrated disease management strategies.

Functional analysis of the novel mitochondrial tRNATrp and tRNASer(AGY) variants associated with type 2 diabetes mellitus.

Mutations in mitochondrial tRNA (mt-tRNA) genes that result in mitochondrial dysfunction play important roles in type 2 diabetes mellitus (T2DM). We pre-viously reported a large Chinese pedigree with maternally inherited T2DM that harbors novel mt-tRNA Trp A5514G and tRNA Ser(AGY) C12237T variants, however, the effects of these mt-tRNA variants on T2DM progression are largely unknown. To assess the potential pathogenicity of T2DM-associated m.A5514G and m.C12237T variants at genetic, molecular, and biochemical levels. Cytoplasmic hybrid (cybrid) cells carrying both m.A5514G and m.C12237T variants, and healthy control cells without these mitochondrial DNA (mtDNA) variants were generated using trans-mitochondrial technology. Mitochondrial features, including mt-tRNA steady-state level, levels of adenosine triphosphate (ATP), mitochondrial membrane potential (MMP), reactive oxygen species (ROS), mtDNA copy number, nicotinamide adenine dinucleotide (NAD+)/NADH ratio, enzymatic activities of respiratory chain complexes (RCCs), 8-hydroxy-deo-xyguanine (8-OhdG), malondialdehyde (MDA), and superoxide dismutase (SOD) were examined in cell lines with and without these mt-tRNA variants. Compared with control cells, the m.A5514G variant caused an approximately 35% reduction in the steady-state level of mt-tRNA Trp (P < 0.0001); however, the m.C12237T variant did not affect the mt-tRNA Ser(AGY) steady-state level (P = 0.5849). Biochemical analysis revealed that cells with both m.A5514G and m.C12237T variants exhibited more severe mitochondrial dysfunctions and elevated oxidative stress than control cells: ATP, MMP, NAD+/NADH ratio, enzyme activities of RCCs and SOD levels were markedly decreased in mutant cells (P < 0.05 for all measures). By contrast, the levels of ROS, 8-OhdG and MDA were significantly increased (P < 0.05 for all measures), but mtDNA copy number was not affected by m.A5514G and m.C12237T variants (P = 0.5942). The m.A5514G variant impaired mt-tRNA Trp metabolism, which subsequently caused mitochondrial dysfunction. The m.C12237T variant did not alter the steady-state level of mt-tRNA Ser(AGY), indicating that it may be a modifier of the m.A5514G variant. The m.A5514G variant may exacerbate the pathogenesis and progression of T2DM in this Chinese pedigree.

Polysaccharides from Alpinia oxyphylla fruit prevent hyperuricemia by inhibiting uric acid synthesis, modulating intestinal flora and reducing renal inflammation

Hyperuricemia (HUA) is one of the most common chronic diseases today, with a prevalence exceeding 14 % in both the United States and China. Current clinical treatments for HUA focus on promoting uric acid (UA) excretion and inhibiting UA production, but often neglect the strain on the liver and kidneys. The fruit of Alpinia oxyphylla (A. oxyphylla) is known to improve renal function, regulate metabolism, and exhibit anti-inflammatory effects; however, its effectiveness and mechanisms in treating HUA are not well understood. In this study, HUA mice induced by potassium oxonate and adenine were treated with A. oxyphylla polysaccharide (AFP) for 21 days. The levels associated with HUA were quantified using assay kits to evaluate the impact of AFP on HUA. Serum metabolomics and 16S rRNA sequencing were used to investigate the mechanisms by which AFP ameliorates HUA. The results showed that AFP treatment reduced abnormal biochemical levels, including UA, blood urea nitrogen, and creatinine, in HUA mice. AFP inhibited UA synthesis by regulating pyrimidine metabolism and the metabolism of alanine, aspartate and glutamate, reduced kidney inflammation, and promoted UA excretion by regulating intestinal flora. Thus, AFP appears to be an effective agent for alleviating HUA symptoms.