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This research aimed to investigate the effects of sodium chloride (NaCl) on the formation of Nε-carboxymethyllysine (CML) and Nε-carboxyethyllysine (CEL), in pork myofibrillar protein (MFP)–reducing sugar (glucose or ribose) systems (with or without oleic acid) during commercial sterilization (121 °C, 10 min). The addition of NaCl (1% or 2%) in all tested model systems (MFP and MFP–glucose systems with or without oleic acid, and MFP–ribose–oleic acid system), except for the MFP–ribose system, resulted in less heat-induced production of CML and CEL. Although the addition of oleic acid in MFP or MFP–sugar systems without NaCl generally promoted the formation of CML and CEL during heating, the heat-induced production of CML and CEL in MFP–sugar–oleic acid–NaCl systems were much lower than that in their MFP–sugar–NaCl counterparts. This implies that the presence of NaCl could alter the general promoting effect of lipid oxidation on the formation of advanced glycation end-products in protein–reducing sugar systems, which may be due to the effects of NaCl on the roles of free radicals playing in the autoxidation of reducing sugars as well as in the Maillard reaction.

This research was aimed to investigate the effects of acetic acid and citric acid (0.5, 1 g/100 g pork) on quality properties of raw pork during storage (0 °C, 0–8 days) and subsequent commercial sterilization (121 °C, 10 and 30 min), and to understand the impacts of the changes of raw pork (with acid added) during storage on the quality properties of their heat-treated counterparts. The addition of acetic/citric acid generally led to decreased levels of thiobarbituric acid reactive substances (TBARS) and Schiff bases and increased intensity of tryptophan fluorescence (ITF) for both raw and sterile pork, implying that acetic/citric acid could inhibit lipid and protein oxidation and promote protein hydrolysis in pork. The levels of trichloroacetic acid solution-soluble peptides, TBARS, Schiff bases, and ITF in sterile pork were highly correlated (r= 0.722–0.887; P< 0.01) with their initial levels in raw pork prior to the heating, indicating that these quality indexes for sterile pork were strong dependence of the conditions of their raw counterparts. The addition of acetic/citric acid led to opposite effects on the cooking loss or area shrinkage of pork heated for 10 min (average increased by 4–23% or 6–27%) and that heated for 30 min (average decreased by 2–16% or 4–8%). Moreover, the quality properties of raw pork prior to the 30 min heating had minor impact on the cooking loss and area shrinkage of subsequently high-heat-treated pork.

For efficiency and cost savings during manufacturing, a fixed wide REBa2Cu3O7-x (REBCO) tape is usually mechanically slit to a width required by varying applications, whereas the multiple edge cracks are induced in this process. In physics, it is widely recognized that the critical current of REBCO tape is degraded with tensile strain, and excessive strain causes crack growth. We are curious about how the edge crack grows under tensile strain, thereby further affecting the critical current performance. Therefore, we first introduced the phase-field method (PFM) to analyze the cracking and fracture behavior. After building a numerical model containing an edge crack and solving it by the finite element method, we presented the propagation path of the edge crack under tension and obtained the quantitative correlation between fracture ratio (crack depth/tape width) and tensile strain. Furthermore, since cracks impede the current flow and make the current shunted to the metal layer, resulting in critical current degradation, we adopted a current shunting model that considers the obstacle of cracks to investigate the relationship between critical current degradation and fracture ratio. Moreover, employing the intermediate variable of fracture ratio, we obtained the ${I}_c - \varepsilon $ curve of REBCO tape. By comparing the calculated ${I}_c - \varepsilon $ curve with previous experimental data, the critical current degradation with tensile strain was very well interpreted by the propagation of the edge crack induced by mechanical slitting.

In this research, soymilk was fermented with Lactobacillus paracaseiL9 (L. paracaseiL9) and a commercial starter containing Lactobacillus bulgaricus(L. bulgaricus) and Streptococcus thermophilus(S. thermophilus). Quality changes including molecular weight distribution of proteins, free and total amino acids, and other quality indicators were evaluated during the fermentation (37 °C, 24 h) of soymilk and the subsequent storage (4 °C, 1–21 d) of the fermented soymilk. Both L. paracaseiL9 and the commercial starter showed good fermentative capability. The viable cell counts in soymilk increased from about 6.5 to over 9 log CFU/mL within 4 or 12 h of fermentation. The fermentation led to the breakage of the highly aggregated proteins (> 1000 kDa) and the production of small peptides. At the same time, there were significant reduction (P< 0.05) of total amino acids, but the total free amino acids of L9SM and CSSM increased by 68% and 134%, respectively. The fermented soymilk products had stable quality with small changes of pH (within 0.05) and viable cell counts (within 0.5 log CFU/mL) during the storage. L. paracaseiL9 exhibited a higher level of ability for degradation of soy proteins during the fermentation of soymilk and later storage of the fermented soymilk.

This study was to understand the formation of protein-bound Nε-carboxymethyllysine (CML) and Nε-carboxyethyllysine (CEL) in raw minced pork during cold storage (0 °C, 1–7 d) as affected by the addition (0.5% w/w) of sodium bicarbonate (SB), sodium tripolyphosphate (STPP) and tetrasodium pyrophosphate (TSPP). The mean levels of TBARS (P= 0.004), the intensity of tryptophan fluorescence (P= 0.000), and TCA-soluble peptides (P= 0.006) in minced pork were significantly affected by the storage duration, implying the occurrence of lipid oxidation, protein oxidation, and protein hydrolysis in minced pork during the storage. However, the storage duration of minced pork in general did not greatly affect their CML and CEL levels. In addition, the average of CML levels (control 1.00 ± 0.53 mg/kg, SB 1.04 ± 0.54 mg/kg, STPP 0.93 ± 0.47 mg/kg, TSPP 0.94 ± 0.48 mg/kg) in minced pork was not affected by the addition of alkaline salt, but all alkaline salt treated pork had higher levels of CEL (control 3.00 ± 0.64 mg/kg, SB 3.79 ± 0.80 mg/kg, STPP 3.92 ± 1.03 mg/kg, TSPP 4.58 ± 0.94 mg/kg) than their control counterparts without alkaline salt. The promoting effects of alkaline salts for CEL formation were not directly tied to their effects on the pH, TBARS, the intensity of tryptophan fluorescence, and TCA-soluble peptides of minced pork.

Contents of protein-bound advanced glycation end-products (AGEs) including Nε-carboxymethyl-lysine (CML) and Nε-carboxyethyl-lysine (CEL), as well as total viable counts (TVC), total volatile basic nitrogen (TVB-N) and thiobarbituric acid reactive substances (TBARS) in vacuum-packed dried salted Spanish Mackerel fillets previously brined with 0‒15% NaCl during storage at 25 °C were determined. In freshly dried fish fillets with increased salt concentration, TVB-N decreased from 39.32 ± 5.39 to 22.35 ± 4.55 mg N/100 g and TBARS increased from 0.18 ± 0.01 to 0.50 ± 0.03 mg MDA/kg. No statistical differences were found in the levels of TVC, CML and CEL of dried fish (P= 0.215‒0.645). TVC and TVB-N levels of the dried salted fish fillets increased during 25 d storage (P= 0.000‒0.036), but reduced with increase of NaCl concentration (P= 0.000‒0.002). TBARS levels of the fillets increased until day15 and then slightly declined (P= 0.000‒0.036). Salt promoted the increase in TBARS in dried fish but showed inhibition effect with higher concentrations (P= 0.000‒0.003). Levels of protein-bound CML and CEL in both unsalted and salted fish fillets increased constantly during storage (P= 0.000‒0.050), whereas NaCl of increased content inhibited the formation of protein-bound CML and CEL in fillets (P= 0.000‒0.119). Positive correlations were observed in the contents of protein-bound CML and CEL with TVC, TVB-N and TBARS (r = 0.445‒0.839, P= 0.000‒0.002), indicating the formation of protein-bound AGEs in dried fish fillets is attributable to protein deterioration and lipid oxidation during storage.

This study was aimed to investigate the cryoprotective effects of collagen peptides (10 g/kg; Collagen A, about 4500 Da; Verisol, 2000–3000 Da) on myofibrillar protein (MP) from silver carp surimi and the effects of collagen peptides on the characteristics of surimi gels during frozen storage (− 18 °C, 3–30 days) in comparison with that of a commercial cryoprotectant (40 g/kg sorbitol and 40 g/kg sucrose), sericin peptides (10 g/kg, about 3500 Da) and the control group (without cryoprotectant). The results indicated that Verisol, a collagen peptide with low molecular weight, was the optimal cryoprotective agent for the frozen storage of silver carp surimi. After 30 days of frozen storage, the MP from frozen surimi with Verisol contained 6.0–33.5% less carbonyls (26.8 ± 0.3 µmol/g), and 0.6–99.6% more free sulfhydryl (84.0 ± 0.4 µmol/g), as well as exhibited 17.6–47.1% greater Ca2+-ATPase activity (0.23 ± 0.02 µmol Pi/mgprot/hour) and 5.8–58.2% more extractable salt-soluble proteins (45.3 ± 1.0%) in comparison with that of the other four treatments. In addition, the water holding capacity (94.2 ± 2.2%), gel strength (314.8 ± 8.0 g·mm) and whiteness (79.1 ± 1.3) of the surimi gels with Verisol were also significantly better (p< 0.05) than that of the other treatments.

Surface-enhanced Raman scattering (SERS) signals of histamine collected via Au–Ag core–shell nanoparticles (Au@Ag NPs; diameter: entire particle, 43 ± 4 – 96 ± 8 nm; Au seed, 20 ± 2 nm) as affected by aggregating agents (AAs; 1 – 106μmol/L) including NaCl, MgCl2, NaClO4, and Mg(ClO4)2were investigated in an attempt to develop a simple and sensitive SERS analytical approach for histamine in squid muscle. Results showed that the effects of AAs on SERS intensities of histamine depended on their types and concentrations, and the addition of an AA at its optimal concentration could result in as much as 139% – 375% increase in the SERS signal of histamine. The incorporation of NaClO4(1 × 104μmol/L) based upon Au@Ag NPs (diameter, 59 ± 6 nm) led to the best SERS amplification effect. As low as 5 mg/kg of histamine in squid muscle could be detected, and the histamine levels in squid muscle (0 – 400 mg/kg, n = 27) predicted by partial least square regression model were highly correlated with their actual levels in squid samples (R2= 0.987). This study showed the possibility of greatly amplifying SERS signals of an analyte via selecting the appropriate type and concentration of an AA.

This study was aimed to investigate the effects of cold storage (0 °C, 1–7 day) and the addition of sodium chloride (0–2.5% w/w) on the amounts of two α-dicarbonyl compounds including glyoxal (GO) and methylglyoxal (MGO) in raw and heated (121 °C, 10 min) white meat of grass carp (Ctenopharyngodon idellus). Although not significantly different, the average amounts of both GO (1 day, 11.86 ± 1.44 mg/kg; 4 days, 13.26 ± 2.42 mg/kg; 7 days, 14.72 ± 2.20 mg/kg) and MGO (1 day, 1.84 ± 0.32 mg/kg; 4 days, 1.88 ± 0.38 mg/kg; 7 days, 2.24 ± 0.83 mg/kg) in raw fish meat increased with storage time, regardless of the addition of salt or not. A longer storage time of fish meat prior to the heat treatment led to the formation of less GO (P= 0.014) and MGO (P= 0.072, although not significant different) or even reduction of GO and MGO due to the heating. The addition of 0.5–2.5% of salt resulted in no obvious effects on the levels of GO and MGO in either raw or sterile fish meat. The total volatile basic nitrogen values (indicating fish freshness) and the levels of GO (R = 0.596, P= 0.002) or MGO (R = 0.480, P= 0.018) for raw fish meat were significantly correlated, but the values of thiobarbituric acid reactive substances (indicating lipid oxidation extent) were not related to the amount of GO or MGO in either raw or heated fish meat.

This research was aimed to evaluate the effects of calcium coagulants (10–40 mmol/L in soymilk) including calcium chloride, calcium lactate, and calcium gluconate on the textural properties (microstructures, large deformation behaviors under uniaxial compression) and some relevant chemical properties of firm tofu. The results showed that the stress–strain behaviors of tofu could be modelled very well with the first order Ogden model (R2= 0.992–0.998), and the effects of three coagulants on the textural properties of tofu followed similar trends. Firstly, the failure strain (0.66–0.73, indicating deformability) of tofu was not significantly affected by the type (P= 0.056) and the concentration (P= 0.399) of calcium added. In addition, as the calcium level increased from 10 to 30 mmol/L, the failure stress (from 5.27–5.38 kPa to 17.09–19.73 kPa) and rigidity (from 3.18–3.36 kPa to 10.16–12.96 kPa) of tofu continuously increased, but a further increase of calcium level to 40 mmol/L led to decreases in both failure stress (14.17–15.51 kPa) and rigidity (9.37–11.39 kPa). Furthermore, the effects of calcium levels on the strength of tofu were consistent with that on the microstructure features, moisture levels, and water holding capacity of tofu. Tofu prepared with all three coagulants were good sources of calcium (123–311 mg/100 g tofu), which may be used to meet consumer needs for diversified products of calcium-rich tofu.

This study was aimed to investigate the cryoprotective effects of silver carp muscle hydrolysate (SCMH, 14% degree of hydrolysis) on frozen dough subjected to 0–6 freeze–thaw cycles, using trehalose as a reference. The results showed that the addition of SCMH in dough led to significant increase (p< 0.05) of the survival rates (79–93%) and relative fermentation capacity (59–95%) of yeast in dough subjected to 2–6 freeze–thaw cycles as compared to the control (survival rates: 43–59%; relative fermentation capacity: 29–58%) and trehalose (survival rates: 61–71%; relative fermentation capacity: 48–73%) treated counterparts. The presence of SCMH in dough also led to an increase in the specific volume of the resulting bread as well as decreases in the hardness, adhesiveness, and chewiness of bread, indicating that SCMH has a good cryoprotective effect on frozen dough. Further analysis found that the freezable water content of frozen dough with SCMH increased by only 5% after 6 freeze–thaw cycles, significantly less (p< 0.05) than the control (increased by 11%) and trehalose treated dough (increased by 9%). Accordingly, frozen dough that included SCMH showed less starch granule deformation and a more complete gluten structure, while the resulting bread was characterized by larger gas-cell density and smaller average cell area. These findings could be interpreted as the consequences of reduced formation and recrystallization of ice crystals due to decreased freezable water content, as well as the subsequent reduction in freeze-damage to the frozen dough.

This study was aimed to investigate effects of acetic acid (0.5%), ethanol (2%), and NaCl (2%) on the formation of protein-bound Nε-carboxymethyllysine (CML), Nε-carboxyethyllysine (CEL) and their precursors, glyoxal (GO) and methylglyoxal (MGO), in ground pork during commercial sterilization (121 °C, 10 min or 30 min). The commercial sterilization led to significant increases of CML and CEL in pork (raw: CML 2.91 ± 0.72, CEL 9.36 ± 3.22; 10 min heating: CML 15.30 ± 3.58, CEL 21.19 ± 5.70; 30 min heating: CML 40.05 ± 16.41, CEL 101.95 ± 42.24; all in mg/kg protein), regardless of the addition of an additive or not. However, all sterile pork (10 min heating) contained 5–31% less GO and 7–34% less MGO (except for the ethanol treated pork) as compared to their raw counterparts (GO: 2.48–4.65 mg/kg; MGO: 1.12–2.33 mg/kg). The addition of acetic acid led to an average of 33% more CML in sterile pork heated for 10 min in comparison with the control counterparts, but an average of 23% less CML and CEL in sterile pork heated for 30 min. The addition of ethanol or NaCl did not result in any obvious difference in the levels of CML and CEL in sterile pork. The amounts of CML and CEL had no direct relationship with the amounts of their precursors (GO and MGO) in sterile pork, which is possibly due to the high reactivity of the two α-dicarbonyl compounds that could rapidly react with other compounds like lysine to form more stable products (such as CML and CEL) during heating.

This study aimed to investigate the effects of pH (5.8, 7.0, and 10.0), linoleic acid, and reheating (heating: 180 °C, 30 min; reheating: 100 °C, 10 min) on the reactants, α-dicarbonyl compounds, and volatile compounds in the glucose-lysine model system, with a specific focus on exploring the influences of volatile compounds. The results showed that under three different pH glucose-lysine model system yielded the detection of 9, 15, and 29 volatile compounds, respectively. Notably, both the number and content of volatile compounds exhibited an upward trend as the pH increased. Reheating treatment led to changes in both the number and content of volatile compounds. The addition of linoleic acid affected the generation of heterocyclic compounds associated with the Maillard reaction. Volatile compounds such as aldehydes and alcohols were generated in systems containing linoleic acid, resulting in a higher number and content of volatile compounds compared to systems without linoleic acid.

Fish gelatin may have unpleasant odor that adversely affects its applications. Powdered activated carbon (PAC, 0.5–2.0% w/v), diatomaceous earth (DE, 0.1–0.5% w/v) and β-cyclodextrin (CD, 1.0–3.0% w/v) were used to purify tilapia (Oreochromis niloticus) skin gelatin at various absorption temperature and time (30–50 °C; 10–60 min). The use of all three materials at optimized conditions could lead to a significant improvement in the clarity of 6.67% (w/w) gelatin solutions: the transmittance was increased from 85 to 92–93% at 640 nm, and the color was changed from light yellow to colorless. There were 18 volatile compounds (11 aldehydes, 4 alcohols, and 3 other compounds) identified from the tilapia gelatin solution, and the number was reduced to 12 after the DE treatment (0.5%, 50 °C, 30 min) and to 9 after the PAC treatment (1.5%, 40 °C, 30 min). The amounts of all detectable compounds were greatly decreased after the PAC (decreased 12–89%) and β-CD treatments (decreased 35–96%), but less satisfied results were obtained with the DE treatments.

Dietary advanced glycation endproducts (AGEs), such as Nε-carboxymethyllysine (CML) and Nε-carboxyethyllysine (CEL), may promote aging and increase the risk for development of various chronic diseases. The effects of beef fat (8.3− 32.4 %), sodium nitrite (50 − 150 mg/kg) and sodium erythorbate (100 − 1000 mg/kg) contents on the formation of free and protein-bound AGEs including CML and CEL in ground beef during heating (100 °C, 0 − 60 min) were investigated. The proportion of free AGEs accounted for 1.1− 8.7 % of the sum of free and protein-bound AGEs, and decreased as the severity of the heat treatments increased. The amounts of free CML and CEL formed in ground beef (based upon protein weight) during heating were not affected by the levels of beef fat or sodium nitrite, but slightly decreased with the addition of 100 − 400 mg/kg erythorbate. The formation of protein-bound CML and CEL during heating was promoted with the addition of beef fat and erythorbate, but inhibited by nitrite. The overall effects of fat, nitrite and erythorbate on AGEs formation were relatively small compared to that of heat treatments. This study provides a greater understanding of the compositional factors involved in the formation of AGEs in complex foods during heating, which may help the optimization of thermal processes so that lower levels of AGEs are formed during food production operation.

A variety of anthropogenic organohalide contaminants generated from industry are released into the environment and thus cause serious pollution that endangers human health. In the present study, we investigated the microbial community composition of industrial saponification wastewater using 16S rRNA sequencing, providing genomic insights of potential organohalide dehalogenation bacteria (OHDBs) by metagenomic sequencing. We also explored yet-to-culture OHDBs involved in the microbial community. Microbial diversity analysis reveals that Proteobacteriaand Patescibacteriaphyla dominate microbiome abundance of the wastewater. In addition, a total of six bacterial groups (Rhizobiales, Rhodobacteraceae, Rhodospirillales, Flavobacteriales, Micrococcales, and Saccharimonadales) were found as biomarkers in the key organohalide removal module. Ninety-four metagenome-assembled genomes were reconstructed from the microbial community, and 105 hydrolytic dehalogenase genes within 42 metagenome-assembled genomes were identified, suggesting that the potential for organohalide hydrolytic dehalogenation is present in the microbial community. Subsequently, we characterized the organohalide dehalogenation of an isolated OHDB, Microbacteriumsp. J1-1, which shows the dehalogenation activities of chloropropanol, dichloropropanol, and epichlorohydrin. This study provides a community-integrated multi-omics approach to gain functional OHDBs for industrial organohalide dehalogenation.

The durability of reinforced concrete structures has always been an important problem in civil engineering because steel rebars rust easily. Therefore, fiber-reinforced polymer (FRP) rebars possessing good corrosion resistance, low weight, and easy construction has become a substitute for reinforcement. However, since FRP rebar has a low elastic modulus and is a brittle failure material, large deflections and cracks occur in the FRP concrete beam with no obvious warning before failure. A hybrid reinforced concrete beam that combines the advantages of steel rebar and FRP rebar is a good structural form. The reliability of hybrid reinforced beams must be analyzed to ensure their safety. A flexural performance test of the hybrid basalt FRP (BFRP)–steel-reinforced beam was performed, the failure mode was explored, and the numerical models were established. The accuracy of the models was verified by comparing them with the test results. The numerical models were used to establish a database (630 cases) that was combined with existing research results (33 cases), to obtain the statistics of the uncertainty of the prediction model. Reliability analysis of a large-scale design space was conducted to calibrate the BFRP. Finally, the average deviation from the target reliability index suggested that the values of the partial coefficient of the materials range from 1.2 to 1.4.

Deterioration of deep frying oils used by street vendors is one of the major food safety concerns. Fourier transform infrared (FTIR) spectroscopy coupled with partial least squares (PLS) regression was applied for rapid evaluation of the quality of deep frying oils collected from different street vendors (n= 109) using various frying processes in Shanghai. The levels of free fatty acids (FFA), total polar compounds (TPC), and viscosity of oils were determined with conventional methods and used as reference values for developing PLS models. The FFA (0.07–1.78 mg (KOH)/g) of all tested frying oils were below the maximum allowed value, while 5.5 % of oils had TPC (3.19–54.47 %) above the maximum allowed value (27 %) based upon the related chinese standards. FTIR coupled with PLS regression resulted in less satisfying results for FFA determination (predictability for soybean oils: R2= 0.709, SEP = 0.14, RPD = 1.83), but showed great promise for rapid determination of viscosity (model predictability: R2= 0.921–0.945, SEP = 0.68–0.71, RPD = 3.54–3.98) and TPC (predictability for soybean oils: R2= 0.790–0.931, SEP = 1.89–2.94, RPD = 2.16–3.55) of frying oils from different commercial settings. Developing separated PLS models for shortening and non-shortening oils improved predictability, particularly for the analysis of TPC.

There is a huge demand to identify the sex of unhatched fertilized eggs for laying industry and to understand the differences between male and female eggs as early as possible. Then the molecular mechanisms of sex determination and sex allocation in chicken were revealed. Therefore, TMT proteomic was applied to characterize the variation of molecular matrix between unhatched male and female egg yolks. A total of 411 proteins were identified and 35 differentially expressed proteins (DEPs), including 375332005, 015809562, 763550308 (upregulated, UPs) and 1337178851, 89000557, 89000581 (downregulated, DPs), etc. were confirmed between them. Gene ontology analyses showed that DEPs were mainly involved in response to stimulus, distributed in the extracellular region and participated in binding; KEGG analyses showed that few DPs were participated in cell growth and death, transport and catabolism, signaling molecules, interaction and were enriched in ubiquitin mediated proteolysis, endocytosis, ferroptosis, etc. metabolic pathways. Moreover, most of the DEPs and related metabolic pathways were associated with sex hormones. More importantly, this study supports maternal sex-allocation theory and extends our understanding of the molecular mechanism of sex determination and differentiation in avian. Which also provides a powerful evidence for ovo sexing of unhatched fertilized domestic chicken eggs by nondestructive approach and will be of great significance to eggs processing and production.

No relevant conflicts of interest to declare.

Objective:To study small interfering RNA (siRNA) targeting EZH2 gene effect on cell proliferation, apoptosis and histone modulation in HL-60 cells line.

Cotton plant is one of the most important economic crops in the world which supplies natural fiber for textile industry. The crucial traits of cotton fiber quality are fiber length and strength, which are mostly determined by the fiber elongation stage. Annexins are assumed to be involved in regulating fiber elongation, but direct evidences remain elusive. Recently, we have investigated the activities of fiber-specific expressed annexins AnGb5/6 and their interacted proteins in cotton. AnGb5 and 6 can interact reciprocally to generate a protein macro-raft in cell membrane. This macro-raft is probably a stabilized scaffold for Actin1 organization. The actin assembling direction and density are correlated with AnGb6gene expression and fiber expanding rate among three fiber length genotypes. These results suggest that annexins may act as the adaptor that linked fiber cell membrane to actin assembling. Due to the strong Ca2+and lipid binding ability of annexins, these results also indicate that annexins complex may function as an intermediate to receive Ca2+or lipid signals during fiber elongation.

Phenyhexyl isothiocyanate (PHI) has been shown to inhibit histone deacetylation in leukemia cell lines in vitro and in animal models. In this study we examined the histone acetylation status in leukemia cells from patients with acute leukemia as well as in non-leukemia cells. From January to September 2006, 19 patients with newly diagnosed acute leukemia were enrolled. 12 patients had AML, 7 had ALL. The age ranged between 2 months to 65 years old. There were 12 males and 7 females among them. 5 non- leukemia patients were also enrolled, with 1 anemia, 1 infection disease, 2 ITP, and 1 healthy volunteer. Mononuclear cells were isolated from patients’ specimen. Apoptosis of the cells and histone acetylation of H3 and H4 were characterized pre- and post- PHI treatment. The expression of acetylated H3 and H4 in acute leukemia cells was decreased, as compared to those in non-leukemia cells. Increased apoptosis was seen in the acute leukemia cells after PHI treatment of the primary culture of the leukemia cells for 3 and 7 hours. The effect of PHI on the leukemia cells from patients was time- and dose-dependent. There was a significant increase of PHI effect on leukemia cells versus non-leukemia cells (p<0.05), suggesting that PHI may have preferential activity against leukemia cells. This study presents the first evidence of PHI with preferential activity against leukemia cells over non-leukemia cells from human subjects. Clinical trial is warranted to study PHI in leukemia patients.

Phenyhexyl isothiocyanate (PHI) has been shown to inhibit histone deacetylation in leukemia cell lines in vitro and in animal models. In this study we examined the histone acetylation status in leukemia cells from patients with acute leukemia as well as in non-leukemia cells. From January to September 2006, 19 patients with newly diagnosed acute leukemia were enrolled. 12 patients had AML, 7 had ALL. The age ranged between 2 months to 65 years old. There were 12 males and 7 females among them. 5 non- leukemia patients were also enrolled, with 1 anemia, 1 infection disease, 2 ITP, and 1 healthy volunteer. Mononuclear cells were isolated from patients' specimen. Apoptosis of the cells and histone acetylation of H3 and H4 were characterized pre- and post- PHI treatment. The expression of acetylated H3 and H4 in acute leukemia cells was decreased, as compared to those in non-leukemia cells. Increased apoptosis was seen in the acute leukemia cells after PHI treatment of the primary culture of the leukemia cells for 3 and 7 hours. The effect of PHI on the leukemia cells from patients was time- and dose-dependent. There was a significant increase of PHI effect on leukemia cells versus non-leukemia cells (p<0.05), suggesting that PHI may have preferential activity against leukemia cells. This study presents the first evidence of PHI with preferential activity against leukemia cells over non-leukemia cells from human subjects. Clinical trial is warranted to study PHI in leukemia patients.

We have shown that phenyhexyl isothiocyanate(PHI) is a novel histone deacetylase inhibitor and can also modulate histone methylation. In this study we investigated the effect of PHI on human Acute Lymphoblastic leukemia cell line Molt-4 in vitro. The viability of Molt-4 cells was determined by MTT method. Apoptosis and cell cycle arrest were measured by flow cytometry. The expression of bcl-2, caspase-9, caspase-8, caspase-3, PRAP protein, acetylated H3 and H4, methylated H3K9 and H3K4 were detected by Western Blotting. The results showed that PHI inhibited the cell growth and decreased viability of Molt-4 cells. Cell cycle analysis indicated an arrest in G0/G1 phase. The expression of bcl-2, caspase-9, caspase-3, and PRAP was inhibited by PHI in a dose and time dependent manner. In contrast, there was no significant change in the expression of Caspase-8. PHI also significantly increased the level of acetylated histone H3 as well as H4. Interestingly, PHI increased the level of methylated H3k4, but decreased methylated H3K9. These data suggest that ALL cells are sensitive to the novel HDAC inhibitor, and PHI may become a novel agent in ALL therapy.

We have shown that phenyhexyl isothiocyanate(PHI) is a novel histone deacetylase inhibitor and can also modulate histone methylation. In this study we investigated the effect of PHI on human Acute Lymphoblastic leukemia cell line Molt-4 in vitro. The viability of Molt-4 cells was determined by MTT method. Apoptosis and cell cycle arrest were measured by flow cytometry. The expression of bcl-2, caspase-9, caspase-8, caspase-3, PRAP protein, acetylated H3 and H4, methylated H3K9 and H3K4 were detected by Western Blotting. The results showed that PHI inhibited the cell growth and decreased viability of Molt-4 cells. Cell cycle analysis indicated an arrest in G0/G1 phase. The expression of bcl-2, caspase-9, caspase-3, and PRAP was inhibited by PHI in a dose and time dependent manner. In contrast, there was no significant change in the expression of Caspase-8. PHI also significantly increased the level of acetylated histone H3 as well as H4. Interestingly, PHI increased the level of methylated H3k4, but decreased methylated H3K9. These data suggest that ALL cells are sensitive to the novel HDAC inhibitor, and PHI may become a novel agent in ALL therapy.