Controlling Solvent Polarity to Regulate Protein Self-Assembly Morphology and Its Universal Insight for Fibrillation Mechanism.

The mechanism of ethanol-induced fibrillation of ß-lactoglobulin (ß-lg) in the acidic aqueous solution upon heating was investigated using various techniques, mainly thioflavin T fluorescence, atomic force microscopy, nonreducing electrophoresis, mass spectrometry, Fourier transform infrared spectroscopy, and circular dichroism spectroscopy. The results showed that fibrillation occurred with a heating time increase, but high ethanol content slowed down the process. At a low ethanol volume fraction, peptides existed after heating for 2 h, with long and straight fibrils formed after 4-6 h, while at a high ethanol volume fraction, the proteins aggregated with very few peptides appeared at the early stage of heating, and short and curved fibrils formed after heating for 8 h. Ethanol weakened the hydrophobic interactions between proteins in the aqueous solution; therefore the latter could not completely balance the electrostatic repulsion, and thus suppressing the fibrillation process. It is believed that the fibrillation of ß-lg in the acidic solution upon heating is mainly dominated by the polypeptide model; however, ethanol inhibited the hydrolysis of proteins, and the self-assembly mechanism changed to the monomer model.

Investigation of the Effect of Rice Bran Content on the Antioxidant Capacity and Related Molecular Conformations of Plant-Based Simulated Meat Based on Raman Spectroscopy.

At present, plant-based simulated meat is attracting more and more attention as a meat substitute. This study discusses the possibility of partial substitution of rice bran (RB) for soybean protein isolate (SPI) in preparing plant-based simulated meat. RB was added to SPI at 0%, 5%, 10%, 15%, and 20% to prepare RB-SPI plant-based simulated meat by the high moisture extrusion technique. RB-SPI plant-based simulated meat revealed greater polyphenol content and preferable antioxidant capacity (DPPH radical scavenging capacity, ABTS scavenging ability, and FRAP antioxidant capacity) compared to SPI plant-based simulated meat. The aromatic amino acids (tryptophan and tyrosine) of RB-SPI plant-based simulated meats tend to be masked first, and then the hydrophobic groups are exposed as RB content increases and the polarity of the surrounding environment increases due to the change in the disulfide conformation of RB-SPI plant-based simulated meats from a stable gauche-gauche-gauche conformation to a trans-gauche-trans conformation.

The MttB superfamily member MtyB from the human gut symbiont Eubacterium limosum is a cobalamin-dependent γ-butyrobetaine methyltransferase.

The production of trimethylamine (TMA) from quaternary amines such as l-carnitine or γ-butyrobetaine (4-(trimethylammonio)butanoate) by gut microbial enzymes has been linked to heart disease. This has led to interest in enzymes of the gut microbiome that might ameliorate net TMA production, such as members of the MttB superfamily of proteins, which can demethylate TMA (e.g., MttB) or l-carnitine (e.g., MtcB). Here, we show that the human gut acetogen Eubacterium limosum demethylates γ-butyrobetaine and produces MtyB, a previously uncharacterized MttB superfamily member catalyzing the demethylation of γ-butyrobetaine. Proteomic analyses of E. limosum grown on either γ-butyrobetaine or dl-lactate were employed to identify candidate proteins underlying catabolic demethylation of the growth substrate. Three proteins were significantly elevated in abundance in γ-butyrobetaine-grown cells: MtyB, MtqC (a corrinoid-binding protein), and MtqA (a corrinoid:tetrahydrofolate methyltransferase). Together, these proteins act as a γ-butyrobetaine:tetrahydrofolate methyltransferase system, forming a key intermediate of acetogenesis. Recombinant MtyB acts as a γ-butyrobetaine:MtqC methyltransferase but cannot methylate free cobalamin cofactor. MtyB is very similar to MtcB, the carnitine methyltransferase, but neither was detectable in cells grown on carnitine nor was detectable in cells grown with γ-butyrobetaine. Both quaternary amines are substrates for either enzyme, but kinetic analysis revealed that, in comparison to MtcB, MtyB has a lower apparent Km for γ-butyrobetaine and higher apparent Vmax, providing a rationale for MtyB abundance in γ-butyrobetaine-grown cells. As TMA is readily produced from γ-butyrobetaine, organisms with MtyB-like proteins may provide a means to lower levels of TMA and proatherogenic TMA-N-oxide via precursor competition.

Diffusion tensor imaging of breast lesions: evaluation of apparent diffusion coefficient and fractional anisotropy and tissue cellularity.

OBJECTIVE: To investigate the apparent diffusion coefficient (ADC) and fractional anisotropy (FA) measured by diffusion tensor imaging (DTI), tissue cellularity and their relationship in breast malignant/benign lesions. METHODS: 88 patients with 88 breast lesions who underwent DTI and dynamic contrast-enhanced MR scanning between November 2013 and December 2014 were retrospectively analyzed. The diagnosis was confirmed pathologically. ADC and FA values as well as histopathological cellularity of different pathological types of lesions were analyzed and compared statistically. The Pearson's correlation between cellularity and ADC and FA was calculated. RESULTS: There were 59 cases of breast cancer and 29 cases of benign lesions included in the study. ADC values of breast cancers were statistically lower than that of benign lesions (p < 0.001). FA and cellularity were higher in cancers than in benign lesions with statistical significance (p < 0.05 and p < 0.001, respectively). The mean FA values in the patients with invasive ductal carcinoma (IDC) were higher than that in the patients with ductal carcinoma in situ (DCIS) without statistical difference (p > 0.05). The ADC and the cellularity in the IDC of grade III were statistically lower (p < 0.05) and higher (p < 0.05) than that in the DCIS and IDC of grade I-II, respectively. ADC was negatively correlated to cellularity (r = -0.8319, p < 0.001) and FA was positively correlated to cellularity (r = 0.4231, p < 0.001). CONCLUSION: ADC and FA values were statistically different between benign and malignant breast lesions and were significantly correlated to tissue cellularity. ADC and FA may help to discriminate malignant from benign breast lesions and to predict cellularity. ADC is helpful in the prediction of the grade of breast cancer. ADVANCES IN KNOWLEDGE: ADC and FA values were statistically different between benign and malignant breast lesions and were significantly correlated to tissue cellularity.

Assessing Detection, Discrimination, and Risk of Breast Cancer According to Anisotropy Parameters of Diffusion Tensor Imaging.

BACKGROUND The aim of this study was to investigate whether the anisotropy parameters are helpful in the detection and discrimination of breast cancers, and to determine its value in predicting the risk of cancers. MATERIAL AND METHODS There were 56 patients with 56 lesions (34 malignant, 22 benign) included in the study. DTI was performed in every patient and apparent diffusion coefficient (ADC), fractional anisotropy (FA), and eigenvalues E1, E2, and E3 were measured in every lesion and the normal breast tissue. RESULTS ADC, FA, and eigenvalues of E1, E2, E3, and E1-E3 in breast cancers were all significantly lower than in normal tissue (P<0.001 for all) with mean reduction of (32 ± 17)%, (24 ± 13)%, (33 ± 19)%, (32 ± 17)%, (31 ± 18)%, and (37 ± 20)% for ADC, FA, E1, E2, E3, and E1-E3, respectively. These parameters were also statistically lower in cancers than in benign lesions (P<0.01 for all), except FA (P>0.05). ADC, E1, E2, and E3 were very similar in discriminating breast cancers and benign lesions, with area under the curve (AUC) 0.885-0.898, sensitivity 73.5-85.3%, and specificity 90.9-100%. CONCLUSIONS ADC, E1, E2, E3, and E1-E3 are much lower in breast cancers than in normal tissue and benign lesions. The reduction of ADC, E1, E2, E3, and E1-E3 of a mass in the breast is highly associated with the risk of breast cancer, but the FA has no utility in breast cancer risk prediction.

Growth arrest and apoptosis of human hepatocellular carcinoma cells induced by hexamethylene bisacetamide.

AIM: To investigate the cellular effects of hybrid polar compound hexamethylene bisacetamide (HMBA) on the growth and apoptosis of human hepatocellular carcinoma cells and to provide the molecular mechanism for potential application of HMBA in the treatment of liver cancer. METHODS: Effects of HMBA on the growth of human hepatocellular carcinoma SMMC-7721 cells were assayed by MTT chronometry. Apoptosis induced by HMBA was detected by phase-contrast microscopy, flow cytometry, propidium iodide staining and immunocytochemical analysis. RESULTS: The growth of SMMC-7721 cells was significantly inhibited by HMBA, and the growth inhibitory rate was 51.1%, 62.6%, 68.7% and 73.9% respectively after treatment with 5.0, 7.5, 10.0 and 12.5 mmol/L of HMBA. In the cells treated with 10 mmol/L of HMBA for 72 h, the population of cells at sub-G(1) phase significantly increased, and the apoptotic bodies and condensed nuclei were detected. Moreover, treatment of SMMC-7721 cells with 10 mmol/L of HMBA down-regulated the expression of Bcl-2 anti-apoptotic protein, while slightly up-regulated the level of pro-apoptotic protein Bax. CONCLUSION: Treatment with 10.0 mmol/L of HMBA can significantly inhibit the growth and induce apoptosis of human hepatocellular carcinoma SMMC-7721 cells by decreasing the ratio of Bcl-2 to Bax.