Lipase Gene Research Articles

RNAi-mediated knockdown of Gastric lipase gene response to spirobudiclofen stress in Panonychus citri

Panonychus citri is a globally distributed pest that is currently controlled using insecticides. However, alternative strategies are needed to minimize the use of chemicals. Given that the enormous potential of RNA interference pesticides in pest control development, lipases also have key roles in underpinning insect reproduction, development, defence from pathogens and oxidative stress, and pheromone signalling.Therefore, studying the function of Gastric lipase as a control measure for P. citri may be a promising approach. In this study, we cloned PcGTGL from the genome and studied its transcriptional response to spirobudiclofen, as well as its response after PcGTGL was knocked down. The results showed that the expression level of PcGTGL was higher during the larval、deutonymph and adult stage compared to the stage of egg. Additionally, under the stress of spirobudiclofen, the expression level decreased with the concentration of insecticides increased. We silenced PcGTGL through RNAi, and the results indicated an increase in mortality rate of P. citri, as well as a lethal phenotype characterized by body folding and darkening. Additionally, we found that treatment with spirobudiclofen on PcGTGL-silenced P. citri not only increased mortality rates and the mRNA expression of hormone-sensitivelipase(HSL), but also led to lower expression levels of PcGTGL、PcVg 、PcVgR and accumulation of TAG. These results indicate that the silencing of PcGTGL effectively inhibits the population growth of P. citri. Following the inhibition of PcGTGL, P. citri helps them obtain the energy needed to resist insecticide stress by triggering HSL action. The results provide a new RNAi-based target for control of P. citri.

A Lipase Gene of Thermomyces lanuginosus: Sequence Analysis and High-Efficiency Expression in Pichia pastoris.

Lipase, a type of enzyme that decomposes and synthesizes triglycerides, plays an important role in lipid processing. In this study, a heat-resisting lipase gene (lip4) from Thermomyces lanuginosus was subcloned into the pPICZαA vector and then transformed into Pichia pastoris X33. The recombinant yeast cell concentration reached the maximum (119.5 g/L) at 144 h, and the lipase (Lip4) activity reached the maximum (3900 U/mL) at 168 h in 10 L bioreactor. Through bioinformatics analysis, S168, as the key site of Lip4, participated in the formation of the catalytic triads S168-D223-H280 and G166-H167-S168-L169-G170. Furthermore, S168 and seven conserved amino acids of G104/288, S105, A195, P196, V225 and I287 constitute the active center of Lip4. Specifically, the structure modeling showed two α-helices of the lid domain, outside the active pocket domain, controlling the entry of the substrate on Lip4. The potential glycosylation of Asn-33 may be involved in exhibiting the high stable temperature for lipase activity. Therefore, the eukaryotic system was constructed to express Lip4 efficiently, and the amino acid sites related to the catalytic efficiency of Lip4 were clarified, providing a new way for its subsequent property research and industrial application.

Molecular approach to the characterization of lipase encoding genes from Moraxella sp. SBE01

Lipase from Moraxella sp. SBE01 is an expression of the gene encoding lipase. Detection and characterization of the Moraxella sp. SBE01 lipase coding gene is necessary for large-scale lipase production through genetic engineering. This study aimed to observe the molecular weight, amino acid sequence, length, and conserved amino acids in the DNA encoding the lipase gene, with the goal of identifying and characterizing the lipase-coding gene from Moraxella sp. SBE01. The primer design process was conducted to amplify the lipase gene from Moraxella sp. SBE01 using specialized software for sequence alignment and phylogenetic analysis. Amplification was carried out using PCR with the designed primer, forward primer (GTC ATG ATG TAC TTC CAY GGN GGN GG), reverse primer (GGT TGC CGC CGG CDS WRT CNC C). PCR was carried out under pre-denatured conditions at 95°C (3 minutes), followed by 30 cycles of denaturation at 95°C, annealing at 66°C (30 seconds), 70°C elongations (1 minute) and final elongation of 70°C (10 minutes). The PCR results were electrophoresed using 1% agarose gel with a 1 kb DNA marker. The PCR results were sequenced and analyzed for gene and amino acid sequences and the type of lipase expressed. Sequencing resulted in 387 bp of the nucleotide sequence. The gene and amino acid sequences from Moraxella sp. SBE01 had high homology with the gene and amino acid sequences from Moraxella sp. strain TA144. The lipase gene encodes a protein consisting of 129 amino acids and contains a conserved HGG (His-Gly-Gly) motif, which is characteristic of lipases in family IV, also known as the hormone-sensitive lipase (HSL) family. This conserved sequence suggests that the lipase shares structural and functional similarities with other enzymes in the HSL family, playing a key role in lipid metabolism.

8182 New Mouse Model Mirrors Human MODY8: Opening Doors to Understanding Exocrine-Endocrine Crosstalk in the Diabetic Pancreas

Abstract Disclosure: K. El Jellas: None. V. Salerno: None. L.S. Katz: None. J. Hu: None. G. Fogarty: None. A. Mandal: None. A. DiStefano-Forti: None. D. Scott: None. M. Lowe: None. A. Molven: None. R.N. Kulkarni: None. Maturity-onset diabetes of the young, type 8 (MODY8) is a dominantly inherited form of monogenic diabetes, caused by heterozygous mutations in the carboxyl ester lipase (CEL) gene expressed in pancreatic acinar cells. MODY8 is characterized by chronic pancreatitis and exocrine dysfunction. Despite the intimate anatomical relationship between the endocrine and exocrine pancreas, little is known about the biological components mediating exocrine-endocrine communication in regulating glucose homeostasis. Here, we report a refined mouse model that bears the mutation present in MODY8 patients on one allele, and the humanized normal variable number of tandem repeat (VNTR) of CEL with 16 repeats (16R) on the other, mimicking the genetic make-up seen in MODY8 patients. Examination of glucose homeostasis revealed that inducing stress (e.g. by high-fat diet) leads to glucose intolerance in MODY8/16R mice and a blunted glucose-stimulated insulin-secretion compared to their control littermates. Histopathological analyses demonstrated features of chronic pancreatitis in some pancreatic lobes, including inflammation, acinar atrophy, acinar-to-ductal metaplasia, fibrosis and fat infiltration, mirroring the morphology of the human disease. Moreover, we applied immunolabeling-enabled three-dimensional imaging of solvent-cleared organs (iDISCO) to intact pancreata and found that beta-cell mass was significantly decreased in the MODY8/16R mice, with a large number of islets located within fatty lobes. These results are coupled with comparative proteomics and lipidomic profiling with the aim of understanding the molecular signatures of exocrine and endocrine cells during pathogenesis. In conclusion, we have generated a mouse that recapitulates several features of the exocrine and endocrine phenotype characteristic of human MODY8. This novel model provides a unique opportunity, not possible in humans, to characterize pancreatic endocrine-exocrine communication in a longitudinal manner as a response to diverse environmental stressors. Presentation: 6/2/2024

A GDSL motif-containing lipase modulates Sclerotinia sclerotiorum resistance in Brassica napus.

Sclerotinia stem rot (SSR) caused by Sclerotinia sclerotiorum (Lib.) De Bary is a devastating disease infecting hundreds of plant species. It also restricts the yield, quality, and safe production of rapeseed (Brassica napus) worldwide. However, the lack of resistance sources and genes to S. sclerotiorum has greatly restricted rapeseed SSR-resistance breeding. In this study, a previously identified GDSL motif-containing lipase gene, Brassica napus GDSL LIPASE-LIKE 1 (BnaC07.GLIP1), encoding a protein localized to the intercellular space, was characterized as functioning in plant immunity to S. sclerotiorum. The BnaC07.GLIP1 promoter is S. sclerotiorum-inducible and the expression of BnaC07.GLIP1 is substantially enhanced after S. sclerotiorum infection. Arabidopsis (Arabidopsis thaliana) heterologously expressing and rapeseed lines overexpressing BnaC07.GLIP1 showed enhanced resistance to S. sclerotiorum, whereas RNAi suppression and CRISPR/Cas9 knockout B. napus lines were hyper-susceptible to S. sclerotiorum. Moreover, BnaC07.GLIP1 affected the lipid composition and induced the production of phospholipid molecules, such as phosphatidylethanolamine, phosphatidylcholine and phosphatidic acid, which were correlated with decreased levels of reactive oxygen species (ROS) and enhanced expression of defense-related genes. A B. napus bZIP44 transcription factor specifically binds the CGTCA motif of the BnaC07.GLIP1 promoter to positively regulate its expression. BnbZIP44 responded to S. sclerotiorum infection, and its heterologous expression inhibited ROS accumulation, thereby enhancing S. sclerotiorum resistance in Arabidopsis. Thus, BnaC07.GLIP1 functions downstream of BnbZIP44 and is involved in S. sclerotiorum resistance by modulating the production of phospholipid molecules and ROS homeostasis in B. napus, providing insights into the potential roles and functional mechanisms of BnaC07.GLIP1 in plant immunity and for improving rapeseed SSR disease-resistance breeding.

The role of monoglyceride lipase gene in promoting proliferation, metastasis, and free fatty acid accumulation in uveal melanoma cells.

Uveal melanoma is a malignant tumor originating from melanocytes in the eye's uvea, often detected during routine ophthalmic examinations due to its typically asymptomatic nature. Despite effective local treatments, up to 50% of patients develop hematogenous metastases, highlighting the need for better prognostic markers and therapeutic targets. In this study, we developed an innovative Metastasis-Related Gene Signature (MERGS) score to classify patients from various cohorts. By establishing this scoring method, we discovered underlying mechanisms responsible for significant differences between samples with high and low MERGS scores. We identified a set of ten genes to construct MERGS, which showed a high predictive accuracy for patient survival. Further, Monoglyceride Lipase (MGLL) emerged as the most important gene in distinguishing uveal melanoma metastasis. Functional studies demonstrated that knocking down MGLL significantly inhibited proliferation, invasion, and migration of uveal melanoma cells in vitro and in vivo, while overexpression of MGLL enhanced these malignant behaviors. Additionally, MGLL modulated free fatty acid (FFA) levels within these cells. Our findings reveal MGLL as a crucial player in uveal melanoma progression and propose it as a novel therapeutic target, potentially leading to improved management and outcomes for patients with this disease.

Plasmid-based and genome-based expression of recombinant T1 lipase in sucrose-utilizing E. coli strain W

Given its thermoalkaliphilic properties, T1 lipase holds significant potential for diverse industrial applications. However, traditional expression methods in Escherichia coli, specifically the plasmid-based system, present challenges of exerting metabolic burden on host cells and elevated costs due to antibiotic usage. This study addresses these issues by pioneering the expression of recombinant T1 lipase in a sucrose-utilizing E. coli strain W, using molasses as an economical carbon source. The gene cassette (KIKO plasmid), containing the T1 lipase gene regulated by tac and trc promoters, was integrated into the E. coli genome via the λ Red recombinase system. T1 lipase was optimally expressed in shake flasks at 16°C and a 3% molasses concentration in M9 medium with 0.8 mM IPTG as inducer, yielding 0.44 U/mL activity in the genome-based system compared to 0.94 U/mL in the plasmid-based system. This study not only underscores the potential of employing sucrose-utilizing E. coli strain for industrial recombinant protein production but also highlights the need for further optimization of genome-based expression systems. It offers an alternative to reduce costs and enhance sustainability in the stable production of industrially relevant enzymes like T1 lipase, without the need for antibiotic supplementation, and has broader implications for leveraging inexpensive carbon sources like molasses in biotechnological applications.

Novel single nucleotide polymorphism G216A of hormone-sensitive lipase gene associated with Awassi sheep reproduction

BackgroundLitter size plays a crucial role in determining profitability in the sheep industry. Breeding sheep with high litter sizes could be enhanced by selecting candidate genes. One gene affecting sheep's reproductive performance is the hormone-sensitive lipase (HSL) gene. As a result, this study investigated whether the HSL gene variation influenced the fertility of Awassi ewes. The genomic DNA was extracted from 52 singleton ewes and 48 twin ewes. The HSL gene exon 9 (278 bp) was amplified using polymerase chain reaction (PCR).ResultsStudy results revealed two genotypes identified in the 278-bp amplicons: GG and GA. Molecular sequence analysis identified a novel mutation in the GA genotype 216G > A. The statistical analysis revealed a significant association between the single nucleotide polymorphism (SNP) 216A > G and reproductive performance. Ewes with the SNP 216G > A genotype exhibited significantly increased litter sizes, twinning rates, lambing rates, and fewer days to lambing compared to ewes with GG genotypes (P ≤ 0.05). The logistic regression analysis results provided strong evidence that the 216G > A mutation significantly increased litter sizes.ConclusionsThis study concluded that variant 216G > A SNP positively impacts Awassi sheep reproduction. There is a higher litter size and more prolificacy in ewes with the 216G > A SNP than in those without the SNP.

Male-derived phospholipase A2 enhances WD46 expression and increases fertility in Ophraella communa.

Successful bisexual reproduction requires interactions between males and females. Male-derived seminal fluid proteins (SFPs) transferred to females during mating profoundly affect females from pre- to post-mating, and the subsequent shift in female physiology enhances their fertility. SFPs have important evolutionary implications for the fitness of many insects. However, little is known about how females respond to male SFPs. In this study, we identified a male-derived SFP-phospholipase A2 (PLA2) in Ophraella communa. PLA2 is a vital enzyme in eicosanoid biosynthesis; however, it has not been identified as an insect SFP. We found that OcPLA2 is specifically expressed in males, especially in the male accessory glands (MAGs); it is transferred to the female during mating and functions as an SFP to enhance fertility. The expression of a female-derived gene encoding the WD repeat-containing protein 46 (WD46) was upregulated when OcPLA2 entered the female reproductive tract, and this contributed to female egg production by increasing triacylglycerol lipase (TGL) gene expression and the triglyceride (TG) content. This is the first study to identify PLA2 as an SFP in insects. Our findings also shed light on the regulatory role of OcPLA2 in beetle reproduction; the expression of OcPLA2 is initially correlated with female WD46 expression and later with the decline in TGL gene expression and the TG content. This represents a unique mechanism of reproductive regulation by an SFP.

Genome-wide identification and mining elite allele variation of the Monoacylglycerol lipase (MAGL) gene family in upland cotton (Gossypium hirsutum L.)

BackgroundMonoacylglycerol lipase (MAGL) genes belong to the alpha/beta hydrolase superfamily, catalyze the terminal step of triglyceride (TAG) hydrolysis, converting monoacylglycerol (MAG) into free fatty acids and glycerol.ResultsIn this study, 30 MAGL genes in upland cotton have been identified, which have been classified into eight subgroups. The duplication of GhMAGL genes in upland cotton was predominantly influenced by segmental duplication events, as revealed through synteny analysis. Furthermore, all GhMAGL genes were found to contain light-responsive elements. Through comprehensive association and haplotype analyses using resequencing data from 355 cotton accessions, GhMAGL3 and GhMAGL6 were detected as key genes related to lipid hydrolysis processes, suggesting a negative regulatory effect.ConclusionsIn summary, MAGL has never been studied in upland cotton previously. This study provides the genetic mechanism foundation for the discover of new genes involved in lipid metabolism to improve cottonseed oil content, which will provide a strategic avenue for marker-assisted breeding aimed at incorporating desirable traits into cultivated cotton varieties.

Expression and characterization of a thermostable monoacylglycerol lipase from thermophilic Geobacillus kaustophilus

Thermophilic Geobacillus kaustophilus HTA426 genome possesses a monoacylglycerol lipase (MAGL) gene. MAGLs can synthesize emulsifiers for use in the food and pharmaceutical industries from fatty acids and glycerol. They can also be used to analyze monoacylglycerol (MAG) levels in serum and food. The MAGL gene from strain HTA426 was artificially synthesized and heterologously expressed in Escherichia coli BL21(DE3). The recombinant His-tag fused MAGL (GkMAGL) was purified using a Ni2+-affinity column. The purified enzyme showed a temperature optimum at 65 °C and was stable up to 75 °C after 30 min incubation. In addition, the enzyme exhibited a pH optimum of 7.5 and was stable from pH 5.0 to 11.0. The enzyme hydrolyzed monoacylglycerols and showed the highest activity toward 1-monolauroylglycerol. The enzyme was stable in the presence of various organic solvents and detergents. The addition of Triton X-100 significantly increased GkMAGL activity. The thermal stability of the enzyme was higher than that of thermostable MAGL from Geobacillus sp. 12AMOR1 (12AMOR1_MAGL). Circular dichroism spectral analysis showed that the conformational stability of the GkMAGL was higher than that of 12AMOR1_MAGL at higher temperatures. These results indicate that the GkMAGL has useful features that can be used for various biotechnological applications.

Dietary supplementation of Astragalus fermentation products improves the growth performance, immunological characteristics, and disease resistance of crucian carp (Carassius auratus)

The fermentation products of Astragalus have been acknowledged for their ability to enhance immune functions. This study assessed the impact of incorporating Astragalus, fermented with Lactobacillus plantarum and Bacillus coagulans, on crucian carp’s growth, disease resistance, and immunological characteristics. The experimental groups were fed with common feed (C), C + Astragalus (A), A + Lactobacillus plantarum (AL), A + Bacillus coagulans (AB), and AL + Bacillus coagulans (ALB). The fermented products were mixed with common feed at a 1:99 ratio, and crucian carp were fed 2% of their body weight for four weeks, with sampling conducted on days 3, 7, 14, 21, and 28. Disease resistance was evaluated using Aeromonas hydrophila (A. hydrophila) at a concentration of 0.2 mL (1.0×107 CFU/mL). The final weights in the AL, AB, and ALB groups significantly increased compared to the C group. The ALB group exhibited elevated serum albumin levels, alkaline phosphatase, intestinal lipase, protease enzyme, C3, and IgM gene expression compared to the C group. At the same time, alanine aminotransferase, aspartate aminotransferase, and glucose contents were significantly reduced. The survival rate significantly increased in all experimental groups after treatment with A. hydrophila. In conclusion, Astragalus products fermented with Lactobacillus plantarum and Bacillus coagulans could effectively improve crucian carp’s growth, disease resistance, and immune response.

GDSL Lipase Gene HTA1 Negatively Regulates Heat Tolerance in Rice Seedlings by Regulating Reactive Oxygen Species Accumulation.

High temperature is a significant environmental stress that limits plant growth and agricultural productivity. GDSL lipase is a hydrolytic enzyme with a conserved GDSL sequence at the N-terminus, which has various biological functions, such as participating in plant growth, development, lipid metabolism, and stress resistance. However, little is known about the function of the GDSL lipase gene in the heat tolerance of rice. Here, we characterized a lipase family protein coding gene HTA1, which was significantly induced by high temperature in rice. Rice seedlings in which the mutant hta1 was knocked out showed enhanced heat tolerance, whereas the overexpressing HTA1 showed more sensitivity to heat stress. Under heat stress, hta1 could reduce plant membrane damage and reactive oxygen species (ROS) levels and elevate the activity of antioxidant enzymes. Moreover, real-time quantitative PCR (RT-qPCR) analysis showed that mutant hta1 significantly activated gene expression in antioxidant enzymes, heat response, and defense. In conclusion, our results suggest that HTA1 negatively regulates heat stress tolerance by modulating the ROS accumulation and the expression of heat-responsive and defense-related genes in rice seedlings. This research will provide a valuable resource for utilizing HTA1 to improve crop heat tolerance.

Diabetes mellitus due to reduced insulin secretion and exocrine pancreatic dysfunction in a patient with a mutation in the carboxyl-ester lipase gene outside the VNTR region

Diabetes mellitus due to reduced insulin secretion and exocrine pancreatic dysfunction in a patient with a mutation in the carboxyl-ester lipase gene outside the VNTR region

Construction of Pseudomonas aeruginosa SDK-6 with synthetic lipase gene cassette and optimization of different parameters using response surface methodology for over-expression of recombinant lipase.

Lipases are industrially important enzymes having vast applications in various fields. Cloning and expression of lipase enzyme-encoding genes in suitable host lead to their widespread use in different fields. The present study represents the first attempt towards the expression of the synthetic lipase gene in Pseudomonas aeruginosa. An alkalophilic lipase gene (GenBankaccession number: NP_388152) from Bacillus subtilis was synthetically designed and introduced in the pJN105 vector and subsequently cloned in Pseudomonas aeruginosa SDK-6. Agarose gel electrophoresis confirmed the transformation of SDK-6, exhibiting a band difference of ~ 700bp between native and recombinant pJN105. Further amplification of cloned lipase gene was confirmed using PCR amplification with Lip 1 and Lip 2 primers respectively, followed by restriction analysis. Approximately 15-fold increase in lipase production was observed in recombinant Pseudomonas as compared to the native strain. One factor at a time (OFAT) analysis revealed L-arabinose, inoculum size (0.5%; v/v), and agitation (120rpm) as significant factors affecting the over-expression of lipase enzyme. Optimization of enzyme induction conditions by central composite design (CCD) led to 1.60-fold increase in the production of lipase at 0.65% (w/v) inducer concentration, OD600-1.075 before induction and 35°C post induction temperature with overall lipase production of 50.50IU/mL. Statistical validation of observed value via ANOVA showed an F-value of 138.70 at p < 0.01 with R2 of 0.9921.

Experimental vaccination against Aeromonas hydrophila in Colossoma macropomum: Bacterial characterization, lethal doses, and mortality

Aeromonas hydrophila is one of the most important pathogens affecting Amazonian fish. In recent years, outbreaks in native aquaculture species displaying hemorrhages, epithelial ulcers, and damaged fins, which often lead to fatalities, have been reported. In the current study, we characterized A. hydrophila isolated from an outbreak on a commercial farm of Colossoma macropomum (gamitana), one of the main native species farmed in Peru and South America. Gram-negative, motile, oxidase-, and catalase-positive bacteria were retrieved from diseased C. macropomum. Aeromonas hydrophila was confirmed by biochemical and molecular characterization. Virulence genes detection showed that A. hydrophila (FO129–26) harbors cytotoxic enterotoxin (act), hemolysin (hly), serine protease (ser), lipase (lip), flagellin (fla), and elastase (ahyB) virulence genes. Then, lethal dose tests (LD10, 50, 90, 99) were performed and stablished at 4.6 × 106; 6.4 × 106; 8.9 × 106; and 1.11 × 107 CFU mL−1 confirming the virulence of the bacterial isolate. Finally, there were used three methods of bacteria inactivation to vaccine gamitanas against this bacterium. 180 gamitanas (C: control; FO: formaldehyde-inactivated group; PH: peroxide-inactivated group; CF: chloroform-inactivated group) were challenged with the pathogen. FO and PH groups had better survival rates (∼ 40 %) than the control group. There were no significant differences (p ≥ 0.05) between the FO and PH groups. Fish from CF presented early mortality after vaccination, probably due to irritation caused by chloroform residues. Finally, this study shows the virulence characteristics of A. hydrophila FO129–26 in an important farmed native fish in Peru and broadens the geographic range of the presence of this pathogen worldwide. Vaccines are a good strategy to improve the resistance of C. macropomum against A. hydrophila. However, further research is necessary to expand our comprehension of the efficacy of this protective technique in Amazonian fish.

Abstract 150: Decoding The Variant-to-function Relationship For LIPA , A Risk Locus For Cad

Background: Genome-wide association studies (GWASs) link genetic variation in the LIPA (lysosomal acid lipase) gene to coronary artery disease (CAD). Quantitative Trait Loci (QTL) studies indicate higher LIPA mRNA and enzyme activity in the monocytes of risk alleles carriers. Yet, the mechanism whereby this locus confers disease etiology has not been fully established. Herein, we aim to establish the variant-to-function relationship and elucidate how increased LIPA impacts atherosclerosis. Methods: Employing post-GWAS pipelines (eQTL, enzyme activity-QTL, Tri-HiC, luciferase assay, allele-specific binding, motif analysis, and mobility shift assay), we connect functional variants to target genes in causal myeloid cells. Myeloid-specific Lipa overexpression mice ( Lipa Tg ) on Ldlr -/- background were generated and characterized. Results: Post-GWAS pipelines support LIPA as the candidate causal gene at this locus. In human monocyte-derived macrophages, risk allele carriers of GWAS CAD variants exhibit higher LIPA mRNA, protein, and enzyme activity. Tri-HiC and luciferase assay identified an intronic region within LIPA , housing GWAS CAD noncoding variants and exhibiting monocyte-specific enhancer activity. Within the enhancer region, 1412445 and rs1320496 were identified as the two potential causal variants, with the risk alleles having increased enhancer activity in luciferase assay and increased binding to PU.1 in mobility shift assay. Collectively, the above data suggest that the risk alleles of CAD variants lead to increased LIPA in monocytes/macrophages. Consistently, Lipa Tg , Ldlr -/- mice fed a Western diet showed significantly increased lesion size and lesion macrophage area compared to Ctl, Ldlr -/- mice. scRNA-seq analysis followed by functional validation demonstrated that Lipa Tg macrophages had higher free cholesterol yet lower neutral lipid accumulation, accompanied by a transcriptomic signature of increased interferon signaling. Lipa overexpression also increased monocyte infiltration into the plaque. Conclusions: We established that the risk alleles of common noncoding variants at the LIPA locus drive increased myeloid LIPA expression and aggravate atherosclerosis.

Immune system parameters in patients with chronic opistorchiasis based on genetic polymorphism associated with carbohydrate and lipid metabolism disorders

A study assessing differences in the immunopathogenesis of chronic opisthorchiasis with or without genetic polymorphisms in patients associated with predisposition to type II diabetes mellitus and lipid metabolism disorders was carried out. Venous blood samples from 89 patients were collected to analyze immunological parameters and genetic polymorphisms by using pyrosequencing. In the presence of mutations affecting regulation of carbohydrate metabolism, patients with opisthorchiasis had less pronouncedly increased phagocytic activity, a lower monocyte count, higher total T-lymphocyte count, lower T-cytotoxic cell and B-lymphocyte counts, lower IgA, IgG but higher IgM concentration. This indicates a moderately compromised nonspecific resistance, imbalanced effector T- and B-immunity, but in most cases (PPARG, TCF7L2 rs12255372, CDKAL1, CDKN2A/2B, SLC30A8) aggravated humoral immune reaction to invasion particularly revealed by lower B-lymphocyte count. The presence of polymorphisms that alter lipid metabolism regulation bidirectionally affects the parameters of immune response. An increased B-lymphocyte count and other indicators of humoral immune activation to invasion, which are detected in groups with mutations in the APOE (rs429358), PCSK9, ABCA1, APOC3 rs2854117 genes, can contribute to a more effective response to sustained antigenic stimulation. Changes in the T-lymphocytes subpopulations, characteristic of opisthorchiasis invasion, are aggravated in the presence of mutations in the PCSK9, ABCA1, and APOC3 rs2854117 genes. Mutations in the genes APOC3 rs5128, LPL rs268 activate patients’ nonspecific resistance, although this effect may also be associated with exhaustion of neutrophil bactericidal reserve. In general, minor alleles of the APOE (rs429358), APOC3 rs2854117, LPL rs268, LPL rs328, PON1 rs662 genes can be considered “protective” for the immunopathogenesis of chronic opisthorchiasis. Thus, patients with chronic opisthorchiasis with different genotypes predisposing to carbohydrate and lipid metabolism disorders had significant differences in immune system parameters, which can also affect the disease course. Mutations in different loci of the PCSK9, ABCA1, APOE, APOC3, and PON1 genes have opposite effects on the analyzed immune parameters. In the presence of mutations in other genes (PPARG, TCF7L2 rs12255372, CDKAL1, CDKN2A/2B, SLC30A8, LPL), opisthorchiasis invasion leads to more pronounced alterations in immune response; mutations in the lipoprotein lipase gene may have some “protective” immune-related effect in opisthorchiasis. The effect of all studied genetic polymorphisms associated with a predisposition to developing type II diabetes mellitus was predominantly negative.

Phenotypic and genetic characterization of a lipolytic Ralstonia mannitolilytica isolate from petroleum-contaminated soil in Iraq.

Lipases play a crucial role in various industrial applications, and microbial lipases, particularly those from bacteria, possess significant properties. With increasing concerns about the environmental and health impacts of hydrocarbons from pipelines and refineries, there is a growing need to mitigate the risks associated with these compounds. In this study, 40 bacterial isolates were recovered from contaminated soil samples collected from multiple refineries across Iraq. Using the Vitek system, bacterial isolates were identified up to the species level, revealing that only 12 isolates exhibited lipase-producing capabilities. Among the lipase-producing isolates, Ralstonia mannitolilytica demonstrated the highest extracellular lipase activity, as determined by an olive oil plate assay supplemented with rhodamine B. Confirmation of the species identity was achieved through 16S rRNA gene sequencing, with the obtained sequence deposited under accession number LC772176.1. Further sequence analysis revealed single nucleotide polymorphisms (SNPs) in the genome of Ralstonia mannitolilytica strain H230303-10_N19_7x_R2 (CP011257.1, positions 1,311,102 and 1,311,457). Additionally, the presence of the lipase gene was confirmed through amplification and sequencing using a thermocycler PCR. Sequence analysis of the gene, aligned using Geneious Prime software, identified SNPs (CP010799, CP049132, AY364601, CP011257, and CP023537), and a phylogenetic tree was constructed based on genetic characterization. Our findings highlight the potential of Ralstonia mannitolilytica as a promising candidate for lipase production and contribute to our understanding of its genetic diversity and biotechnological applications in hydrocarbon degradation and industrial processes.

Excessive fat expenditure in MCT-induced heart failure rats is associated with BMAL1/REV-ERBα circadian rhythmic loop disruption

Fat loss predicts adverse outcomes in advanced heart failure (HF). Disrupted circadian clocks are a primary cause of lipid metabolic issues, but it's unclear if this disruption affects fat expenditure in HF. To address this issue, we investigated the effects of disruption of the BMAL1/REV-ERBα circadian rhythmic loop on adipose tissue metabolism in HF.50 Wistar rats were initially divided into control (n = 10) and model (n = 40) groups. The model rats were induced with HF via monocrotaline (MCT) injections, while the control group received equivalent solvent injections. After establishing the HF model, the model group was further subdivided into four groups: normal rhythm (LD), inverted rhythm (DL), lentivirus vector carrying Bmal1 short hairpin RNA (LV-Bmal1 shRNA), and empty lentivirus vector control (LV-Control shRNA) groups, each with 10 rats. The DL subgroup was exposed to a reversed light–dark cycle of 8 h: 16 h (dark: light), while the rest adhered to normal light–dark conditions (light: dark 12 h: 12 h). Histological analyses were conducted using H&E, Oil Red O, and Picrosirius red stains to examine adipose and liver tissues. Immunohistochemical staining, RT-qPCR, and Western blotting were performed to detect markers of lipolysis, lipogenesis, and beiging of white adipose tissue (WAT), while thermogenesis indicators were detected in brown adipose tissue (BAT). The LD group rats exhibited decreased levels of BMAL1 protein, increased levels of REV-ERBα protein, and disrupted circadian circuits in adipose tissue compared to controls. Additionally, HF rats showed reduced adipose mass and increased ectopic lipid deposition, along with smaller adipocytes containing lower lipid content and fibrotic adipose tissue. In the LD group WAT, expression of ATGL, HSL, PKA, and p-PKA proteins increased, alongside elevated mRNA levels of lipase genes (Hsl, Atgl, Peripilin) and FFA β-oxidation genes (Cpt1, acyl-CoA). Conversely, lipogenic gene expression (Scd1, Fas, Mgat, Dgat2) decreased, while beige adipocyte markers (Cd137, Tbx-1, Ucp-1, Zic-1) and UCP-1 protein expression increased. In BAT, HF rats exhibited elevated levels of PKA, p-PKA, and UCP-1 proteins, along with increased expression of thermogenic genes (Ucp-1, Pparγ, Pgc-1α) and lipid transportation genes (Cd36, Fatp-1, Cpt-1). Plasma NT-proBNP levels were higher in LD rats, accompanied by elevated NE and IL-6 levels in adipose tissue. Remarkably, morphologically, the adipocytes in the DL and LV-Bmal1 shRNA groups showed reduced size and lower lipid content, while lipid deposition in the liver was more pronounced in these groups compared to the LD group. At the gene/protein level, the BMAL1/REV-ERBα circadian loop exhibited severe disruption in LV-Bmal1 shRNA rats compared to LD rats. Additionally, there was increased expression of lipase genes, FFA β oxidation genes, and beige adipocyte markers in WAT, as well as higher expression of thermogenic genes and lipid transportation genes in BAT. Furthermore, plasma NT-proBNP levels and adipose tissue levels of NE and IL-6 were elevated in LV-Bmal1 shRNA rats compared with LD rats. The present study demonstrates that disruption of the BMAL1/REV-ERBα circadian rhythmic loop is associated with fat expenditure in HF. This result suggests that restoring circadian rhythms in adipose tissue may help counteract disorders of adipose metabolism and reduce fat loss in HF.