Increasing Glycerol Concentration Research Articles

Sorghum starch: Extraction, characterization and film development

Sorghum starch was extracted from sorghum grain (24.27 % average yield) and had a high carbohydrate content (86.66 %), indicating its high purity. This starch was used for the production of films with different concentrations of sorghum starch (2, 3 and 4 %) and glycerol (20, 30 and 40 g/100 g of starch). Most films were homogeneous and practically amorphous; however, the micrographs revealed the presence of some starch granules. These films also had relevant barrier properties to ultraviolet light, with infrared spectrum characteristic of sorghum starch, high lightness (L* = 87.77–90.19) and a tendency of color for the pink, but with low intensity of hue (h° = 297.06–315.20; chroma = 7.00–9.47). Thickness varied from 0.048 to 0.081 mm and the water solubility from 27 % to 48 %. Thermogravimetric analysis revealed the existence of three thermal events, correspondents to the elimination of water, decomposition of glycerol and starch, respectively. The glass transition temperature decreased with increasing glycerol content, keeping in range of 9–15°C. The film with the best mechanical performance was the one composed of 4 % starch and 30 g/100 g glycerol, as it presented a tensile strength of 3.63 MPa and elongation of 44.10 %. Water vapor permeability varied from 5.504 a 14.234 g.mm.m−2.dia−1.kPa−1, due to the increase in the concentrations of starch and glycerol. The characterization directed the usage of such films to pack vegetables and, therefore, proved that the sorghum starch is a promising source for the production of biodegradable films.

Effect of different proportions of glycerol and D-mannitol as plasticizer on the properties of extruded corn starch.

In this study, thermoplastic starch (TPS) was prepared by melt extrusion process, in which glycerol and/or D-mannitol were used as plasticizers, and the effect of different glycerol/D-mannitol ratios (4:0, 3:1, 2:2, 1:3, and 0:4) on the physicochemical properties of the extruded starch samples was investigated. The short-range molecular order, crystallization, gelatinization, thermal stability, and thermal properties of the TPS samples were analyzed through attenuated total reflection-Fourier transform infrared (ATR-FTIR) spectroscopy, X-ray diffraction (XRD), rapid visco analysis (RVA), differential scanning calorimetry (DSC), and thermogravimetric analysis (TGA). The results showed that the crystallinity and short-range molecular order of the TPS samples increased with increasing glycerol content. Conversely, the water absorption index (WAI) and water solubility index (WSI) of the TPS samples decreased with increasing glycerol content. In addition, the TPS samples with higher glycerol content exhibited higher gel and thermal stabilities. This study provides a theoretical basis for starch extrusion and plasticization in the preparation of TPS-based materials with specific properties.

The strengthening mechanism of mechanical performance for polyurethanes with micro-content crosslinkers

Chemical crosslinking is one widely used method to endow polyurethane (PU) with desirable properties. However, the introduction of crosslinkers in PU elastomers can often lead to the destruction of phase separation, resulting in weakened mechanical properties. Herein, a series of PUs with micro-content crosslinker were synthesized from poly(oxytetramethylene) glycol (PTMG), diphenylmethane-4,4′-diisocyanate (MDI), and chain extender of butane-1,4-diol (BDO) containing varied amounts of glycerol. The phase separation of PUs decreases with increased glycerol content, while the tensile strength increases. To elucidate this enhancement in mechanical properties, morphology investigation with capability of direct observation of components distribution is employed. An increased thickness of interphase between the domain of hard segment (HS) and soft segment (SS) is observed, which indicates that more HS is moved to the interphase upon the introduction of glycerol. The results of electron energy loss spectroscopy (EELS) further disclosed that glycerol formed aggregations located at the interphase. These aggregations increase the crosslinking density of the interphase, which strengthens HS domains and prevents them from being destroyed under tension, thereby increasing the tensile strength. It is the first time of disclosing distribution feature of micro-content crosslinker of PU as well its strengthening mechanism in tensile strength, which could assist in developing PU with desirable mechanical properties.

Antifungal mechanism of Angelica sinensis essential oil against Penicillium roqueforti and its application in extending the shelf life of bread

There are a variety of reports on the application of Angelica sinensis essential oil (ASEO) in the biomedical field. However, the antifungal mechanism of ASEO has not been reported. In this study, the antifungal mechanism of ASEO against Penicillium roqueforti was investigated by proteomics and genomics. ASEO can increase the permeability of P. roqueforti cell membrane and decrease the content of lipid and trehalose. With the increase of glycerol content, the HOG signaling pathway can be upregulated. Consistent with the above phenotypic changes, proteomics confirmed that ASEO treatment inhibited the steroid synthesis pathway of P. roqueforti. The significant down-regulation of ERG4, ERG6, ERG25, SMT1, and FDFT1 gene expression confirmed this conclusion. Cluster+activates the MAPK and UPP signaling pathways and ultimately leads to cell apoptosis. The bread shelf life experiment showed that ASEO could extend the shelf life of bread up to day 7. This study provides new evidence for the antifungal activity of ASEO against P. roqueforti and will promote the use of ASEO in the preservation of food and agricultural products.

Effect of film properties on droplet impact on suspended films

Understanding the dynamics and underlying mechanisms of droplet impact on suspended films is crucial for controlling droplet motion. In this study, we conducted experiments to investigate droplet impact on suspended films and examined the typical phenomena and corresponding Weber number domains. We explored the effects of changing sodium dodecyl sulfate (SDS) concentration and glycerol content on the surface tension and dynamic viscosity of the films. Additionally, we elucidated the characteristics of film deformation and droplet trajectory. An energy analysis was performed, considering the droplet kinetic energy (Ek0), surface energy increment of the film (ΔEfs), and viscous dissipation (Evis). The results demonstrate that as the SDS concentration increases, the upper critical Weber number (Wecru) between bouncing and passing decreases, while the lower critical Weber number (Wecrl) between coalescence and bouncing first decreases and then increases. For droplet bouncing, increasing the SDS concentration makes the films more susceptible to deformation, whereas increasing the glycerol content enhances film rigidity. Moreover, increasing the SDS concentration and glycerol content leads to higher energy losses, resulting in a reduction in the maximum bouncing height (hbdmax). Regarding droplet passing, the passing velocity (Vpd) increases with increasing SDS concentration. For films without SDS, Vpd increases with increasing glycerol content, while for films with SDS, Vpd decreases. The different impact modes are categorized based on the dimensionless energy parameter E* = Ek0/(ΔEfs + Evis). We have successfully predicted the critical height thresholds (Hd0crl and Hd0cru) and established a relationship of We* = f (Re*, Bo*) that aligns well with experimental results.

Effect of glycerol concentration on levels of toxicants emissions from water-pipe tobacco smoking (WTS)

Glycerol, flavorings and sweeteners constitute approximately 70% of water-pipe tobacco smoking (WTS) mixtures. Tobacco mixture combustion produces smoke toxins (e.g. carbonyl compounds), of which the type and amount are highly dependable on tobacco mixture formula. While glycerol in tobacco mixture contribute to enhanced smoking experience, its’ combustion produces toxicants such as acrolein. According to WHO, there are no approved international upper limits regulations on WTS ingredients. This study aims to assess toxicant emission levels corresponding to increasing glycerol concentration in WTS mixtures, which may aid in developing tobacco regulations towards harm reduction.MethodsLaboratory experimental study. Using laboratory water-pipe smoking machine, levels of toxicant emissions in the smoke from WTS mixture samples containing varying glycerol concentrations were measured using High-performance Liquid Chromatography (HPLC). Smoke from 5 consecutive smoking cycles with 35 puffs each (ISO 22486 standard) was led through a trapping system as described in the Cooperation Centre for Scientific Research Relative to Tobacco (CORESTA) recommended method No. 74 (Determination of selected carbonyls in mainstream cigarette smoke by HPLC). Trapped carbonyls were then analysed by HPLC with a DAD detector.ResultsAcrolein emission is associated with glycerol addition in WTS mixture indicated by lab-made samples throughout all glycerol concentrations (10%, 20%, 40% and 60%), and brand samples with glycerol concentrations 10% to 20%. However, brand samples showed no increase in acrolein emission corresponding to the increase in glycerol concentrations from 20% to 60%.ConclusionThe effect of glycerol addition in waterpipe tobacco on acrolein emission varies between products. Tobacco fillers, additives and contents quality and other factors may affect toxicant emission levels. Therefore, regulatory recommendations towards defining upper limits of content concentrations require further investigations regarding potential confounders in acrolein emissions and health effects of market-available glycerol concentrations in waterpipe tobacco smoking.

A terpyridyl functionalized rhodamine dye for viscosity sensing with fluorescence enhancement and mitochondria-targeting imaging

Exploring new fluorescent organic dyes with mitochondria-targeted and viscosity-sensitive functions attracts extensive attention because they may provide access to a better understanding of the microenvironments of the subcellular level via biological imaging. Herein a terpyridyl functionalized rhodamine dye (TRMe) has been facilely designed and prepared, which shows a highly selective fluorescence enhancement response to viscosity (1.0–1238.0 cp) in water-glycerol mixtures with the increased glycerol content. In addition, TRMe can respond to viscosity enhancement over a wide pH range (1.0–10.0) and presents low cell toxicity. Besides, a good linearity between the viscosity and fluorescence lifetime of TRMe can be observed. Significantly, TRMe shows excellent mitochondria-targeted performance with a Pearson's colocalization coefficient of 0.958 and can be employed as a tracer for viscosity imaging in Nystatin-stimulated living HeLa cells.

Development of Silk Fibroin-Based Non-Crosslinking Thermosensitive Bioinks for 3D Bioprinting.

Three-dimensional (3D) bioprinting holds great promise for tissue engineering, allowing cells to thrive in a 3D environment. However, the applicability of natural polymers such as silk fibroin (SF) in 3D bioprinting faces hurdles due to limited mechanical strength and printability. SF, derived from the silkworm Bombyx mori, is emerging as a potential bioink due to its inherent physical gelling properties. However, research on inducing thermosensitive behavior in SF-based bioinks and tailoring their mechanical properties to specific tissue requirements is notably lacking. This study addresses these gaps through the development of silk fibroin-based thermosensitive bioinks (SF-TPBs). Precise modulation of gelation time and mechanical robustness is achieved by manipulating glycerol content without recourse to cross-linkers. Chemical analysis confirms β-sheet conformation in SF-TPBs independent of glycerol concentration. Increased glycerol content improves gelation kinetics and results in rheological properties suitable for 3D printing. Overall, SF-TPBs offer promising prospects for realizing the potential of 3D bioprinting using natural polymers.

Versatile hydrolyzed polyacrylamide-polyethyleneimine gels reinforced with glycerol for plugging and abandonment of oil wells

Temporary abandonment of oil wells requires isolation protocols to ensure safety of the environment and operators, for subsequent reactivation. For this, barrier materials are used to prevent influx of fluids from the rock formation to the wellbore. Polymeric gels are a promising alternative due to their ability to form a three-dimensional structure in situ with adequate elastic properties. With this goal, solid-free gels of hydrolyzed polyacrylamide and polyethyleneimine with glycerol as reinforcement material were developed. Furthermore, diethylenetriamine and diethanolamine were added to the formulations as anti-syneresis agents. The gelation kinetics and gels stability were evaluated through bottle tests and rheological measurements. The increase in glycerol content combined with DETA and DEA in the gels resulted in a reduction of gelation time varying according to the concentration of anti-syneresis agents added. This combination also enabled the increase on thermal stability of the gels with significant delay of 110% in the syneresis time. Furthermore, the storage and loss modulus and tan delta results showed that the new HPAM-PEI-Glycerol-DETA/DEA gel formulation had superior elastic properties compared to the HPAM-PEI gel.

Interfacial polymerization process based on diffusion control: Role of chemical composition and morphology on fouling resistance

Nanofiltration, as a technology with low energy consumption, no secondary pollution, and continuous and compact operation, is widely used in the fields of seawater desalination, resource recovery and reuse, and drinking water purification, etc. However, “trade-off” effect and membrane fouling are the main problems limiting the application of nanofiltration membranes. Herein, glycerol was introduced into the aqueous reactive solution in interfacial polymerization (IP) for regulating chemical composition and membrane structure simultaneously. As glycerol raised the viscosity of the reactive solution which slowing down the diffusion rate of amine monomer, crumpled structure generated by the diffusion-driven instability. At the same time, the addition of glycerol varied the oxygen content in the skin layer of the membrane. At 60% glycerol content, the as-prepared membrane achieved a pure water permeance of 21.14 L·m−2·h−1·bar−1 with sodium sulfate rejection of 95.6%. However, the oxygen content in the chemical composition of the membrane showed a minimum value with the increase of glycerol concentration indicating a competitive reaction between the amine monomer and glycerol. Through the anti-fouling analysis of membranes with different chemical compositions and structures, it was found that the increase in roughness of nanofiltration membranes with similar chemical composition resulted in an increase of irreversible pollution. The nanofiltration membrane with similar membrane structure and higher oxygen content had stronger anti-fouling ability to humic acid. It is concluded that the rational design of the crumpled membrane with high oxygen-chemical composition is expected to obtain a high flux nanofiltration membrane with anti-fouling performance.

Simultaneous Hydrogen and Ethanol Production from Crude Glycerol by a Microbial Consortium Using Fed-Batch Fermentation

Simultaneous bioproduction of hydrogen and ethanol from cheaper waste feedstock has the potential for the development of a more cost-effective biofuel generation process. Crude glycerol (CG), a by-product of the biodiesel industry, is a renewable resource, abundant, sold at low prices and available worldwide. However, the main CG limitations in fermentation processes are mainly related to the presence of impurities and the lack of nitrogen sources, both acting on microbial activity. In this study, a fermentation process with CG was improved using a highly specific microbial consortium called GlyCeroL (GCL). The process was developed in fed-batch fermentation mode using not diluted substrate and carried out under non-sterile conditions and at increasing amounts of the substrate (from 20 to 80 gL−1 of glycerol). The results showed higher H2 (from 6 to 8 LL−1) and EtOH (from 13 to 20 gL−1) production by increasing glycerol concentration from 20 to 40 gL−1. On the other hand, a decrease in glycerol degradation efficiency (from 75 to 56%) was observed. Then, the nitrogen sparging strategy was applied. Using CG of 40 gL−1, process improvement was achieved, leading to the increased production of hydrogen (10 LL−1) but not that of ethanol (20 gL−1). A further increase to 60 gL−1 of glycerol produced a slight increment of EtOH (21 gL−1) and H2 (11 gL−1) but a sharp decrease in glycerol degradation efficiency (41%). Acetate, as the main impurity of CG, was an additional carbon source for GCL microorganisms contributing to EtOH production and increasing that of lactic acid to restore the redox balance. The Denaturing Gradient Gel Electrophoresis (DGGE) fingerprint at the end of all fed-batch fermentations supported the robustness of GCL functional units and their adaptability to fermentation conditions.

Toward a Circular Bioeconomy: Exploring Pineapple Stem Starch Film as Protective Coating for Fruits and Vegetables.

In order to reduce our dependence on nonrenewable plastics and solve the problem of non-biodegradable plastic waste, there has been much attention paid to the development of biodegradable plastics from natural resources. Starch-based materials have been widely studied and developed for commercial production, primarily from corn and tapioca. However, the use of these starches could generate food security problems. Therefore, the use of alternative starch sources, such as agricultural waste, would be of great interest. In this work, we investigated the properties of films prepared from pineapple stem starch, which has a high amylose content. Pineapple stem starch (PSS) films and glycerol-plasticized PSS films were prepared and characterized using X-ray diffraction and water contact angle measurements. All films exhibited some degree of crystallinity, making them water-resistant. The effect of glycerol content on mechanical properties and gas (oxygen, carbon dioxide and water vapor) transmission rates was also studied. The tensile modulus and tensile strength of the films decreased with increasing glycerol content, while gas transmission rates increased. Preliminary studies showed that coatings made from PSS films could slow down the ripening process of bananas and extend their shelf life.

Rapid wetting of shear-thinning fluids

By studying the spreading behavior of aqueous glycerol and polymer solutions on smooth surfaces, we found that shear-thinning solutions behave similarly to water in the initial spreading phase, while glycerol solutions show significantly slower spreading. For the glycerol solutions, an increase in glycerol concentration effectively increases the contact-line friction, resulting in increased resistance to wetting. For the polymeric solutions, however, an increase in polymer concentration does not modify contact-line friction. As a consequence, the energy dissipation at the contact line can not be controlled by varying the amount of additives for shear-thinning fluids.

Insight into the Effect of Glycerol on Dielectric Relaxation and Transport Properties of Potassium-Ion-Conducting Solid Biopolymer Electrolytes for Application in Solid-State Electrochemical Double-Layer Capacitor.

The increased interest in the transition from liquid to solid polymer electrolytes (SPEs) has driven enormous research in the area polymer electrolyte technology. Solid biopolymer electrolytes (SBEs) are a special class of SPEs that are obtained from natural polymers. Recently, SBEs have been generating much attention because they are simple, inexpensive, and environmentally friendly. In this work, SBEs based on glycerol-plasticized methylcellulose/pectin/potassium phosphate (MC/PC/K3PO4) are investigated for their potential application in an electrochemical double-layer capacitor (EDLC). The structural, electrical, thermal, dielectric, and energy moduli of the SBEs were analyzed via X-ray diffractometry (XRD), Fourier transforms infrared spectroscopy (FTIR), electrochemical impedance spectroscopy (EIS), transference number measurement (TNM), and linear sweep voltammetry (LSV). The plasticizing effect of glycerol in the MC/PC/K3PO4/glycerol system was confirmed by the change in the intensity of the samples' FTIR absorption bands. The broadening of the XRD peaks demonstrates that the amorphous component of SBEs increases with increasing glycerol concentration, while EIS plots demonstrate an increase in ionic conductivity with increasing plasticizer content owing to the formation of charge-transfer complexes and the expansion of amorphous domains in polymer electrolytes (PEs). The sample containing 50% glycerol has a maximal ionic conductivity of about 7.5 × 10-4 scm-1, a broad potential window of 3.99 V, and a cation transference number of 0.959 at room temperature. Using the cyclic voltammetry (CV) test, the EDLC constructed from the sample with the highest conductivity revealed a capacitive characteristic. At 5 mVs-1, a leaf-shaped profile with a specific capacitance of 57.14 Fg-1 was measured based on the CV data.

Ultrafast fluorescence depolarisation in green fluorescence protein tandem dimers as hydrophobic environment sensitive probes.

Advances in ultra-fast photonics have enabled monitoring of biochemical interactions on a sub nano-second time scale. In addition, picosecond dynamics of intermolecular energy transfer in fluorescent proteins has been observed. Here, we present the development of a genetically encoded fluorescent sensor that can detect changes in hydrophobicity by monitoring ultrafast fluorescence depolarisation. Our sensor is composed of a pair of dimeric enhanced green fluorescent proteins (dEGFPs) linked by a flexible amino-acid linker. We show dimerisation is perturbed by the addition of glycerol which interferes with the hydrophobic interaction of the two proteins. Time-resolved fluorescence anisotropy revealed a systematic attenuation of ultrafast fluorescence depolarisation when the sensor was exposed to increasing glycerol concentrations. This suggests that as hydrophobicity increases, dEGFP pairing decreases within a tandem dimer. Un-pairing of the protein fluorophores dramatically alters the rate of energy transfer between the proteins, resulting in an increase in the limiting anisotropy of the sensor.

Starmerella bacillaris Released in Vineyards at Different Concentrations Influences Wine Glycerol Content Depending on the Vinification Protocols.

Starmerella bacillaris is a non-Saccharomyces yeast proposed for must fermentation together with Saccharomyces cerevisiae because of its high glycerol and moderate volatile acidity production. Furthermore, it was demonstrated that the same S. bacillaris strains that possess interesting technological properties exhibited antifungal activity against Botrytis cinerea, suggesting the release of this yeast in the vineyard. To obtain a positive effect during the following winemaking process, the maintenance of suitable concentrations of S. bacillaris is essential. Therefore, to obtain information on the survival of S. bacillaris, a small-scale field trial was performed. One week before the harvest, two different concentrations of S. bacillaris (106 and 107 cells/mL) were sprayed on Pinot grigio bunches, and the strain concentration was monitored by means of qPCR during the subsequent fermentation process. In addition, the combined effect of different winemaking techniques was evaluated, i.e., the vinification of juice, juice with marc and cryomaceration treatment. Results demonstrated that, under the tested conditions, S. bacillaris released in the vineyard remained viable for one week on grape bunches and increased glycerol content during the subsequent fermentation process. Different vinification protocols influenced cell concentrations. In particular, the cryomaceration treatment, due to the use of low temperature, supported S. bacillaris growth due to its cryotolerant aptitude. The collected data open new perspectives on the control of alcoholic fermentation, involving both vineyard and cellar management.

Green extraction of bioactive compounds from Azadirachta indica in aqueous glycerol and modelling and optimisation by response surface methodology

Abstract Development of efficient and green methods for extracting bioactive phytochemicals has great industrial value. Increasing environmental sensitivity at the global level has tremendously enhanced the demand for such methods. Azadirachta indica is a well-known medicinal tree. As glycerol has emerged as a green and safe extraction solvent for bioactive phytochemicals, this study aimed to investigate the efficacy of a glycerol–water solvent system to extract bioactive compounds from A. indica leaves. Modelling and optimisation were carried out by using response surface methodology (RSM) as per the Box–Behnken design with three variables, namely, solvent concentration, time and temperature. The responses were total phenolic content (TPC), total flavonoid content (TFC), 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity and metal chelating activity (MCA). The optimum conditions found by numerical optimisation were a solvent concentration of 69.713%, a time period of 38.328 min and a temperature of 32.114 °C with the predicted values of TPC, TFC, %DPPH and %MCA as 5.27 mg gallic acid equivalents · g−1 DW (dry weight), 9.869 mg rutin equivalents · g−1 DW, 73.8% and 54.366%, respectively. The validation experiments showed almost the same results for each response with very low% errors (5.431–7.661). Increasing glycerol concentration in the extracting medium favoured the extraction of TPC, TFC and antioxidant phytochemicals, but for MCA, the trend was the opposite. In conclusion, 70% aqueous glycerol is an effective medium for the extraction of polyphenolic and antioxidant phytochemicals from A. indica leaves. Extraction models suggested by RSM have high prospects to be used on a large industrial scale.

PDMS 표면에서 충돌 액적의 점도에 따른 거동의 주기성

Periodicity behavior was observed after droplet collision in viscous solutions with controlled glycerol concentrations onto a PDMS surface. No periodic movement was observed in the droplets at glycerol concentrations of 50% or more. In contrast, the vertical diameter of the droplets increased and decreased periodically at glycerol solution concentrations of 40% or less. Moreover, there was little change in the periodicity of the impacting droplet movement, and the vibration frequency was measured at approximately 80 to 98 Hz in the entire range. The maximum droplet spreading factor after collision decreased significantly with increasing glycerol concentrations of 40% or more (Ohnesorge number 1.4 × 10SUP-2/SUP or more). The results suggest that the effect of viscosity became greater than that of the solution surface tension with increasing glycerol concentrations.

An Approach for Incorporating Glycerol as a Co-Substrate into Unconcentrated Sugarcane Bagasse Hydrolysate for Improved Lipid Production in Rhodotorula glutinis

Sugarcane bagasse is a potential raw material for microbial lipid production by oleaginous yeasts. Due to the limited sugar concentrations in bagasse hydrolysate, increasing carbon the concentration is necessary in order to improve lipid production. We aimed to increase carbon concentration by incorporating glycerol as a co-substrate into unconcentrated bagasse hydrolysate in the cultivation of Rhodotorula glutinis TISTR 5159. Cultivation in hydrolysate without nitrogen supplementation (C/N = 42) resulted in 60.31% lipid accumulation with 11.45 ± 0.75 g/L biomass. Nitrogen source supplementation increased biomass to 26.29 ± 2.05 g/L without losing lipid accumulation at a C/N of 25. Yeast extract improved lipid production in the hydrolysate due to high growth without altering the lipid content of the cells. Mixing glycerol up to 10% v/v into the unconcentrated hydrolysate improved biomass and lipid production. A further increase in glycerol concentrations drastically decreased growth and lipid accumulation by the yeast. By maintaining C/N at 27 using yeast extract as the sole nitrogen source, hydrolysate mixed with 10% v/v glycerol resulted in the highest lipid yield, at 19.57 ± 0.53 g/L with 50.55% lipid content, which was a 2.8-fold increase compared to using the hydrolysate alone. In addition, yeast extracts were superior for promoting growth and lipid production compared to inorganic nitrogen sources.

Tumor necrosis factor-α promotes lipolysis and reduces insulin sensitivity by activating nuclear factor kappa B and c-Jun N-terminal kinase in primary bovine adipocytes.

Sustained lipolysis and insulin resistance increase the risk of metabolic dysfunction in dairy cows during the transition period. Proinflammatory cytokines are key regulators of adipose tissue metabolism in nonruminants, but biological functions of these molecules in ruminants are not well known. Thus, the objective of this study was to investigate whether tumor necrosis factor-α (TNF-α) could affect insulin sensitivity and lipolysis in bovine adipocytes as well as the underlying mechanisms. Bovine adipocytes (obtained from the omental and mesenteric adipose depots) isolated from 5 Holstein female calves (1 d old) with similar body weight (median: 36.9 kg, range: 35.5-41.2 kg) were differentiated and used for (1) treatment with different concentrations of TNF-α (0, 0.1, 1, or 10 ng/mL) for 12 h; (2) pretreatment with 10 μM lipolytic agonist isoproterenol (ISO) for 3 h, followed by treatment with or without 10 ng/mL TNF-α for 12 h; and (3) pretreatment with the c-Jun N-terminal kinase (JNK) inhibitor SP600125 (20 μM for 2 h) and nuclear factor kappa B (NF-κB) inhibitor BAY 11-7082 (10 μM for 1 h) followed by treatment with or without 10 ng/mL TNF-α for 12 h. The TNF-α increased glycerol content in supernatant, decreased triglyceride content and insulin-stimulated phosphorylation of protein kinase B suggesting activation of lipolysis and impairment of insulin sensitivity. The TNF-α reduced cell viability, upregulated mRNA abundance of Caspase 3 (CASP3), an apoptosis marker, and increased activity of Caspase 3. In addition, increased phosphorylation of NF-κB and JNK, upregulation of mRNA abundance of interleukin-6 (IL-6), TNFA, and suppressor of cytokine signaling 3 (SOCS3) suggested that TNF-α activated NF-κB and JNK signaling pathways. Furthermore, ISO plus TNF-α-activated NF-κB and JNK signaling pathway to a greater extent than TNF-α alone. Combining TNF-α and ISO aggravated TNF-α-induced apoptosis, insulin insensitivity and lipolysis. In the absence of TNF-α, inhibition of NF-κB and JNK did not alter glycerol content in supernatant, triglyceride content or insulin-stimulated phosphorylation of protein kinase B. In the presence of TNF-α, inhibition of NF-κB and JNK alleviated TNF-α-induced apoptosis, insulin insensitivity and lipolysis. Overall, TNF-α impairs insulin sensitivity and induces lipolysis and apoptosis in bovine adipocytes, which may be partly mediated by activation of NF-κB and JNK. Thus, the data suggested that NF-κB and JNK are potential therapeutic targets for alleviating lipolysis dysregulation and insulin resistance in adipocytes.