ABSTRACT
Calosoma maximoviczi, a predatory pest beetle, poses a significant threat to wild silk farm production due to its predation on wild silkworms. Given the coexistence of this species with beneficial silkworms in the farm orchards, chemical pesticides are not an ideal solution for controlling its population. In this study, we employed a comprehensive multi-target RNA interference (RNAi) approach to disrupt the olfactory perception of C. maximoviczi through independently silencing 16 odorant receptors (ORs) in the respective genders. Specifically, gene-specific siRNAs were designed to target a panel of ORs, allowing us to investigate the specific interactions between odorant receptors and ligands within this species. Our investigation led to identifying four candidate siOR groups that effectively disrupted the beetle's olfactory tracking of various odorant ligands associated with different trophic levels. Furthermore, we observed sex-specific differences in innate RNAi responses reflected by subsequent gene expression, physiological and behavioral consequences, underscoring the complexity of olfactory signaling and emphasizing the significance of considering species/sex-specific traits when implementing pest control measures. These findings advance our understanding of olfactory coding patterns in C. maximoviczi beetles and establish a foundation for future research in the field of pest management strategies.
Subject(s)
Coleoptera , Receptors, Odorant , Animals , Female , Male , Coleoptera/metabolism , RNA, Small Interfering/genetics , RNA, Small Interfering/metabolism , Receptors, Odorant/genetics , Receptors, Odorant/metabolism , Predatory Behavior , Smell/genetics , LigandsABSTRACT
BACKGROUND: The black carabid beetle Calosoma maximoviczi is a successful predator that serves as both a beneficial insect and a severe threat to economic herbivores. Its hunting technique relies heavily on olfaction, but the underlying mechanism has not been studied. Here, we report the electrophysiological, ecological and molecular traits of bioactive components identified from a comprehensive panel of natural odorants in the beetle-prey-plant system. The aim of this work was to investigate olfactory perceptions and their influence on the behaviours of C. maximoviczi. RESULTS: Among the 200 identified volatiles, 18 were concentrated in beetle and prey samples, and 14 were concentrated in plants. Insect feeding damage to plants led to a shift in the emission fingerprint. Twelve volatiles were selected using successive electrophysiological tests. Field trials showed that significant sex differences existed when trapping with a single chemical or chemical mixture. Expression profiles indicated that sex-biased catches were related to the expression of 15 annotated CmaxOBPs and 40 CmaxORs across 12 chemosensory organs. In silico evaluations were conducted with 16 CmaxORs using modelling and docking. Better recognition was predicted for the pairs CmaxOR5-(Z)-3-hexenyl acetate, CmaxOR6-ß-caryophyllene, CmaxOR18-(E)-ß-ocimene and CmaxOR18-tetradecane, with higher binding affinity and a suitable binding pocket. Lastly, 168Y in CmaxOR6 and 142Y in CmaxOR18 were predicted as key amino acid residues for binding ß-caryophyllene and tetradecane, respectively. CONCLUSION: This work provides an example pipeline for de novo investigation in C. maximoviczi baits and the underlying olfactory perceptions. The results will benefit the future development of trapping-based integrated pest management strategies and the deorphanization of odorant receptors in ground beetles. © 2022 Society of Chemical Industry.
Subject(s)
Coleoptera , Receptors, Odorant , Animals , Coleoptera/genetics , Coleoptera/metabolism , Insect Proteins/metabolism , Odorants , Plants/metabolism , Receptors, Odorant/chemistry , SmellABSTRACT
Breast cancer stem cells are a group of undifferentiated cells with self-renewal and multidifferentiation potential. Chemotherapeutic and radiotherapeutic resistance, hypoxic resistance, high tumorigenicity, high cell invasion, and metastatic abilities are characteristics of these cells, which are responsible for breast cancer recurrence. Therefore, the correct sorting and identification of breast cancer stem cells is a primary step for research in this field. This article briefly describes the recent progress on sorting and identification technologies for breast cancer stem cells. Sorting technologies include the side population technique, technologies that depend on cell surface markers, ALDEFLUOR assays, and in situ detection. Identification technologies include mammosphere cultures, limited dilution in vitro, and in-vivo animal models. This review provides an important reference for breast cancer stem cell research, which will explore new methods for the treatment of patients with breast cancer.
Subject(s)
Breast Neoplasms/pathology , Membrane Proteins/metabolism , Neoplastic Stem Cells/pathology , Side-Population Cells/cytology , AC133 Antigen , ATP Binding Cassette Transporter, Subfamily G, Member 2 , ATP-Binding Cassette Transporters/metabolism , Aldehyde Dehydrogenase/metabolism , Aldehyde Dehydrogenase 1 Family , Animals , Antigens, CD/metabolism , Female , Flow Cytometry/methods , Glycoproteins/metabolism , Humans , Hyaluronan Receptors/metabolism , Integrin alpha6/metabolism , Integrin beta1/metabolism , Integrin beta3/metabolism , Isoenzymes/metabolism , Neoplasm Proteins/metabolism , Neoplastic Stem Cells/metabolism , Peptides/metabolism , Retinal Dehydrogenase/metabolism , Side-Population Cells/metabolismABSTRACT
OBJECTIVE: To investigate the expression of angiopoietin (Ang)-1 and Ang-2 in colorectal tumors and its relations to microvessel density (MVD) in tumor tissue. METHODS: Ang-1, Ang-2 and factor VIII-related antigen were stained immunohistochemically in 91 cases of primary colorectal adenocarcinoma, 20 cases of colorectal adenoma and 24 cases of normal colorectal mucosal tissue, and MVD was also assayed in above tissue specimens. RESULT: (1) A significantly higher Ang-1 (7.07+/-2.00) was observed in normal tissue compared with 1.75 +/-1.98 in the adenoma and 1.40 +/- 1.22 in the adenocarcinoma (P<0.01). (2) Ang-2 protein positive rate in adenocarcinoma was significantly higher than that in normal tissue and adenoma (P<0.01). The expression of Ang-2 in adenocarcinoma was closely associated with poor differentiation and vessel invasion. (3) There were significant correlations between Ang-1 and Ang-2 (r=-0.338, P<0.01), Ang-1 and MVD (r=-0.388, P<0.01), Ang-2 and MVD (r=0.594, P<0.01) in the 135 cases. CONCLUSION: The overexpression of Ang-2 may play an important role in angiogenesis of colorectal adenocarcinoma. It can be regarded as an index for malignancy and prognosis in colorectal adenocarcinoma.
Subject(s)
Angiopoietin-2/biosynthesis , Capillaries/pathology , Colorectal Neoplasms/blood supply , Adenocarcinoma/blood supply , Adenocarcinoma/metabolism , Adult , Aged , Aged, 80 and over , Angiopoietin-1/biosynthesis , Angiopoietin-1/genetics , Angiopoietin-2/genetics , Biomarkers, Tumor , Colorectal Neoplasms/metabolism , Female , Humans , Male , Middle Aged , Prognosis , von Willebrand Factor/biosynthesis , von Willebrand Factor/geneticsABSTRACT
OBJECTIVE: To investigate the effect of nitric oxide (NO) on cytotoxicity of Lomustine (CCNU) in vitro. METHODS: CCNU was used to treat human glioma cell line BT-325 with different concentration of cytokines or NO donors, NO levels was measured by Griess assay and cell survival was evaluated by MTT assay. RESULT: (1) Pretreatment with IL-1 beta and LPS markedly suppressed CCNU cytotoxicity in BT-325 cells with a significant increase in NO production (P<0.05). This function could be inhibited by L-NAME. (2) DETA NONOate suppressed cytotoxicity of CCNU to BT -325 cells in a dose-dependent manner (P<0.05). (3) CCNU co-cultured with SNAP for 24 h showed higher cytotoxic to BT-325 cells(P<0.05). CONCLUSION: NO partly suppresses cytotoxicity of Lomustine, which might be associated with chemoresistance of BT-325 cells against CCNU in vitro. NO can also slow the degradation of CCNU in water solution.