Investigation of the Seroprevalence of Brucella Antibodies and Characterization of Field Strains in Immunized Dairy Cows by B. abortus A19.

(1) Background: One method of eradicating brucellosis is to cull cattle that test positive for antibodies 12 months after being vaccinated with the 19-strain vaccine. Variations in immunization regimens and feeding practices may contribute to differences in the rate of persistent antibodies. We conducted this study to investigate the real positive rate of Brucella antibody in field strains of Brucella spp. after immunization over 12 months in dairy cows. This research aims to provide data to support the development of strategies for preventing, controlling, and eradicating brucellosis. (2) Method: We employed the baseline sampling method to collect samples from cows immunized with the A19 vaccine for over 12 months in Lingwu City from 2021 to 2023. Serological detection was conducted using the RBPT method. An established PCR method that could distinguish between 19 and non-19 strains of Brucella was utilized to investigate the field strains of Brucella on 10 dairy farms based on six samples mixed into one using the Mathematical Expectation strategy. (3) Results: We analyzed the rates of individual seropositivity and herd seropositive rates in dairy cattle in Lingwu City from 2021 to 2023 and revealed that antibodies induced by the Brucella abortus strain A19 vaccine persist in dairy herds for more than 12 months. We established a PCR method for identifying both Brucella A19 and non-A19 strains, resulting in the detection of 10 field strains of Brucella abortus from 1537 dairy cows. By employing a Mathematical Expectation strategy, we completed testing of 1537 samples after conducting only 306 tests, thereby reducing the workload by 80.1%. (4) Conclusions: There was a certain proportion of cows with a persistent antibody titer, but there was no evidence that all of these cattle were naturally infected with Brucella. The established PCR method for distinguishing between Brucella abortus strain 19 and non-19 strains can be specifically utilized for detecting natural Brucella infection in immunized cattle. We propose that relying solely on the detection of antibodies in cattle immunized with the A19 vaccine more than 12 months previously should not be solely relied upon as a diagnostic basis for brucellosis, and it is essential to complement this approach with PCR analysis to specifically identify field Brucella spp. Brucella abortus was the predominant strain identified in the field during this study. Detection based on the Mathematical Expectation strategy can significantly enhance detection efficiency.

Iron-catalyzed C-7 selective NH2 amination of indoles.

7-Aminoindoles are important synthetic intermediates to a broad range of bioactive molecules. Transition metal-catalyzed directed C-H amination is among the most straightforward route for their synthesis, whereas methods that could directly incorporate an NH2 group in a highly selective manner remains elusive. Moreover, there is still high demand for the development of earth-abundant metal catalysis for such attractive reactivity. We present here the first C-7 selective NH2 amination of indoles through a directed homolytic aromatic substitution (HAS) with iron-aminyl radical. The reaction exhibits broad substrate scope, tolerates variety of functional groups, and is readily scalable with catalyst loading down to 0.1 mol% and turnover number (TON) up to 4500.

Potentially suitable habitats of Daodi goji berry in China under climate change.

Introduction: Goji berry (Lycium barbarum L.) is a famous edible and medicinal herb worldwide with considerable consumption. The recent cultivation of goji berries in the Daodi region was seriously reduced due to increased production costs and the influence of policy on preventing nongrain use of arable land in China. Consequently, production of Daodi goji berry was insufficient to meet market demands for high-quality medicinal materials. Searching for regions similar to the Daodi region was necessary. Methods: The MaxEnt model was used to predicted the current and future potential regions suitable for goji berry in China based on the environmental characteristics of the Daodi region (including Zhongning County of Zhongwei prefecture-level city, and its surroundings), and the ArcGIS software was used to analyze the changes in its suitable region. Results: The results showed that when the parameters were FC = LQHP and RM = 2.1, the MaxEnt model was optimal, and the AUC and TSS values were greater than 0.90. The mean temperature and precipitation of the coldest quarter were the most critical variables shaping the distribution of Daodi goji berries. Under current climate conditions, the suitable habitats of the Daodi goji berry were 45,973.88 km2, accounting for 0.48% of China's land area, which were concentrated in the central and western Ningxia Province (22,589.42 km2), and the central region of Gansu Province (18,787.07 km2) bordering western Ningxia. Under future climate scenarios, the suitable area was higher than that under current climate conditions and reached the maximum under RCP 6.0 (91,256.42 km2) in the 2050s and RCP 8.5 (82,459.17 km2) in the 2070s. The expansion regions were mainly distributed in the northeast of the current suitable ranges, and the distributional centroids were mainly shifted to the northeast. The moderately and highly suitable overlapping habitats were mainly distributed in Baiyin (7,241.75 km2), Zhongwei (6,757.81 km2), and Wuzhong (5, 236.87 km2) prefecture-level cities. Discussion: In this stduy, MaxEnt and ArcGIS were applied to predict and analyze the suitable habitats of Daodi goji berry in China under climate change. Our results indicate that climate warming is conducive to cultivating Daodi goji berry and will not cause a shift in the Daodi region. The goji berry produced in Baiyin could be used to satisfy the demand for high-quality medicinal materials. This study addresses the insufficient supply and guides the cultivation of Daodi goji berry.

Hydroamination of Unactivated Alkenes with Aliphatic Azides.

We report here an efficient and highly diastereoselective intermolecular anti-Markovnikov hydroamination of unactivated alkenes with aliphatic azides in the presence of silane. The system tolerates a wide range of azides and alkenes and operates with alkene as limiting reagent. Mechanistic studies suggest a radical chain pathway that involves aminium radical formation, radical addition to alkenes and HAT from silane to ß-aminium alkyl radical. The use of sterically bulky silane is proposed to contribute to the excellent diastereoselectivity for HAT. Computational analysis uncovers the reaction pathway of aliphatic azide activation with silyl radical for aminyl radical formation.

An NHC-catalyzed, stereoselective α-functionalization of α,ß-unsaturated carboxylic acids through in situ activation.

An N-heterocyclic carbene (NHC)-catalyzed α-functionalization of the in situ activated α,ß-unsaturated carboxylic acids bearing γ-H was realized through formal [4 + 2] annulations with o-quinone methides, which paved a new avenue for the assembly and modification of the dihydrocoumarin scaffold in good yields with excellent diastereo- and enantioselectivities.

NHC-Catalyzed Aldol-Like Reactions of Allenoates with Isatins: Regiospecific Syntheses of γ-Functionalized Allenoates.

An N-heterocyclic carbene (NHC) catalyzed γ-specific aldol-like reaction between allenoates and isatins has been achieved under mild conditions, giving trisubstituted allene derivatives bearing isatin moiety in moderate to good yields with high diastereoselectivity and excellent atom efficiency. The DFT computations indicated that the formation of the γ-adduct was more energetically favorable than that of the α-adduct. The result reported herein opens a new route for NHC-promoted allenoate-involved reaction.

Investigation of Chinese Wolfberry (Lycium spp.) Germplasm by Restriction Site-Associated DNA Sequencing (RAD-seq).

Chinese wolfberry (Lycium spp.) is an important edible and medicinal plant, with a long cultivation history. The genetic relationships among wild Lycium species and landraces have been unclear for a number of reasons, which has hindered the breeding of modern Chinese wolfberry cultivars. In this study, we collected 19 accessions of Chinese wolfberry germplasm, and constructed the genetic relationship based on RAD-seq markers. We obtained 30.32 Gb of clean data, with the average value of each sample being 1.596 Gb. The average mapping rate was 85.7%, and the average coverage depth was 6.76 X. The phylogeny results distinguished all accessions clearly. All the studied landraces shared their most recent common ancestor with L. barbarum, which indicated that L. barbarum may be involved in cultivation of these landraces. The relationship of some landraces, namely the 'Ningqi' series, 'Qingqi-1' and 'Mengqi-1,' has been supported by the phylogeny results, while the triploid wolfberry was shown to be based on a hybrid between 'Ningqi-1' and a tetraploid wolfberry. This study uncovered the genetic background of Chinese wolfberry, and developed the foundation for species classification, accession identification and protection, and the production of hybrid cultivars of wolfberry.

NHC-Catalyzed Hetero-Diels-Alder Reaction of Allenoate with Chalcone: Synthesis of Polysubstituted Pyranyl Carboxylate.

An NHC-catalyzed hetero-Diels-Alder and isomerization process of chalcones with allenoates was discovered, which furnished highly functionalized multisubstituted pyranyl carboxylates successfully. This method features a convergent assembly, mild reaction conditions, moderate to good yields, and high atom economy.

An integrated metabonomics and transcriptomics approach to understanding metabolic pathway disturbance induced by perfluorooctanoic acid.

Perfluorooctanoic acid (PFOA) is one of the most representative perfluorinated compounds and liver is the major organ where PFOA is accumulated. Although the multiple toxicities had been reported, its toxicological profile remained unclear. In this study, a systems toxicology strategy integrating liquid chromatography/mass spectrometry-based metabonomics and transcriptomics analyses was applied for the first time to investigate the effects of PFOA on a representative Chinese normal human liver cell line L-02, with focusing on the metabolic disturbance. Fifteen potential biomarkers were identified on metabolic level and most observations were consistent with the altered levels of gene expression. Our results showed that PFOA induced the perturbations in various metabolic processes in L-02 cells, especially lipid metabolism-related pathways. The up-stream mitochondrial carnitine metabolism was proved to be influenced by PFOA treatment. The specific transformation from carnitine to acylcarnitines, which showed a dose-dependent effect, and the expression level of key genes involved in this pathway were observed to be altered correspondingly. Furthermore, the down-stream cholesterol biosynthesis was directly confirmed to be up-regulated by both increased cholesterol content and elevated expression level of key genes. The PFOA-induced lipid metabolism-related effects in L-02 cells started from the fatty acid catabolism in cytosol, fluctuated to the processes in mitochondria, extended to the cholesterol biosynthesis. Many other metabolic pathways like amino acid metabolism and tricarboxylic acid cycle might also be disturbed. The findings obtained from the systems biological research provide more details about metabolic disorders induced by PFOA in human liver.

Perfluorooctanoic acid induces gene promoter hypermethylation of glutathione-S-transferase Pi in human liver L02 cells.

Perfluorooctanoic acid (PFOA) is one of the most commonly used perfluorinated compounds. Being a persistent environmental pollutant, it can accumulate in human tissues via various exposure routes. PFOA may interfere in a toxic fashion on the immune system, liver, development, and endocrine systems. In utero human exposure had been associated with cord serum global DNA hypomethylation. In light of this, we investigated possible PFOA-induced DNA methylation alterations in L02 cells in order to shed light into its epigenetic-mediated mechanisms of toxicity in human liver. L02 cells were exposed to 5, 10, 25, 50 or 100 mg/L PFOA for 72h. Global DNA methylation levels were determined by LC/ESI-MS, glutathione-S-transferase Pi (GSTP) gene promoter DNA methylation was investigated by methylation-specific polymerase chain reaction (PCR) with bisulfite sequencing, and consequent mRNA expression levels were measured with quantitative real-time reverse transcriptase PCR. A dose-related increase of GSTP promoter methylation at the transcription factor specificity protein 1 (SP1) binding site was observed. However, PFOA did not significantly influence global DNA methylation; nor did it markedly alter the promoter gene methylation of p16 (cyclin-dependent kinase inhibitor 2A), ERα (estrogen receptor α) or PRB (progesterone receptor B). In addition, PFOA significantly elevated mRNA transcript levels of DNMT3A (which mediates de novo DNA methylation), Acox (lipid metabolism) and p16 (cell apoptosis). Considering the role of GSTP in detoxification, aberrant methylation may be pivotal in PFOA-mediated toxicity response via the inhibition of SP1 binding to GSTP promoter.

[Effects of NaCl stress on Hippophae rhamnoides and Shepherdia argentea seedlings growth and photosynthetic characteristics].

With two-year old seedlings of Hippophea rhamnoides and Shepherdia argentea as test materials, this paper studied their growth and photosynthetic characteristics under the stress of different concentration (0, 200, 400 and 600 mmol x L(-1)) NaCl. The results showed that the biomass and total leaf area per plant of H. rhamnoides and S. argentea seedlings decreased significantly with increasing NaCl concentration. Comparing with the control, the root/shoot ratio of H. rhamnoides and S. argentea seedlings under NaCl stress increased obviously, while the leaf mass per area (LMA) decreased slightly. When the NaCl concentration increased and the stress time prolonged, the net photosynthetic rate (P(n)), transpiration rate (T(r)), and stomatal conductance (G(s)) of H. rhamnoides and S. argentea seedlings declined markedly, the intercellular CO2 concentration (C(i)) increased after an initial decrease, whereas the water use efficiency (WUE) and stomatal limiting value (L(s)) decreased after an initial increase. The dynamic changes of G(s), C(i) and L(s) indicated that the decline of P(n) was mainly caused by the stomatal limitation in a short-term stress, and by non-stomatal limitation in a long-term stress. The poorer the salt tolerance of tree species and the higher the NaCl concentration, the earlier the transition from stomatal limitation to non-stomatal limitation would occur. As for H. rhamnoides, its morphological symptoms of salt injury appeared on the 10th day, and all of its seedlings were died on the 22th day under 600 mmol NaCl x L(-1) stress. In contrast, S. argentea could tolerate 600 mmol NaCl x L(-1) stress for above 30 days, illustrating that S. argentea, as an introduced tree species, had higher salt tolerance than H. rhamnoides, and could be planted widely in saline regions of China.