IL-6/IL-10 and IL-1ß/IL-4 ratios associated with poor ovarian response in women undergoing in-vitro fertilization.

The aim of this work was to evaluate whether serum cytokines levels are associated with ovarian response in IVF. 149 patients were included in a retrospective study. Cytokines IL-2, IL-4, IL- 6, IL-8, IL-10, VEGF, IFNγ, TNFα, IL-1α, IL-1ß, MCP-1 and EGF were measured by sandwich immunoassay previously to ovarian stimulation. Performing hierarchical cluster analysis, we observed that the antral follicle count, the total oocytes recovered and the MII recovered are grouped in the same cluster as the cytokines IL-2-4-6-10-1α-1ß, IFNγ y TNFα. Then, we found that the ratio between IL and 6 and IL-10 was higher in low responder women (2.15 versus 1.55; p = 0.035). If we establish 0.9 as a cut-off for the IL-6/IL-10, we observed that above this value the risk of having a low response to ovarian stimulation was more than 3 times greater than below this value (22.9 % versus 6.0 %; p = 0.007). Also, the ratio IL-1ß/IL-4 was higher in patients with normal or suboptimal response (0.62 versus 0.34; p = 0.034) and any patient with low response had a value greater than 0.7 (p = 0.003). As a conclusion, the IL-6/IL-10 and IL-1ß/IL-4 ratios showed differences between normoresponder women and patients with low ovarian response.

Identification of vaginal microbiome associated with IVF pregnancy.

The factors that cause a preterm birth (PTB) are not completely understood up to date. Moreover, PTB is more common in pregnancies achieved by in-vitro fertilization (IVF) than in spontaneous pregnancies. Our aim was to compare the composition of vaginal microbiome at 12 weeks of gestation between women who conceived naturally or through IVF in order to study whether IVF PTB-risk could be related to vaginal microbiome composition. We performed an observational, prospective and multicentre study among two public hospitals and a fertility private clinic in Spain. Vaginal swabs from 64 pregnant women at 12 weeks of gestation were collected to analyse the microbiome composition by sequencing the V3-V4 region of the 16S rRNA. Our results showed that the vaginal microbiome signature at 12 weeks of pregnancy was different from women who conceived naturally or through IVF. The beta diversity and the genus composition were different between both cohorts. Gardnerella, Neisseria, Prevotella, and Staphylococcus genus were enriched genus in the vaginal microbiome from the IVF group, allowing us to create a balance model to predict both cohorts. Moreover, at species level the L. iners abundance was higher and L. gasseri was lower in the IVF group. As a conclusion, our findings were consistent with a proposed framework in which IVF pregnancy are related to risk for preterm birth (PTB) suggesting vaginal microbiome could be the reason to the relation between IVF pregnancy and risk for PTB.

Unravelling potential biomedical applications of the edible flower Tulbaghia violacea.

Traditionally, edible flowers have been used as foods and for therapeutic purposes, today they have gained importance due to their bioactive compounds such as flavonols, anthocyanins or other phenolic compounds, which give them potential for biomedical applications. This work evaluated a methanolic extract of Tulbaghia violacea. Eleven individual phenolic compounds were found and quantified by mass spectrometry in the extract. Antioxidant activity tests (TEAC, FRAP and DPPH) and other characterization parameters were assayed (total phenolic content and total flavonoid content). In vitro studies showed antitumoral activity against ovarian tumoral cells mediated by the induction of non-dependent caspase cell death and by the activation of reactive oxygen species. The effect of the extract against features of Alzheimer disease was in vivo assayed in Caenorhabditis elegans. Tulbaghia extract led to a reduction in the 1-42 beta amyloid peptide formation and prevented oxidative stress. These results suggested that Tulbaghia violacea could be a new source of phenolic compounds for nutraceuticals and functional food development.

Evolution of the capsular gene locus of Streptococcus pneumoniae serogroup 6.

Streptococcus pneumoniae expressing serogroup 6 capsules frequently causes pneumococcal infections and the evolutionary origins of the serogroup 6 strains have been extensively studied. However, these studies were performed when serogroup 6 had only two known members (serotypes 6A and 6B) and before the two new members (serotypes 6C and 6D) expressing wciN(ß) were found. We have therefore reinvestigated the evolutionary origins of serogroup 6 by examining the profiles of the capsule gene loci and the multilocus sequence types (MLSTs) of many serogroup 6 isolates from several continents. We confirmed that there are two classes of cps locus sequences for serogroup 6 isolates. In our study, class 2 cps sequences were limited to a few serotype 6B isolates. Neighbour-joining analysis of cps sequence profiles showed a distinct clade for 6C and moderately distinct clades for class 1 6A and 6B sequences. The serotype 6D cps profile was found within the class 1 6B clade, suggesting that it was created by recombination between 6C and 6B cps loci. Interestingly, all 6C isolates also had a unique wzy allele with a 6 bp deletion. This suggests that serotype switching to 6C involves the transfer of a large (>4 kb) gene segment that includes both the wciN(ß) allele and the 'short' wzy allele. The MLST studies of serotype 6C isolates suggest that the 6C cps locus is incorporated into many different pneumococcal genomic backgrounds but that, interestingly, 6C cps may have preferentially entered strains of the same genomic backgrounds as those of serotype 6A.

Reductive Stress, Bioactive Compounds, Redox-Active Metals, and Dormant Tumor Cell Biology to Develop Redox-Based Tools for the Treatment of Cancer.

Significance: Cancer is related to redox biology from many points of view, such as initiation and promotion, metabolism and growth, invasion and metastasis, vascularization, or through the interaction with the immune system. In addition, this extremely complex relationship depends on the redox homeostasis of each cellular compartment, which might be used to fight cancer. Recent Advances: New ways of modulating specific and little explored aspects of redox biology have been revealed, as well as new delivery methods or uses of previously known treatments against cancer. Here, we review the latest experimental evidence regarding redox biology in cancer treatment and analyze its potential impact in the development of improved and more effective antineoplastic therapies. Critical Issues: A critical issue that deserves particular attention is the understanding that both extremes of redox biology (i.e., oxidative stress [OS] and reductive stress) might be useful or harmful in relation to cancer prevention and treatment. Future Directions: Additional research is needed to understand how to selectively induce reductive or OS adequately to avoid cancer proliferation or to induce cancer cell death.

Compostability assessment of nano-reinforced poly(lactic acid) films.

Nanomaterials can provide plastics with great advantages on mechanical and active properties (i.e. release and capture of specific substances). Therefore, packaging is expected to become one of the leading applications for these substances by 2020. There are some applications already in the market. Nevertheless, there is still some areas under development. A key issue to be analyzed is the end-of-life of these materials once they become waste, and specifically when nanomaterials are used in biodegradable products. The present study evaluated the disintegration, biodegradability, and ecotoxicity of poly(lactic acid) films reinforced with the three following nanomaterials: (1) montmorillonite modified with an ammonium quaternary salt, (2) calcium carbonate and (3) silicon dioxide. Results on disintegration showed that films completely disintegrated into visually indistinguishable residues after 6-7weeks of incubation in composting environment. Moreover, no differences were observed in the evolution of the bioresidue with respect to color, aspect, and odor in comparison with the control. It was also observed that nanomaterials did not significantly reduce the level of biodegradability of PLA (p>0.05). In fact, biodegradation was higher, without finding significant differences (p>0.05), in all the nano-reinforced samples with respect to PLA after 130days in composting (9.4% in PLA+Nano-SiO2; 34.0% in PLA+Clay1; 48.0% in PLA+Nano-CaCO3). Finally, no significant differences (p>0.05) in ecotoxicity in plants were observed as a result of the incorporation of nanoparticles in the PLA matrix.

The influence of printed electronics on the recyclability of paper: a case study for smart envelopes in courier and postal services.

The aim of this paper is to analyse the effects of the presence of printed electronics on the paper waste streams and specifically on paper recyclability. The analysis is based on a case study focussed on envelopes for postal and courier services provided with these intelligent systems. The smart printed envelope of the study includes a combination of both conventional (thin flexible batteries and resistors) and printed electronic components (conductive track layout based on nanosilver ink). For this purpose, a comparison between envelopes with and without these components (batteries, resistors and conductive track layouts) was carried out through pilot scale paper recycling tests. The generation of rejects during the recycling process as well as the final quality of the recycled paper (mechanical and optical properties) were tested and quantitatively evaluated. The results show that resistors are retained during the screening process in the sieves and consequently they cannot end up in the final screened pulp. Therefore, mechanical and optical properties of the recycled paper are not affected. Nevertheless, inks from the conductive track layouts and batteries were partially dissolved in the process water. These substances were not totally retained in the sieving systems resulting in slight changes in the optical properties of the final recycled paper (variations are 7.2-7.5% in brightness, 8.5-10.7% in whiteness, 1.2-2.2% in L(∗) values, 3.3-3.5% in opacity and 16.1-27% in yellowness). These variations are not in ranges able to cause problems in current paper recycling processes and restrict the use of recycled paper in current applications. Moreover, real impacts on industrial recycling are expected to be even significantly lower since the proportion of paper product with printed circuits in the current paper waste streams are much lower than the ones tested in this work. However, it should be underlined the fact that this situation may change over the next years due to the future developments in printed electronics and the gradual penetration of these types of devices in the market.

Colonizing and invasive strains of Streptococcus pneumoniae in Uruguayan children: type distribution and patterns of antibiotic resistance.

For the first time in Uruguay the frequency of moderate to heavy colonization of the upper respiratory tract by Streptococcus pneumoniae was investigated in children with acute respiratory infections (n = 717) and in healthy controls (n = 564). Serotypes of S. pneumoniae were identified as colonizing and invasive strains and their susceptibility to antibiotics was determined. Semiquantitative cultures of nasopharyngeal aspirates yielded 42.1% of positives in ambulatory patients and 15.2% in controls. Throat swabs from hospitalized children and matched controls revealed, respectively, 18.4 and 11.5% colonization. Different sampling and culture procedures were evaluated. Seasonal variations in colonization were also detected. Geographic variations in serotype frequency and distribution were assessed. Serotype 14 was predominant among invasive and colonizing strains in patients. Ten serotypes, included in the 23-valent S. pneumoniae vaccine, were not detected. Intermediate resistance to penicillin was seldom observed, but 37.2% of the invasive strains were resistant to cotrimoxazole, indicating the need for a permanent surveillance of isolates to antibiotic susceptibility.

Evolution of Streptococcus pneumoniae serotypes and penicillin susceptibility in Latin America, Sireva-Vigía Group, 1993 to 1999. PAHO Sireva-Vigía Study Group. Pan American Health Organization.

BACKGROUND: Since 1993 the Pan American Health Organization has coordinated a surveillance network with the National Reference Laboratories of Argentina, Brazil, Chile, Colombia, Mexico and Uruguay aimed at monitoring capsular types and antimicrobial susceptibility of Streptococcus pneumoniae causing invasive disease in children <6 years of age. METHODS: The surveillance system included children 6 years of age and younger with invasive disease caused by S. pneumoniae. The identification, capsular typing and susceptibility to penicillin of the isolates were conducted using a common protocol, based on standard methodologies. RESULTS: By June, 1999, 4,105 invasive pneumococcal isolates had been collected mainly from pneumonia (44.1%) and meningitis (41.1%) cases. Thirteen capsular types accounting for 86.1% of the isolates (14, 6A/6B, 5, 1, 23F, 19F, 18C, 19A, 9V, 7F, 3, 9N and 4) remained the most common types during the surveillance period. Diminished susceptibility to penicillin was detected in 28.6% of the isolates, 17.3% with intermediate and 11.3% with high level resistance. Resistance varied among countries and increased during this period in Argentina, Colombia and Uruguay. Serotypes 14 and 23F accounted for 66.6% of the resistance. CONCLUSION: These surveillance data clearly demonstrate the potential impact of the introduction of a conjugate vaccine on pneumococcal disease and the need for more judicious use of antibiotics to slow or reverse the development of antimicrobial resistance.

The apparent importation of penicillin-resistant capsular type 14 Spanish/French clone of Streptococcus pneumoniae into Uruguay in the early 1990s.

Since 1987, Streptococcus pneumoniae (SPN) has been monitored in pediatric and adult populations. In 1994, the SIREVA/PAHO surveillance allowed the assessment of a dramatic increase of penicillin resistance, mainly associated to SPN type 14 also resistance to trimethoprim/sulfamethoxazole. To determine the genetic relatedness of the resistant isolates, analysis of SmaI digests of 61 isolates was performed by pulsed field gel electrophoresis (PFGE). Twelve different profiles were identified. The type 14 resistant SPN (n = 47) displayed a common B pattern, with 15 genetically related subtypes (1-6 bands of difference), with B1 the predominant one (51%). One of the subtypes (B12) was indistinguishable from a French isolate (M13P). This strain represented a South European clone, identified in the late 1980s, that also included serogroup 9 isolates. The type 23F isolates (n = 3) were identical with a representative strain of the well-characterized intercontinental type 23F clone. Neither the type 14 penicillin-susceptible (n = 4) nor the resistant SPN of other serotypes (n = 10) belonged to the B pattern. In summary, the results suggest that the dramatic increase of penicillin resistance in Uruguay depends on the introduction and spread of a type 14 clone, apparently imported from the south of Europe. Follow-up of this phenomenon is mandatory from the point of view of epidemiology and is also a priority for biologic research.

Capsular type distribution and susceptibility to antibiotics of Streptococcus pneumoniae clinical strains isolated from Uruguayan children with systemic infections. Pneumococcus Study Group.

Children under 24 months of age are at high risk for serious infection with Streptococcus pneumoniae but they do not elicit effective immune responses to the currently available capsular polysaccharide vaccines. A polysaccharide protein conjugated vaccine involving the most frequent types has become an urgent need. To produce such a vaccine for Latin America, information on type distribution is required. Recently, Uruguay was 1 of the 6 countries in Latin America where surveillance for invasive pneumococcal infections in children under the age of 5 years was carried out. Seventy percent of the 182 invasive S. pneumoniae isolates were recovered from patients under 24 months of age, and 19% were recovered from infants under 6 months. The 7 most frequent types were 14, 5, 1, 6B, 3, 7F, and 19A; representing 80% of invasive isolates. Twenty-one types were identified, 16 in pneumonia and 14 in meningitis. Resistance to penicillin increased during the study period, from 29% in 1994, to 40% in 1995-1996, mainly because of the spread of type 14 strains resistant to penicillin and trimethoprim/sulphamethoxazol (89% of resistant isolates). The high proportion of systemic pneumococcal infections recorded in patients under 24 months of age and the increasing resistance of these agents to first-choice antibiotics point to an urgent need for a capsular polysaccharide protein conjugated vaccine.

Antibiotic resistance in Streptococcus pneumoniae in six Latin American countries: 1993-1999 surveillance.

The impact of invasive pneumococcal invasive disease is increased by the emergence of antibiotic resistance. We report regional and temporal variations in antibiotic resistance for 4,105 invasive Streptococcus pneumoniae isolates collected from Latin American children <5 years, between 1993 and 1999. Reduced susceptibility to penicillin was detected in 1,182 isolates (28.8%); 36% of these were resistant (> or = 2 microg/ml), including 12.6% with MIC > or = 4 microg/ml, occurring primarily in serotypes 14 and 23F. Reduced susceptibility to third-generation cephalosporins was detected in 12.1% of the collection. Mexico had the highest proportion of reduced susceptibility to penicillin (51.6%) and to third-generation cephalosporins (22%), whereas Brazil had the lowest at 20.9% and 0.7%, respectively. Isolates cultured from patients with pneumonia were more likely to have reduced susceptibility to third-generation cephalosporins than isolates from patients with meningitis (p < 0.0001). Susceptibility to trimethoprim-sulfamethoxazole, chloramphenicol, erythromycin, and vancomycin was tested by disk diffusion for 2.899 isolates. Reduced susceptibility was observed for 45.6%, 11.5%, 6.9%, and 0%, respectively. Thirty-one percent of the strains were resistant to > or = 2 drugs. High levels of antibiotic resistance in Latin America emphasize the need for the development of and adherence to rational antibiotic use guidelines. On-going surveillance will monitor the impact of these programs.

Ten-year review of invasive pneumococcal diseases in children and adults from Uruguay: clinical spectrum, serotypes, and antimicrobial resistance.

OBJECTIVES: Since 1987, the Reference Laboratory of the Ministry of Health of Uruguay has been monitoring infections due to Streptococcus pneumoniae in patients under 5 years of age, in those between 5 to 14 years of age, and in adults. The purpose of the present study was to retrospectively analyze a 10-year collection of invasive S. pneumoniae isolates from children 5 to 14 years of age and adults. METHODS: The Reference Children's Hospital, Pasteur Hospital, and two private hospitals in Montevideo as well as four hospitals located in other representative areas of the country participated in the pneumococcal surveillance program. Based on the information available at the Microbiology Department of the Central Public Health Laboratory (demographic data, date and site of isolate, and clinical diagnosis), all patients with an invasive pneumococcal disease were recorded. Pneumonia was clinically and radiologically diagnosed and etiology was assessed by isolation of S. pneumoniae from blood or pleural fluid. All specimens were collected at the Emergency Service. Capsular serotyping and antimicrobial susceptibilities were determined for each isolate. RESULTS: During the 10-year period, 228 invasive S. pneumoniae were identified and included in the study (blood, n = 129; cerebrospinal fluid [CSF], n = 73; pleural fluid, n = 20; peritoneal fluid, n = 3; synovial fluid, n = 1; pericardic fluid, n = 1; abscess, n = 1). The most frequent clinical presentations were pneumonia (n = 71) and meningitis (n = 69). Thirty-five adults had an underlying condition including, four with malignancies, four with lupus, two with human immunodeficiency virus (HIV)-infected, and two patients in hemodialysis among others. Eighteen of the 228 patients died (7.9% fatality rate), but only four of these had an underlying condition. Eleven fatal cases were attributable to meningitis (2 children, 9 and 11 years old; 9 adults, mean age, 59 y). Four patients with pneumonia and three with sepsis died, including a splenectomized woman. Nine different capsular serotypes (1, 5, 7, 9, 12, 15, 19A, 20, and 23A) were identified among the 18 fatal cases. Resistance to penicillin, generally combined with trimethoprim-sulfamethoxazole, fluctuated annually, not surpassing 10%. CONCLUSIONS: The study results indicated that 96% of the serotypes involved in severe pneumococcal diseases were included in the 23-valent vaccine and that S. pneumoniae resistance to penicillin was moderate.

Acute respiratory infections in children from a deprived urban population from Uruguay.

To obtain base line data on incidence, duration, clinical characteristics and etiology of acute respiratory infections (ARI), 276 children from deprived families living in Montevideo were followed during 32 months. The target population was divided into two groups for the analysis of the results: children aged less than 12 months and those older than this age. During the follow-up period 1.056 ARI episodes were recorded. ARI incidence was 5.2 per child/year. It was 87% higher in infants than in the older group, as was the duration of the episodes. Most of the diseases were mild. Tachypnea and retractions were seldom observed, but 12 children were referred to the hospital, and 2 infants died. Viral etiology was identified in 15.3% of the episodes. RSV was the predominant agent producing annual outbreaks. Moderate to heavy colonization of the upper respiratory tract by Streptococcus pneumoniae (32.3%) and Hemophilus sp. (18.9%) was recorded during ARI episodes. This community-based study furnish original data on ARI in Uruguay. It enabled to assess the impact of these infections on childhood.

Etiology and severity of community acquired pneumonia in children from Uruguay: a 4-year study.

The 4-year study (1987-1990) covered the major clinical-epidemiological characteristics of pneumonia in children as diagnosed at the emergency service of the Children's Hospital, as well as etiologies, and factors involved in the most severe cases. Etiology was determined in 47.7% of the 541 pneumonia cases, involving 283 pathogens of which 38.6% were viruses and 12.6% bacteria. Viral and mixed etiologies were more frequent in children under 12 months of age. Bacteria predominated in ages between 6 and 23 months. Among the viruses, respiratory syncytial virus predominated (66%). The bacterial pneumonias accounted for 12.2% of the recognized etiologies. The most important bacterial agents were S. pneumoniae (64%) and H. influenzae (19%). H. influenzae and mixed infections had a relevant participation during the 1988 season, pointing to annual variations in the relative participation of pathogens and its possible implication in severity of diseases. Correlation of severity and increased percentage of etiological diagnosis was assessed: patients with respiratory rates over 70 rpm, or pleural effusion and/or extensive pulmonary parenchyma compromise yielded higher positive laboratory results. Various individual and family risk factors were recognized when comparing pneumonia children with healthy controls.

Recent measles viral activity in Uruguai: serological and genetic approaches.

In the summer 1999, a measles outbreak occurred in Uruguai. During this outbreak 58 cases were recorded, 36 of which were laboratory confirmed as positive for measles virus (MV) IgM. The cases occurred in touristic places (Montevideo and Maldonado) predominantly among health facilities and tourist service personnel. Urine specimens collected between days 1 and 4 after the onset of the rash from seven cases were analyzed by reverse transcription-polymerase chain reaction (RT-PCR) and nested PCR with primers specific for the carboxyl-terminal region of the nucleoprotein (N) gene. Three of these specimens/cases were positive for MV. Sequencing of 300 nucleotides (nt) of PCR products corresponding to a part of the carboxyl-terminal region of the MV N gene detected in these specimens MV of D6 genotype. The same nucleotide sequences and the same genotype were also previously observed for MV isolates from the 1997 epidemic in Brazil and the 1998 epidemic in Argentina, demonstrating that the D6 genotype was, and may be still circulating in South America.

Difficulties encountered in a community-based study of acute respiratory infections in Uruguay.

A cohort study on acute respiratory infections, involving 270 children observed by pediatricians in their homes every 10 days over a period of 32 months, gave the opportunity to experience logistic and methodological problems seldom described in the literature. The purpose of this article is to alert researchers as to the difficulties faced when performing community-based studies in developing countries. Although a carefully planned project was undertaken, problem areas included the establishment of the target population, population dynamics, field related problems, laboratory aspects and data management. It is hoped that other investigators may benefit from the extensive experience gained from our program in foreseeing and coping with the difficulties involved.

Serological evidence of human Hantavirus infection in Argentina, Bolivia and Uruguay.

A serological survey was conducted in 1985-1987 to determine the presence of infection for Hantavirus in the general population in Argentina, Uruguay, Paraguay and Bolivia, as well as among rodent-exposed laboratory workers in Argentina. Out of 748 individuals tested by immunofluorescence 20 proved positive for Hantaan virus 76/118 strain of whom 16 also reacted against Seoul virus 80/39 strain and 2 against Puumala virus Sotkamo strain. Ten out of 72 Argentine laboratory workers were positive for the first 2 viruses by ELISA, immunofluorescence and/or plaque reduction neutralization test, in 4 of whom recent infection was demonstrated by IgM antibody presence. Inapparent Hantavirus infection was thus demonstrated for the first time in 2.7% of regional inhabitants, together with 13.9% among rodent-exposed laboratory workers. Our data established the existence of human Hantavirus infection nearly 10 years before the recognition of clinical cases of hemorrhagic fever with renal syndrome and hantavirus pulmonary syndrome in Argentina.

[Determination of bactericidal antibodies directed against vibrio cholerae in an adult population in Montevideo]. / Determinación de anticuerpos bactericidas dirigidos contra Vibrio cholerae en una población de adultos de Montevideo.

Uruguay is the only Latinoamerican country that remains free of cholera. Thus, to obtain a baseline for future diagnosis vibriocidal antibodies were investigated in 100 sera from blood donors serogroup "O". The reaction was carried out in microplates with live antigen Ogawa VC 12. In 95% of sera, titers were below 160, suggesting a high degree of susceptibility to V. cholerae in the population.

[Comparison between immunofluorescence and immunoenzymatic assay for the rapid diagnosis of the respiratory syncytial virus in nasopharyngeal secretions]. / Comparación entre la inmunofluorescencia y el ensayo inmunoenzimático para el diagnóstico rápido del virus respiratorio sincicial en secreciones nasofaríngeas.

An enzyme immunoassay, RSV-EIA Abbot, was evaluated by comparison with indirect immunofluorescence. Nasopharyngeal secretions obtained from 95 infants and young children with acute respiratory infections were examined for the presence of respiratory syncytial virus antigens with both methods. Specimens were stored at -70 degrees C before being tested by EIA. Out of 60 samples positive by indirect immunofluorescence, 46 were also positive by RSV-EIA (sensitivity 78.7%) and 34 out of 35 immunofluorescence negative specimens were negative by RSV-EIA (specificity 97.1%). Therefore, the EIA appears to be an acceptable test for the rapid detection of RSV as an alternative for indirect immunofluorescence.