Brain-Derived Neurotrophic Factor-Loaded Low-Temperature-Sensitive liposomes as a drug delivery system for repairing podocyte damage.

Podocytes, cells of the glomerular filtration barrier, play a crucial role in kidney diseases and are gaining attention as potential targets for new therapies. Brain-Derived Neurotrophic Factor (BDNF) has shown promising results in repairing podocyte damage, but its efficacy via parenteral administration is limited by a short half-life. Low temperature sensitive liposomes (LTSL) are a promising tool for targeted BDNF delivery, preserving its activity after encapsulation. This study aimed to improve LTSL design for efficient BDNF encapsulation and targeted release to podocytes, while maintaining stability and biological activity, and exploiting the conjugation of targeting peptides. While cyclic RGD (cRGD) was used for targeting endothelial cells in vitro, a homing peptide (HITSLLS) was conjugated for more specific uptake by glomerular endothelial cells in vivo. BDNF-loaded LTSL successfully repaired cytoskeleton damage in podocytes and reduced albumin permeability in a glomerular co-culture model. cRGD conjugation enhanced endothelial cell targeting and uptake, highlighting an improved therapeutic effect when BDNF release was induced by thermoresponsive liposomal degradation. In vivo, targeted LTSL showed evidence of accumulation in the kidneys, and their BDNF delivery decreased proteinuria and ameliorated kidney histology. These findings highlight the potential of BDNF-LTSL formulations in restoring podocyte function and treating glomerular diseases.

Global availability of parenteral nutrition: Pre- and post-COVID-19 pandemic surveys.

OBJECTIVE: Parenteral nutrition represents a therapeutic option for patients with type 3 intestinal failure. If used exclusively, parenteral nutrition has to be complete to provide all essential nutrients. The aim was to assess the availability of parenteral nutrition in all parts of the world, to better comprehend the global situation, and to prepare an action plan to increase access to parenteral nutrition. METHODS: An international survey using an electronic questionnaire was conducted in August 2019 and repeated in May 2022. An electronic questionnaire was sent to 52 members or affiliates of the International Clinical Nutrition Section of the American Society for Parenteral and Enteral Nutrition. Questions addressed the availability of parenteral nutrition admixtures and their components, reimbursement, and prescribing pre- and post-COVID-19 pandemic. All participating countries were categorized by their economic status. RESULTS: Thirty-six country representatives responded, answering all questions. Parenteral nutrition was available in all countries (100%), but in four countries (11.1%) three-chamber bags were the only option, and in six countries a multibottle system was still used. Liver-sparing amino acids were available in 18 (50%), kidney-sparing in eight (22.2%), and electrolyte-free in 11 (30.5%) countries (30.5%). In most countries (n = 28; 79.4%), fat-soluble and water-soluble vitamins were available. Trace elements solutions were unavailable in four (11.1%) countries. Parenteral nutrition was reimbursed in most countries (n = 33; 91.6%). No significant problems due to the coronavirus pandemic were reported. CONCLUSIONS: Despite the apparent high availability of parenteral nutrition worldwide, there are some factors that may have a substantial effect on the quality of parenteral nutrition admixtures. These shortages create an environment of inequality.

Implementing a tobacco-free workplace program at a substance use treatment center: a case study.

BACKGROUND: People with substance use disorders smoke cigarettes at much higher rates than the general population in the United States and are disproportionately affected by tobacco-related diseases. Many substance use treatment centers do not provide evidence-based tobacco cessation treatment or maintain comprehensive tobacco-free workplace policies. The goal of the current work is to identify barriers and facilitators to a successful and sustainable implementation of a tobacco-free workplace program, which includes a comprehensive tobacco-free policy and evidence-based cessation treatment services, in a substance use treatment center. METHODS: This study is based on an ethnographic approach and uses a qualitative case study design. Data were collected via interviews with staff (n = 6) and clients (n = 16) at the substance use treatment center and site visits (n = 8). Data were analyzed using thematic analysis guided by the extended Normalization Process Theory designed to inform the implementation of innovations in healthcare practice. RESULTS: Staff at the substance use treatment center supported the implementation of the program and shared a good understanding of the purpose of the intervention and its potential benefits. However, the study identified significant challenges faced by the center during implementation, including widespread tobacco use among clients, contributing to attitudes among staff that tobacco cessation was a low-priority problem due to a perceived lack of interest in quitting and inability to quit among their clients. We identified several factors that contributed to changing this attitude, including provision of tobacco training to staff, active leadership support, low number of staff members who smoked, and access to material resources, including nicotine replacement products. The implementation and active enforcement of a comprehensive tobacco-free workplace program contributed to a gradual change in attitudes and improved the provision of evidence-based tobacco cessation care at the substance use treatment center. CONCLUSIONS: Substance use treatment centers can integrate tobacco cessation practices in their daily operations, despite multiple challenges they face due to the complex behavioral health and socioeconomic needs of their clients. With proper support, substance use treatment centers can provide much needed tobacco cessation care to their clients who are disproportionately affected by tobacco-related health conditions and systemic health inequities.

Design of fluorinated stealth poly(ε-caprolactone) nanocarriers.

The covalent functionalization of polymers with fluorinated moieties represents a promising strategy for the development of multimodal systems. Moreover, polymer fluorination often endows the resulting nanocarriers with improved colloidal stability in the biological environment. In this work, we developed fluorinated pegylated (PEG) biodegradable poly(ε-caprolactone) (PCL) drug nanocarriers showing both high colloidal stability and stealth properties, as well as being (19F)-Nuclear Magnetic Resonance (NMR) detectable. The optimized nanocarriers were obtained mixing a PEG-PCL block copolymer with a nonafluoro-functionalized PCL polymer. The role of PEGylation and fluorination on self-assembly and colloidal behavior of the obtained nanoparticles (NPs) was investigated, as well as their respective role on stealth properties and colloidal stability. To prove the feasibility of the developed NPs as potential 19F NMR detectable drug delivery systems, a hydrophobic drug was successfully encapsulated, and the maintenance of the relevant 19F NMR properties evaluated. Drug-loaded fluorinated NPs still retained a sharp and intense 19F NMR signal and good relaxivity parameters (i.e., T1 and T2 relaxation times) in water, which were not impaired by drug encapsulation.

Influence of UO2 crystal orientation on laser ablation performance.

Crystallographic orientation dependence deteriorates the performance of surface analysis methods such as secondary ion mass spectrometry (SIMS) and focused ion beam (FIB). This study explores the corresponding potential challenges of laser ablation (LA) as a powerful sampling tool for inductively coupled plasma-mass spectrometry (ICP-MS). To this end, three UO2 single crystals of different orientation as well as polycrystalline UO2 were produced and characterized. Subsequently, a ns-laser ablation system was employed to study laser-matter interaction in detail. Firing the laser continuously at 1 Hz with various single shot fluence (2, 4, 6, 8, 12 J cm-2) for diverse periods created LA craters impacted by cumulative fluence between 50 and 650 J cm-2. Repeated LA experiments on the (100) plane of a UO2 single crystal at the beginning and end of the entire study revealed highly reproducible (<3%) LA rates, only limited by the fluctuation of the laser energy output of the ns-LA system. After thorough cleaning of the ablated samples, surface roughness and average depth of LA craters were determined using confocal laser scanning profilometry. Both LA rate and average depth of craters decreased exponentially with increasing single shot fluence independently of the crystal orientation. Surface roughness increased linearly with increasing cumulative fluence having largest intensification for lowest single shot fluence. Scanning electron microscope (SEM) images not only revealed the conical silhouette of LA craters, but also identified a convex meniscus at its bottom. This particular shape of the crater bottom with a deeper ring surrounding the central region is a result of melted and re-solidified UO2 generated during the LA process and the main limiting factor for the achievable depth resolution. The rapid re-solidification of the liquid phase after each single laser shot created tiles of different shape and orientation, depending on UO2 crystal orientation. Three different types of ejected particles radially distributed around the LA craters were identified by SEM, providing profound insights into laser-UO2 interaction.

Abrupt loss and uncertain recovery from fires of Amazon forests under low climate mitigation scenarios.

Tropical forests contribute a major sink for anthropogenic carbon emissions essential to slowing down the buildup of atmospheric CO2 and buffering climate change impacts. However, the response of tropical forests to more frequent weather extremes and long-recovery disturbances like fires remains uncertain. Analyses of field data and ecological theory raise concerns about the possibility of the Amazon crossing a tipping point leading to catastrophic tropical forest loss. In contrast, climate models consistently project an enhanced tropical sink. Here, we show a heterogeneous response of Amazonian carbon stocks in GFDL-ESM4.1, an Earth System Model (ESM) featuring dynamic disturbances and height-structured tree-grass competition. Enhanced productivity due to CO2 fertilization promotes increases in forest biomass that, under low emission scenarios, last until the end of the century. Under high emissions, positive trends reverse after 2060, when simulated fires prompt forest loss that results in a 40% decline in tropical forest biomass by 2100. Projected fires occur under dry conditions associated with El Niño Southern Oscillation and the Atlantic Multidecadal Oscillation, a response observed under current climate conditions, but exacerbated by an overall decline in precipitation. Following the initial disturbance, grassland dominance promotes recurrent fires and tree competitive exclusion, which prevents forest recovery. EC-Earth3-Veg, an ESM with a dynamic vegetation model of similar complexity, projected comparable wildfire forest loss under high emissions but faster postfire recovery rates. Our results reveal the importance of complex nonlinear responses to assessing climate change impacts and the urgent need to research postfire recovery and its representation in ESMs.

An effective algorithm to detect the possibility of being MSI phenotype in endometrial cancer given the BMI status and histological subtype: a statistical study.

PURPOSE: In endometrial cancer, the incidence of mutations in mismatch repair genes (MMR) is estimated at 17-30%. Patients with alterations at this level (MSI) are known to have different clinical and anatomopathological characteristics than those without this genetic alteration (MSS). In this study, we aim to identify the MSI phenotype in patients who underwent hysterectomy for endometrial cancer. We assessed the correlation of this phenotype with anatomoclinical parameters such as obesity and histological subtype. METHODS/PATIENTS: Clinical and anatomopathological data were collected from 147 patients diagnosed with endometrial cancer and an immunohistochemical study of MMR system proteins was performed. PMS2 and MSH6 proteins were evaluated as primary screening and subsequent evaluation of MLH1 and MSH6, respectively, if the former were negative. Statistical association between the anatomopathological data and the immunohistochemical result was analyzed. RESULTS AND CONCLUSIONS: 22.4% of our patients were MSI phenotype. We obtained statistically significant differences by multivariate analysis between endometrioid subtype and higher FIGO classification grade with MSI phenotype and obesity with MSS phenotype. Given these statistical results, we propose a function for predicting the probability of being MSI phenotype taking into account the histological subtype (endometrioid/non-endometrioid carcinoma) and FIGO grade as well as obesity. This prediction may be useful prior to hysterectomy, for genetic study of the MLH1 promoter and subsequent genetic counseling.

Distinct roles of the dystrophin-glycoprotein complex: α-dystrobrevin and α-syntrophin in the maintenance of the postsynaptic apparatus of the neuromuscular synapse.

α-syntrophin (α-syn) and α-dystrobrevin (α-dbn), two components of the dystrophin-glycoprotein complex, are essential for the maturation and maintenance of the neuromuscular junction (NMJ) and mice deficient in either α-syn or α-dbn exhibit similar synaptic defects. However, the functional link between these two proteins and whether they exert distinct or redundant functions in the postsynaptic organization of the NMJ remain largely unknown. We generated and analyzed the synaptic phenotype of double heterozygote (α-dbn+/-, α-syn+/-), and double homozygote knockout (α-dbn-/-; α-syn-/-) mice and examined the ability of individual molecules to restore their defects in the synaptic phenotype. We showed that in double heterozygote mice, NMJs have normal synaptic phenotypes and no signs of muscular dystrophy. However, in double knockout mice (α-dbn-/-; α-syn-/-), the synaptic phenotype (the density, the turnover and the distribution of AChRs within synaptic branches) is more severely impaired than in single α-dbn-/- or α-syn-/- mutants. Furthermore, double mutant and single α-dbn-/- mutant mice showed more severe exercise-induced fatigue and more significant reductions in grip strength than single α-syn-/- mutant and wild-type. Finally, we showed that the overexpression of the transgene α-syn-GFP in muscles of double mutant restores primarily the abnormal extensions of membrane containing AChRs that extend beyond synaptic gutters and lack synaptic folds, whereas the overexpression of α-dbn essentially restores the abnormal dispersion of patchy AChR aggregates in the crests of synaptic folds. Altogether, these data suggest that α-syn and α-dbn act in parallel pathways and exert distinct functions on the postsynaptic structural organization of NMJs.

Smoking Intervention Practices in Texas Healthcare Centers with Sexual and Gender Minority Patients.

Objectives: This study evaluated the use of evidence-based practices (EBPs) for smoking cessation in centers providing behavioral healthcare for patient populations that included some proportion of sexual and gender minorities (SGMs). Methods: Healthcare providers from 75 healthcare centers across Texas serving SGMs with behavioral health needs participated in a survey assessing their center's tobacco control policies and practices. Results: Nearly half (N = 36) of participating centers had a comprehensive tobacco-free workplace policy, 30.67% employed ≥1 tobacco treatment specialist, 73.91% employed ≥1 prescriber, 80.82% mandated screening for patient tobacco use at intake, and 57.53% provided a template for tobacco use assessments. Overall, 70.67% of providers asked patients about smoking status, 69.33% advised patients to quit, 64.00% assessed patients' interest in quitting, 58.67% assisted patients with quit attempts, and 36.00% arranged follow-up. Providers' ability to tailor interventions for special populations like SGMs ranged from very low/0 to very high/10 (M = 4.63 ± 2.59). Conclusions: There are opportunities to improve policy implementation, standardization and usage of evidence-based interventions, and intervention tailoring within settings providing care to SGM patients in Texas to better address their tobacco use inequities.

Physical Model for Investigating Intracranial Pressure with Clinical Pressure Sensors and Diagnostic Ultrasound: Preliminary Results.

INTRODUCTION: Intracranial pressure (ICP) is a commonly collected neurocritical parameter, but accurate signal modelling remains challenging. The goal of this project was to mimic clinical ICP waveforms using a physical model. MATERIALS AND METHODS: A physical head model was developed. The skull was segmented from a head computed tomography (CT) scan, remodelled, 3D-printed, and filled with a brain tissue mimicking material and a pressure generator. Pressure measurements and tissue displacement around an attached pressure sensor were explored. RESULTS: Analysis of the measured pressure demonstrated that the waveform did not perfectly resemble that of the clinical ICP. Through iterative improvements and using a revised second pressure generator, subpeaks could be seen in the waveform. A speckle image recorded using ultrasound during pressure application enabled visualization of tissue displacement around the pressure sensor. Comparison with measured ICP signals revealed that minuscule patterns were not distinct in the displacement images. DISCUSSION: We present the first steps towards mimicking clinical ICP using a physical head phantom model. The physical model enabled pressure tests and visualization of tissue displacement and will be foundational for further improvements.

Empowered Choices: African-American Women's Breast Reconstruction Decisions.

Objectives: Breast reconstruction (BR) potentially can improve quality of life in postmastectomy breast cancer survivors (BCS); however, African-American women are less likely to undergo BR than Caucasian women. This qualitative study was undertaken to explore individual, sociocultural, and contextual factors influencing African-American women's BR decision-making processes and preferences. Methods: Postmastectomy African-American BCS with and without BR participated in semi-structured interviews. We adopted a grounded theory approach using the constant comparison method to understand the contexts and processes informing participants' BR decision-making. Results: Twenty-three women participated, of whom 17 elected BR and 6 did not. Whereas women's primary reasons for deciding for or against BR differed, our core category, "empowered choices ," describes both groups' decision-making as a process focused on empowering themselves physically and/or psychologically, through self-advocacy, informed and shared decision-making, and giving back/receiving communal and spiritual support from church and African-American survivor groups. Socioeconomic factors influenced women's access to BR. Women preferred autologous BR and expressed the need for greater culturally-matched resources and support to inform treatment and shared BR decision-making. Conclusions: Understanding and supporting African-American women's BR preferences and empowerment is essential to ensuring equal access, and culturally-relevant, high-quality, and informed patient-centered care.

Perceptions and Barriers to Physical Activity in Childhood and Adulthood Among Latinas.

OBJECTIVE: The purpose of this qualitative study was to identify perceptions and barriers to physical activity in childhood and adulthood among Latina adults. METHODS: Three focus groups, 2 dyadic interviews, and an individual interview were conducted using semi-structured interview guides with 23 Latina women aged 21-35. A thematic analysis approach employing inductive and deductive coding was utilized to code, categorize, and summarize data into themes. RESULTS: The themes that emerged focused on: (1) physical activity is enjoyable; (2) family influenced physical activity; (3) different lifestyle in the US influenced physical activity; (4) physical activity is important for health; and (5) responsibilities (eg, work, caregiving) as barriers to physical activity in adulthood. CONCLUSIONS: Perceptions and barriers to physical activity experienced in both childhood and adulthood should be considered in the promotion of physical activity among Latinas.

Racial disparities in COVID-19 mortality across Michigan, United States

Black populations in the US are disproportionately affected by the COVID-19 pandemic, but the increased mortality burden after accounting for health and demographic characteristics is not well understood. We evaluated COVID-19 mortality in Michigan using individual-level death certificate and surveillance data from the Michigan Department of Health and Human Services from March 16 to October 26, 2020. Among the 6,065 COVID-19-related deaths, Black individuals experienced 3.6 times the mortality rate as White individuals. Black individuals under 65 years without comorbidities had a mortality rate 12.6 times that of their White counterparts. After accounting for age, sex, and comorbidities, we found that Black individuals in all strata are at higher risk of COVID-19 mortality than their White peers. We demonstrate that inequities in mortality are driven by ongoing systemic racism, as opposed to comorbidity burden or older age, and further highlight how underlying disparities across the race are compounded in crises.

Adapting and Evaluating Implementation of a Tobacco-Free Workplace Program in Behavioral Health Centers.

Objectives: State-administered community behavioral health centers (CBHCs) rarely treat tobacco dependence, despite high client tobacco use. Using a mixed-methods approach we examine the adaptation and implementation of an evidence-based tobacco-free workplace (TFW) program in 2 CBHCs (17 individual clinics). Methods: Varied data collection included pre- and post-implementation leader, clinician, and staff surveys; pre-, mid-, and post-implementation staff and client focus groups; and monthly implementation logs. The RE-AIM framework guided translation of behavioral interventions into sustainable practice. Results: Pre- to post-implementation increases were seen in training receipt among clinicians and employees. Both CBHCs adopted a 100% TFW policy, integrated tobacco screenings into routine practice, and delivered evidence-based practices (EBPs). Qualitative methods enlisted key stakeholders contributing towards adapting program strategies to local contexts, addressing barriers, adjusting tobacco screening administration, and understanding reasons for success or failure to implement specific components. Conclusions: Program implementation at both CBHCs increased organizational capacity in the provision of EBPs to treat tobacco dependence through successfully meeting the majority of our RE-AIM targets. Findings contribute to the development of flexible strategies and interventions responsive to variable implementation contexts and barriers; enhancing the effectiveness and sustainability of a TFW program.

Tobacco-free Workplace Program at a Substance Use Treatment Center.

Objectives: About 65%-87% of substance use disorder patients smoke cigarettes, compared to 14% of the general adult population. Few substance use treatment centers (SUTCs) have comprehensive tobacco-free workplace (TFW) policies or offer tobacco interventions. Taking Texas Tobacco Free (TTTF) implements an evidence-based TFW program in SUTCs, including at the Billy T. Cattan Recovery Outreach Center (BTC). We present a mixed methods case study of BTC's TTTF implementation, success factors, and challenges. Methods: TTTF provided policy development assistance, training, treatment resources, and technical assistance over ∼9 months. Implementation was tailored using mixed methods. Quantitative data included surveys to stakeholders (Nmax = 7), a pre- and post-training questionnaire assessing knowledge gain, and reported quantities of tobacco use assessments (TUAs) administered and nicotine replacement therapy (NRT) provided. Qualitative data included stakeholder focus groups and interviews (18 participants). Results: All employees reported TFW policy compliance. Employees exhibited a 20% knowledge gain. Clinicians increased self-report of NRT provision and tobacco cessation counseling. During implementation, BTC administered TUAs to 171 patients and dispensed NRT to 70 of 110 tobacco-using patients. Conclusion: Qualitative findings contextualized quantitative outcomes. TTTF implementation changed clinician attitudes, knowledge, and practices regarding tobacco treatment, facilitating patient quit attempts.

Impact of Gender and Relationship Status on Young People's Autonomy and Psychological Wellbeing.

This study uses scales of autonomy and psychological wellbeing to determine whether young people's gender and romantic relationship status give rise to differences in relation to a series of specific dimensions. To this end, we used Ryff's Model of Psychological Wellbeing, which comprises several dimensions: self-acceptance, positive relations with others, autonomy, environmental mastery, personal growth, and purpose in life; and our own Transition to Adulthood Autonomy Scale (EDATVA), whose dimensions are: self-organization, understanding context, critical thinking, and socio-political engagement. As a result, a quantitative study was performed with 1,148 young people aged 16-21 from Madrid, Spain and Bogotá, Colombia, of whom 60.2% were female and 39.8% were male. The findings show that in the gender variable there are differences between males and females in the dimensions of positive relations with others, personal growth (wellbeing questionnaire), and understanding context (autonomy questionnaire); the female sample obtained the highest scores. In the relationship variable, differences were found in environmental mastery and purpose in life; higher scores were obtained by young people in a romantic relationship. However, no differences were found in the different dimensions in the autonomy questionnaire between young people in a relationship and those not.

Clinician Training in the Adaptation of a Comprehensive Tobacco-Free Workplace Program in Agencies Serving the Homeless and Vulnerably Housed.

Tobacco use is exceedingly high among those who are homeless or at risk of homelessness but not commonly addressed by clinicians. Taking Texas Tobacco Free (TTTF) is a tobacco control program that addresses known clinician barriers to intervention (e.g., low training receipt, limited resources). Here, we examine the process and outcomes of TTTF's adaptation within four agencies that provide housing or other services to individuals who are homeless or vulnerably housed. Pre- and post-implementation data were collected from clinicians (N = 68) to assess changes in training receipt, knowledge, and intervention behaviors, relative to program goals. Results indicated significant gains in clinicians' receipt of training in 9 (of 9) target areas (p's ≤ 0.0042) and a 53% knowledge gain (p < 0.0001). From pre- to post-implementation, there were mean increases in the use of the 5As (ask, advise, assess, assist, and arrange) and other evidence-based interventions for tobacco cessation, with significant gains seen in assisting residents/clients to quit, arranging follow-ups, and providing or referring for non-nicotine medications (p's ≤ 0.0491). All program goals, except gains related to advising smokers to quit and the use of specific interventions (behavioral counseling), were met. Overall, TTTF improved clinicians' capacity to address tobacco use among homeless and vulnerably housed individuals and can serve as a model for tobacco control efforts in similar agencies.

Impact of mixing layer height variations on air pollutant concentrations and health in a European urban area: Madrid (Spain), a case study.

The occurrence of local high-pollution episodes in densely populated urban areas, which have huge fleets of vehicles, is currently one of the most worrying problems associated with air pollution worldwide. Such episodes are produced under specific meteorological conditions, which favour the sudden increase of levels of air pollutants. This study has investigated the influence of the mixing layer height (MLH) on the concentration levels of atmospheric pollutants and daily mortality in Madrid, Spain, during the period 2011-2014. It may help to understand the causes and impact of local high-pollution episodes. MLH at midday over Madrid was daily estimated from meteorological radio soundings. Then, days with different MLH over this urban area were characterized by meteorological parameters registered at different levels of an instrumented tower and by composite sea level pressure maps, representing the associated synoptic meteorological scenarios. Next, statistically significant associations between MLH and levels of PM10, PM2.5, NO, NO2, CO and ultra-fine particles number concentrations registered at representative monitoring stations were evaluated. Finally, associations between all-natural cause daily mortality in Madrid, MLH, and air pollutants were estimated using conditional Poisson regression models. The reduction of MLH to values below 482 m above-ground level under strong atmospheric stagnation conditions was accompanied by a statistically significant increase in levels of NO, NO2, CO, PM2.5 and ultra-fine particle number concentrations at urban-traffic and suburban monitoring sites. The decrease of the MLH was also associated to a linear increase of the daily number of exceedances of the UE NO2 hourly limit value (200 µg/m3) and levels of air pollutants at hotspot urban-traffic monitoring stations. Also, a statistically significant association of the MLH with all-natural cause daily mortality was obtained. When the MLH increased by 830 m, the risk of mortality decreased by 2.5% the same day and by 3.3% the next day, when African dust episodic days were excluded. They were also higher in absolute terms than the increases in risk of mortality that were determined for the exposition to any other air pollutant. Our results suggest that when the prediction models foresee values of MLH below 482 m above-ground level in Madrid, the evolution of high-contamination episodes will be very favourable. Therefore, short-term policy measures will have to be implemented to reduce NO, NO2, CO, PM2.5 and ultra-fine particle emissions from anthropogenic sources in this southern European urban location.

Concordance of Giardia duodenalis assemblages determined by different PCR methodologies in three observational studies in Cuba.

Giardia duodenalis is one of the most important intestinal parasites globally, especially in children, and in Cuba is the leading cause of chronic paediatric diarrhoea in this population. G. duodenalis is composed of eight genetic groups (or assemblages), two of which (A and B) are apparently zoonotic, occurring in both humans and other animals. However, consensus on the most appropriate genotyping scheme for optimal characterization of G. duodenalis isolates is lacking. In this article we present the results of three descriptive observational studies conducted in Havana, Cuba between 2010 and 2013, with the aim of comparing the results from molecular (PCR) approaches targeting different genes in order to assign with confidence 224 isolates of G. duodenalis to the correct assemblages. In each sub-study, following DNA isolation by the phenol/chloroform/isoamyl alcohol extraction method, PCR targeting the triose phosphate isomerase (tpi) gene was used for molecular characterization, as well as one additional PCR-method targeting another gene or pair of genes. DNA amplification was obtained in 87%, 83%, and 80% in the three sub-studies. Although excellent agreement (kappa index = 1) was recorded between results from some pairs of genes, for other combinations only moderate or substantial agreement was achieved. These results highlight the importance of interpretation of genotyping data, especially when single genetic markers are used. From the results of our studies, PCR targeting a combination of the tpi gene and the intergenic spacer region of rDNA may be a useful approach for the molecular characterization of G. duodenalis isolates.