Biomedical subjects

M Ramirez

Publications and source records attributed to M Ramirez.

At least 19 recordsLinked to original sources

PubMed2006

DNA methylase activity as a marker for the presence of a family of phage-like elements conferring efflux-mediated macrolide resistance in streptococci.

Recently, two related chimeric genetic elements (Tn1207.3 and Phi10394.4) were shown to carry the macrolide efflux gene mef in Streptococcus pyogenes (group A streptococci [GAS]). The dissemination of elements belonging to the Tn1207.3/Phi10394.4 family in recent isolates of GAS, Streptococcus dysgalactiae subsp. equisimilis, Streptococcus pneumoniae, and Streptococcus agalactiae recovered in Portugal was surveyed. In total, 149 GAS, 18 S. pneumoniae, 4 S. dysgalactiae subsp. equisimilis, and 5 S. agalactiae isolates from infections, presenting the M phenotype of macrolide resistance and containing the mef gene, were screened for the presence of Tn1207.3/Phi10394.4 by PCR targeting open reading frames (ORFs) specific for these related elements. All the GAS isolates tested and one of the S. dysgalactiae subsp. equisimilis isolates carried Tn1207.3. However, neither of these elements was found in the isolates of the other streptococcal species. It was also noted that the DNAs of the isolates carrying Tn1207.3 were resistant to cleavage by the endonuclease SmaI. Cloning and expression of ORF12 of Tn1207.3 in Escherichia coli showed that it encoded a methyltransferase that rendered DNA refractory to cleavage by SmaI (M.Spy10394I). Using this characteristic as a marker for the presence of the Tn1207.3/Phi10394.4 family, we reviewed the literature and concluded that these genetic elements are widely distributed among tetracycline-susceptible GAS isolates presenting the M phenotype from diverse geographic origins and may have played an important role in the dissemination of macrolide resistance in this species.

Anti-Bacterial Agents↗

PubMed2006

Heterogeneity of pneumococcal phase variants in invasive human infections.

BACKGROUND: Streptococcus pneumoniae can be carried asymptomatically in the nasopharynx of its human host but can also cause a wide range of infections. A role for pneumococcal phase variants in the different lifestyles of this bacterium has been suggested but no systematic survey of the colony phenotypes of isolates associated with human infections has been undertaken. RESULTS: We report the colony opacity phenotypes of a genetically diverse set of 304 invasive isolates representing 10 serotypes. Over half of the isolates (52%) presented the opaque phenotype whereas transparent variants accounted for only 26% of the total. However, the frequency of recovery of each phase variant was not uniform, while serotypes 1, 4, 12B and 23F presented the opaque phenotype more frequently than expected by chance, serotypes 3 and 14 where less frequently associated with this phenotype. CONCLUSION: The opaque phenotype was the most frequent phenotype found among invasive isolates. An unexpected and equally important finding is the variability of the dominant opacity phenotype found among serotypes. This observation highlights the heterogeneity of opacity phenotypes in invasive isolates and lends further support to the proposal that other factors, in addition to the site of isolation, determine the opacity phenotype of a given isolate. The association between serotype and colonial opacity could help explain epidemiological differences observed among pneumococcal serotypes such as a higher invasive disease potential.

Animals↗

PubMed2006

Emergence of optochin resistance among Streptococcus pneumoniae in Portugal.

In most clinical microbiology laboratories optochin susceptibility is used in the screening and identification of Streptococcus pneumoniae. We report the characterization of 32 optochin-resistant S. pneumoniae strains from 10 laboratories that constituted 3.2% of all isolates recovered in 2005 in 30 laboratories in Portugal. Resistant isolates consisted of bile-soluble optochin-susceptible and optochin-resistant subpopulations with identical antimicrobial susceptibility patterns, capsular types and pulsed-field gel electrophoresis (PFGE) profiles. The most frequent serotypes--1, 19A, 11A, 3, 8, and 15A--were all common serotypes present in infection and colonization isolates in the country. The PFGE profiles of the 32 isolates corresponded to those of previously identified clones and confirmed that the emergence of these strains could not be attributed to clonal expansion. Clinical laboratories must be aware that optochin-resistant pneumococci are presently circulating in the community. Because accurate identification of S. pneumoniae is essential for correct diagnosis and adequate therapy of patients, we recommend that at least the bile solubility test should be routinely performed in cases of suspected pneumococcal etiology, even if the isolates are optochin-resistant.

Anti-Bacterial Agents↗

PubMed2006

Identification of macrolide-resistant clones of Streptococcus pyogenes in Portugal.

Although the overall level of macrolide resistance (27%) has remained stable in Portugal, a rapid inversion in the dominant phenotypes has been noted, with a sharp decrease in the MLS(B) phenotype paralleled by an increase in the M phenotype. To gain further insight into these changes, 325 macrolide-resistant isolates were characterised using a combination of pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). The use of Cfr9I, an isoschizomer of SmaI, to digest M phenotype isolates that were refractory to SmaI digestion allowed direct comparison of MLS(B) and M isolates. The results from PFGE and MLST were highly concordant and identified eight major clones, accounting for 92% of the isolates, each of which was associated exclusively with a single macrolide resistance phenotype. Two major clones were found among MLS(B) isolates, characterised by sequence types (ST) 46 (T12/emm22) and ST52 (T28/emm28), whereas clones characterised by ST39 (T4/emm4) and ST28 (T1/emm1) dominated among M isolates. The clone defined by ST52 corresponded to a bacitracin-resistant clone circulating in Europe, and a novel variant expressing other surface antigens (T12/emm22) was detected. The presence of the four major clones has been reported previously in other European countries, suggesting Europe-wide dissemination of a few macrolide-resistant lineages.

Bacitracin↗

PubMed2006

Evaluation of disk diffusion method for determining posaconazole susceptibility of filamentous fungi: comparison with CLSI broth microdilution method.

The disk diffusion method was evaluated for determining posaconazole susceptibility against 78 strains of molds using two culture media in comparison with the CLSI (Clinical Laboratory Standards Institute) broth microdilution method (M38-A). A significant correlation between disk diffusion and microdilution methods was observed with both culture media.

Antifungal Agents↗

PubMed2006

Illustration of a common framework for relating multiple typing methods by application to macrolide-resistant Streptococcus pyogenes.

The studies that correlate the results obtained by different typing methodologies rely solely on qualitative comparisons of the groups defined by each methodology. We propose a framework of measures for the quantitative assessment of correspondences between different typing methods as a first step to the global mapping of type equivalences. A collection of 325 macrolide-resistant Streptococcus pyogenes isolates associated with pharyngitis cases in Portugal was used to benchmark the proposed measures. All isolates were characterized by macrolide resistance phenotyping, T serotyping, emm sequence typing, and pulsed-field gel electrophoresis (PFGE), using SmaI or Cfr9I and SfiI. A subset of 41 isolates, representing each PFGE cluster, was also characterized by multilocus sequence typing (MLST). The application of Adjusted Rand and Wallace indices allowed the evaluation of the strength and the directionality of the correspondences between the various typing methods and showed that if PFGE or MLST data are available one can confidently predict the emm type (Wallace coefficients of 0.952 for both methods). In contrast, emm typing was a poor predictor of PFGE cluster or MLST sequence type (Wallace coefficients of 0.803 and 0.655, respectively). This was confirmed by the analysis of the larger data set available from http://spyogenes.mlst.net and underscores the necessity of performing PFGE or MLST to unambiguously define clones in S. pyogenes.

Anti-Bacterial Agents↗

PubMed2006

Clonal relationships between invasive and noninvasive Lancefield group C and G streptococci and emm-specific differences in invasiveness.

Lancefield group G and group C streptococci (GGS and GCS, respectively) are pathogens responsible for a number of life-threatening infections. A collection of 116 recent (1998 to 2004) invasive (n = 28) and noninvasive (n = 88) GGS and GCS clinical isolates from Portugal were characterized. All isolates were identified as Streptococcus dysgalactiae subsp. equisimilis and characterized by emm typing and DNA macrorestriction profiling using pulsed-field gel electrophoresis (PFGE). emm typing revealed the presence of 22 distinct types, including 3 novel types. PFGE identified 14 clones with more than two isolates, but over half of the isolates were concentrated in 3 large clones. Individual clones and emm types showed a low level of association, since the majority of the clones included more than one emm type and the same emm type was found among diverse genetic backgrounds. Two emm types, stg2078 and stg10, were significantly more frequent among invasive isolates, and another two, stg6792 and stg166b, were present only in noninvasive isolates, suggesting a correlation between emm type and invasive disease potential.

Bacterial Typing Techniques↗

PubMed2005

Copper bioavailability in a coastal environment of Northern Chile: comparison of bioassay and analytical speciation approaches.

An integrated approach including chemical speciation analyses and microalgal bioassays was used to assess the impact of copper from copper mining on a coastal area in Northern Chile. Dissolved copper ranged from <1 microg l(-1) at reference sites to 48 microgl(-1) at sites close to the mine discharge. Dissolved copper at sites closest to the discharge always exceeded seawater complexing capacities determined by anodic stripping voltammetry (ASV), and consequently labile copper was always detected (1-37 microg l(-1)). Agreement between ASV-labile copper and copper retained by cation exchange (ALSA) columns was excellent. Measured labile copper also accurately predicted bioavailable copper determined by growth inhibition of Nitzschia closterium and enzyme inhibition in Dunaliella tertiolecta. Seawater from Caleta La Lancha had the highest dissolved and labile copper and was the most toxic to micro-algal growth and enzyme activity. Previous studies at this site confirmed it had the lowest level of biodiversity, suggesting that copper may have both direct and indirect effects on these communities.

Biological Assay↗

PubMed2005

Dendritic cell immunizations alone or combined with low doses of interleukin-2 induce specific immune responses in melanoma patients.

Dendritic cell (DC)-based therapy has proved to be effective in patients with a variety of malignancies. However, an optimal immunization protocol using DCs and the best means for delivering antigens has not yet been described. In this study, 20 patients with malignant melanoma in stages III or IV were vaccinated with autologous DCs pulsed with a melanoma cell lysate, alone (n = 13) or in combination with low doses of subcutaneous (s.c.) interleukin (IL)-2 injections (n = 7), to assess toxicity, immunological and clinical responses. Monocyte-derived DCs were morphological, phenotypic and functionally characterized in vitro. Peripheral blood mononuclear cells (PBMC), harvested from patients either prior to and after the treatment, were analysed using enzyme-linked immunosorbent spot (ELISPOT). After vaccination, 50% of the patients tested (seven of 13) from the first group and (three of seven) from the second, showed an increase in interferon (IFN)-gamma production in response to allogeneic melanoma cell lines but not to controls. Four of five tested human leucocyte antigen (HLA)-A2(+) patients with anti-melanoma activity also showed specific T cell responses against peptides derived from melanoma-associated antigens. Delayed type IV hypersensitivity reaction (DTH) against melanoma cell lysate was observed in six of 13 patients from the group treated with DC vaccines only and four of seven from the group treated with the combination of DCs and IL-2. Significant correlations were found between DTH-positive responses against tumour lysate and both disease stability and post-vaccination survival on the stage IV patients. There were no toxicities associated with the vaccines or evidence of autoimmunity including vitiligo. Furthermore, no significant enhancement was observed as a result of combining DC vaccination with IL-2. Our data suggest that autologous DCs pulsed with tumour lysate may provide a standardized and widely applicable source of melanoma specific antigens for clinical use. It is safe and causes no significant side effects and has been demonstrated to be partially efficient at triggering effective anti-melanoma immunity.

Antigen Presentation↗

PubMed2005

Rapid inversion of the prevalences of macrolide resistance phenotypes paralleled by a diversification of T and emm types among Streptococcus pyogenes in Portugal.

In Portugal erythromycin resistance of 26.6% (n = 352) remained constant during 1998 to 2003, however in 1998 the MLS(B) phenotype dominated (85%), whereas in 2003 the M phenotype prevailed (77%). A decline in T12/emm22 MLS(B) isolates could partially explain the drop in this phenotype, but the rise of the M phenotype was not due to clonal expansion.

Anti-Bacterial Agents↗

PubMed2005

Characterization of the genetic lineages responsible for pneumococcal invasive disease in Portugal.

The availability of a conjugate vaccine has the potential to reduce the disease burden of pneumococci and to alter the serotype frequency in the disease-causing population through immunoselection. These changes will probably be reflected in the distributions of individual genetic lineages within the population. We present a characterization of a collection of recent (1999 to 2002) invasive isolates from Portugal (n = 465) by macrorestriction profiling with pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing. During this time, serotypes 14, 1, 3, 4, 8, 9V, 23F, 7F, 19A, and 12B were the 10 most prevalent overall by decreasing rank order. By combining the PFGE data with the sequence types (STs) of 104 isolates, we were able to identify the genetic lineages of the majority of the isolates. We found 66 STs, including 20 novel STs, corresponding to 47 different lineages by e-BURST analysis. We found in our collection a number of previously identified internationally disseminated lineages, especially among macrolide-resistant and penicillin-resistant isolates, and these accounted for most of the isolates. Most of the major lineages (17 of 25) were identified in all years of the study, suggesting that the pneumococcal population associated with invasive disease was stable. This study provides a characterization of the pneumococcal population associated with invasive disease that will be useful for detecting potential selective effects of the novel conjugate vaccine.

Alleles↗

PubMed2005

Results of a national study on anti-mycobacterial drug resistance in El Salvador.

SETTING: The DOTS strategy was introduced in El Salvador in 1997 and had become fully implemented countrywide by 2001. Previously, many deficiencies were identified in the management of patients with tuberculosis (TB). OBJECTIVE: To ascertain the prevalence of Mycobacterium tuberculosis resistant to first-line drugs. DESIGN: A national prospective survey was carried out in 2001 using the standardised World Health Organization/ International Union Against Tuberculosis and Lung Disease model. RESULTS: A total of 711 cultures were analysed (59% of total smear-positive cases); 611 were never treated and 100 were previously treated patients. The study showed resistance rates to isoniazid and rifampicin, either alone or combined (multidrug resistance, MDR), of 1.3%, 1.1% and 0.3%, respectively, in never treated patients, and 12%, 13% and 7%, respectively, in previously treated patients. CONCLUSION: The low rates of MDR-TB in El Salvador are puzzling, as out-patient DOTS was introduced only recently and fixed-dose combination tablets have not been used.

Age Distribution↗

PubMed2004

A prospective randomized study of clinical and economic consequences of using G-CSF following autologous peripheral blood progenitor cell (PBPC) transplantation in children.

This prospective and randomized study was conducted to evaluate clinical and economic consequences of using granulocyte colony-stimulating factor (G-CSF) following autologous peripheral blood progenitor cell (PBPC) transplantation in children. Between January 1999 and December 2003, 117 patients underwent autologous PBPCT: 51 patients received G-CSF following PBPCT, while 66 patients did not receive G-CSF. Median time to absolute neutrophil count > 0.5 x 10(9)/l was 10 days in the treatment group and 11 days in the control group (P < 0.009). The median time to platelets >20 x 10(9)/l was 12 days in both groups (P = NS). The median time to platelets >50 x 10(9)/l was 15 days in the G-CSF group and 14 days in the control group (P<0.005). In patients who received <5 x 10(6)/kg CD34+ cells, the median time to platelets >20 x 10(9)/l and >50 x 10(9)/l was similar with or without G-CSF (12 and 15 days, respectively). Platelet transfusion requirements were lower in the control group (2 vs 3 U in G-CSF group). There was a trend towards higher total costs with G-CSF: 8146.82 Euros and 7873.34 Euros with and without G-CSF, respectively (P = 0.1). Our data suggest that there is no indication of the standard application of G-CSF in children following PBPC transplantation. The only possible indication is the group of patients with a lower yield of CD34+ cells.

Adolescent↗

PubMed2004

Streptococcus agalactiae in a large Portuguese teaching hospital: antimicrobial susceptibility, serotype distribution, and clonal analysis of macrolide-resistant isolates.

Group B streptococci are emerging as a cause of serious infection worldwide. The capsular polysaccharides are not only important virulence factors but also the target of vaccine development efforts. Serotypes III (24.6%), V (23.4%), Ia (17.8%), and II (16.3%) were the most prevalent among 252 Streptococcus agalactiae isolates collected during 1999-2002 in the largest hospital of Lisbon, Portugal. The substantial proportion of bacteremic patients (17 neonates and 21 adults) in this period illustrates the present importance of S. agalactiae as a cause of invasive disease. All isolates were fully susceptible to penicillin (MIC(50) = 0.064 microg/ml; MIC(90) = 0.094 microg/ml, range 0.008-0.094), cefotaxime, chloramphenicol, ofloxacin, and vancomycin. Resistance was found to tetracycline (75.4%), erythromycin (10.7%), and clindamycin (9.9%). Of the 27 erythromycin-resistant isolates, 70.4% had the cMLS(B), 22.2% the iMLS(B), and 7.4% the M phenotype. All isolates presenting the M phenotype carried the mef(A) gene, whereas the erm(B) gene was found in a large fraction of MLS(B) isolates (n = 17) and only a small proportion (n = 7) the erm(A) gene [erm(TR) variant]. All isolates carried a single macrolide-resistance determinant. Macrolide resistance was not attributable to a single clone as evidenced by distinct serotype and pulsed-field gel electrophoretic profiles. Careful surveillance of S. agalactiae invasive infections in Portugal is essential, and the treatment or intrapartum prophylaxis of patients who are allergic to penicillin should be guided by contemporary resistance patterns observed in the country.

Anti-Bacterial Agents↗

PubMed2004

Invasive Streptococcus pneumoniae from Portugal: implications for vaccination and antimicrobial therapy.

The distribution of pneumococcal serotypes among 465 invasive isolates recovered from 1999 to 2002 in Portugal was analysed by age group. Serotype 14 was either the most prevalent or the second most prevalent in all age groups. Among children aged < 2 years, serotypes 6B and 23F, which are usually associated with children, together with serotypes 19A and 14, accounted for more than half of the isolates. In contrast, in older adults (> or = 60 years), serotypes 3, 14, 1, 8 and 4 were the most prevalent. The potential coverage of the seven-valent conjugate vaccine is 63.2% among infants, and does not change significantly if children aged < 6 years are considered, which is a lower coverage than in other European countries. The potential coverage of the 23-valent polysaccharide vaccine is high in all age groups, particularly among older adults (80.7%). All isolates were tested for their susceptibility to penicillin, cefuroxime, cefotaxime, vancomycin, erythromycin, clindamycin, levofloxacin, gatifloxacin, moxifloxacin, linezolid, quinupristin-dalfopristin, tetracycline, chloramphenicol and trimethoprim-sulphamethoxazole. Most isolates collected from children aged < 6 years had decreased susceptibility to at least one antibiotic class, whereas isolates from patients aged > or = 6 years were mostly susceptible to all antimicrobial agents tested. Overall, 23% of isolates showed reduced susceptibility to penicillin. Most (98.5%) isolates remained fully susceptible to cefotaxime, and a single isolate was resistant to quinolones.

Age Distribution↗

PubMed2004

Disability and risk of school related injury.

OBJECTIVE: Approximately six million children with disabilities attend school in the United States. Cognitive and physical limitations may compromise their ability to handle environmental hazards and hence increase their risk for injury. The objective of this study was to describe the epidemiology of school related injury among children enrolled in 17 special education schools in one large, urban school district. DESIGN: Altogether 6769 schoolchildren with disabilities were followed up from 1994-98. Injury and population data were collected from pupil accident reports and existing school records. Associations were estimated through generalized estimating equations. RESULTS: A total of 697 injuries were reported for a rate of 4.7/100 students per year. Children with multiple disabilities had a 70% increased odds of injury compared with the developmentally disabled (odds ratio (OR) 1.7, 95% confidence interval (CI) 1.3 to 2.3). The physically disabled (OR 1.4, 95% CI 1.0 to 1.9) had a modest increased odds of injury. Cuts, bruises, and abrasions composed almost three fourths of all injuries; almost half of these injuries were to the face. Falls (34%) and insults by other students (31%) were the most common external causes. More than a fourth of injuries were sports related, and 21% occurred on the playground/athletic field. Injury patterns differed across disabilities. CONCLUSIONS: Although limited to one school district, the population studied is the largest cohort thus far of schoolchildren with disabilities. With this large study base, potentially high risk groups were identified and circumstances of injury described. This information is imperative for developing and improving school based injury prevention measures.

Accidental Falls↗

PubMed2004

Effects of culture and education on neuropsychological testing: a preliminary study with indigenous and nonindigenous population.

We analyzed the influence of education and of culture on the neuropsychological profile of an indigenous and a nonindigenous population. The sample included 27 individuals divided into four groups: (a) seven illiterate Maya indigenous participants, (b) six illiterate Pame indigenous participants, (c) seven nonindigenous participants with no education, and (d) seven Maya indigenous participants with 1 to 4 years of education . A brief neuropsychological test battery developed and standardized in Mexico was individually administered. Results demonstrated differential effects for both variables. Both groups of indigenous participants (Maya and Pame) obtained higher scores in visuospatial tasks, and the level of education had significant effects on working and verbal memory. Our data suggested that culture dictates what it is important for survival and that education could be considered as a type of subculture that facilitates the development of certain skills.

Adult↗

PubMed2003

Survivorship analysis of Cotrel-Dubousset instrumentation in idiopathic scoliosis.

This study presents a survivorship analysis of Cotrel-Dubousset instrumentation in the surgical treatment of idiopathic scoliosis. Between 1987 and 1995, a total of 133 patients with idiopathic scoliosis received posterior spine fusion and instrumentation with the CD system at our center. The patients' mean age at surgery was 16.5 years (range 11-43 years). The magnitude of the thoracic scoliosis averaged 62.7 degrees (range 40 degrees -125 degrees ) and that of the lumbar curve was 58.8 degrees (range 40 degrees -100 degrees ). On average, 12.2 segments were fused (range 8-17) and, excluding the rods, 14.1 implants were set for each patient (range 10-21). Survivorship analysis was carried out using the Kaplan-Meier method. Implant removal was considered the terminal event, or "death". The effect of several variables on survival rate was determined with the Cox regression method. The patients remained in the study for 56.7 months (range 2-120 months). One-hundred and ten patients were withdrawn ("censored"): 90 "alive" (did not require repeat surgery and attended follow-up control in 1997) and 20 "lost" (did not attend control in 1997). Twenty-three patients attained the terminal event of implant removal for a variety of reasons: acute infection (three cases), late infection (ten cases), implant failure requiring revision (six cases) and local pain (four cases). The survival rate was 95.5% at 3 months, 94.7% at 6 months, 93.9% at 1 year, 91.5% at 2 years, 82.2% at 5 years and 76.5% at 10 years. The magnitude of the curves, total number of implants and number of fused segments did not correlate with survival probability. A positive correlation was found between survival rate and correction loss between surgery and last control. A survival rate of 76.5% at 10 years is unexpectedly low. Current data suggest that the incapacity to maintain correction after initial surgery plays a major roll in the long-term evolution of Cotrel-Dubousset instrumentation.

Adolescent↗