Dissemination of antibiotic resistance genes from aboveground sources to groundwater in livestock farms.

Antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs) are prevalent in various environments on livestock farms, including livestock waste, soil, and groundwater. Contamination of groundwater by ARB and ARGs in livestock farms is a growing concern as it may have potentially huge risks to human health. However, the source of groundwater-borne ARB and ARGs in animal farms remains largely unknown. In this study, different types of samples including groundwater and its potential contamination sources from aboveground (pig feces, wastewater, and soil) from both working and abandoned swine feedlots in southern China were collected and subjected to metagenomic sequencing and ARB isolation. The source tracking based on metagenomic analysis revealed that 56-95 % of ARGs in groundwater was attributable to aboveground sources. Using metagenomic assembly, we found that 45 ARGs predominantly conferring resistance to aminoglycosides, sulfonamides, and tetracyclines could be transferred from the aboveground sources to groundwater, mostly through plasmid-mediated horizontal gene transfer. Furthermore, the full-length nucleotide sequences of sul1, tetA, and TEM-1 detected in ARB isolates exhibited the close evolutionary relationships between aboveground sources and groundwater. Some isolated strains of antibiotic-resistant Pseudomonas spp. from aboveground sources and groundwater had the high similarity (average nucleotide identity > 99 %). Notably, the groundwater-borne ARGs were identified as mainly carried by bacterial pathogens, potentially posing risks to human and animal health. Overall, this study underscores the dissemination of ARGs from aboveground sources to groundwater in animal farms and associated risks.

Inter-plasmid transfer of antibiotic resistance genes accelerates antibiotic resistance in bacterial pathogens.

Antimicrobial resistance is a major threat for public health. Plasmids play a critical role in the spread of antimicrobial resistance via horizontal gene transfer between bacterial species. However, it remains unclear how plasmids originally recruit and assemble various antibiotic resistance genes (ARGs). Here, we track ARG recruitment and assembly in clinically relevant plasmids by combining a systematic analysis of 2420 complete plasmid genomes and experimental validation. Results showed that ARG transfer across plasmids is prevalent, and 87% ARGs were observed to potentially transfer among various plasmids among 8229 plasmid-borne ARGs. Interestingly, recruitment and assembly of ARGs occur mostly among compatible plasmids within the same bacterial cell, with over 88% of ARG transfers occurring between compatible plasmids. Integron and insertion sequences drive the ongoing ARG acquisition by plasmids, especially in which IS26 facilitates 63.1% of ARG transfer events among plasmids. In vitro experiment validated the important role of IS26 involved in transferring gentamicin resistance gene aacC1 between compatible plasmids. Network analysis showed four beta-lactam genes (blaTEM-1, blaNDM-4, blaKPC-2, and blaSHV-1) shuffling among 1029 plasmids and 45 clinical pathogens, suggesting that clinically alarming ARGs transferred accelerate the propagation of antibiotic resistance in clinical pathogens. ARGs in plasmids are also able to transmit across clinical and environmental boundaries, in terms of the high-sequence similarities of plasmid-borne ARGs between clinical and environmental plasmids. This study demonstrated that inter-plasmid ARG transfer is a universal mechanism for plasmid to recruit various ARGs, thus advancing our understanding of the emergence of multidrug-resistant plasmids.

Effects of a chitosan nanoparticles encapsulation on the properties of litchi polyphenols.

Litchi polyphenols have very specific biological activities. Nevertheless, the low and inconsistent oral bioavailability and instability hinder the further application of litchi polyphenols in food systems. This work prepared litchi polyphenols loaded chitosan nanoparticles (LP-CSNPs) by ionic gelation method to enhance the encapsulation on the properties of litchi polyphenols. The optimum conditions of formation via single factors and the Box-Behnken design were chitosan (CS) concentration 1.065 mg/mL, sodium tripolyphosphate (TPP) concentration 0.975 mg/mL, and the mass ratios of polyphenols and CS 1:1 with encapsulation efficiency (EE%) of 45.53%. LP-CSNPs presented the nanosized range of particle size (mean 170 nm), excellent polydispersity index (PDI) (0.156 ± 0.025), and zeta potential values (+ 35.44 ± 0.59). The in vitro release in simulated gastric fluid (pH 1.2) and intestinal fluid (pH 6.8) during 100 h was 58.34% and 81.68%, respectively. LP-CSNPs could effectively improve the storage stability and had great antibacterial activity compared with unencapsulated litchi polyphenols.

Synthesis, and biological evaluation of pyrazole matrine derivatives as an insecticide against Spodoptera frugiperda.

As one of the major threats to global food security, Spodoptera frugiperda (S. frugiperda) is highly gaining consideration due to its severe damage. Matrine is a widely and effectively used botanical insecticide in controlling S.frugiperda but lacks a rapidly available effect. To further improved the insecticidal activity of matrine based on combination principles, this work synthesized five new pyrazole matrine derivatives (PMDs) using Michael addition and investigated insecticidal activity against 2nd instar larvae of S. frugiperda(in vivo) and its isolated cell(in vitro). Our result demonstrated that PMDs show higher pesticidal activity than that matrine in both in vitro and in vivo assays. The most toxic derivatives in vitro and in vivo are PMD-3 and PMD-1, with IC50 of 2.49 mM and LC50 of 22.76 mg/L respectively. This research also investigates the anti-proliferation mechanism of PMDs based on isolated cells. PMDs decrease mitochondria membrane potential, arrested cell cycle at the G2/M phase, and upregulated Caspase 3, Caspase 9, and Apaf-1 to induce Caspase-dependent apoptosis. For Caspase-independent apoptosis, AIF and Endo G were found to be upregulated. Besides, pro-apoptotic factors like p53, IBM-1, and anti-apoptotic factors like IAP were upregulated. Moreover, we supposed that there was a linkage between lysosomes and PMD-induced apoptosis according to increased apoptosis rate, activated lysosomes, and upregulated Cathepsin B. This research provides new ideas for the synthesis of matrine derivatives and further demonstrated the anti-proliferation mechanism of PMDs.

How generic is iodide-tagging photoelectron spectroscopy: An extended investigation on the Gly·X- (Gly = glycine, X = Cl or Br) complexes.

We report a joint negative ion photoelectron spectroscopy (NIPES) and quantum chemical computational study on glycine-chloride/bromide complexes (denoted Gly·X-, X = Cl/Br) in close comparison to the previously studied Gly·I- cluster ion. Combining experimental NIPE spectra and theoretical calculations, various Gly·X- complexes were found to adopt the same types of low-lying isomers, albeit with different relative energies. Despite more congested spectral profiles for Gly·Cl- and Gly·Br-, spectral assignments were accomplished with the guidance of the knowledge learned from Gly·I-, where a larger spin-orbit splitting of iodine afforded well-resolved, recognizable spectral peaks. Three canonical plus one zwitterionic isomer for Gly·Cl- and four canonical conformers for Gly·Br- were experimentally identified and characterized in contrast to the five canonical ones observed for Gly·I- under similar experimental conditions. Taken together, this study investigates both genericity and variations in binding patterns for the complexes composed of glycine and various halides, demonstrating that iodide-tagging is an effective spectroscopic means to unravel diverse ion-molecule binding motifs for cluster anions with congested spectral bands by substituting the respective ion with iodide.

Global Increase of Antibiotic Resistance Genes in Conjugative Plasmids.

Antibiotic resistance is propagating worldwide, but the predominant dissemination mechanisms are not fully understood. Here, we report that antibiotic resistance gene (ARG) abundance in conjugative plasmids that are recorded in the National Center for Biotechnology Information (NCBI) RefSeq plasmid database is increasing globally, which is likely a key factor in the propagation of resistance. ARG abundance in plasmids increased by 10-fold on a global scale from the year 2000 to the year 2020 (from 0.25 to 2.93 ARG copies/plasmid), with a more pronounced increase being observed in low-to-middle income countries. This increasing trend of plasmid-borne ARGs was corroborated by bootstrap resampling from each year of the NCBI RefSeq plasmid database. The results of a correlation analysis imply that if antibiotic consumption keeps growing at the current rates, a 2.7-fold global increase in the ARG abundance of clinically relevant plasmids may be reached by 2030. High sequence similarities of clinically relevant, conjugative plasmids that are isolated both from clinics and from the environment raise concerns about the environmental resistome serving as a potential ARG maintenance reservoir that facilitates transmission across these ecological boundaries. IMPORTANCE Antibiotic resistance propagation is a significant concern due to its projected impacts on both global health and the economy. However, global propagation mechanisms are not fully understood, including regional and temporal trends in the abundance of resistance plasmids that facilitate antibiotic resistance gene (ARG) dissemination. This unprecedented study reports that ARG abundance in the conjugative plasmids that are recorded in the National Center for Biotechnology Information (NCBI) database and harbor ARGs is increasing globally with antibiotic consumption, especially in low-to-medium income countries. Through network and comparative genomic analyses, we also found high sequence similarities of clinically relevant conjugative resistance plasmids that were isolated from clinical and environmental sources, suggesting transmission between these ecological boundaries. Therefore, this study informs the One Health perspective to develop effective strategies by which to curtail the propagation of plasmid-borne antibiotic resistance.

RecT Affects Prophage Lifestyle and Host Core Cellular Processes in Pseudomonas aeruginosa.

Pseudomonas aeruginosa is a notorious pathogen that causes various nosocomial infections. Several prophage genes located on the chromosomes of P. aeruginosa have been reported to contribute to bacterial pathogenesis via host phenotype transformations, such as serotype conversion and antibiotic resistance. However, our understanding of the molecular mechanism behind host phenotype shifts induced by prophage genes remains largely unknown. Here, we report a systematic study around a hypothetical recombinase, Pg54 (RecT), located on a 48-kb putative prophage (designated PP9W) of a clinical P. aeruginosa strain P9W. Using a ΔrecT mutant (designated P9D), we found that RecT promoted prophage PP9W excision and gene transcription via the inhibition of the gene expression level of pg40, which encodes a CI-like repressor protein. Further transcriptomic profiling and various phenotypic tests showed that RecT modulated like a suppressor to some transcription factors and vital genes of diverse cellular processes, providing multiple advantages for the host, including cell growth, biofilm formation, and virulence. The versatile functions of RecT hint at a strong impact of phage proteins on host P. aeruginosa phenotypic flexibility. IMPORTANCE Multidrug-resistant and metabolically versatile P. aeruginosa are difficult to eradicate by anti-infective therapy and frequently lead to significant morbidity and mortality. This study characterizes a putative recombinase (RecT) encoded by a prophage of a clinical P. aeruginosa strain isolated from severely burned patients, altering prophage lifestyle and host core cellular processes. It implies the potential role of RecT in the coevolution arm race between bacteria and phage. The excised free phages from the chromosome of host bacteria can be used as weapons against other sensitive competitors in diverse environments, which may increase the lysogeny frequency of different P. aeruginosa subgroups. Subsequent analyses revealed that RecT both positively and negatively affects different phenotypic traits of the host. These findings concerning RecT functions of host phenotypic flexibility improve our understanding of the association between phage recombinases and clinical P. aeruginosa, providing new insight into mitigating the pathogen infection.

Synthesis of Halopyrazole Matrine Derivatives and Their Insecticidal and Fungicidal Activities.

Matrine is a traditional botanical pesticide with a broad-spectrum biological activity that is widely applied in agriculture. Halopyrazole groups are successfully introduced to the C13 of matrine to synthesize eight new derivatives with a yield of 78-87%. The insecticidal activity results show that the introduction of halopyrazole groups can significantly improve the insecticidal activity of matrine on Plutella xylostella, Mythimna separata and Spodoptera frugiperda with a corrected mortality rate of 100%, which is 25-65% higher than matrine. The fungicidal activity results indicate that derivatives have a high inhibitory effect on Ceratobasidium cornigerum, Cibberella sanbinetti, Gibberrlla zeae and Collectot tichum gloeosporioides. Thereinto, 4-Cl-Pyr-Mat has the best result, with an inhibition rate of 23-33% higher than that of matrine. Therefore, the introduction of halogenated pyrazole groups can improve the agricultural activity of matrine.

Characteristics of Wild Bird Resistomes and Dissemination of Antibiotic Resistance Genes in Interconnected Bird-Habitat Systems Revealed by Similarity of blaTEM Polymorphic Sequences.

Wild birds are known to harbor and discharge antibiotic-resistant bacteria (ARB) and their associated antibiotic resistance genes (ARGs). However, assessments of their contribution to the dissemination of antibiotic resistance in the environment are limited to culture-dependent bacterial snapshots. Here, we present a high-throughput sequencing study that corroborates extensive ARG exchange between wild bird feces and their habitats and implies the need to scrutinize high-mobility birds as potential vectors for global propagation of ARGs. We characterized the resistome (281 ARGs) and microbiome of seven wild bird species and their terrestrial and aquatic habitats. The resistomes of bird feces were influenced by the microbial community structure, mobile genetic elements (MGEs), and residual antibiotics. We designated 33 ARGs found in more than 90% of the bird fecal samples as core ARGs of wild bird feces, among which 16 ARGs were shared as core ARGs in both wild bird feces and their habitats; these genes represent a large proportion of both the bird feces (35.0 ± 15.9%) and the environmental resistome (29.9 ± 21.4%). One of the most detected ß-lactam resistance genes (blaTEM, commonly harbored by multidrug resistant "superbugs") was used as molecular marker to demonstrate the high interconnectivity of ARGs between the microbiomes of wild birds and their habitats. Overall, this work provides a comprehensive analysis of the wild bird resistome and underscores the importance to consider genetic exchange between animals and the environment in the One Health approach.

Observation and Exploitation of Spin-Orbit Excited Dipole-Bound States in Ion-Molecule Clusters.

We report an observation of spin-orbit excited dipole-bound states (DBSs) in arginine-iodide complexes (Arg·I-) by using temperature-dependent, wavelength-resolved "iodide-tagging" negative ion photoelectron spectroscopy. The observed DBSs are bound to the spin-orbit excited I(2P1/2) level of the neutral Arg·I complex in zwitterionic conformations and identified based on the resonant enhancement due to spin-orbit electronic autodetachment from the I(2P1/2) DBS to the I(2P3/2) neutral ground state. The observed DBS binding energies are correlated to the dipole moments of neutral Arg·I isomers and tautomers. This work thus demonstrates a new and generic spectroscopic approach to identify ion-molecule cluster conformations based on their distinguishable dipole moments.

Observation of Conformational Simplification upon N-Methylation on Amino Acid Iodide Clusters.

This Letter reports a counterintuitive observation that methylation of the glycine-iodide cluster leads to fewer conformations and spectroscopic simplicity. Cryogenic "iodide-tagging" negative ion photoelectron spectroscopy (NIPES) is used to probe specific binding sites of three N-methylated glycine derivatives, i.e., N-methylglycine (sarcosine), N,N-dimethylglycine, and N,N,N-trimethylglycine (glycine betaine). NIPES reveals a progressive spectral simplification of the iodide clusters with increasing methylation due to fewer contributing structures. Low energy conformers and tautomers of each cluster are computationally identified, and those observed in the experiments are assigned based on excellent agreement between the NIPE spectra and theoretical simulations. Zwitterionic cluster structures are found to be less stable than their canonical forms and do not contribute to the observed spectra. This work demonstrates the power of iodide-tagging NIPES in probing conformations of amino acid-iodide clusters and provides a molecular level understanding on the effect of methyl substitution on amino acid binding sites.

Determinant Factor for Thermodynamic Stability of Sulfuric Acid-Amine Complexes.

Atmospheric amines are thought to play significant roles in the nucleation of sulfuric acid-mediated aerosol particles. Their enhancing effects on the stabilization of the related complexes have formerly been correlated with the amine base strength, but there are a few exceptions reported. In this work, the influence of seven alkylamines on the thermodynamic stability of sulfuric acid-amine complexes has been theoretically investigated, e.g., ethylamine, propylamine, isopropylamine, tert-butylamine, dimethylamine, ethylmethylamine, and trimethylamine. For all primary and secondary amine-mediated complexes, a dual hydrogen bond configuration is generally suggested in the most stable isomer. The stabilization of this special structure predicted by the electrostatic potential distribution on the molecular surface of amines exactly agrees with the base strength sequence, providing crucial evidence for the previous deduction of correlation between the base strength and the enhancing effect. Meanwhile, the considerable van der Waals interactions are found between the free hydroxyl of sulfuric acid and the ß-methyl group of amine, resulting in the extra stability for sulfuric acid-dimethylamine and sulfuric acid-ethylmethylamine complexes. Therefore, the electrostatic potential distribution of amines is the essential determinant factor for the thermodynamic stability of the relevant complexes. Our conclusions provide new insight into a way to evaluate the enhancing abilities of amines in aerosol particle nucleation.

Synthesis, characterization of two matrine derivatives and their cytotoxic effect on Sf9 cell of Spodoptera frugiperda.

The invasion of Spodoptera frugiperda has imposed a serious impact on global food security. Matrine is a botanical pesticide with a broad spectrum of insecticidal activity which was recommended for controlling Spodoptera frugiperda. In order to discover effective insecticide for Spodoptera frugiperda, two matrine derivatives modified with carbon disulfide and nitrogen-containing groups were systhesized. And their inhibition activities on Sf9 cell were evaluated. The structural configuration of compounds were characterized by IR, HPLC, MS, NMR and XRD, with yields of 52% and 65%, respectively. The IC50 of the two newly synthesized compounds on Sf9 cell reduced to 0.648 mmol/L and 1.13 mmol/L, respectively, compared with that of matrine (5.330 mmol/L). In addition, microscopic observation of Sf9 cell treated with the compounds showed that the number of adherent cells decreased, the cells shrunk, vacuolated and apoptotic bodies appeared. The two newly synthesized compounds exhibited better inhibitory effect on Sf9 cell than that of the parent matrine, suggesting that the positive effect of the introduction of 1-pyrrolidinecarbodithioate and diethylcarbamodithioate groups to matrine. The morphological observation of Sf9 cell induced by derivatives indicated that apoptosis induction may be a mechanism that inhibits insect cell proliferation and exerts insecticidal effect.

Spectroscopic evidence for intact carbonic acid stabilized by halide anions in the gas phase.

This work shows elusive carbonic acid being effectively stabilized in the gas phase by interacting with halide anions X- (X = F, Cl, Br, and I). The formed H2CO3·X- complexes, characterized by negative ion photoelectron spectroscopy and ab initio calculations, all contain intact trans-trans carbonic acid binding onto the respective halide via two identical strong ionic O-HX- hydrogen bonds. For X = Cl, Br, and I, the complex spectra exhibit the corresponding X- signature by simply shifting to the higher binding energy side, while an extremely 2 eV wide broader band is observed for X = F. This spectroscopic evidence indicates that an excess electron is removed from each halide in the former case, while a proton is transferred from carbonic acid to fluoride upon electron detachment for the latter. The above H2CO3·X- structures as well as those of the previously studied H2SO4·X- along the homologous halogen series cannot be explained using the proton affinity (PA) argument. Instead, a qualitative correlation is found between these structural motifs and the constituent acid pKa values, strongly suggesting that pKa is a more suitable factor to predict correct acid-base chemistry between these diprotic oxyacids and halides.

Cryogenic "Iodide-Tagging" Photoelectron Spectroscopy: A Sensitive Probe for Specific Binding Sites of Amino Acids.

This work showcases cryogenic and temperature-dependent "iodide-tagging" photoelectron spectroscopy to probe specific binding sites of amino acids using the glycine-iodide complex (Gly·I-) as a case study. Multiple Gly·I- isomers were generated from ambient electrospray ionization and kinetically isolated in a cryogenic ion trap. These structures were characterized with temperature-dependent "iodide-tagging" negative ion photoelectron spectroscopy (NIPES), where iodide was used as the "messenger" to interpret electronic energetics and structural information of various Gly·I- isomers. Accompanied by theoretical computations and Franck-Condon simulations, a total of five cluster structures have been identified along with their various binding motifs. This work demonstrates that "iodide-tagging" NIPES is a powerful general means for probing specific binding interactions in biological molecules of interest.

Semi-synthesis and characterization of some new matrine derivatives as insecticidal agents.

BACKGROUND: Matrine is an important traditional plant-derived insecticide with broad-spectrum activity. However, due to its moderate activity, matrine is mainly applied in combination with other pesticides. In order to discover new potential natural-product-based crop protection agents, a series of matrine derivatives characterized by cyclohexylamine group were synthesized to screen their insecticidal activity against seven typically agricultural pests. RESULTS: The structural configurations of compounds were characterized by IR, 1 H NMR, 13 C NMR, MS and XRD, with the pure yields of 42%, 65% and 71%, respectively. Although all compounds showed poor insecticidal activity against five lepidoptera pests, the compounds 2 and 4 displayed remarkable insecticidal activities against Lipaphis erysimi and Mulberry Root-Knot Nematode with a concentration-dependent manner within 0.5~1.5 mg/ mL. Compared with matrine (60%), compounds 2 and 4 exhibited potent insecticidal activities against L. erysimi, with a corrected mortality of 83.3% and 89.7%, respectively. They also showed excellent control effects on Mulberry Root-Knot Nematode, with corrected mortality as high as 88% and 80%, respectively. CONCLUSION: All four synthesized matrine derivatives showed poor insecticidal activity against five lepidoptera pests, but the compounds 2 and 4 exhibited much stronger insecticidal activities against L. erysimi and Mulberry Root-Knot Nematode than matrine. Combined with the structural characteristics of compounds 1~4, we conclude that 4-methylcyclohexylamine, not the carbon disulfide group or cyclohexylamine group alone, mainly contributed to the improvement of insecticidal activities of matrine derivatives against these two agricultural pests. This work provides a direction and foundation for structural optimization of the matrine pesticides in the future. © 2020 Society of Chemical Industry.

Identification of Chinese Herbal Compounds with Potential as JAK3 Inhibitors.

The Janus kinases (JAKs) consist of four similar tyrosine kinases and function as key hubs in the signaling pathways that are implicated in both innate and adaptive immunity. Among the four members, JAK3 is probably the more attractive target for treatment of inflammatory diseases because its inhibition demonstrates the greatest immunosuppression and most profound effect in the treatment of such disorders. Although many JAK3 inhibitors are already available, certain shortcomings have been identified, mostly acquired drug resistance or unwanted side effects. To discover and identify new promising lead candidates, in this study, the structure of JAK3 (3LXK) was obtained from the Protein Data Bank and used for simulation modeling and protein-ligand interaction analysis. The ~36,000 Chinese herbal compounds obtained from TCM Database@Taiwan were virtually screened by AutoDock Vina docking program and filtered with Lipinski's Rules and ADME/T virtual predictions. Because of high occurrence of fake hits during docking, we selected 12 phytochemicals which have demonstrated modulating JAKs expressions among the top 50 chemicals from docking results. To validate whether these compounds are able to directly mediate JAK3 kinase, we have investigated the inhibitory activity using enzymatic activity assays, western blot, and HEK 293 cell STAT5 transactivity assays. The molecular analysis included docking and molecular dynamics (MD) simulations in order to investigate structural conformations and to explore the key amino acids in the interaction between JAK3 kinase and its putative ligands. The results demonstrated that Cryptotanshinone, Icaritin, and Indirubin exhibited substantial inhibitory activity against JAK3 kinase in vitro. The results also provide binding models of the protein-ligand interaction, detailing the interacting amino acid residues at the active ATP-binding domains of JAK3 kinase. In conclusion, our work discovered 3 potential natural inhibitors of JAK3 kinase and could provide new possibilities and stimulate new insights for the treatment of JAK3-targeted diseases.

Cl-Loss dynamics in the dissociative photoionization of CF3Cl with threshold photoelectron-photoion coincidence imaging.

The dissociative photoionization of CF3Cl was investigated in the photon energy range of 12.30-18.50 eV. The low-lying electronic states of CF3Cl+ cations were prepared by the method of threshold photoelectron-photoion coincidence (TPEPICO). The threshold photoelectron spectrum and the coincident time-of-flight mass spectra at the specific photon energies were recorded. Only a CF3+ fragment was observed at lower energy, while a CF2Cl+ fragment appeared for C2E and D2E states. As Cl-loss from the ground ionic state is statistical, the total kinetic energy release distribution (KERD) is represented as a Boltzmann profile, and a 0 K appearance energy of AP0 =12.79 ± 0.02 eV is derived from the statistical modelling of the breakdown diagram from 12.60 to 12.85 eV without taking into account the kinetic shift. For the A2A1 and B2A2 states of CF3Cl+ cations, the total KERDs are bimodal, where a parallel faster dissociation appears together with the statistical distribution. At higher energies like the C2E and D2E ionic states, a bimodal distribution similar to that of the A2A1 and B2A2 states is also observed for the KERD. With the aid of the calculated Cl-loss potential energy curves, the dissociative mechanisms of internal energy-selected CF3Cl+ cations are proposed.

[Non-covalent albumin conjugates of silicon (IV) phthalocyanines axially substituted with nucleoside: preparation and in vitro photodynamic activities].

The interactions of bovine serum albumin (BSA) with five novel silicon (N) phthalocyanines(SiPcl-5) axially modified by nucleosides (cytidine, 5-N-cytidine, methyl cytidine, uridine and methyl uridine) derivatives were studied by fluorescence spectroscopy. The results show that there are strong interactions between these silicon phthalocyanines and BSA with a binding constant of (4.90-83.18) x 10(5) mol(-1) x L. Therefore, the non-covalent BSA conjugate of bis(2', 3'-O-isopropyl-cytidine-oxy) phthalocyaninatosilicon(IV) (SiPc1) was further been prepared. The molar ratio of phthalocyanine to albumin was found to be 1:1 for the obtained SiPcl-BSA conjugate. The absorption spectra of SiPc1 and SiPc1-BSA in the visible region have no significant difference, both showing an Q-band maximum at about 686 nm. It indicates that the spectroscopic characteristics of SiPc1 are not affected by binding to albumin. The SiPcl-BSA conjugate exhibits high photodynamic activity against human hepatoma cell line HepG2 with an IC50 value of 3.0 x 10(-7) mol x L(-1). By comparsion, SiPc1-BSA has a higher photodynamic activity than SiPc1 (in PBS formation, IC50 = 7.0 x 10(-7) mol x L(-1)), which can be attributed to its higher cellular uptake.

Endoscopic biopsy in gastrointestinal neuroendocrine neoplasms: a retrospective study.

BACKGROUND: Gastrointestinal neuroendocrine neoplasms (GI-NENs) are often located in the deep mucosa or submucosa, and the efficacy of endoscopic biopsy for diagnosis and treatment of GI-NENs is not fully understood. OBJECTIVE: The current study analyzed GI-NENs, especially those diagnosed pathologically and resected endoscopically, and focused on the biopsy and cold biopsy forceps polypectomy (CBP) to analyze their roles in diagnosing and treating GI-NENs. METHODS: Clinical data of all GI-NENs were reviewed from January 2006 to March 2012. Histopathology was used to diagnose GI-NENs, which were confirmed by immunohistochemistry. RESULTS: 67.96% GI-NENs were diagnosed pathologically by endoscopy. Only 26.21% were diagnosed pathologically by biopsies before treatment. The diagnostic rate was significantly higher in polypoid (76.47%) and submucosal lesions (68.75%), than in ulcerative lesions (12.00%). However, biopsies were only taken in 56.31% patients, including 51.52% of polypoid lesions, 35.56% of submucosal lesions and 100.00% of ulcerative lesions. Endoscopic resection removed 61.76% of GI-NENs, including six by CBP, 14 by snare polypectomy with electrocauterization, 28 by endoscopic mucosal resection (EMR) and 15 by endoscopic submucosal dissection (ESD). 51.52% polypoid GI-NENs had infiltrated the submucosa under microscopic examination. CBP had a significantly higher rate of remnant (33.33%) than snare polypectomy with electrocauterization, EMR and ESD (all 0.00%). CONCLUSIONS: Biopsies for all polypoid and submucosal lesions will improve pre-operative diagnosis. The high rate of submucosal infiltration of polypoid GI-NENs determined that CBP was inadequate in the treatment of GI-NENs. Diminutive polypoid GI-NENs that disappeared after CBP had a high risk of remnant and should be closely followed up over the long term.