Structure and antimicrobial activity of phloroglucinol derivatives from Achyrocline satureioides.

The new prenylated phloroglucinol α-pyrones 1-3 and the new dibenzofuran 4, together with the known 23-methyl-6-O-demethylauricepyrone (5), achyrofuran (6), and 5,7-dihydroxy-3,8-dimethoxyflavone (gnaphaliin A), were isolated from the aerial parts of Achyrocline satureioides. Their structures were determined by 1D and 2D NMR spectroscopic studies, while the absolute configuration of the sole stereogenic center of 1 was established by vibrational circular dichroism measurements in comparison to density functional theory calculated data. The same (S) absolute configuration of the α-methylbutyryl chain attached to the phloroglucinol nucleus was assumed for compounds 2-6 based on biogenetic considerations.

Multicomponent synthesis of antibacterial dihydropyridin and dihydropyran embelin derivatives.

A series of dihydropyran and dihydropyridin embelin derivatives were synthesized through a novel and straightforward one-pot protocol based on a three-component reaction with embelin, aldehydes, and cyclic enaminones as synthetic imputs. The type of substituent on the nitrogen atom of the β-enaminone is key to obtain nitrogenated or oxygenated rings. The obtained compounds were active against Gram-positive bacteria, including multiresistant Staphylococcus aureus clinical isolates.

Molecular epidemiology and characterization of virulence genes of community-acquired and hospital-acquired methicillin-resistant Staphylococcus aureus isolates in Colombia.

Objective: To determine the molecular epidemiology and presence of virulence genes in community-acquired (CA) and hospital-acquired (HA) methicillin-resistant Staphylococcus aureus (MRSA) isolates and their relationship to clinical outcomes.

Methods: An observational and prospective study of infections caused by MRSA was conducted between June 2006 and December 2007 across seven hospitals in three Colombian cities. MRSA isolates were analyzed for SCCmec.

Molecular epidemiology of plasmid-mediated high-level mupirocin resistance in methicillin-resistant Staphylococcus aureus in four Spanish health care settings.

Mupirocin is used for the decolonization of methicillin-resistant Staphylococcus aureus (MRSA). High-level mupirocin resistance (Hi-Mup(R)) is of concern, having been associated with therapeutic failure. Our main objective was to assess the emergence and mode/s of spread of Hi-Mup(R) in the MRSA population recovered between 2002 and 2009 in four health care settings in the Pontevedra province, northwest Spain.

Achyrofuran is an antibacterial agent capable of killing methicillin-resistant vancomycin-intermediate Staphylococcus aureus in the nanomolar range.

Currently, there is a pressing need for novel antibacterial agents against drug-resistant bacteria, especially those which have been common in our communities and hospitals, such as methicillin-resistant Staphylococcus aureus (MRSA). The South American plant Achyrocline satureioides ("Marcela") has been widely used in traditional medicine for a number of diseases, including infections. Several crude extracts from this plant have shown good antimicrobial activities in vitro.

Genetic diversity of community-associated methicillin-resistant Staphylococcus aureus isolated from Tenerife Island, Spain.

With the recent detection of MRSA (methicillin-resistant Staphylococcus aureus) infections in patients lacking health care-related risk factors, the term community-acquired MRSA (CA-MRSA) has been become widely recognised. Many cases of CA-MRSA spreading to the community have been described worldwide. The aim of this study was to determine the features of CA-MRSA isolates from Tenerife Island.

New Staphylococcus aureus genetic cluster associated with infectious osteomyelitis.

Diverse genotyping methods, including multiple-locus variable-number tandem-repeat (VNTR) analysis (MLVA), pulsed field gel electrophoresis (PFGE), and multilocus sequence typing (MLST), were used for genotyping Staphylococcus aureus in samples recovered from a clinical case of osteomyelitis. An unexpected genetic diversity of strains was determined, including four new sequence types (ST 1521, 1522, 1628 and 1629) belonging to the same genetic lineage, implying the appearance of a new subgroup derived from clonal complex CC121 isolated from that hospital. A close phylogenetic relationship among the STs was demonstrated, reflecting a possible diversifying evolution process.

PCR-based amplification of heterogeneous IS257-ileS2 junctions for molecular monitoring of high-level mupirocin resistance in staphylococci.

Objectives: Plasmids encoding the ileS2 gene are responsible for the wide spread of high-level mupirocin resistance in staphylococci, and consequent clinical and epidemiological problems. We investigated the location of insertion sequence IS257 flanking ileS2 in different plasmids and developed a method for molecular typing of the IS257-ileS2 spacer regions.

Methods: Nine ileS2-encoding plasmids (i.

Evaluation of multiple-locus variable-number tandem-repeat analysis for typing a polyclonal hospital-acquired methicillin-resistant Staphylococcus aureus population in an area where such infections are endemic.

Multiple-locus variable-number tandem-repeat analysis (MLVA) was performed with 292 methicillin-resistant Staphylococcus aureus (MRSA) isolates previously characterized by pulsed-field gel electrophoresis, multilocus sequence typing, and staphylococcal cassette chromosome mec typing. Quantitative correspondence analyses showed the best correlation between data when an >or=80% cutoff was applied to MLVA. We confirmed the validity of MLVA for identification of related strains in a polyclonal MRSA population.

Complete nucleotide sequence and comparative analysis of pPR9, a 41.7-kilobase conjugative staphylococcal multiresistance plasmid conferring high-level mupirocin resistance.

We have sequenced the conjugative plasmid pPR9, which carries the ileS2 gene, which had contributed to the dissemination of high-level mupirocin resistance at our institution. The plasmid backbone shows extensive genetic conservation with plasmids belonging to the pSK41/pGO1 family, but comparative analyses have revealed key differences that provide important insights into the evolution of these medically important plasmids and high-level mupirocin resistance in staphylococci and highlight the role of insertion sequence IS257 in these processes.

Nosocomial infections caused by community-associated methicillin-resistant Staphylococcus aureus in Colombia.

Background: Community-associated methicillin-resistant Staphylococcus aureus strains (CA-MRSA) have emerged as the causative agent of health care-associated infections.

Methods: An observational and prospective study was carried out in 5 hospitals in Bogotá, Colombia; severe MRSA infections were identified, and their origin led to classification as health care-associated (HA-MRSA), community-associated, or nosocomial infections. MRSA isolates were analyzed by SCCmec, pulsed-field gel electrophoresis, multilocus sequence typing, and virulence factors.

Molecular characterization of Alternaria alternata causing ocular infection: detection of IGS-RFLP intraspecific polymorphism.

A case of fungal keratitis is presented in which corneal scrapings were obtained for microbiological studies, including morphological identification and molecular characterization of the etiologic agent. Comparative sequence analyses of the Internal Transcribed Spacer domain of 5.8S and 26S regions of nuclear rDNA showed 100% identity with different species of Alternaria and PCR-RFLP analysis of Intergenic Spacer regions revealed intraspecific variation.

Spatial heterogeneity of bacterial populations in monomictic Lake Estanya (Huesca, Spain).

Bacterial population changes were investigated in the monomictic Lake Estanya by combining microscopic analysis and two molecular methods involving the amplification of 16S rDNA genes using primers for the domain Bacteria and subsequent restriction fragment length polymorphism (PCR-RFLP) and denaturing gradient gel electrophoresis (PCR-DGGE). Both approaches revealed the vertical distribution of predominant microbial morphotypes and phylotypes in both holomictic and stratified periods, respectively, and showed that variations in structure and composition of bacterial populations are occurring in this lake as a function of depth and time. Through principal component analysis (PCA), these shifts could be related to different physicochemical parameters with temperature, oxygen concentration, and the incident light being of paramount importance as structuring variables.

High-level mupirocin resistance within methicillin-resistant Staphylococcus aureus pandemic lineages.

The methicillin-resistant Staphylococcus aureus (MRSA) population in the Hospital Universitario Nuestra Señora de Candelaria over a 5-year period (1998 to 2002) was marked by shifts in the circulation of pandemic clones. Here, we investigated the emergence of high-level mupirocin resistance (Hi-Mup(r)). In addition to clonal spread, transfer of ileS2-carrying plasmids played a significant role in the dissemination of Hi-Mup(r) among pandemic MRSA lineages.

Seasonal microbial ribotype shifts in the sulfurous karstic lakes Cisó and Vilar, in northeastern Spain.

Spatio-temporal changes in two sulfurous lakes from the karstic area of Banyoles (Girona, Spain), holomictic lake Cisó and meromictic lake Vilar, were studied over one year. Samples were collected at different depths from the two lakes on the same days, during each of the four seasons, and several physico-chemical variables (temperature, light, pH, conductivity, sulfide, oxygen concentration, pigment concentrations, etc.) were measured.

PCR protocol for specific identification of Candida nivariensis, a recently described pathogenic yeast.

Candida nivariensis is a recently described pathogenic yeast closely related to Candida glabrata. We developed a specific set of oligonucleotide primers based on the internal transcribed spacer regions of the rRNA gene for the rapid identification of C. nivariensis.

Phenotypic and molecular characterization of Candida nivariensis sp. nov., a possible new opportunistic fungus.

The new species Candida nivariensis, isolated from the clinical samples of three patients in Spain over a 3-year period, is presented here. This species can be easily differentiated from Candida glabrata, the closest genetic species, by different colony color on CHROMagar and by its ability to ferment trehalose. The analyses of the internal transcribed spacer region and the D1-D2 region of the 26S rRNA gene sequences support a new species designation.

Outbreak of Shigella sonnei in a rural hotel in La Gomera, Canary Islands, Spain.

Shigella sonnei is a significant cause of gastroenteritis in both developing and industrialized countries. Knowledge of the diversity and antimicrobial susceptibility of the bacterium may be helpful in the management of both individual cases and outbreaks. This study was undertaken to evaluate the molecular epidemiology of an outbreak of diarrhea due to S.

Tracking methicillin-resistant Staphylococcus aureus clones during a 5-year period (1998 to 2002) in a Spanish hospital.

Three hundred seventy-five consecutive methicillin-resistant Staphylococcus aureus (MRSA) clinical isolates recovered between 1998 and 2002 at the Nuestra Señora de Candelaria University Hospital in Tenerife, Spain, were analyzed by molecular fingerprinting techniques to determine MRSA clonal types and their prevalence over time. After determining antibiotic susceptibility, we used SmaI-digested genomic DNA separated by pulsed-field gel electrophoresis (PFGE) to characterize MRSA isolates and to establish PFGE types. Additionally, several selected isolates representative of each major PFGE type were tested by multilocus sequence typing (MLST) and by a multiplex PCR method capable of identifying the structural type of the staphylococcal cassette chromosome mec (SCCmec), generating the corresponding sequence type (ST)-SCCmec types.

Simultaneous PCR detection of ica cluster and methicillin and mupirocin resistance genes in catheter-isolated Staphylococcus.

Recent data show that more than 50% of catheter-associated bloodstream infections are caused by staphylococci. Staphylococcal infections produced by intercellular-adhesion cluster (ica) carriers can be even more problematic due to the presence of methicillin and mupirocin resistance genes. In the present study, a multiplex PCR protocol that allows the simultaneous identification of staphylococci and detection of both the ica and methicillin and/or mupirocin resistance genes was developed.

Biofilm: the microbial "bunker" for intravascular catheter-related infection.

Catheter-related infection in cancer patients remains an important health-care problem with major financial implications. During the last few years a better understanding of the pathogenesis of catheter-related infections and the interaction between microorganisms and catheter surfaces has emerged. Recently the influence of biofilm formation in catheter-related infections has been established.

Characterization of the first VanB vancomycin-resistant Enterococcus faecium isolated in a Spanish hospital.

We report the detection and characterization of the first vancomycin-resistant VanB-type Enterococcus faecium to be isolated in a Spanish hospital. Sequence analysis of the vanB gene showed that this isolate belonged to subtype vanB2. Moreover, PCR amplification analysis indicated that the vanB gene cluster was linked to a Tn5382-like transposon.

Factors affecting interpretation of restriction fragment length polymorphism (RFLP) patterns from PCR-amplified bacterial 16S rRNA genes: operon number and primer mismatching.

PCR methods have been shown to be biased by several factors. In the present study, we have developed a theoretic and practical approximation to elucidate how the presence of mismatches at the primers annealing regions and the different number of rDNA operons per cell can influence PCR and subsequent restriction fragment length polymorphism (RFLP) analyses from bacterial populations. We have performed RFLP analyses of 16S rRNA genes amplified by PCR from mixed bacterial cultures showing different primer identities and number of rDNA operons.

[Multiplex PCR in clinical microbiology].

The introduction of molecular biology methods in clinical microbiology laboratories brings important insights to obtain sensitive and specific diagnoses as fast as possible. These methods are not intended for replacement but for complement of the already applied microbiologic methods. The integrated analyses of all of them is bringing to the most feasible and efficient results.