BACKGROUND:Afzelia africana is a tropical plant species with numerous documented ethnobotanical and medicinal properties. AIM/OBJECTIVE:The study focused on the isolation, purification, characterization and biological activity of betulinic acid from the bark of A. africana. METHODOLOGY:The series of methods employed include column fractionation and purification, characterization (MS, 1H, 13C, DEPT-135, HSQC, COSY and HMBC), antimicrobial (Micro-broth dilution, and synergistic effects), and antiplasmodial activity (MSF assay). RESULTS:Repeated column chromatographic fractionation of the methanol (extract eluent: n-hexane-ethyl acetate (75:25)) and TLC profiling afforded a 15 mg white crystal characterized as betulinic acid. The compound reported good antiplasmodial activity against 3D7 strain of P. falciparum (IC50: 12.89 ± 0.52 µg/mL). The compound exhibited varying degrees of bactericidal activities (MBC/MIC ≤ 4) against S. mutans, C. albicans 2, B. subtilis, K. pneumonia and bacteriostatic activities (MBC/MIC ≥ 4) against S. aureus, E. coli, methicillin-resistant S. aureus, P. aeruginosa, S. typhi and C. albicans 1. The compound further reported synergistic effects (FIC ≤ 0.5) against E. coli, K. pneumonia, S. aureus and S. mutans; partial synergism (FIC > 0.5 and < 1) against S. typhi, methicillin-resistant S. aureus, B. subtilis and indifferent effects (FIC > 1 and < 4) against P. aeruginosa when combined with tetracycline. The compound with Ciprofloxacin showed synergistic effect against Klebsiella pneumoniae, Streptococcus mutans, and Salmonella typhi. Additionally, it demonstrated further synergism with tetracycline against Escherichia coli, Klebsiella pneumoniae, and Staphylococcus aureus. In the antifungal synergistic studies, the combination of the compound with fluconazole and nystatin, ciprofloxacin exhibited partial synergism (FIC > 0.5 and < 1) against one strain of Candida albicans and full synergism (FIC ≤ 0.5) against a second strain of Candida albicans. When combined with fluconazole and nystatin. CONCLUSION:Betulinic acid has been isolated for the first time from the bark of Afzelia africana. It demonstrated good antiplasmodial activity against P. falciparum, as well as various bactericidal and bacteriostatic effects against multiple microbial strains. Additionally, betulinic acid demonstrated synergistic effects when combined with tetracycline, ciprofloxacin, fluconazole, and nystatin.
OBJECTIVES:Telomere length, the markers of biological aging, can be influenced by risk factors that may lead to health issues like chronic non-communicable diseases. This study explored the paternal telomere length influenced by diseases like diabetes and hypertension with age and its association with newborn telomere length (TL) and the telomerase gene. METHODS:In this cross-sectional study, 204 father-newborn dyads were recruited. The qPCR was used for the quantification of TL (T/S ratio), and Sanger sequencing was done for telomerase (TERT) genotype identification. Statistical Package for Social Sciences (SPSS) and GraphPad Prism Software were used for data analysis. The Spearman correlation was used to find an association between father and newborn TL. The Kruskal-Wallis and Mann-Whitney test was used to find differences among disease groups and TL. The P<0.05 was considered statistically significant. RESULTS:A positive correlation (r=0.39) (P<0.0001) was seen among fathers and newborn TL. The mean TL (T/S ratio) was found to be longer in healthy fathers and their newborns (1.67±1.18, 2.36±1.39), whereas significantly shorter TL (1.41±0.98, 2.02±1.58) (P=0.000) was seen in fathers suffering from diseases. The healthy fathers and their newborns TL (2.94±0.72, 3.10±0.61) were seen longer in the 15-20 (yrs) age category. Moreover, newborns of fathers (41-45 yrs) with both diabetes and hypertension had significantly longer telomere length (3.20±2.92) compared to their fathers (1.15±1.65) (P=0.006). The TERT risk genotype AC (rs2736100) was the most prevalent among the newborn girls. CONCLUSION:A positive association with shorter TL in newborns was found in fathers having chronic diseases such as diabetes and hypertension, highlighting the contribution of fathers in reprogramming newborns' telomere biology.
BACKGROUND:Paracetamol is a widely used over-the-counter drug for pain relief and fever management. However, its misuse through chronic overuse or acute overdose presents significant risks to human health, primarily causing hepatotoxicity and systemic oxidative stress. METHODOLOGY:This study evaluated the hepatoprotective, antioxidant, and anti-inflammatory effects of aqueous leaf extracts of Celosia trigyna and Euphorbia hirta in mitigating paracetamol-induced liver damage in male Wistar rats. RESULTS:Paracetamol administration (150 mg/kg) significantly elevated liver function markers (ALT, AST, ALP, and bilirubin), oxidative stress parameters (MDA), and inflammatory cytokines (IL-6 and TNF-α), while depleting antioxidant defenses (SOD and GSH). Disrupted lipid profiles were also observed in the paracetamol-only group. Pretreatment with Celosia trigyna and Euphorbia hirta extracts (125 mg/kg and 250 mg/kg) effectively ameliorated these effects by normalizing liver function markers, reducing oxidative stress and inflammation, and restoring lipid profiles. Molecular docking identified bioactive compounds such as rutin, quercetin, and kaempferol as potent inhibitors of Glutathione-S-Transferase, Tumor Necrosis Factor-alpha, and Cytochrome P450, with binding affinities of -9.3, -7.2, and -8.3 kcal/mol, respectively. These interactions underpin the antioxidant and anti-inflammatory activities observed in vivo. CONCLUSION:These findings suggest that Celosia trigyna and Euphorbia hirta have the potential to serve as natural prophylactic or therapeutic agents for mitigating paracetamol toxicity. Further research is required to isolate their active compounds and explore their synergistic potential with conventional treatments. This study bridges traditional medicine and modern pharmacology, offering innovative approaches to managing drug-induced liver.
Edible oils and ghee are vital parts of our daily culinary practices. In recent years, owing to heightened demand in the domestic and global markets, consistent reports regarding the adulteration of edible oils and ghee with substandard ingredients have been reported. Adulteration in edible oils is widespread, with distinctive contaminants, including cottonseed, mineral, and lower-cost oils like palm olein. In the case of ghee, it is repeatedly combined with animal fats, synthetic materials, or vanaspati. The consumption of contaminated oils and trans-fats within the human diet has resulted in adverse health effects, including cardiovascular diseases, digestive disorders, and even cancer. The review aims to summarize various adulterants found in edible oil (mustard oil) and ghee, their detection techniques, and harmful effects. This review provides an overview of the contemplation linked to the adulteration of edible oils and ghee, the compliance with relevant regulations, and the technological approaches available for detection. The detection technologies for identifying adulteration in edible oils are chromatography and spectroscopy biochemical methods and the use of high-precision analytical instruments. The presence of adulterants in edible oil and ghee undermines our societal integrity and our ethical standards. Awareness among consumers is essential for effectively combating these adulterations.
OBJECTIVE:Lately, various scientists have been paying a lot of consideration to the design of operational antimicrobial agents due to the rise of multiple drug-resistant strains. Therefore, this work is aimed at discovering the biochemical behavior of the analyzed polypeptides in relation to glutamine amidotransferase GatD (pdb id: 5n9m) for gram positive bacteria and beta-lactamase class A (pdb id: 5fqq) for gram negative bacteria. Additionally, this study aims to identify the specific atoms involved in the observed biochemical interactions between the studied complexes using computational methods. METHODS:In this work, five polypeptides were studied using insilico approach via Spartan 14 software, molecular operating environment, ADMETSar, and Gromacs. RESULTS:The descriptors obtained revealed the activities of the studied compounds, the molecular interaction between the studied ligands as well as glutamine amidotransferase GatD (pdb id: 5n9m) and beta-lactamase class A (pdb id: 5fqq) which thereby exposed compound 1 and 5 to be the compounds with greatest ability to inhibit the studied targets among other studied compounds. CONCLUSION:Our discoveries may open door for the design of collection of proficient polypeptide-based drug-like compounds as potential anti-microbial agents.
Marine environments harbor a wealth of bioactive peptides with potential anticancer properties, sourced from diverse organisms like tunicates, sea sponges, and mollusks. Through isolation, identification, and modification, peptides such as Stylisin and Papuamides have shown enhanced activity and progressed to clinical trials, underscoring their therapeutic promise. Enzymatic hydrolysis emerges as a favored method for peptide extraction from marine proteins, with sponges identified as particularly rich sources. Compounds like Jaspamide and Homophymins exhibit potent cytotoxic activity against cancer cells, highlighting their therapeutic potential. Additionally, peptides from ascidians and mollusks, including Aplidine and Kahalalide F, demonstrate significant anticancer properties. The study delves into peptides affecting apoptosis, microtubule dynamics, and angiogenesis inhibition, offering insights into potential cancer treatment mechanisms. Marine-derived peptides hold great promise as valuable candidates for novel anticancer therapies, with ongoing research aimed at unlocking their full therapeutic benefits.
The family of fibroblast growth factors (FGFs) consists of 22 members with diverse biological functions in cells, from cellular development to metabolism. The family can be further categorized into three subgroups based on their three modes of action. FGF19, FGF21, and FGF23 are endocrine FGFs that act in a hormone-like/endocrine manner to regulate various metabolic activities. However, all three members of the endocrine family require both FGF receptors (FGFRs) and klotho co-receptors to elicit their functions. α-klotho and β-klotho act as scaffolds to bring endocrine FGFs closer to their receptors (FGFRs) to form active complexes. Numerous novel studies about metabolic FGFs' structures, mechanisms, and physiological insights have been published to further understand the complex molecular interactions and physiological activities of endocrine FGFs. Herein, we aim to review the structures, physiological functions, binding mechanisms to cognate receptors, and novel biomedical applications of endocrine FGFs in recent years.
Cadmium (Cd) is a heavy metal pollutant widely distributed in the environment due to industrial activities, mining, and agricultural practices. Cadmium-induced Toxicity exerts profound effects on ER functioning through multiple mechanisms, leading to cellular dysfunction and pathological consequences. Cadmium disrupts protein folding and activates the unfolded protein response (UPR). Cd exposure leads to the accumulation of misfolded proteins, triggering UPR pathways mediated by critical ER transmembrane sensors: IRE1, PERK, and ATF6. The subsequent UPR aims to restore ER homeostasis but can also induce apoptosis under severe stress conditions. Cd disrupts ER calcium homeostasis by inhibiting the SERCA pump, further exacerbating ER stress. The generation of reactive oxygen species (ROS also plays a critical role in Cd toxicity, damaging ER-resident proteins and amplifying UPR activation). Cadmium also affects the lipid metabolism. This review examines the mechanisms by which Cd toxicity impairs ER functioning, disruption of protein folding and quality control mechanisms, and dysregulation of calcium signaling and lipid metabolism. The subsequent cellular consequences, including oxidative stress, apoptosis, and inflammation, are discussed in the context of Cd-induced pathogenesis of diseases such as Cancer and neurodegenerative and cardiovascular disorders. Finally, potential therapeutic strategies must be explored to mitigate the adverse effects of Cd on ER functioning and human health.
In this paper, we explore marine bioactive peptides with anticancer potential sourced from various marine organisms, including tunicates, sea sponges, and mollusks. Peptides like Stylisin and Papuamides have been isolated, identified, and modified to enhance their activity, with many advancing to clinical trials due to their diverse biological activities, promising prospects in medicine. Enzymatic hydrolysis is a favored method for extracting peptides from marine proteins, particularly from sponges known for their rich bioactive compounds. Compounds such as Jaspamide and Homophymins exhibit potent cytotoxic activity against cancer cells, underscoring their therapeutic potential. Additionally, peptides from ascidians and mollusks, such as Aplidine and Kahalalide F, demonstrate significant anticancer properties. This study also explores peptides influencing apoptosis, microtubule dynamics, and angiogenesis, providing insights into potential mechanisms for cancer treatment. While peptides like Neovastat and mycothiazole target known pathways, others such as patellamides act through unknown mechanisms, highlighting the intricate interactions of marine peptides with cancer cells. Overall, marine-derived peptides show promise as valuable candidates for developing novel anticancer therapies.
OBJECTIVES:The current study aimed to correlate OCT1 (organic cation transporter 1) polymorphisms with metformin response variability in Iraqi women with PCOS (polycystic ovarian syndrome) and determine the impact of OCT1 polymorphism. PCOS, an endocrine metabolic disorder, can seriously impact female health including infertility. Although its cause is unclear, it is usually known to be associated with hormonal imbalances. OCT1 is essential for metformin absorption in the liver. Recent research shown that OCT1 polymorphisms affects metformin responsiveness. METHODS:In the present work, a prospective case-control study was conducted at Department of Infertility, Karbala Teaching Hospital for Obstetrics and Gynecology. 100 PCOS patients and 50 healthy controls aged 20-40 were enrolled. Consultant gynecologist diagnosed PCOS patients using Rotterdam criteria and recommended metformin 500 mg twice daily for 3 months. At the start of the trial and after 3 months, all patients and healthy controls underwent hormonal, biochemical and genetic tests. RESULTS:The similar allelic frequencies of OCT1 polymorphism in PCOS and control groups was observed. Most patients with reference wild type alleles (C) showed considerable hormonal and metabolic responses to metformin, while those with mutant alleles (T) showed non-significant responses. CONCLUSION:FSH, prolactin and testosterone hormonal levels may be considered as candidate biomarkers for PCOS detection and metformin related biomedical respond.
INTRODUCTION:Human epidemiological studies have shown that diets rich in plant polyphenols have beneficial effects on various diseases including cancer. Epigallocatechin Gallate, a flavonoid polyphenol molecule, has been shown to be both chemotherapeutic and chemo-preventive in the treatment of several forms of cancer, including lung cancer. 80% of cancers of the lungs are non-small cell lung cancers. OBJECTIVE:The study was carried out to see the effects of Epigallocatechin Gallate in non-small cell lung cancer cells (A549) using in-vitro studies. MATERIALS AND METHODS:Cell Viability Assay was performed using MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay. Wound Healing assay was also performed at different concentrations of the compound. Dexamethasone and Doxorubicin, the drugs with anti-cancer properties served as control. A549 cell lines were used. RESULTS:In the current study, it was demonstrated using Cell viability assay and Wound Healing assay that Epigallocatechin gallate exhibits anti-proliferative activity on A549 lung cancer cells and inhibits cancer cell proliferation in a concentration and time-dependent manner. It was observed that Epigallocatechin gallate (P = 0.0016, P = 0.0018) could significantly inhibit the growth of lung cancer cells with IC50 values 60.55 ± 1.0 μM. The result of wound Healing assay suggests that Epigallocatechin gallate can inhibit the proliferation and migration of A549 cells with concentrations near or higher to 50 μM. CONCLUSION:Epigallocatechin gallate's protective effect has been shown in A549 lung adenocarcinoma cells in a time and dose-dependent manner. This suggests the implication of Epigallocatechin gallate for the prevention and therapy for lung cancer.
INTRODUCTION:Glomerular podocytes are specialized epithelial cells localized to the blood-urine interface of the kidney. Podocyte slit-diaphragm (SD), a size-and-charge-selective junction, is instrumental in blood ultrafiltration and the formation of protein-free urine. The SD consists of macromolecular complexes of several proteins, such as nephrin, podocin, and CD2-associated protein (CD2AP). CD2AP is an adapter protein and is considered to be crucial for the integrity of SD. Mutations in the SD proteins cause nephrotic syndrome (NS), characterized by proteinuria. SD proteins' structural features must be elucidated to understand the mechanism of proteinuria in NS. In this study, we expressed, purified, and biophysically characterized heterologously expressed human CD2AP.METHODS:Codon-optimized human CD2AP was expressed in E. coli Rosetta cells. The recombinant protein was induced with 1 mM IPTG and purified by Ni-NTA affinity chromatography. Analytical size-exclusion chromatography, blue native-PAGE, circular dichroism, and fluorescence spectroscopy were performed to decipher the oligomeric nature, secondary structural content, and tertiary packing of CD2AP.RESULTS:Our analysis revealed that CD2AP adopts a predominantly disordered secondary structure despite exhibiting moderate tertiary packing, characterized by low helical and β-sheet content. CD2AP readily assembles into homo-oligomers, with octamers and tetramers constituting the primary population. Interestingly, the inherent flexibility of CD2AP's secondary structural elements appears resistant to thermal denaturation. Frameshift mutation (p.K579Efs*7) that leads to loss of the coiled-coil domain promotes aberrant oligomerization of CD2AP through SH3 domains.CONCLUSION:We successfully expressed full-length human CD2AP in a heterologous system, wherein the secondary structure of CD2AP is predominantly disordered. CD2AP can form higher-order oligomers, and the significance of these oligomers and the impact of mutations in the context of size-selective permeability of SD needs further investigation.
BACKGROUND:Cancer chemotherapy with doxorubicin (DOX) has been linked to serious testicular damage and spermatotoxicity due to the induction of oxidative stress, inflammation, and apoptosis. Thus, the current study was carried out to assess the potential ameliorative impact of diosmin, an antioxidant drug, against DOX-mediated spermatoxicity and testicular injury in rats.MATERIAL AND METHODS:In the experimental protocol, rats were grouped into 4: Group 1 received vehicle and saline for 8 weeks while group 2 received diosmin and saline concomitantly for 8 weeks. Group 3 was given 3 mg/kg intraperitoneal DOX once every 7 days for 8 weeks. Group 4 was given 40 mg/kg of diosmin orally for 56 days followed by DOX diosmin administration after one hour. After 56 days of treatment, sperm quality, hormonal testing, biochemical parameters, and histological alterations in the testes were evaluated.RESULTS:DOX-induced reduce spermatogenic function, testicular 3- and 17β-Hydroxysteroid dehydrogenases, and serum follicle stimulating hormone, luteinizing hormone, and testosterone. It also enhanced inflammation, testicular oxidative damage, and apoptosis. The histopathologic examinations corroborated the biochemical results obtained. Significantly, diosmin treatment reduced DOX-induced injury, as evidenced by restored testicular architecture, increased steroidogenesis, preservation of spermatogenesis, suppression of oxide-inflammatory response, and apoptosis.CONCLUSION:It was found that through diosmin antioxidant, anti-apoptotic, and anti-oxido-inflammatory it presents a possible therapeutic alternative for protecting testicular tissue against DOX's harmful effects.
BACKGROUND:Diabetic nephropathy (DN) is a prevalent and chronic, severe complication of diabetes, representing a serious global health concern. Early detection of DN is essential for initiating timely and effective therapeutic interventions and accurately assessing prognosis. Neutrophil Gelatinase-Associated Lipocalin (NGAL), a low molecular weight protein, has emerged as a potential biomarker for DN due to its association with renal injury and its ability to provide early indications of kidney damage. NGAL levels in both serum and urine are elevated in individuals with renal damage, making it a valuable biomarker for detecting early signs of kidney impairment in the context of diabetes. This study aims to investigate the utility of NGAL as an early biomarker for DN and explore its correlation with various clinical parameters associated with the disease. Understanding the relationship between NGAL levels and clinical parameters such as glycemic control, renal function, blood pressure, and duration of diabetes is crucial for comprehensively evaluating the potential of NGAL as a diagnostic and prognostic tool for DN. Furthermore, assessing the sensitivity and specificity of NGAL in detecting early-stage DN will provide valuable insights into its clinical applicability and reliability.METHODOLOGY:A planned meta-analysis was conducted following PRISMA and MOOSE guidelines. The PubMed database was searched from January 2016 to June 2023 for English-language studies on DN and NGAL. Fifteen eligible studies were included as per the criteria. Data on serum NGAL levels in DN patients and healthy controls were analyzed using Stata 16.0 software.RESULT:The study revealed a significantly higher mean serum NGAL level in DN patients (168.08 ng/ml, 95% CI: 105.50-230.67) compared to healthy controls (75.02 ng/ml, 95% CI: 43.02-107.03), demonstrating NGAL's potential as a biomarker (P=0.01).CONCLUSION:NGAL offers a powerful tool for DN diagnosis, staging, and monitoring, surpassing traditional markers in sensitivity. Challenges include defining universal threshold values and ensuring consistent test performance across diverse clinical settings. The study underscores NGAL's potential in transforming DN diagnosis and management.
OBJECTIVES:Accumulative effects of heavy metals and polycyclic aromatic hydrocarbon could result in various toxicities. This study evaluated the effects of long-term exposure to low doses of nickel and benzo [a] anthracene on the kidney of rats, simulating human exposure through food. METHODS:Thirty-six (36) Male rats weighing between 80-100 g were assigned into six groups of 6 animals each; Group A (normal), Group B1 and B2 (fed nickel contaminated feed for 12 and 24 weeks), Group C1 and C2 (fed benzo [a] anthracene contaminated feed for 12 and 24 weeks). Blood and kidney of the rats were harvested after animal sacrifice. Serum creatinine and urea concentration and renal Superoxide Dismutase (SOD) activity, GSH, MDA, protein carbonyl, and total protein concentration by spectrophotometric methods. While the concentration of 8-oxodeoxyguanosine in kidney was determined by ELISA method and protein carbonyl by colorimetric method. Renal histological analysis was done with H and E staining. Statistical analysis was performed with Statistical Package for Social Sciences (SPSS) and statistical significance was accepted 95 percent confidence level. RESULT:From the results, urea concentration increased significantly (P<0.05) in the nickel exposed group after 24 weeks exposure whereas creatinine concentration increased significantly (P<0.05) after 12 weeks of exposure when compared with the control. Comparison of the serum urea and creatinine level of the benzo [a] anthracene exposed group with the control showed no significant (P>0.05) difference. Histological observations indicate glomerular atrophy and widened capsular space haemorrhagic areas, visceral and parietal layer of the Bowman's capsule, the proximal convoluted tubule in the nickel exposed group while the kidney of benzo (a) anthracene exposed rats showed deviation in the histo-architecture of the renal parenchyma as evidenced by glomerular atrophy and widened Bowman's capsular space and focal haemorrhagic areas. Protein thiol level and Superoxide dismutase activity was significantly (P<0.05) depleted in the benzo [a] anthracene exposed groups. The levels of total protein, protein carbonyl, and 8-oxodeoxyguanosine were significantly (P<0.05) elevated in the nickel and benzo [a] anthracene exposed groups. CONCLUSION:This study demonstrated the oxidative stress causing effects of benzo [a] anthracene and nickel in the kidney. It also shows that consistent exposure to low doses of the contaminants for a lifetime might result in renal oxidative stress with consequential loss of renal function.
OBJECTIVES:Jumonji C domain-containing (JMJD) 2B (JMJD2B) is a transcriptional cofactor and histone demethylase that is involved in prostate cancer formation. However, how its function is regulated by posttranslational modification has remained elusive. Hence, we examined if JMJD2B would be regulated by lysine methylation.METHODS:Through in vitro methylation assays and Western blotting with methyl-lysine specific antibodies, we analyzed lysine methylation within JMJD2B. Identified methylated lysine residues were mutated to arginine residues and the respective impact on JMJD2B transcriptional activity measured with a reporter gene assay in human LNCaP prostate cancer cells.RESULTS:We discovered that JMJD2B is methylated on up to six different lysine residues. Further, we identified the suppressor of variegation 3-9/enhancer of zeste/trithorax (SET) domain-containing protein 7/9 (SET7/9) as the methyltransferase being responsible for this posttranslational modification. Mutating the methylation sites in JMJD2B to arginine residues led to diminished coactivation of the Ju-nana (JUN) transcription factor, which is a known oncogenic protein in prostate tumors. In contrast, methylation of JMJD2B had no impact on its ability to coactivate another transcription factor associated with prostate cancer, the DNA-binding protein E26 transformation-specific (ETS) variant 1 (ETV1). Consistent with a potential joint action of JMJD2B, SET7/9 and JUN in prostate cancer, the expression of JMJD2B in human prostate tumors was positively correlated with both SET7/9 and JUN levels.CONCLUSIONS:The identified SET7/9-mediated methylation of JMJD2B appears to impact its cooperation with selected interacting transcription factors in prostate cancer cells. Given the implicated roles of JMJD2B beyond prostate tumorigenesis, SET7/9-mediated methylation of JMJD2B possibly also influences the development of other cancers, while its impairment might have relevance for obesity or a global developmental delay that can be elicited by reduced JMJD2B activity.
BACKGROUND:Phospholipids are highly diverse molecules with pleiotropic biological roles, from membrane components and signaling molecules, whose composition can change in response to both endogenous and external stimuli. Recent lipidomic studies on edible bivalve mollusks were focused on lipid nutritional value and growth requirements. However, no data are available on phospholipid profiles during bivalve larval development. In the model marine bivalve Mytilus galloprovincialis, early larvae (up to 48 hours post fertilization-hpf) undergo dramatic molecular and functional changes, including shell biogenesis and neurogenesis, that are sustained by egg lipid reserves. Changes in phospholipid composition may also occur participating in the complex processes of early development.OBJECTIVE:The lipidome of M. galloprovincialis eggs and early larval stages (24 and 48 hpf) was investigated in order to identify possible changes in phospholipid classes and related metabolic pathways that may play a role in key steps of development.MATERIALS AND METHODS:Lipidomic analysis were performed by NMR spectroscopy and liquid chromatography-mass spectrometry (LC-MS), with focus on phospholipids. Shifts in relative species composition of phosphatidylcholine, phosphatidylethanolamine, plasmalogen, and ceramide aminoethylphosphonate-CAEP, the bivalve analogue of the main mammalian ceramide sphingomyelin, were observed. Expression of genes involved in ceramide homeostasis was also modulated from eggs to early larval stages.RESULTS:The results represent the first data on changes in phospholipid composition in bivalve larvae and suggest a functional role of phospholipids in mussel early development.CONCLUSION:The results underline the importance of lipidomic studies in bivalve larvae, in both physiological conditions and in response to environmental stress.
BACKGROUND:The diverse nature of carbohydrate structures and linkages requires a variety of enzymes responsible for sugar degradation. The E. coli periplasmic protein encoded by the bglX gene has been assigned to glycoside hydrolase family 3 and is predicted to function as a β-glucosidase.OBJECTIVES:We investigated the catalytic properties of the E. coli protein BglX and identified two functionally important amino acid residues.METHODS:The bglX gene was cloned into a pET20b(+) vector, and three mutants, D111N, D287G, and E293Q, were generated using site-directed mutagenesis. Kinetic studies were performed on the wild-type and mutant enzymes.RESULTS:Substrate specificity tests indicated that the BglX enzyme hydrolyzes β-glycosidic bonds in nitrophenyl-β-glycosides and demonstrates greater activity towards galactose-containing substrates compared to glucose derivatives. Monomeric glucose and galactose inhibit enzyme activity to a different degree in a substrate-dependent manner. In addition, BglX can hydrolyze lactose but not cellobiose, maltose, or laminarin. Subsequently, E. coli cells overexpressing active BglX have a growth advantage on minimal media supplemented with lactose as a carbon source. Mutation of D287 or D111 residues negatively affected the activity of BglX indicating their involvement in catalysis. Overexpression of BglX by E. coli cells did not increase biofilm formation.CONCLUSIONS:The low activity towards glucose-containing substrates and significantly elevated activity towards galactosides suggests that β-glucosidase activity may not be the primary function of the BglX enzyme.
Type 1 diabetes mellitus (T1DM), associated with autoimmune destruction of pancreatic β cells, is observed in children and adolescents. OBJECTIVE:We investigated the potential association of the apolipoprotein M (APOM) polymorphisms rs707921, rs805264, rs805296, rs805297, and rs9404941 in childhood-onset T1DM (n = 144) and compared them to those in healthy (mostly Euro-Brazilian) children (n = 168). METHODS:This project was approved by the Ethics Committee of the Federal University of Parana (CAAE 24676613.6.0000.0102). Genotyping was performed using PCR-restriction fragment length polymorphisms (rs805296 and rs9404941) and TaqMan probes (rs707921, rs805264, and rs805297). RESULTS:All polymorphisms were in Hardy-Weinberg equilibrium. In the codominant model, no significant differences (P > 0.05) were observed in genotype and allele frequencies between healthy controls and children with T1DM. The minor allele frequencies (95% CI) for healthy subjects were rs707921 (A, 10.7%; 7-14%), rs805264 (A, 6.5%; 4-9%), rs805296 (C, 3.6%; 2-6%), rs805297 (A, 22.6%; 22-31%), and rs9404941 (C, 2.7%; 1-4%). The frequencies of the rs805297 A allele and rs805296 C allele were similar to those of other Caucasian populations; both the rs707921 and rs805264 A alleles were similar to American and Latin American populations, whereas that of the rs9404941 C allele was lower than that observed in the Caucasian and Asian populations. CONCLUSIONS:Haplotype analysis suggests that rs805297-C, rs9404941-T, rs805296-T, rs805264-G, and rs707921-C conferred risk (OR: 4.25; 95% CI: 1.81-10.1) to childhood-onset T1DM in the Euro-Brazilian population.
