Design, synthesis and pharmacological evaluation of 4-(3-chloro-4-(3-cyclopropylthioureido)-2-fluorophenoxy)-7-methoxyquinoline-6-carboxamide (WXFL-152): a novel triple angiokinase inhibitor for cancer therapy

Angiokinases, such as vascular endothelial-, fibroblast- and platelet-derived growth factor receptors (VEGFRs, FGFRs and PDGFRs) play crucial roles in tumor angiogenesis. Anti-angiogenesis therapy using multi-angiokinase inhibitor has achieved great success in recent years. In this study, we presented the design, synthesis, target identification, molecular mechanism, pharmacodynamics (PD) and pharmacokinetics (PK) research of a novel triple-angiokinase inhibitor WXFL-152. WXFL-152, identified from a series of 4-oxyquinoline derivatives based on a structure-activity relationship study, inhibited the proliferation of vascular endothelial cells (ECs) and pericytes by blocking the angiokinase signals VEGF/VEGFR2, FGF/FGFRs and PDGF/PDGFR simultaneously . Significant anticancer effects of WXFL-152 were confirmed in multiple preclinical tumor xenograft models, including a patient-derived tumor xenograft (PDX) model. Pharmacokinetic studies of WXFL-152 demonstrated high favourable bioavailability with single-dose and continuous multi-dose by oral administration in rats and beagles. In conclusion, WXFL-152, which is currently in phase Ib clinical trials, is a novel and effective triple-angiokinase inhibitor with clear PD and PK in tumor therapy.

National production of certified reference fungal cultures / Producción nacional de cultivos fúngicos de referencia certificados

Reference fungal cultures (RFCs) are essential for the internal quality control of laboratories. The production of these cultures requires standardized procedures (IRAM 14950:2016 and ISO 17034:2016 standards) carried out by a recognized and accredited laboratory. The aim of this work was to produce RFC in paper disks of autochthonous strains, characterized by two, homogeneous and stable reference methods traceable at species level. RFC were produced using 14 regional species (7 yeasts and 7 filamentous fungi) from the fungal culture collection (DMic). Paper disks were impregnated with a culture suspension, dried and packed. Homogeneity, viability, identity and purity were verified. Short-and long-term stability at different temperatures and storage times were studied. Characterization of each strain allowed to confirm its identity and to ensure its traceability at international level. Produced batches were homogeneous and stable at -20 ±5 °C for 30 months. This method of production was adequate to produce homogeneous and stable RFC with phenotypic and genotypic characteristics correctly defined and internationally traceable. Standardized procedures were developed for the production of certified RFC that could be transferred to other microorganisms. Providing RFC that represent regional strains allows laboratories to produce more reliable results with a favorable impact on medical diagnosis, the environment or the food industry.

Los cultivos microbianos de referencia (CR) son imprescindibles para el control de calidad interno de los laboratorios. Asegurar su producción requiere de procedimientos estandarizados (IRAM 14950:2016 e ISO 17034:2016) realizados en un laboratorio reconocido y acreditado. El objetivo de este estudio fue producir cultivos fúngicos de referencia en discos de papel, a partir de un panel de cultivos autóctonos caracterizados por dos métodos de referencia, trazables a nivel taxonómico de especie, homogéneos y estables. Se produjeron CR de 14 especies circulantes en Argentina (7 de levaduras y 7 de hongos miceliales), depositadas en la colección de hongos de interés médico (DMic). Los discos de papel fueron embebidos con una suspensión del cultivo por producir, secados y envasados. Se verificó la homogeneidad, viabilidad, identidad y pureza de cada lote. Se evaluó la estabilidad a corto y largo plazo a distintas temperaturas y tiempos de almacenamiento. La caracterización de cada CR nos permitió confirmar su identidad y asegurar su trazabilidad a nivel internacional. Los lotes producidos fueron homogéneos y estables durante 30 meses conservados a -20 ±5 °C. Este método resultó adecuado para producir CR homogéneos y estables, con características fenotípicas y genotípicas correctamente definidas y trazables a nivel internacional. Los procedimientos estandarizados desarrollados en este trabajo pueden ser transferidos para producir CR certificados de otros microorganismos. La provisión de CR que represente cepas regionales permite a los laboratorios producir resultados más confiables con un impacto favorable en el diagnóstico médico, los estudios ambientales y la industria alimenticia.

Molecular Analysis of Eight American Type Culture Collection Gonococcal Strains by Neisseria gonorrhoeae Multiantigen Sequence Typing and PorB Sequence Typing

BACKGROUND: Molecular epidemiological typing of Neisseria gonorrhoeae is crucial for monitoring the spread of resistant strains. As reference strains can be used for laboratory internal quality control, we genetically characterised the American Type Culture Collection (ATCC) gonococcal strains by Neisseria gonorrhoeae multiantigen sequence typing (NG-MAST) and porB sequence typing using public multilocus sequence typing (PubMLST). METHODS: Eight ATCC gonococcal reference strains (ATCC 19424, ATCC 31426, ATCC 35541, ATCC 43069, ATCC 43070, ATCC 49226, ATCC 49926, and ATCC 49981) from Culti-Loops (Thermo Fisher Scientific, USA) were cultured. After DNA extraction, porB and tbpB were amplified and sequenced. Sequence types (STs) and allele numbers were each determined by NG-MAST (http://www.ng-mast.net) and porB sequence typing using PubMLST (http://pubmlst.org/neisseria/porB/). RESULTS: ATCC 19424 was identified as ST 266 by NG-MAST, and as Allele 946 by PubMLST. ATCC31426 was assigned a novel ST by NG-MAST, and was assigned Allele 958 with 1.2% mismatch by PubMLST. ATCC 35541 was identified as ST 12 by NG-MAST, and as Allele 624 by PubMLST. ATCC 43069 and ATCC 43070 were both identified as ST 681 by NG-MAST, and as Allele 984 by PubMLST. ATCC 49226 was identified as ST 1572 by NG-MAST, and as Allele 2110 by PubMLST. ATCC 49926 and ATCC 49981 were both identified as ST 16496 by NG-MAST, and as Allele 928 by PubMLST. CONCLUSIONS: The ST data obtained for ATCC gonococcal reference strains by NG-MAST and porB sequence typing using PubMLST can be used for quality assurance of molecular epidemiological typing in clinical microbiological laboratories.

Dual-targeting and microenvironment-responsive micelles as a gene delivery system to improve the sensitivity of glioma to radiotherapy

Dbait is a small double-stranded DNA molecule that has been utilized as a radiosensitizer to enhance the sensitivity of glioma to radiotherapy (RT). However, there is no effective drug delivery system to effectively overcome the blood-brain barrier (BBB). The aim of this study was to develop a gene delivery system by using the BBB and glioma dual-targeting and microenvironment-responsive micelles (ch-K(s-s)R8-An) to deliver Dbait into glioma for RT. Angiopep-2 can target the low-density lipoprotein receptor-related protein-1 (LRP1) that is overexpressed on brain capillary endothelial cells (BCECs) and glioma cells. In particular, due to upregulated matrix metalloproteinase 2 (MMP-2) in the tumor microenvironment, we utilized MMP-2-responsive peptides as the enzymatically degradable linkers to conjugate angiopep-2. The results showed that ch-K(s-s)R8-An micelles maintained a reasonable size (80-160 nm) with a moderate distribution and a decreased mean diameter from the cross-linking as well as exhibited low critical micelle concentration (CMC) with positive surface charge, ranging from 15 to 40 mV. The ch-K5(s-s)R8-An/pEGFP showed high gene transfection efficiency , improved uptake in glioma cells and good biocompatibility and . In addition, the combination of ch-K5(s-s)R8-An/Dbait with RT significantly inhibited the growth of U251 cells . Thus, ch-K5(s-s)R8-An/Dbait may prove to be a promising gene delivery system to target glioma and enhance the efficacy of RT on U251 cells.

Antimicrobials and Antimicrobial Resistant Superbacteria / 이화의대지

Antimicrobials were one of the great invention of modern era. However, the abuse of antimicrobial both in human and animals has led to a high rate of occurrence of antimicrobial resistant microbes. Disease treatment caused by antimicrobial resistant microbes including superbacteria has emerged as critical issue worldwide. Communication and cooperation among researchers in diverse fields are needed to solve the resistance to antimicrobials. Culture Collection of Antimicrobial Resistant Microbes (CCARM) has taken a leadership role an intermediary among various research fields by providing certified antimicrobial resistant microbes with their information since 1999. CCARM collects antimicrobial resistant microbes from clinical, agricultural animals and products, and environmental fields, and classifies and stores them according to their origins, species and antimicrobial resistance mechanisms. CCARM is performing the roles (collection, deposit, preservation, distribution, service, and consulting) of Biological Resource Center designated by Organisation for Economic Co-operation and Development.

Organización, mantenimiento y preservación de la Colección de Cultivos Bacterianos del Instituto de Ciencias Biológicas de la Universidad de Ciencias y Artes de Chiapas (UNICACH), México / Organization, maintenance, and preservation of the Bacterial Culture Collection of the Biological Sciences Institute, University of Science and Arts of Chiapas (UNICACH), Mexico

Los ceparios o colecciones de microorganismos son fuentes de recursos genéticos cuyo propósito es la preservación de la diversidad biológica, garantizando su disponibilidad para actividades de docencia, investigación y comerciales. En este trabajo se verificó la viabilidad, pureza y características biológicas de las bacterias que conforman el cepario del Instituto de Ciencias Biológicas de la Universidad de Ciencias y Artes de Chiapas, y se organizó y estructuró la información obtenida en un portal virtual, para propiciar la cooperación académica. El cepario cuenta con 33 microorganismos, la mayoría del género Streptococcus y Escherichia (45,1 y 21,2%, respectivamente). Del primer género, se confirmó la identificación de S. pyogenes (40%), exhibiendo la mayoría genes que codifican para DNAsas. Con respecto al segundo género, un 58,3% de las bacterias fueron confirmadas taxonómicamente como E. coli. De esta especie, la colección cuenta con las cepas prototipo causantes de diarrea y que han preservado sus rasgos genéticos por más de cinco años. Dicho acervo ha impulsado actividades de docencia e investigación, a nivel local e internacional. Es importante que los ceparios sean fuentes sustentables de recursos biológicos, para la adquisición y suministro de especies bacterianas, con la finalidad de fomentar la interacción con la comunidad académica.

Strain collections or bacterial culture collections are genetic resources whose purpose is the preservation of biological diversity, ensuring their availability for teaching, research and trade activities. In this work viability, purity and biological characteristics of bacteria from the bacterial collection of the Institute of Biological Sciences, University of Science and Arts of Chiapas were studied. Information was structured and organized in a virtual site, to promote academic cooperation. The strain bank includes 33 microorganisms, most of the genus Streptococcus and Escherichia (45.1 and 21.2%, respectively). For Streptococcus, the identification of S. pyogenes (40%) was confirmed, by determination of most DNAses encoding genes. For Escherichia 58.3% were taxonomically confirmed as E. coli. For this species, the collection includes typical strains that produce diarrhea and their genetic traits have been preserved for more than five years. This bacterial culture collection has stimulated teaching and research activities at local and international levels. Strain collections are important sources of biological material which can provide bacterial species, in order to encourage interaction with the academic community.

Design and development of management information system for bacteria (virus) preservation / 军事医学

Objective To establish a management information system for bacteria ( virus ) preservation in order to improve the management efficiency and quality of bacteria ( virus) preservation organizations to the largest possible extent . Methods Client/Server(C/S) structure was adopted to design the system with Visual Basic 6.0 and SQL Server2000 as the platform of development .A bacteria ( virus) information database was established with the E-R model and 3NF.The system was developed by face to object language program .Results Devices needed were supplied .Functional modules of bacteria ( virus ) information management , scientific research project management , user management and system maintenance were constructed .Visiting permission was installed on the platform and database to maximize the protection of bacteria ( virus ) information safety .Conclusion Bacteria ( virus ) information flow management , involving storage , classification, preservation, collection and record, is achieved.All the resources of the preservation organization are integrated.Bacteria (virus) management efficiency is increased and bacteria (virus) information safety is ensured.

Viability and molecular authentication of Coccidioides spp. isolates from the Instituto de Medicina Tropical de São Paulo culture collection, Brazil / Viabilidade e autenticação molecular de isolados de Coccidioides spp. da coleção de culturas do Instituto de Medicina Tropical de São Paulo, Brasil

Coccidioidomycosis is an emerging fungal disease in Brazil; adequate maintenance and authentication of Coccidioides isolates are essential for research into genetic diversity of the environmental organisms, as well as for understanding the human disease. Seventeen Coccidioides isolates maintained under mineral oil since 1975 in the Instituto de Medicina Tropical de São Paulo (IMTSP) culture collection, Brazil, were evaluated with respect to their viability, morphological characteristics and genetic features in order to authenticate these fungal cultures. Only five isolates were viable after almost 30 years, showing typical morphological characteristics, and sequencing analysis using Coi-F and Coi-R primers revealed 99% identity with Coccidioides genera. These five isolates were then preserved in liquid nitrogen and sterile water, and remained viable after two years of storage under these conditions, maintaining the same features.

Coccidioidomicose é uma doença emergente no Brasil; a manutenção adequada e autenticação de isolados de Coccidioides spp são essenciais para a pesquisa em diversidade genética de micro-organismos, bem como para a compreensão da doença em humanos. Dezessete isolados de Coccidioides preservados em óleo mineral desde 1975 na coleção de culturas do Instituto de Medicina Tropical de São Paulo (IMTSP) foram avaliados com relação à viabilidade, características morfológicas e genéticas, com o objetivo de autenticação das culturas fúngicas. Dos 17 isolados, apenas cinco foram viáveis após quase 30 anos mantidos em óleo mineral, apresentando características morfológicas e moleculares típicas do gênero, o sequenciamento utilizando os oligonucleotídeos Coi-F e Coi-R revelou identidade de 99% com isolados de Coccidioides. Estes cinco isolados foram preservados em nitrogênio líquido e água destilada esterilizada, e permaneceram viáveis após dois anos de armazenamento sob estas condições, mantendo as mesmas características.

Potencial antagônico de Trichoderma spp. originários de diferentes agroecossistemas contra Sclerotinia sclerotiorum e Fusarium solani / Antagonist potential of Trichoderma spp. from distinct agricultural ecosystems against Sclerotinia sclerotiorum and Fusarium solani

A partir de 40 amostras de solo, provenientes dos estados do Rio Grande do Sul, Minas Gerais, São Paulo, Bahia, Goiás e Tocantins, uma coleção de 230 isolados monospóricos de Trichoderma spp. foi estabelecida, usando o meio seletivo TSM. Com o objetivo de selecionarem-se isolados com potencial para controle biológico de doenças, foram conduzidos testes de pareamento de culturas em meio BDA, a 20 ºC para Sclerotinia sclerotiorum e a 25 ºC para Fusarium solani f.sp. phaseoli. Antagonismo contra os dois patógenos foi observado em 10 por cento dos isolados. Avaliações ao microscópio eletrônico de varredura de sete isolados selecionados in vitro mostraram que nem todos promoveram o hiperparasitismo dos patógenos, sugerindo a existência de outros mecanismos de antagonismo, como antibiose ou competição.

From 40 soil samples collected in the Rio Grande do Sul, Minas Gerais, São Paulo, Bahia, Goiás and Tocantins states, Brazil, a collection of 230 monosporic isolates of Trichoderma spp. was established using TSM selective media. In order to select efficient isolates for biological control, dual culture tests were carried out on PDA media at 20 ºC for Sclerotinia sclerotiorum and at 25 ºC for Fusarium solani f.sp. phaseoli. From the whole collection, 50 isolates presented antagonism against F. solani and 111 isolates to S. sclerotiorum. The antagonism against both pathogens was found in only 10 percent of the isolates. Scanning electron microscopy assessments with seven in vitro selected isolates showed that not all promoted hyperparasitism on the pathogens, suggesting the existence of other mechanisms of antagonism, as antibiosis or competition.