Relationship of multidrug-resistant gene and extended-spectrum carbapenem-resistance in <i>Staphylococcus aureus</i>

The aim of this study was to determine the relationship between phenotypic antimicrobial susceptibilitypatterns and extended-spectrum, carbapenem-resistance genes. A total of 109 clinical Staphilococcus aureus strains weresubjected to 19 antimicrobial susceptibility tests. Resistance to methicillin (mecA), penicillin (blaTEM), and tetracycline(tetM) was detected. We compared the presence of the blaTEM genes with extended-spectrum, carbapenem-relatedgenes and identified the types of SCCmec genes. Of 109 clinical S. aureus strains, 62 (56.88%) had methicillin resistanceand 60 strains carried mecA. The prevalence of blaTEM and tetM genes was 81.65% and 37.61%, respectively. The mostpredominant SCCmec type was SCCmec type II 28/60 (46.67%), in 60 mecA-positive methicillin-resistant S. aureus(MRSA) isolates. The SCCmec prevalence rates were type IVA 30.00% (18/60), type IVb 8.33% (5/60), type IVd 6.67%(4/60), and non-typable 8.33% (5/60). Sixty of the 109 (55.05%) MRSA isolates were positive for extended-spectrumcarbapenems (31/60) (51.67%), cephalosporins 40/60 (66.67%) and carbapenems 31/60 (51.67%). The predominantSCCmec type II demonstrated more carbapenem-resistance than the IVA, IVb and IVd types.     

Tanshinone IIA protects intestinal epithelial cells from ferroptosis through the upregulation of <i>GPX4</i> and <i>SLC7A11</i>

Background: Inflammatory bowel disease (IBD) is a chronic inflammatory disease of the gastrointestinal tract. The destruction of the intestinal epithelial barrier is one of the major pathological processes in IBD pathology. Growing evidence indicated that epithelial cell ferroptosis is linked to IBD and is considered a target process. Methods: RAS-selective lethal 3 (RSL3) was used to induce ferroptosis in intestinal epithelial cell line No. 6 (IEC-6) cells, and cell ferroptosis and the effects of tanshinone IIA (Tan IIA) were determined by cell counting kit-8 (CCK-8), reactive oxygen species (ROS) staining, Giemsa staining and transmission electron microscope (TEM). The cell viability of natural product library compounds was determined by CCK-8. The expression of ferroptosis-related genes were detected by real-time quantitative polymerase chain reaction (RT-qPCR) and western blot. Results: Treatment of IEC-6 cells results in the accumulation of ROS and typical morphological characteristics of ferroptosis. RSL3 treatment caused rapid cellular cytotoxicity which could be reversed by ferrostatin-1 (Fer-1) in IEC-6 cells. Natural product library screening revealed that Tan IIA is a potent inhibitor of IEC-6 cell ferroptosis. Tan IIA could significantly protect the RSL3-induced ferroptosis of IEC-6 cells. Furthermore, the ferroptosis suppressors, glutathione peroxidase 4 (GPX4), solute carrier family 7 member 11 (SLC7A11), and miR-17-92 were found to be early response genes in RSL3-treated cells. Treatment of IEC-6 cells with Tan IIA resulted in upregulation of GPX4, SLC7A11, and miR-17-92. Conclusion: Our study demonstrated that Tan IIA protects IEC-6 cells from ferroptosis through the upregulation of GPX4, SLC7A11, and miR-17-92. The findings might provide a theoretical grounding for the future application of Tan IIA to treat or prevent IBD.     

Expression analysis of <i>OsSERK</i>, <i>OsLEC1</i> and <i>OsWOX4</i> genes in rice (<i>Oryza</i> sativa L.) callus during somatic embryo development

Somatic embryogenesis is an asexual reproduction process that occurs in many plant species, including rice. This process contains several totipotency markers such as Somatic Embryogenesis Receptor-like Kinase (SERK), Leafy Cotyledon1 (LEC1) and WUSCHEL-Related Homeobox4 (WOX4) and also a helpful model for embryo development and clones and transformations. Here, we report the gene expression during somatic embryo development correlates with regeneration frequency in 14 Javanica rice (pigmented and non-pigmented) using modifified N6 media supplemented with Kinetin (2.0 mg/L) and NAA (1.0 mg/L). Although there have been advances in understanding the genetic basis of somatic embryogenesis in other varieties, rice is still unexplored, especially during somatic embryo development. Moreover, for the formation of callus induction from immature embryos, 2,4-D (2.0 mg/L, 3.0 mg/L) was used. This study analysed the gene expression of OsSERK, OsWOX4 and OsLEC1 genes through RT-PCR analysis. Higher expression of the OsLEC1 gene indicates that their function may correlate in the in vitro with the high response of rice after transfer to regeneration media. This study found that rice varieties of pigmented rice (MS Pendek and Gogoniti II) and non-pigmented rice (Pandan Ungu) showed high regeneration frequency, showing higher OsLEC1 expression than other varieties because OsLEC1 promotes the maturation of somatic embryos in plant regeneration on day 14. However, the contrast with Genjah nganjuk may be effective because of other regulatory genes. RT-PCR analysis showed OsSERK had less expression level than OsLEC1 and OsWOX4 in the varieties, which correlate with the percentage of plant regeneration, but not for Gogoniti II. In conclusion, the higher percentage of plant regeneration correlates with the higher expression level of OsLEC1 at day 14 of media regeneration of rice.     

Response of <i>MaHMA2</i> gene expression and stress tolerance to zinc stress in mulberry (<i>Morus alba</i> L.)

HMA2 (heavy metal ATPase 2) plays a crucial role in extracellular and intracellular Zn²⁺ transport across biomembranes, maintaining ion homeostasis, and playing an important role in the normal physiological metabolism, growth, and development of plants. In our study, a novel HMA2 gene, named MaHMA2, was isolated and cloned from white mulberry (Morus alba L.). The gene sequence obtained was 1,342 bp long, with an open reading frame of 1,194 bp, encoding a protein of 397 amino acids, with a predicted molecular mass of 42.852 kD and an isoelectric point of 7.53. This protein belonged to the PIB-type ATPase transport protein family. We analyzed the expression of the MaHMA2 gene by quantitative real-time PCR. The results showed that the level of MaHMA2 gene expression decreased to a Zn concentration of 800 mg/kg. Malondialdehyde and proline levels increased and responded to increasing Zn when the MaHMA2 gene was silenced, whereas the activities of peroxidase and superoxide dismutase tended to increase in response to increasing Zn²⁺ ion stress concentrations but were lower in the gene-silenced plants. These findings suggested that the MaHMA2 gene played an active role in the tolerance response of mulberry to Zn stress.     

Ultrastructural analysis of sperm from the genus <i>Idarnes</i> (Hymenoptera: Chalcidoidea,Sycophaginae)

In this study, the sperm ultrastructure of three species of Idarnes genus was investigated using light and transmission electron microscopy. Spermatozoon morphology of the three species was similar to that of most Chalcidoidea, with helicoidally twisted nucleus and flagellum. The head region consists of an acrosome and a nucleus; the nucleus-flagellum transition region characterized by the presence of mitochondrial derivatives and the centriolar adjunct; a flagellum region, which includes the axoneme with microtubular arrangement 9 + 9 + 2 and two mitochondrial derivatives. However, the sperm of these three species exhibit features that discriminate one species from each other: (1) only one species, Idarnes sp. 2 (carme group) exhibited an extracellular sheath surrounding the anterior portion of the nucleus, which extends to the anterior region of the flagellum, but it did not present filaments; (2) the acrosome in the three species was quite different, Idarnes sp. 1 and Idarnes sp. 2 (carme group) has two compartments (acrosomal and subacrosomal vesicles) while Idarnes sp. 3 (flavicollis group) has a third compartment (perforatorium); (3) the centriolar adjunct elongated and its location among the mitochondrial derivatives is similar for the three species analyzed; (4) mitochondrial derivatives differ between the species, with triangular (Idarnes sp. 1 and sp. 3) and elongated or flat shaped (Idarnes sp. 2) appearance. These data shows that sperm structure may differ within the same genus and confirms the potential of these cells in phylogenetic and taxonomic analyses in the Chalcidoidea superfamily, as well as in Hymenoptera in general.     

Genome-wide identification of <i>U-box</i> gene family and expression analysis under abiotic stresses in <i>Salvia miltiorrhiza</i>

Plant U-box (PUB) E3 ubiquitin ligases play important roles in hormone signaling pathways and in response to different abiotic stresses, but little is known about U-box genes in Danshen (root of Salvia miltiorrhiza Bunge). Here, we identified and characterized 70 SmPUB genes based on its genome sequence. Phylogenetic analysis of U-box genes from S. miltiorrhiza and Arabidopsis suggested that they can be clustered into seven subgroups (I–VII). Typical U-box domains were found in all identified SmPUB genes through the analysis of conserved motifs. Moreover, qRT-PCR was applied to analyze the relative expression levels of U-box genes in S. miltiorrhiza roots and leaves under PEG-induced water deficit and salt stresses. Results revealed that the SmPUB genes exhibited stronger response to drought than to salt stress. To the best of our knowledge, this report is the first to perform genome-wide identification and analysis of the U-box gene family in S. miltiorrhiza, and the results provide valuable information for better understanding of the function of U-box in S. miltiorrhiza.     

Genome-wide identification,characterization, and expression analysis of aluminum-activated malate transporter genes (ALMTs) in <i>Gossypium hirsutum</i> L.

Aluminum-activated malate transporters (ALMT) are widely involved in plant growth and metabolic processes, including adaptation to acid soils, guard cell regulation, anion homeostasis, and seed development. Although ALMT genes have been identified in Arabidopsis, wheat, barley, and Lotus japonicus, little is known about its presence in Gossypium hirsutum L. In this study, ALMT gene recognition in diploid and tetraploid cotton were done using bioinformatics analysis that examined correlation between homology and evolution. Differentially regulated ALMT genetic profile in G. hirsutum was examined, using RNA sequencing and qRT-PCR, during six fiber developmental time-points, namely 5 d, 7 d, 10 d, 15 d, 20 d, and 25 d. We detected 36 ALMT genes in G. hirsutum, which were subsequently annotated and divided into seven sub-categories. Among these ALMT genes, 34 had uneven distribution across 14/26 chromosomes. Conserved domains and gene structure analysis indicated that ALMT genes were highly conserved and composed of exons and introns. The GhALMT gene expression profile at different DPA (days post anthesis) in different varieties of G. hirsutum is indicative of a crucial role of ALMT genes in fiber development in G. hirsutum. This study provides basis for advancements in the cloning and functional enhancements of ALMT genes in enhancing fiber development and augmenting high quality crop production.     

Phytochemistry and ethnomedicinal qualities of metabolites from <i>Phyllanthus emblica</i> L.: A review

Phyllanthus emblica or Indian gooseberry is an integrated part of Ayurvedic and Traditional Chinese Medicines. For several decades, the well-known ancient herb has been extensively utilized in traditional medicine to cure diseases like fever, diabetes, constipation, jaundice, ulcers, biliousness, anemia, anorexia, and dyspepsia. In the traditional system, Indian gooseberry has various ethnomedicinal applications. In the Ayurvedic system, different methods of administration (anupan) have shown different ethnomedicinal properties of Indian gooseberry. Seventy well-known chemical components in Indian gooseberry have been identified through phytochemical evaluation, among which the flavonoids and phenols are most prominent. From the toxicity perspective, it is considered a safe herb in India, and is taken as a food supplement in European countries. The wide-spectrum pharmacological activities of the crude extracts and isolates of Indian gooseberry are attributed to the predominance of phenols and flavonoids. Thus, it is important to study the exact mechanism of the activity of the phytochemicals in Indian gooseberry, especially in anti-cancer activities. Extract of Indian gooseberry enhances proliferation in several cancer cells in vitro, including stem cells like ovarian cancer (OC) cells, and also has been observed to possess anti-proliferative characteristics in vivo. This review intends to explore the therapeutic potential of Indian gooseberry based on scientific reports and attempts to find the gaps for future research.     

Polymorphic information and genetic diversity in <i>Brassica</i> species revealed by RAPD markers

Randomly amplified polymorphic DNA (RAPD) is a tremendously convenient approach used to discriminatebetween Brassica species owing to its accuracy and speed. RAPD primers generate adequate genetic information that canbe used in the primer-marker system. In this work, twenty RAPD-PCR based markers were executed to generatepolymorphic data, like polymorphic information content (PIC), mean resolving power (MRP), resolving power (RP),effective multiplex ratio (EMR), and marker index (MI) for the first time and genetic distance among and between sixBrassica species were calculated. Our results indicated that 20 primers produced a total of 231 scored band andgenerated 87% polymorphic bands. Average PIC, MRP, RP, MI, and EMR values were 0.088, 0.65, 6.7, 0.78, and 8.9,respectively. PIC showed an overall negative correlation with MRP, RP, MI, and EMR, whereas MRP, RP, and EMR,were positively correlated with each other. Genetic identities ranged from 41.99% (between Brassica napus andBrassica oleracea) to 68.83% (between Brassica campestris and Brassica oleracea). Dendrogram results showed noclustering between species except between Brassica campestris and Brassica nigra. Nevertheless, these results will behelpful to acquire useful information about the markers and their use to determine the genomic structures of Brassicaspecies. Further, based on genetic distance and polymorphic information, new hybrids can be developed for effectiveoilseed production.     

Structural characterization of four <i>Rhododendron</i> spp. chloroplast genomes and comparative analyses with other azaleas

Azalea is a general designation of Rhododendron in the Ericaceae family. Rhododendron not only has highornamental value but also has application value in ecological protection, medicine, and scientific research. In thisstudy, we used Illumina and PacBio sequencing to assemble and annotate the entire chloroplast genomes (cpgenomes) of four Rhododendron species. The chloroplast genomes of R. concinnum, R. henanense subsp. lingbaoense,R. micranthum, and R. simsii were assembled into 207,236, 208,015, 207,233, and 206,912 bp, respectively. Allchloroplast genomes contain eight rRNA genes, with either 88 or 89 protein-coding genes. The four cp genomes werecompared and analyzed by bioinformatics, and the phylogenetic analysis based on chloroplast genomes of 26 speciesof Ericaceae, Actinidiaceae, and Primulaceae under Ericales was conducted. A comparison of the linear structure of cpgenomes of four Rhododendron showed that there were substantial sequence similarities in coding regions, but highdifferences in non-coding regions. A phylogenetic analysis, based on chloroplast whole genome sequences, showedthat all Rhododendron species are in the clade Ericaceae. This study provides valuable genetic information for thestudy of population genetics and evolutionary relationships in Rhododendron and other azalea species.     

Detection of new antibiotic resistance gene profile in <i>Escherichia coli</i> associated with avian leukosis virus infection from broiler chickens

The Escherichia coli (E. coli) is prevailing worldwide, but the epidemiology of E. coli infections feature regionaldistribution characteristics to some extent. E. coli, as a zoonotic pathogen, can be transferred from animals to humansthrough food chain or via contact with wounds, causing a public health risk. We reported the swelling ofproventriculus and tracheal bleeding following the death in two broiler chickens (Gallus gallus domesticus) fromBeijing, China. To investigate whether a virus was involved in the infection, Madin Darby Bovine Kidney (MDCK)cells were co-cultured with supernatants of proventriculus, trachea and spleen homogenates. The avian leucosis viruswas detected in the samples of proventriculus and trachea, but the avian influenza virus, the Newcastle disease virusand the avian infectious laryngotracheitis virus were not detected. E. coli isolates were resistant to almost all theantimicrobial as tested except for the combinations of amoxicillin/clavulanic acid and sulfamethoxazole/trimethoprim.PCR tests demonstrated the presence of antibiotic resistance genes in these E. coli isolates and further researchrevealed a novel gene profile with the presence of CTX-M-1, gyrA, gyrB, oqxA, oqxB, parC and Sul2 antibioticresistance genes in a strain isolated from a proventriculus sample. These results demonstrated that the presence ofantibiotic resistant E. coli would not necessarily cause outbreak of large-scale disease. However, when the bacteriacarrying new antibiotic resistance genes enter the environment, it may result in the development of more virulentstrains which will potentially impact human and animal health.     

Co-ordinated combination of Embden-Meyerhof-Parnas pathway and pentose phosphate pathway in <i>Escherichia coli</i> to promote L-tryptophan production

In this study, phosphoenolpyruvate and erythrose-4-phosphate are efficiently supplied by collaborative design of Embden-Meyerhof-Parnas (EMP) pathway and pentose phosphate (PP) pathway in Escherichia coli, thus increasing the L-tryptophan production. Firstly, the effects of disrupting EMP pathway on L-tryptophan production were studied, and the results indicated that the strain with deletion of phosphofructokinase A (i.e., E. coli JW-5 ΔpfkA) produced 23.4 ± 2.1 g/L of L-tryptophan production. However, deletion of phosphofructokinase A and glucosephosphate isomerase is not conducive to glucose consumption and cell growth, especially deletion of glucosephosphate isomerase. Next, the carbon flux in PP pathway was enhanced by introduction of the desensitized glucose-6-phosphate dehydrogenase (zwf) and 6-phosphogluconate dehydrogenase (gnd) and thus increasing the L-tryptophan production (i.e., 26.5 ± 3.2 g/L vs. 21.7 ± 1.3 g/L) without obviously changing the cell growth (i.e., 0.41 h⁻¹ vs. 0.44 h⁻¹) as compared with the original strain JW-5. Finally, the effects of co-modifying EMP pathway and PP pathway on L-tryptophan production were investigated. It was found that the strain with deletion of phosphofructokinase A as well as introduction of the desensitized zwf and gnd (i.e., E. coli JW-5 zwf243 gnd361 ΔpfkA) produced 31.9 ± 2.7 g/L of L-tryptophan, which was 47.0% higher than that of strain JW-5. In addition, the glucose consumption rate of strain JW-5 zwf243 gnd361 ΔpfkA was obviously increased despite of the bad cell growth as compared with strain JW-5. The results of this study have important reference value for the following application of metabolic engineering to improve aromatic amino acids producing strains.