Fever-like temperature impacts on Staphylococcus aureus and Pseudomonas aeruginosa interaction, physiology, and virulence both in vitro and in vivo.

BACKGROUND: Staphylococcus aureus (SA) and Pseudomonas aeruginosa (PA) cause a wide variety of bacterial infections and coinfections, showing a complex interaction that involves the production of different metabolites and metabolic changes. Temperature is a key factor for bacterial survival and virulence and within the host, bacteria could be exposed to an increment in temperature during fever development. We analyzed the previously unexplored effect of fever-like temperatures (39 °C) on S. aureus USA300 and P. aeruginosa PAO1 microaerobic mono- and co-cultures compared with 37 °C, by using RNAseq and physiological assays including in vivo experiments. RESULTS: In general terms both temperature and co-culturing had a strong impact on both PA and SA with the exception of the temperature response of monocultured PA. We studied metabolic and virulence changes in both species. Altered metabolic features at 39 °C included arginine biosynthesis and the periplasmic glucose oxidation in S. aureus and P. aeruginosa monocultures respectively. When PA co-cultures were exposed at 39 °C, they upregulated ethanol oxidation-related genes along with an increment in organic acid accumulation. Regarding virulence factors, monocultured SA showed an increase in the mRNA expression of the agr operon and hld, pmsα, and pmsß genes at 39 °C. Supported by mRNA data, we performed physiological experiments and detected and increment in hemolysis, staphyloxantin production, and a decrease in biofilm formation at 39 °C. On the side of PA monocultures, we observed an increase in extracellular lipase and protease and biofilm formation at 39 °C along with a decrease in the motility in correlation with changes observed at mRNA abundance. Additionally, we assessed host-pathogen interaction both in vitro and in vivo. S. aureus monocultured at 39οC showed a decrease in cellular invasion and an increase in IL-8-but not in IL-6-production by A549 cell line. PA also decreased its cellular invasion when monocultured at 39 °C and did not induce any change in IL-8 or IL-6 production. PA strongly increased cellular invasion when co-cultured at 37 and 39 °C. Finally, we observed increased lethality in mice intranasally inoculated with S. aureus monocultures pre-incubated at 39 °C and even higher levels when inoculated with co-cultures. The bacterial burden for P. aeruginosa was higher in liver when the mice were infected with co-cultures previously incubated at 39 °C comparing with 37 °C. CONCLUSIONS: Our results highlight a relevant change in the virulence of bacterial opportunistic pathogens exposed to fever-like temperatures in presence of competitors, opening new questions related to bacteria-bacteria and host-pathogen interactions and coevolution.

Regenerative potential of the sural neuroadipofascial flap in the treatment of chronic ulcers of the lower third of the leg.

OBJECTIVE: Surgical reconstruction of soft tissue defects in the distal third of the lower limbs has always been challenging for surgeons. The adipofascial flaps are made up of vascularized adipose tissue, which plastically conforms to the site to be reconstructed with its rich presence of stem cells. In this study, we resumed our case history of reverse neuroadipofascial sural flap, and we evaluated the stability of the scar to verify if this type of reconstruction can ensure long-term results and its regenerative power. PATIENTS AND METHODS: In this retrospective cohort study, we analyzed 32 patients who had undergone lower limb reconstruction with the sural neuroadipovenous flap. RESULTS: A total of 32 patients were included in the study. The average age was 62.2 years. Chronic skin ulcer was the cause of the defects in 13 (40.6%) patients, chronic wound after trauma in 7 (21.9%) patients, wheel bedsores in 4 (12.5%) patients, osteomyelitis in 5 (15.6%) patients, exposed internal hardware in 2 (6.3%) patients, dog bite in 1 (3.1%). The site defect comprised 11 heels (34.4%), 4 external malleoli (12.5%), three medial malleoli (9.4%), 12 lower third of the leg (37.5%), and two dorsa of the foot (6.3%). In all cases, defects were covered with reverse sural island flap. The dimension of the flap ranged from 5 to 9 cm in length and from 3 to 7 in width. Six patients showed early or later postoperative complications. CONCLUSIONS: The neuroadipofascial sural flap is versatile, fast, and easy-to-perform to cover a chronic defect in the distal part of the lower limb, including both the malleoli and the heel, due to its long vascular pedicle. Consistent with the studies carried out in other districts, the present study confirms the remarkable regenerative power of the vascularized adipose tissue at the level of complex wounds of the lower third of the leg.

Fever-like temperature impacts on Staphylococcus aureus and Pseudomonas aeruginosa interaction, physiology, and virulence both in vitro and in vivo.

Background: Staphylococcus aureus and Pseudomonas aeruginosa cause a wide variety of bacterial infections and coinfections, showing a complex interaction that involves the production of different metabolites and metabolic changes. Temperature is a key factor for bacterial survival and virulence and within the host, bacteria could be exposed to an increment in temperature during fever development. We analyzed the previously unexplored effect of fever-like temperatures (39°C) on S. aureus USA300 and P. aeruginosa PAO1 microaerobic mono- and co-cultures compared with 37°C, by using RNAseq and physiological assays including in-vivo experiments. Results: In general terms both temperature and co-culturing had a strong impact on both PA and SA with the exception of the temperature response of monocultured PA. We studied metabolic and virulence changes on both species. Altered metabolic features at 39°C included arginine biosynthesis and the periplasmic glucose oxidation in S. aureus and P. aeruginosa monocultures respectively. When PA co-cultures were exposed at 39°C they upregulated ethanol oxidation related genes along with an increment in organic acid accumulation. Regarding virulence factors, monocultured SA showed an increase in the mRNA expression of the agr operon and hld, pmsα and pmsß genes at 39°C. Supported by mRNA data, we performed physiological experiments and detected and increment in hemolysis, staphylxantin production and a decrease in biofilm formation at 39°C. On the side of PA monocultures, we observed increase in extracellular lipase and protease and biofilm formation at 39°C along with a decrease in motility in correlation with changes observed at mRNA abundance. Additionally, we assessed host-pathogen interaction both in-vitro and in-vivo . S. aureus monocultured at 39°C showed a decrease in cellular invasion and an increase in IL-8 -but not in IL-6- production by A549 cell line. PA also decreased its cellular invasion when monocultured at 39°C and did not induce any change in IL-8 or IL-6 production. PA strongly increased cellular invasion when co-cultured at 37°C and 39°C. Finally, we observed increased lethality in mice intranasally inoculated with S. aureus monocultures pre-incubated at 39°C and even higher levels when inoculated with co-cultures. The bacterial burden for P. aeruginosa was higher in liver when the mice were infected with co-cultures previously incubated at 39°C comparing with 37°C. Conclusion: Our results highlight a relevant change in the virulence of bacterial opportunistic pathogens exposed to fever-like temperatures in presence of competitors, opening new questions related to bacteria-bacteria and host-pathogen interactions and coevolution.

Maternal stress and hair cortisol among pregnant women following hurricane Florence.

Natural disasters represent major stressors, resulting in psychological distress and physiological responses such as increased cortisol. During pregnancy, this impacts not only maternal well-being, but also fetal development. In 2018, Hurricane Florence caused extensive damage across the eastern United States. Studies indicated that compared to married pregnant women, unmarried pregnant women had higher risk of distress. Here we assess hair cortisol among a subsample of participants, and variations based on marital status. METHODS: We analyzed multiple stress measures among 37 participants who were pregnant during Hurricane Florence. We used questionnaires modeled on previous studies to assess hardship associated with the hurricane, psychological distress, sociodemographic characteristics, social support, and food security. We analyzed cortisol concentrations in proximal and distal hair sections, representing stress around the time of the disaster (distal) and 3-4 months following the disaster (proximal). We used linear regression to test relationships between hair cortisol and self-report stress measures, and variations based on marital status. RESULTS: Self-report measures of distress and hardship were similar among married and unmarried participants. Mean cortisol levels in distal and proximal sections were higher among unmarried participants. Controlling for confounding variables, hardship was not associated with hair cortisol. Distress predicted cortisol in distal sections (ß = .482, p = .018), with a trend for proximal sections (ß = .368, p = .055). Marital status was a significant predictor of distal (ß = .388, p = .027) and proximal (ß = .333, p = .047) hair cortisol, explaining 8.6%-11.7% of unique variance. CONCLUSIONS: Preexisting and intersecting risk factors likely place unmarried pregnant individuals at risk of stress during and following a disaster.

SAFETY AND EFFICACY OF THYMIC PEPTIDES IN THE TREATMENT OF HOSPITALIZED COVID-19 PATIENTS IN HONDURAS.

Coronavirus disease 2019 (Covid-19) active cases continue to demand the development of safe and effective treatments. This is the first clinical trial to evaluate the safety and efficacy of oral thymic peptides. ; We conducted a nonrandomized phase 2 trial with a historic control group to evaluate the safety and efficacy of a daily 250-mg oral dose of thymic peptides in the treatment of hospitalized Covid-19 patients. Comparisons based on standard care from registry data were performed after propensity score matching. The primary outcomes were survival, time to recovery, and number of participants with treatment-related adverse events or side effects by day 20. ; A total of 44 patients were analyzed in this study: 22 in the thymic peptide group and 22 in the standard care group. There were no deaths in the intervention group compared to 24% mortality in standard care by day 20 (log-rank P=0.02). Kaplan-Meier analysis showed a significantly shorter time to recovery by day 20 in the thymic peptide group than in the standard care group (median, 6 days vs. 12 days; hazard ratio for recovery, 2.75 [95% confidence interval, 1.34 to 5.62]; log-rank P=0.002). No side effects or adverse events were reported. ; In patients hospitalized with Covid-19, the use of thymic peptides resulted in no side effects, adverse events, or deaths by day 20. Compared with the registry data, a significantly shorter time to recovery and mortality reduction were measured.

QTL analysis reveals quantitative resistant loci for Phytophthora infestans and Tecia solanivora in tetraploid potato (Solanum tuberosum L.).

Late blight and Guatemalan potato tuber moth caused by Phytophthora infestans and Tecia solanivora, respectively, are major phytosanitary problems on potato crops in Colombia and Ecuador. Hence, the development of resistant cultivars is an alternative for their control. However, breeding initiatives for durable resistance using molecular tools are limited due to the genome complexity and high heterozygosity in autotetraploid potatoes. To contribute to a better understanding of the genetic basis underlying the resistance to P. infestans and T. solanivora in potato, the aim of this study was to identify QTLs for resistance to P. infestans and T. solanivora using a F1 tetraploid potato segregant population for both traits. Ninety-four individuals comprised this population. Parent genotypes and their progeny were genotyped using SOLCAP 12K potato array. Forty-five percent of the markers were polymorphic. A genetic linkage map was built with a length of 968.4 cM and 1,287 SNPs showing good distribution across the genome. Severity and incidence were evaluated in two crop cycles for two years. QTL analysis revealed six QTLs linked to P. infestans, four of these related to previous QTLs reported, and two novel QTLs (qrAUDPC-3 and qrAUDPC-8). Fifteen QTLs were linked to T. solanivora, being qIPC-6 and qOPA-6.1, and qIPC-10 and qIPC-10.1 stable in two different trials. This study is one of the first to identify QTLs for T. solanivora. As the population employed is a breeding population, results will contribute significantly to breeding programs to select resistant plant material, especially in countries where P. infestans and T. solanivora limit potato production.

Economía, política y economía política para el acceso y la cobertura Universal de salud en Uruguay / Economics, politics and political economy for access and coverage Universal health in Uruguay

El presente libro se inserta en esta estrategia de cooperación al país y tiene por objetivo ordenar, registrar y diseminar estudios, investigaciones, reflexiones y debates construidos en el período reciente, en el ámbito del equipo de la OPS/OMS en Uruguay y/o promovidos, incentivados y financiados por la Organización. Los artículos combinan metodologías clásicas de la ciencia económica con otras heterodoxas y con perspectivas innovadoras de la ciencia políticaaportadas por el institucionalismo histórico, buscando avanzar en un necesariamente largo proceso de síntesis dialéctica de múltiples disciplinas indispensable a la consolidación del campo de saber de la Economía Política del Sector Salud, sin abundar en electicismo metodológico.

Viral load of human bocavirus-1 in stools from children with viral diarrhoea in Paraguay.

Since their discovery, four species of human bocavirus (HBoV) have been described in patients with respiratory and gastrointestinal diseases. However, a clear causal association between HBoV-1 and gastroenteritis has not been demonstrated. In this study, we describe the detection and quantification of HBoV-1 in stools from children with acute non-bacterial gastroenteritis using quantitative polymerase chain reaction. HBoV-1 genome was detected in 10.6% of stools with frequent association with rotavirus and norovirus. The median of HBoV-1 viral load was 1.88 × 104 genome/ml, lower than previously shown in secretions of patients with respiratory infections, without any obvious association between high viral load and presence of HBoV as single agent. Thus, although HBoV-1 was frequently detected in these patients, there is no clear causal association of this agent with diarrhoea. Indeed, HBoV-1 DNA in stools of patients with gastroenteritis without respiratory symptoms may be a remnant of previous infections or associated with prolonged shedding of virus in the respiratory or digestive tracts.

Reconstruction of the thumb with a modified wrap-around flap in a patient suffering from ß-thalassemia minor.

Thalassemia is a congenital hemolytic disorder caused by a partial or complete deficiency of α- or ß-globin chain synthesis. It has been seen that thalassemic patients exhibit an increased frequency of thrombotic events. The article presents the first case of thumb reconstruction with a modified wrap-around flap in a patient suffering from ß-thalassemia minor.

Classification of morphological features extracted from intracranial pressure recordings in the diagnosis of Normal Pressure Hydrocephalus (NPH).

The intracranial pressure (ICP) monitoring is a common procedure in neuro-intensive care for pathologies such traumatic brain injuries or hemorrhages, but also for chronic ones as the Normal Pressure Hydrocephalus (NPH). The only available treatment for NPH is the surgical implantation of a shunt with the aim of routing cerebrospinal fluid (CSF) away from the brain to another part of the body. In this study, using the classification software WEKA, an intensive investigation of ICP signals has been conducted. In particular we studied 14 ICP recordings of different patients who underwent an infusion test, with the aim of investigating the presence of NPH through the ICP recording. More precisely, 20 morphological features are extracted from the ICP pulsed wave, the trend have been computed and, for each one, 9 statistical functions determined. The 180 features have been selected and passed for the classification. The results obtained shows how, among the 14 patients, a number of 12 out of 14 (85.7%) have been correctly classified, looking at just 3 features. In particular 8 out of 9 not-NPH-affected patients were correctly identified (88.89%) while 4 out of 5 NPH-affected patients were correctly identified (80%).

R-point detection for noise affected ECG recording through signal segmentation.

In this work we propose a novel approach for filtering noise-affected electrocardiogram (ECG) signals. The proposed method, mainly based on signal approximation by means of linear segments, has been applied for R-peaks recognition and has been compared with both cardiologists' manual marking and the automatic Laguna's method. The obtained results show that when compared to the Laguna's method the proposed algorithm provides a smaller mean error and a better error distribution.

Analysis of intracranial pressure recordings: comparison of PCA and signal averaging based filtering methods and signal period estimation.

Intracranial pressure monitoring is a common used approach for neuro-intensive care in cases of brain damages and injuries or to investigate chronic pathologies. Several types of noises and artifacts normally contaminate ICP recordings. They can be sorted in 2 classes, i.e. high-frequency noises (due to measurement and amplifier devices or electricity supply presence) and low-frequency noises (due to unwanted patient's movement, speeches, coughing during the recording and quantization noise). Thus, deep investigations on ICP components aimed to extract features from ICP signal, require a denoised signal. For this reason the authors have addressed a study upon the most common filtering techniques. On each ICP recording we have performed 4 configurations of filters, which involve the use of a FIR filter together with Signal Averaging filters or PCA based filters. Next step is period estimation for absolute minima detection. The results obtained by the algorithm for automatic ICP marking are compared to those ones obtained from manual marking (peaks are manually identified and annotated by a brain surgeon). The procedure is repeated varying the filters sliding window size to minimize the mean square error. The results show how the configurations FIR filter + Signal averaging provides smaller mean squared error (MSE=118.84[sample(2)]) than the others 3 configurations FIR filter + PCA filter based (MSE=135.29-147.15[sample(2)]).

Short communication: Biological and genetic characterization of HIV type 1 subtype B and nonsubtype B transmitted viruses: usefulness for vaccine candidate assessment.

Due to the extraordinary degree of genetic diversity of HIV-1 and the structural complexity of its envelope glycoproteins, designing an effective vaccine is difficult, requiring the development of viral reagents to assess vaccine-elicited neutralizing antibodies. The aim of this study was to improve on our previously developed panel of HIV-1 strains of different genetic forms, focusing on strains from acute and recently acquired infections as the most representative of the transmitted viruses. HIV-1 primary isolates were expanded in peripheral blood mononuclear cells. Viral stocks of 40 ml each were produced. Syncytium-inducing (SI) phenotype, coreceptor use, and TCID(50)/ml were determined. Near full-length HIV-1 genomes were amplified by RT-nested PCR in four overlapping segments. Phylogenetic analyses were performed with neighbor-joining trees and bootscanning. Forty-four HIV-1 strains were included in the panel. Twenty-four (54.1%) strains were from early infections (16 acute and 8 recent); of them, 21 (87%) were sexually transmitted. NSI/R5 phenotype was detected in 37 (84.1%) viruses and SI/R5,X4 in another 7 (15.9%). TCID(50)/ml ranged between 10(4) and 10(6.6). Twelve different genetic forms constituted this panel: subtypes A1, B, C, F1, and G; circulating recombinant forms CRF02_AG, CRF14_BG, and CRF24_BG; and unique recombinant forms CRF02_AG/A3, BF1, CRF12_BF/B, and DF1G. In conclusion, in this study, we report the development of a comprehensive and well-characterized panel of HIV-1 isolates for assessing neutralization in HIV vaccine research. This panel is available for distribution through the Programme EVA Centre for AIDS Reagents, National Institute for Biological Standard and Control (NIBSC).

A preliminary study for investigating idiopatic normal pressure hydrocephalus by means of statistical parameters classification of intracranial pressure recordings.

The objective of this study is to investigate Id-iopatic Normal Pressure Hydrocephalus (INPH) through a multidimensional and multiparameter analysis of statistical data obtained from accurate analysis of Intracranial Pressure (ICP) recordings. Such a study could permit to detect new factors, correlated with therapeutic response, which are able to validate a predicting significance for infusion test. The algorithm developed by the authors computes 13 ICP parameter trends on each of the recording, afterward 9 statistical information from each trend is determined. All data are transferred to the datamining software WEKA. According to the exploited feature-selection techniques, the WEKA has revealed that the most significant statistical parameter is the maximum of Single-Wave-Amplitude: setting a 27 mmHg threshold leads to over 90% of correct classification.

Incidence of non-B subtypes of HIV-1 in Galicia, Spain: high frequency and diversity of HIV-1 among men who have sex with men.

An increase in HIV transmission among men who have sex with men (MSM) has been reported in eight regions of Spain from 2003 to 2007. In order to study the incidence of HIV-1 genetic forms in Galicia, northwest of Spain, in particular the spread of HIV-1 variants among MSM, 93 newly diagnosed HIV-1 patients, including those with acute and recently acquired infections, were studied for a year from August 2008 to August 2009. Thirty eight (41%) were MSM. Of them, nine (24%) were infected by non-B viruses, including seven different genetic forms. The analysis of transmission clusters showed that 23 (60%) MSM grouped in different clusters and mostly in large clusters. Resistance mutations were detected in six (16%) MSM.

Environmental contributors to the achievement gap.

Extensive research shows that blacks, those of low socioeconomic status, and other disadvantaged groups continue to exhibit poorer school performance compared with middle and upper-class whites in the United States' educational system. Environmental exposures may contribute to the observed achievement gap. In particular, childhood lead exposure has been linked to a number of adverse cognitive outcomes. In previous work, we demonstrated a relationship between early childhood lead exposure and end-of-grade (EOG) test scores on a limited dataset. In this analysis, data from the North Carolina Childhood Lead Poisoning Prevention Program surveillance registry were linked to educational outcomes available through the North Carolina Education Research Data Center for all 100 counties in NC. Our objectives were to confirm the earlier study results in a larger population-level database, determine whether there are differences in the impact of lead across the EOG distribution, and elucidate the impact of cumulative childhood social and environmental stress on educational outcomes. Multivariate and quantile regression techniques were employed. We find that early childhood lead exposure is associated with lower performance on reading EOG test scores in a clear dose-response pattern, with the effects increasingly more pronounced in moving from the high end to the low end of the test score distribution. Parental educational attainment and family poverty status also affect EOG test scores, in a similar dose-response fashion, with the effects again most pronounced at the low end of the EOG test score distribution. The effects of environmental and social stressors (especially as they stretch out the lower tail of the EOG distribution) demonstrate the particular vulnerabilities of socioeconomically and environmentally disadvantaged children. Given the higher average lead exposure experienced by African American children in the United States, lead does in fact explain part of the achievement gap.

Development of a panel of well-characterized human immunodeficiency virus type 1 isolates from newly diagnosed patients including acute and recent infections.

The aim of this study was the development of a panel constituted by well-defined HIV-1 strains of different genetic forms, with a particular focus on isolates from acute and recent infections. Fourteen HIV-1 isolates, including four from acute and five from recent infections, were expanded in peripheral blood mononuclear cells. SI phenotype, coreceptors use, and TCID(50)/ml were determined. V3 net charge was calculated. Near full-length genomes were amplified by RT-nested PCR in four overlapping segments. Phylogenetic analyses were performed with neighbor-joining trees and bootscanning. Analysis of cysteine residues, lengths of variable regions, and potential N-linked glycosylation sites in gp120 and gp41 was performed. Viral stocks were produced. Thirteen strains were NSI/R5 and one SI/R5,X4. TCID(50)/ml ranged between 10(4.6) and 10(6). V3 net charge was <+5 in 12 sequences and +5 in two sequences. Near full-length HIV-1 genomes analysis identified viruses of the following genetic forms: eight subtype B, three subtype C, two CRF02_AG, and one subtype G. Cysteine residues that form the V1,V2,V3, and V4 loops were highly conserved. The number of potential N-linked glycosylation sites in gp120 and gp41 ranged between 24-29 and 4-6, respectively. Seven potential N-linked glycosylation sites in gp120 and three in gp41 were conserved. V1, V2, V4, and V5 variable regions exhibited substantial length variation. In addition, an analysis of transmitted and natural resistance to current antiretroviral drugs in these strains was performed. It is worth mentioning that the 13S mutation in the V3 sequence, associated with resistance to maraviroc, was observed in a subtype B strain that harbored resistance mutations to nucleoside reverse transcriptase inhibitors and to T20. The availability of a panel including strains from acute and recent infections should be a valuable resource for optimizing and standardizing vaccine candidate assessment. Near full-length genome characterization may be necessary for evaluating clade-specific reactivities.