Lipid Content Of Cells Research Articles

LncRNA lncLLM Facilitates Lipid Deposition by Promoting the Ubiquitination of MYH9 in Chicken LMH Cells.

The liver plays an important role in regulating lipid metabolism in animals. This study investigated the function and mechanism of lncLLM in liver lipid metabolism in hens at the peak of egg production. The effect of lncLLM on intracellular lipid content in LMH cells was evaluated by qPCR, Oil Red O staining, and detection of triglyceride (TG) and cholesterol (TC) content. The interaction between lncLLM and MYH9 was confirmed by RNA purification chromatin fractionation (CHIRP) and RNA immunoprecipitation (RIP) analysis. The results showed that lncLLM increased the intracellular content of TG and TC and promoted the expression of genes related to lipid synthesis. It was further found that lncLLM had a negative regulatory effect on the expression level of MYH9 protein in LMH cells. The intracellular TG and TC content of MYH9 knockdown cells increased, and the expression of genes related to lipid decomposition was significantly reduced. In addition, this study confirmed that the role of lncLLM is at least partly through mediating the ubiquitination of MYH9 protein to accelerate the degradation of MYH9 protein. This discovery provides a new molecular target for improving egg-laying performance in hens and treating fatty liver disease in humans.

Chronic UVB exposure induces hepatic injury in mice: Mechanistic insights from integrated multi-omics

Chronic UVB exposure poses a significant threat to both skin and visceral health. In recent years, the adverse role of chronic UVB exposure in liver health has been suggested but not fully elucidated. This study aims to comprehensively investigate the effects of chronic UVB exposure on liver health in male SKH-1 hairless mice and clarify potential mechanisms through multi-omics approaches. The findings suggested that 10-week chronic skin exposure to UVB not only triggers hepatic inflammation and oxidative stress but also, more importantly, results in lipid metabolism abnormalities in the liver. Hepatic transcriptomic analysis revealed significant alterations in various signaling pathways and physiological processes associated with inflammation, oxidative stress, and lipid metabolism. Further lipidomic analysis illustrated significant changes in the metabolism of glycerolipids, sphingolipids, and glycerophospholipids in the liver following chronic UVB exposure. The 16S rRNA sequencing analysis indicated that chronic UVB exposure disrupts the structure and function of the microbiota. In search of potential mechanisms used by the microbiome to regulate the hepatic disease morphology, we filtered mouse fecal supernatants and cultured the supernatants with HepG2 cells. Fecal supernatant from UVB-exposed mice induced increased secretion of the inflammatory cytokine IL-8, accumulation of MDA, reduced SOD activity, and decreased lipid content in normal hepatic cells. In summary, skin chronic exposure to UVB induces multiple liver injuries and gut microbiota dysbiosis in mice and gut microbiota metabolites may be one of the contributing factors to hepatic injury caused by chronic UVB exposure. These discoveries deepen the comprehension of the health risks associated with chronic UVB exposure.

ANTI-ATHEROSCLEROSIS EFFECT OF THE HERB SPERANSKIA TUBERCULATA (BUNGE) BAILL

The article describes the inhibitory effect of extracts of the herb Speranskia Tuberculata (Bunge) Baill on atherosclerosis of vessels. Crude extracts were prepared using petroleum ether (PE extracts). Subsequently, the dried extracts were dissolved in dimethyl sulfoxide to prepare working solutions with 12.5 to 75 μg/mL concentrations. RAW 264.7 cells, a macrophage cell line, were used as the research object. Atherosclerosis was induced in the cells by incubating oxidised low-density lipoproteins. The study of the influence of PE extract concentration was carried out in two experiments. The first included oil red O staining on macrophages with different concentrations of PE extract added, followed by a spectrophotometric examination of the solutions. Thus, the effect of PE extract concentration on lipid content in cells was investigated. The second method was based on flow cytometry and ended by measuring the intensity of fluorescent cholesterol in the cells of various samples. Cell samples additionally containing only oxidised low-density lipoproteins, as well as cells treated with PE extract and caffeic acid, were compared. PE extracts of Speranskia Tuberculata (Bunge) Baill effectively inhibit the transition from macrophages to foam cells, as shown by labelling foam cells with neutral fat using Oil red O staining. The processes of cholesterol uptake by macrophages were investigated by flow cytometry using fluorescently labelled cholesterol. PE extract minimises cholesterol deposition in macrophages by inhibiting macrophage phagocytosis of cholesterol while promoting the effect of intracellular cholesterol outflux. Thus, the role of PE extracts in regulating the cholesterol balance in macrophages and inhibiting the transformation of macrophages into foam cells has been experimentally proven. Possible mechanisms for realising these effects are proposed.

Sustainable cultivation of Porphyridium cruentum via agro-industrial by-products: A study on biomass and lipid enhancement

This study explored the cultivation of Porphyridium cruentum using beet molasses and corn steep liquor (CSL) as alternative agri-waste substrates. The objective was to enhance the biomass and lipid production of P. cruentum, known for its potential as an industrial EPA producer. Using a Box-Behnken Design (BBD) as part of the response surface methodology (RSM), we investigated the effects of beet molasses, CSL, and F/2 nutrients on the microalga's cultivation. The results demonstrated a significant influence of these factors on the final cell count and lipid content over an 8-day cultivation period. Specifically, optimal growth conditions were identified at approximately 1.78 g/L of molasses and 1.89 g/L of CSL, yielding a cell count of 12.1 × 106 cells/mL and a lipid content of 24.48%. Validation experiments reaffirmed these findings, with observed results closely aligning with predicted values. Optimized media achieved also a significant higher content of carotenoids and phycobiliproteins compared to the standard media. This research highlights the potential of using agro-industrial by-products for large-scale cultivation of P. cruentum, offering a sustainable approach for enhancing lipid and pigment production.

Cryptococcosis, tuberculosis, and a kidney cancer fail to fit the atherosclerosis paradigm for foam cell lipid content.

Foam cells are dysfunctional, lipid-laden macrophages associated with chronic inflammation of diverse origin. The long-standing paradigm that foam cells are cholesterol-laden derives from atherosclerosis research. We previously showed that, in tuberculosis, foam cells surprisingly accumulate triglycerides. Here, we utilized bacterial ( Mycobacterium tuberculosis ), fungal ( Cryptococcus neoformans ), and human papillary renal cell carcinoma (pRCC) models to address the need for a new explanation of foam cell biogenesis. We applied mass spectrometry-based imaging to assess the spatial distribution of storage lipids relative to foam-cell-rich areas in lesional tissues, and we characterized lipid-laden macrophages generated under corresponding in vitro conditions. The in vivo data and the in vitro findings showed that cryptococcus-infected macrophages accumulate triglycerides, while macrophages exposed to pRCC- conditioned-medium accumulated both triglycerides and cholesterol. Moreover, cryptococcus- and mycobacterium-infected macrophages accumulated triglycerides in different ways. Collectively, the data show that the molecular events underlying foam cell formation are specific to disease and microenvironment. Since foam cells are potential therapeutic targets, recognizing that their formation is disease-specific opens new biomedical research directions.

Regulation of PPARγ2 Stability and Activity by SHP-1.

The protein tyrosine phosphatase Src homology region 2 domain-containing phosphatase-1 (SHP-1) plays an important role in modulating glucose and lipid homeostasis. We previously suggested a potential role of SHP-1 in the regulation of peroxisome proliferator-activated receptor γ2 (PPARγ2) expression and activity but the mechanisms were unexplored. PPARγ2 is the master regulator of adipogenesis, but how its activity is regulated by tyrosine phosphorylation is largely unknown. Here, we found that SHP-1 binds to PPARγ2 primarily via its N-terminal SH2-domain. We confirmed the phosphorylation of PPARγ2 on tyrosine-residue 78 (Y78), which was reduced by SHP-1 invitro resulting in decreased PPARγ2 stability. Loss of SHP-1 led to elevated, agonist-induced expression of the classical PPARγ2 targets FABP4 and CD36, concomitant with increased lipid content in cells expressing PPARγ2, an effect blunted by abrogation of PPARγ2 phosphorylation. Collectively, we discovered that SHP-1 affects the stability of PPARγ2 through dephosphorylation thereby influencing adipogenesis.

Individually Cultured Bovine Zygotes Successfully Develop to the Blastocyst Stage in an Extremely Confined Environment

The possibility of detecting the developmental competence of individually cultured embryos through analysis of spent media is a major current trend in an ART setting. However, individual embryo culture is detrimental compared with high-density group culture due to the reduced concentration of putative embryotropins. The main aim of this study was to identify an individual culture system that is not detrimental over high-density group culture in the bovine model. Blastocyst rates and competence were investigated in a conventional (GC) group, semi-confined group (MG), and individual culture (MS) in a commercial microwell device. Main findings showed that: (1) individual embryos can be continuously cultured for 7 days in ~70 nL microwells (MS) without detrimental effects compared with the GC and MG; (2) MS and MG blastocysts had a reduced number of TUNEL-positive cells compared to GC blastocysts; (3) though blastocyst mean cell numbers, mitochondrial activity, and lipid content were not different among the three culture conditions, MS blastocysts had a higher frequency of small-sized lipid droplets and a reduced mean droplet diameter compared with GC and MG blastocysts. Overall, findings open the way to optimize the development and competence of single embryos in an ART setting.

Effects of salinity on the growth, physiological and biochemical components of microalga Euchlorocystis marina

Euchlorocystis marina, a new marine species of the genus Euchlorocystis discovered in 2022, has the potential to improve the water quality in mariculture ponds. However, the effects of salinity on the growth, physiology, and biochemical composition of these algae are not well understood. In this study, changes in physiological and biochemical indices such as cell density, photosynthetic pigment, polysaccharide, and lipid content of E. marina under different salinity treatments were analyzed. The results showed that the highest cell density was observed at a salinity of 15‰. The lowest photosynthetic pigment content was observed at a salinity of 60‰, and the highest polysaccharide and lipid content was observed at a salinity of 60‰. These results indicated that lower salinity was more conducive to E. marina reproduction and growth. E. marina can accumulate polysaccharides and lipids in high salinity environments. This study provides new information for understanding the salinity adaptation strategies of E. marina and has practical significance for its development and utilization.

Palmitic acid induces lipid droplet accumulation and senescence in nucleus pulposus cells via ER-stress pathway

Intervertebral disc degeneration (IDD) is a highly prevalent musculoskeletal disorder affecting millions of adults worldwide, but a poor understanding of its pathogenesis has limited the effectiveness of therapy. In the current study, we integrated untargeted LC/MS metabolomics and magnetic resonance spectroscopy data to investigate metabolic profile alterations during IDD. Combined with validation via a large-cohort analysis, we found excessive lipid droplet accumulation in the nucleus pulposus cells of advanced-stage IDD samples. We also found abnormal palmitic acid (PA) accumulation in IDD nucleus pulposus cells, and PA exposure resulted in lipid droplet accumulation and cell senescence in an endoplasmic reticulum stress-dependent manner. Complementary transcriptome and proteome profiles enabled us to identify solute carrier transporter (SLC) 43A3 involvement in the regulation of the intracellular PA level. SLC43A3 was expressed at low levels and negatively correlated with intracellular lipid content in IDD nucleus pulposus cells. Overexpression of SLC43A3 significantly alleviated PA-induced endoplasmic reticulum stress, lipid droplet accumulation and cell senescence by inhibiting PA uptake. This work provides novel integration analysis-based insight into the metabolic profile alterations in IDD and further reveals new therapeutic targets for IDD treatment.

Classifying Raman Spectra of Colon Cells Based on Machine Learning Algorithms

Colorectal cancer is very widespread in developed countries. Its diagnosis partly depends on pathologists’ experience and their laboratories’ instrumentation, producing uncertainty in diagnosis. The use of spectroscopic techniques sensitive to the cellular biochemical environment could aid in achieving a reliable diagnosis. So, we used Raman micro-spectroscopy, combined with a spectral analysis by means of machine learning methods, to build classification models, which allow colon cancer to be diagnosed in cell samples, in order to support such methods as complementary tools for achieving a reliable identification of colon cancer. The Raman spectra were analyzed in the 980–1800 cm−1 range by focusing the laser beam onto the nuclei and the cytoplasm regions of single FHC and CaCo-2 cells (modelling healthy and cancerous samples, respectively) grown onto glass coverslips. The comparison of the Raman intensity of several spectral peaks and the Principal Component Analysis highlighted small biochemical differences between healthy and cancerous cells mainly due to the larger relative lipid content in the former cells with respect to the latter ones and to the larger relative amount of nucleic acid components in cancerous cells compared with healthy ones. We considered four classification algorithms (logistic regression, support vector machine, k nearest neighbors, and a neural network) to associate unknown Raman spectra with the cell type to which they belong. The built machine learning methods achieved median values of classification accuracy ranging from 95.5% to 97.1%, sensitivity values ranging from 95.5% to 100%, and specificity values ranging from 93.9% to 97.1%. The same median values of the classification parameters, which were estimated for a testing set including unknown spectra, ranged between 93.1% and 100% for accuracy and between 92.9% and 100% for sensitivity and specificity. A comparison of the four methods pointed out that k nearest neighbors and neural networks better perform the classification of nucleus and cytoplasm spectra, respectively. These findings are a further step towards the perspective of clinical translation of the Raman technique assisted by multivariate analysis as a support method to the standard cytological and immunohistochemical methods for diagnostic purposes.

Optimation of the cell density, biomass production, lipid, and carbohydrate content of Nannochloropsis oculata with fluorescein and pH manipulation

Nannochloropsis oculata is a potential marine microalga. However, the mass production of N. oculata encounters several challenges, such as the high costs of resources, contamination, and light absorption optimization. These problems can be solved by pH manipulation and fluorescein induction on culture media. Various studies have shown that pH values manipulation can cause fluctuations in cell density, dry biomass, and carbohydrate and lipid production of microalgae. On the other hand, fluorescein can act as a molecular antenna that improves light absorption. This research is novel because of its focus on its unique attempt to utilize pH manipulation combined with fluorescein induction to enhance the growth and metabolite production of N. oculata. The implications of this research may provide cost-effective, viable, and sustainable development of the algal industry in general. In this research, cultivation was carried out for seven days with four levels of pH (7 – 10) combined with three variations of fluorescein addition (0, 0.15, and 0.3 mL). Every treatment combination had three replications. Cell density and biomass as growth analysis parameters were calculated during the cultivation using the Haemocytometer and gravimetric methods. Bligh & Dyer and phenol sulfate methods were used to analyze lipid and carbohydrate content. Based on the result, the best treatment combination, pH 9 with 0.15 mL of fluorescein, increases cell density and carbohydrate content of N. oculata by 31.45 and 4.4%, albeit not statistically significant.

Phytoremediation capability of Anabaena sp. for high organic and chromium-loaded wastewater in membrane bioreactors.

The efficiency of Anabaena sp. was analyzed for the phytoremediation of wastewater loaded with organic matter and heavy metals like chromium. Simulated wastewater was contaminated with chromium. A side-stream membrane bioreactor was used for the treatment of wastewater. A feed tank of 20 L capacity was used with a stirring arrangement. A ceramic microfiltration membrane composed of clay and alumina was obtained from Johnson & Johnson. The removal efficiency of chemical oxygen demand, biochemical oxygen demand, and chromium was evaluated. The process was used for algae harvesting and wastewater treatment. About 92% of chemical oxygen demand (COD), 98% chromium, and oil and grease were completely removed. Membrane fouling was explained by the pore blocking and cake resistance model. Stress in algal cells was determined from the superoxide dismutase (SOD) and catalase (CAT) analysis. The lipid content of algal cells was measured.

Analysis of cell growth, photosynthetic behavior and the fatty acid profile in Tetraselmis subcordiformis under different lighting scenarios

Tetraselmis has been investigated as a potential source of lipids. This microalga possesses good growth characteristics and can be used to develop viable platforms for fatty acid production. This research aims to evaluate the effect of high photon flux density with light-dark cycles and light wavelength on biomass production and fatty acid profile in Tetraselmis subcordiformis. A low light control and treatments with high photon flux density with different light-dark cycles (24:0 h, 12:12 h, 1:1 h, and 15:15 min) and different light wavelength (white, green, red, and blue) were evaluated to determine cell concentration, nutrient consumption, chlorophyll content, photosynthetic yields, lipid content, and fatty acid profile. Significant differences were found in all variables, except for phosphate consumption. High photon flux density promotes cell growth with T. subcordiformis reaching biomass productivities of 0.10 g L-1 day-1 when continuous white light is used. However, no differences were observed in biomass productivities and lipid content for all high photon flux density treatments. On the other hand, red light resulted in higher cell growth, with a productivity of 0.12 g L-1 day-1, and the highest lipid content was achieved under white light. There was a significant effect on the fatty acid profile under different light conditions, with palmitic acid, oleic acid, and eicosapentaenoic acid being the most abundant. This study demonstrated that cellular growth and fatty acid profiles in T. subcordiformis can be influenced by different lighting schemes in the cultivation.

Removal of sulfamethoxazole and production of cyanobacterial lipid promoted by the construction of a consortium containing a non-toxic cyanobacterium and sewage bacteria

This study evaluated the possibility of constructing a consortium containing sewage bacteria and cyanobacterial cells immobilized by Ca-alginate for the advanced treatment of antibiotic-polluted sewage coupled to cyanobacterial lipid production. Two non-toxic cyanobacterial species (Synechococcus sp. and Chroococcus sp.) were exposed to 0.5–20.0 μg/L of sulfamethoxazole (SMZ) for 4 days. Comparison between suspended and immobilized conditions indicated that immobilization increased (p < 0.05) SMZ removal percentage without reducing cyanobacterial growth performance (indicated by OD680), chlorophyll-a content, dry cell weight, lipid content and lipid weight/productivity of each cyanobacterium. Chroococcus sp. presented better antibiotic removal and lipid production ability than Synechococcus sp. After the addition of immobilized Chroococcus sp. into sewage, sewage bacteria attached to the surface and filled the pores of cyanobacterial beads. Immobilized Chroococcus sp. promoted the succession of sewage microbial community towards the dominance of Cellvibrio, Flavobacterium, Formivibrio and Pseudomonas, which participated in denitrification, phosphate accumulation and antibiotic removal. In the sewage microbial community, various functional genes upregulated by immobilized Chroococcus sp. directly showed negative correlation (p < 0.05) with sewage contaminants, including gltB, glnA and napA responsible for nitrogen removal; pstS responsible for phosphorus removal; nqrF, cbhA, glgC, malZ, pulA, uxaA and maeA responsible for COD removal; frmA, katG, maeA, uxaA, carB and cbhA responsible for SMZ removal. Coexisting sewage bacteria elevated (p < 0.05) lipid productivity of immobilized Chroococcus sp. to 29.13–36.52 mg/L/day through the upregulation of various genes related to metabolite exchange and signal transduction. Cyanobacterial cells degraded SMZ in collaboration with sewage bacteria, through deamination, nitration, bond cleavage, hydroxylation, desulfonation and nitrosation. The cyanobacteria-bacteria consortium presented 4-day removal percentages of 54.58–68.39%, 66.14–68.69%, 95.77–96.31% and 81.09–82.35% for SMZ, TN, TP and COD, respectively. In general, the cyanobacteria-bacteria consortium achieved a good combination of cyanobacterial lipid productivity and sewage treatment efficiency.

Recent Advancement Of Algal Biofuel Production And Its Potential Uses

The utilization of diverse energy sources is linked to the growth of industry, agriculture, and transportation. After the third generation of biodiesel production began, it was anticipated that algae would emerge as the most advantageous source because it not only can accumulate large amounts of lipids but also has the potential to reduce the amount of agricultural land required for the production of biofuels and enhance air quality by capturing carbon dioxide. In contrast to the price of making biodiesel from crops, the cost of algal biomass cultivation and processing poses a significant issue. The first step is to use genetic engineering to boost the lipid content of algal cells. The promotion of enhanced lipid accumulation by stressing algae is related to the second way. The third direction entails looking for new, promising strains of algae that will differ from the currently recognized strains in their ability to collect biomass more quickly, have greater Triacylglycerols [TAG] concentrations, and accumulate saturated and unsaturated fatty acids in the right ratios. By defining the evolutionary relationships within the main taxonomic groups of algae, a new technique in the search for biotechnologically relevant strains of algae has recently been developed. These investigations have resulted in a significant cost decrease for the manufacture of biofuel based on algae biomass. All of this research aimed at improving algae biofuel production and exploring additional biofuel sources is crucial.

Poster Session II: Can a drug for liver disease be used to treat Age-Related Macular Degeneration?

Age-related macular degeneration (AMD) is one of the most common degenerative eye diseases among the older population. One of the primary pathological features of AMD is the accumulation of fatty deposits known as drusen between the retinal pigment epithelium (RPE) and Bruch's membrane in the eye. Few options exist that can slow AMD-associated retinal degeneration. One may be to enhance the RPE's ability to oxidize fatty acids to delay or prevent drusen accumulation. Firsocostat is a small molecule that increases fatty acid oxidation in the liver. We determined the ability of Firsocostat to increase fatty acid oxidation and to decrease lipid contents in human induced pluripotent stem cells (iPSC) differentiated into RPE. In iPSC-RPE, firsocostat boosts fatty acid oxidation, remodels lipid profiles, and reduces apolipoprotein release. These data suggest that firsocostat may alleviate a pathological increase in drusen deposits and can help to preserve vision in AMD patients.

Beneficial Effect of Metformin, Quercetin, and Resveratrol Combination on High Glucose-Induced lipogenesis in HepG2 Cells

Objectives: It has been reported that Metformin (MET), Resveratrol (RSV), and Quercetin (QRS) possess anti-lipogenic effects. This study aimed to investigate the combined effects of these compounds on lipid accumulation in HepG2 cells treated with high glucose (HG).&#x0D; Methods: HepG2 cells were treated with HG (33 mM), and different concentrations of MET, QRS, and RSV. The cytotoxic effects of these compounds were determined by an MTT assay. Changes in total lipid content and triglyceride (TG) levels were measured using Oil Red O staining and a triglyceride assay kit, respectively. The expression of fatty acid synthetase (FAS) and sterol regulatory element-binding protein 1c (SREBP1c) was evaluated by quantitative real-time PCR.&#x0D; Results: MET at doses 1, 2, and 5 mM, QRS at doses 5, 10, and 20 µM, and RSV at doses 25 and 50 µM could decrease total lipid content and triglyceride levels in HepG2 cells. The EC50 (half maximal effective concentration) from Oil red O staining results were MET: 1.786 mM, QRS: 8.132 μM, and RSV: 10.9 μM. The combination of MET (mM), QRS (µM), and RSV (µM) at ratios of (2:20:50), (1:10:25), and (0.5:5:10), could reduce lipogenesis greater than that observed with each of the individual compounds of MET, QRS, or RSV or the double combinations of MET+QRS or MET+RSV. In addition, combined treatment of MET (0.5mM), QRS (5 µM), and RSV (10 µM) was able to decrease SREBP-1c and FAS genes expression in HG-treated cells.&#x0D; Conclusion: The combination of MET, QRS, and RSV could inhibit lipid accumulation in HepG2 cells by reducing total lipid content, triglyceride levels, and the expression of the genes involved in lipogenesis.

10‑Gingerol, a novel ginger compound, exhibits antiadipogenic effects without compromising cell viability in 3T3‑L1 cells.

Obesity is defined as excessive fat accumulation that can be detrimental to health and currently affects a large part of the global population. Obesity arises from excessive energy intake along with a sedentary lifestyle and leads to adipocytes with aggravated hypertrophy. Strategies have been designed to prevent and treat obesity. Nutrigenomics may serve a role in prevention of obesity using bioactive compounds present in certain foods with anti-obesogenic effects. Ginger (Zingiber officinale Roscoe) contains gingerols, key bioactive compounds that inhibit hypertrophy and hyperplasia of adipocytes. The present study aimed to evaluate the antiadipogenic activity of 10-gingerol (10-G) in the 3T3-L1 cell line. Three study groups were formed: Negative (3T3-L1 preadipocytes) and positive control (mature 3T3-L1 adipocytes) and 10-G (3T3-L1 preadipocytes stimulated with 10-G during adipogenic differentiation). Cell viability and lipid content were evaluated by MTT assay and Oil Red O staining, respectively. mRNA expression of CCAAT enhancer-binding protein α (C/ebpα), peroxisome proliferator-activated receptor γ (Pparγ), mechanistic target of rapamycin complex (Mtor), sterol regulatory element binding transcription factor 1 (Srebf1), acetyl-coenzyme A carboxylase (Acaca), fatty acid binding protein 4 (Fabp4), and 18S rRNA (Rn18s), was quantified by quantitative PCR. The protein expression of C/EPBα was analyzed by western blot. In the 10-G group, lipid content was decreased by 28.83% (P<0.0001) compared with the positive control; notably, cell viability was not affected (P=0.336). The mRNA expression in the 10-G group was higher for C/ebpα (P<0.001) and lower for Acaca (P<0.001), Fabp4 (P<0.001), Mtor (P<0.0001) and Srebf1 (P<0.0001) compared with the positive control group, while gene expression of Pparγ did not present significant changes. The presence of 10-G notably decreased C/EBPα protein levels in 3T3-L1 adipocytes. In summary, the antiadipogenic effect of 10-G during the differentiation of 3T3-L1 cells into adipocytes may be explained by mRNA downregulation of adipogenic transcriptional factors and lipid metabolism-associated genes.

Metabolic interpretation of NaCl stress-induced lipid accumulation in microalgae for promising biodiesel production with saline wastewater

The rising use of saline wastewater for biodiesel production through oleaginous microalgae demands fundamental studies on the metabolic response of microalgal cells to salt stress. Thus, we investigated how salt stress promotes lipid accumulation inside Botryococcus sp. NJD-1 by tracking critical metabolites and proteases as well as analyzing the cellular redox balance. The results showed that the lipid content of microalgal cells followed a negative binomial correlation with intracellular reactive oxygen species (ROS), while the neutral lipid content increased linearly with salinity in range of 0–20 g/L NaCl. The optimal lipid production was achieved as 54.5 % and 110.5 mg/L d-1 at 10 g/L NaCl. Further comparing the metabolomics and proteomics data at 0 and 10 g/L NaCl, the adenosine triphosphate (ATP) and nicotinamide adenine nucleotide phosphate (NADPH) were found significantly increased to quench the Na+-induced ROS. The created NADPH and ATP accord the enhanced activity of the upper Pyruvate-Malate cycle and tricarboxylic acid cycle (TCA) cycle. Therefore, ROS detoxification is proposed to be the essential mechanism for microalgal lipid accumulation under salt stress. The cellular triacylglycerol (TAG) synthesis triggered by salinity is mainly mediated by an acetyl-CoA-dependent Kennedy pathway. These findings will deepen our understanding of the nexus of ambient salt, intracellular ROS and lipid metabolism for microalgae and advance the exploit of saline wastewater for microalgal lipid production.

Localization and phenotyping of tuberculosis bacteria using a combination of deep learning and SVMs

Successful treatment of pulmonary tuberculosis (TB) depends on early diagnosis and careful monitoring of treatment response. Identification of acid-fast bacilli by fluorescence microscopy of sputum smears is a common tool for both tasks. Microscopy-based analysis of the intracellular lipid content and dimensions of individual Mycobacterium tuberculosis (Mtb) cells also describe phenotypic changes which may improve our biological understanding of antibiotic therapy for TB. However, fluorescence microscopy is a challenging, time-consuming and subjective procedure. In this work, we automate examination of fields of view (FOVs) from microscopy images to determine the lipid content and dimensions (length and width) of Mtb cells. We introduce an adapted variation of the UNet model to efficiently localising bacteria within FOVs stained by two fluorescence dyes; auramine O to identify Mtb and LipidTox Red to identify intracellular lipids. Thereafter, we propose a feature extractor in conjunction with feature descriptors to extract a representation into a support vector multi-regressor and estimate the length and width of each bacterium. Using a real-world data corpus from Tanzania, the proposed method i) outperformed previous methods for bacterial detection with a 8% improvement (Dice coefficient) and ii) estimated the cell length and width with a root mean square error of less than 0.01%. Our network can be used to examine phenotypic characteristics of Mtb cells visualised by fluorescence microscopy, improving consistency and time efficiency of this procedure compared to manual methods.