The potential for synthesized invasive plant biochar with hydroxyapatite to mitigate allelopathy of Solidago canadensis.

Few studies tried to explore the mitigation effect and underlying mechanisms of biochar and their complex for negative allelopathy from invasive plants, which may provide a new way in the invasive plant management. Herein, an invasive plant (Solidago canadensis)-derived biochar (IBC) and its composite with hydroxyapatite (HAP/IBC) were synthesized by high temperature pyrolysis, and characterized by scanning electron microscopy, energy dispersion spectrometer, X-ray diffraction, Fourier transform infrared spectroscopy, and X-ray photoelectron spectroscopy. Then, both the batch adsorption and pot experiments were conducted to compare the removal effects of kaempferol-3-O-ß-D-glucoside (C21 H20 O11 , kaempf), an allelochemical from S. canadensis, on IBC and HAP/IBC, respectively. HAP/IBC showed a stronger affinity for kaempf than IBC due to its higher specific surface area, more functional groups (P-O, P-O-P, PO4 3- ), stronger crystallization [Ca3 (PO4 )2 ]. The maximum kaempf adsorption capacity on HAP/IBC was six times higher than on IBC (10.482 mg/g > 1.709 mg/g) via π-π interactions, functional groups, and metal complexation. The kaempf adsorption process could be fitted best by both pseudo-second-order kinetic and Langmuir isotherm models. Furthermore, HAP/IBC addition into soils could enhance and even recover the germination rate and/or seedling growth of tomato inhibited by negative allelopathy from the invasive S. canadensis. These results indicate that the composite of HAP/IBC could more effectively mitigate the allelopathy from S. canadensis than IBC, which may be a potential efficient approach to control the invasive plant and improve invaded soils.

Gastrointestinal digestibility of micellar casein dispersions: Effects of caprine vs bovine origin, and partial colloidal calcium depletion using in vitro digestion models for the adults and elderly.

In vitro coagulation and digestion of caprine and bovine micellar casein concentrate (MCC) with or without partial colloidal calcium depletion (deCa) were studied under simulated adult and elderly conditions. Gastric clots were smaller and looser for caprine than bovine MCC, and were further looser with deCa and under elderly condition for both caprine and bovine MCC. Casein hydrolysis and concomitant formation of large peptides was faster for caprine than bovine MCC, and with deCa and under adult condition for caprine and bovine MCC. Formation of free amino groups and small peptides were faster for caprine MCC, and with deCa and under adult condition. Upon intestinal digestion, proteolysis occurred rapidly, and was faster under adult condition, but showed less differences with increasing digestion between caprine and bovine MCC, and with and without deCa. These results suggested weakened coagulation and greater digestibility for caprine MCC and MCC with deCa under both conditions.

Glutamate receptor-interacting protein 1 in D1- and D2-dopamine receptor-expressing medium spiny neurons differentially regulates cocaine acquisition, reinstatement, and associated spine plasticity.

Background: The nucleus accumbens (NAc) is involved in the expression of cocaine addictive phenotypes, including acquisition, extinction, and reinstatement. In the NAc, D1-medium spiny neurons (MSNs) encode cocaine reward, whereas D2-MSNs encode aversive responses in drug addiction. Glutamate receptor-interacting protein 1 (GRIP1) is known to be associated with cocaine addiction, but the role of GRIP1 in D1-MSNs and D2-MSNs of the NAc in cocaine acquisition and reinstatement remains unknown. Methods: A conditioned place preference apparatus was used to establish cocaine acquisition, extinction, and reinstatement in mouse models. GRIP1 expression was evaluated using Western blotting. Furthermore, GRIP1-siRNA and GRIP1 overexpression lentivirus were used to interfere with GRIP1 in the NAc. After the behavioral test, green fluorescent protein immunostaining of brain slices was used to detect spine density. Results: GRIP1 expression decreased during cocaine acquisition and reinstatement. GRIP1-siRNA enhanced cocaine-induced CPP behavior in acquisition and reinstatement and regulated associated spine plasticity. Importantly, the decreased GRIP1 expression that mediated cocaine acquisition and reinstatement was mainly driven by the interference of the GRIP1-GluA2 interaction in D1-MSNs and could be blocked by the interference of the GRIP1-GluA2 interaction in D2-MSNs. Interference with the GRIP1-GluA2 interaction in D1- and D2-MSNs decreased spine density in D1- and D2-MSNs, respectively. Conclusion: GRIP1 in D1- and D2-MSNs of the NAc differentially modulates cocaine acquisition and reinstatement. GRIP1 downregulation in D1-MSNs has a positive effect on cocaine acquisition and reinstatement, while GRIP1 downregulation in D2-MSNs has a negative effect. Additionally, GRIP1 downregulation in D1-MSNs plays a leading role in cocaine acquisition and reinstatement.

Characterization of metabolic pathways for biosynthesis of the flavor compound 3-methylbutanal by Lactococcus lactis.

3-Methylbutanal is a key volatile compound that imparts a nutty flavor to Cheddar cheese. Lactococcus lactis has been successfully applied as a starter to increase the level of 3-methylbutanal produced during the ripening of cheese. However, the mechanism of action and genetic diversity of this bacterium for 3-methylbutanal biosynthesis remains unclear. In this study, we investigated the association between the L. lactis genotype and phenotype in the biosynthesis of 3-methylbutanal via both direct and indirect pathways. Fourteen strains of L. lactis were screened for the capacity to produce 3-methylbutanal, and strain 408 (>140 µM) produced the highest among all tested strains, which exhibited both α-keto acid decarboxylase and α-ketoacid dehydrogenase activities. Furthermore, the results of a sodium meta-arsenite inhibition experiment showed that the 3-methylbutanal-producing capacities of each strain declined to various degrees. The kdcA gene, which encodes the direct pathway component α-ketoacid decarboxylase, was detected in 4 of the 14 strains, of which only strain 408 contained the full-length gene. We then characterized the genes associated with the indirect pathway by detecting the expression levels of the pdh gene cluster, ack, and pta, which were expressed at relatively higher levels in a high-yield strain than in a low-yield strain. As a result, these L. lactis strains were divided into 3 categories according to gene diversity, gene expression, and 3-methylbutanal production. The results of this study refine our knowledge of the genetic determinants of 3-methylbutanal biosynthesis in L. lactis and explain the effect of both synthesis pathways on 3-methylbutanal production.

Anatomic Measurement and Variability Analysis of the Anterior Talofibular Ligament and Calcaneofibular Ligament of the Ankle.

BACKGROUND: The anterior talofibular ligament (ATFL) and calcaneofibular ligament (CFL) contribute greatly to the overall stability of the ankle joint; however, ATFL and combined ATFL-CFL sprains are common. Anatomic reconstruction of the lateral collateral ligament with grafts has been proposed for patients with poor tissue quality or inadequate local tissue. Anatomic reconstruction of the lateral ankle ligaments requires a good understanding of their anatomic location. PURPOSE: To describe the anatomy of the ATFL and CFL ligaments quantitatively and qualitatively and explore the relationship of some morphological parameters. STUDY DESIGN: Descriptive laboratory study. METHODS: A total of 66 adult ankle specimens were analyzed for ATFL band type, origin, length, width, thickness, and angle between the ATFL and CFL, and 73 adult ankle specimens were used for measuring the origin of the CFL. The coefficient of variation was used to describe and compare the respective variability of angle, length, width, and thickness. The origin of the ATFL was labeled as point A, and the leading edge of the CFL intersection with the articular surface of the calcaneus was considered point B. RESULTS: The ATFL had a variable number of bands. A high degree of variability (coefficient of variation >0.2) was seen for most morphological measurements of the ATFL. In addition, the length of distance AB also varied. The CFL originated at the tip of the fibula in only 9% of specimens. It was found more commonly at the anterior border of the lateral malleolus (4.94 ± 1.70 mm from the tip). The angle between the ATFL and CFL was consistent at 100° to 105º. CONCLUSION: A fair amount of variability of ATFL length, width, and thickness were found in our study, with less variability in the ATFL-CFL angle. Most CFLs attached anterior to the tip of the fibula. CLINICAL RELEVANCE: Providing relevant anatomic data of ATFL and CFL is important in ensuring proper surgical treatment of ankle joint injuries.

Evaluation of the Perceptual Interactions among Aldehydes in a Cheddar Cheese Matrix According to Odor Threshold and Aroma Intensity.

To evaluate the contributions of 3-methylbutanal, 2-methylbutanal, 2-methylpropanal, and benzaldehyde in cheddar cheese models, the threshold values, optimal concentration ranges, and perceptual actions of these compounds were determined at various concentrations. The thresholds for 3-methylbutanal, 2-methylbutanal, 2-methylpropanal, and benzaldehyde in the cheese matrix were 150.31, 175.39, 150.66, and 500.21 µg/kg, respectively, which were significantly higher than the corresponding values in water. The optimal concentration ranges of these aldehydes were determined as 150-300, 175-325, 150-350, and 500-1500 µg/kg, respectively. Based on the results of the threshold method and Feller's model, five binary mixtures were found to have synergistic effects, and only the pair of 2-methylpropanal and benzaldehyde was determined to have a masking effect. In addition, the synergistic olfactory effects between the four ternary mixtures and the quaternary mixture of these aldehydes were also assesSsed using Feller's model. In a σ-τ plot analysis, synergism was usually observed when these odor pairs were at their threshold levels. In summary, the results suggested that perceptual interactions among these aldehydes exist in a cheese model variably with different concentrations and threshold ratios. This study will be helpful to a further understanding of the nutty aroma and improving the aroma quality of cheddar cheese.

Functional identification and expressional responses of large yellow croaker (Larimichthys crocea) interleukin-8 and its receptor.

Interleukin-8 (IL-8 or chemokine (C-X-C motif) ligand 8, CXCL8) is a chemokine produced by multiple cell types. It promotes chemotaxis and phagocytosis via interaction with chemokine receptors CXCR1 and CXCR2. Using published data, IL-8 gene (LcIL-8) of the large yellow croaker (Larimichthys crocea) was cloned into the pcDNA3.1 plasmid, and an interleukin-8 receptor (LcCXCR2) was cloned into the pEGFP-N1 plasmid. Secratory expression of LcIL-8 in HEK293T cells was carried out, and product in culture medium was collected for LcCXCR2 stimulation in HEK293 cells. Following receptor internalization observation and intracellular signaling detection, the functional interaction of LcIL-8 and LcCXCR2 was further determined and the ERK phosphorylation signal activation mediated by LcCXCR2 was demonstrated. Quantitative real-time PCR analysis was used to analyze transcription level regulation of LcIL-8 and LcCXCR2 in various tissues of large yellow croaker. Expression of LcIL-8 and LcCXCR2 was elevated in the spleen, head kidney, and liver after Vibrio parahemolyticus challenge. Results illustrated the functional interaction between LcIL-8 and LcCXCR2 in mediating intracellular ERK1/2 phosphorylation signaling and suggested that the LcIL-8 and LcCXCR2 system is part of the immune response induced by V. Parahemolyticus in L. crocea.

Construction and application of the vectors to identify genes encoding exported proteins of Escherichia coli.

In order to clone genes having signal sequences of Escherichia coli, four vectors with or without Lac or Ara promoter were constructed using a leaderless ß-lactamase as reporter. Fragments of tetracycline resistance gene (Tet) with or without promoter were used to confirm the vectors' ability to clone and report signal sequences. The minimum inhibitory concentration of ampicillin of the transformants was measured to detect the expression and secretion efficiency of the vectors. The results showed that the ß-lactamase could be co-expressed and secreted with Tet protein. The Lac or Ara promoter in the vectors could be regulated by different inducers, and the Ara promoter showed higher regulative efficiency than the Lac. The best induction dose of L-arabinose for the Ara promoter is 1.25 %. All the four vectors were stably maintained in host after being inoculated for 20 passages in antibiotics-free media. Genomic library of an avian pathogenic strain, E. coli O2, was constructed using the pMB-Ara-T vector we developed. 318 clones were obtained from the genomic library of E. coli strain O2, and the inserts in these clones represented 276 genes based on sequence analysis. Among the 276 cloned fragments, only 128 had complete promoter sequence. For the 128 fragments with promoter, only 27 could be expressed under LB culture condition without inducer, the other 101 were only expressed under induction. The results showed our constructed vectors could efficiently capture all kinds of exported protein genes in vitro, including the ones without promoter or with inactive promoter.