The potential role of collagen type VII in breast cancer proliferation.

BACKGROUND: Breast cancer is the most common cancer in women. Cancer cells can persist in a prolonged dormant state for years without any clinical evidence of disease creating an urgent need to better understand the molecular mechanisms leading to relapse. This study aimed to identify extracellular matrix (ECM) components associated with hypoxia-induced breast cancer dormancy. The effects of selected ECM proteins on breast cancer cell proliferation were analyzed, along with their correlation with established prognostic markers in human breast cancer tissue. MATERIALS AND METHODS: Screening of extracellular matrix proteins was performed in hypoxia-induced dormant MCF-7 breast cancer cells. Proliferation of MCF-7 cells in vitro was subsequently determined in the presence of recombinant ColVII. Adipose tissue-derived mesenchymal stem cells (AdMSCs) subpopulation overexpressing ColVII were indirectly isolated by ColVII receptor integrin-α6 specific antibodies. AdMSCs- MCF-7 3D spheroid cultures were generated to model solid tumour conditions. In addition, the association between ColVII and various prognostic markers was evaluated in clinical samples of human breast cancer tissue. RESULTS: Dormant MCF-7 cells showed an elevated expression of ColVII while MCF-7 cells cultured on ColVII exhibited reduced proliferation in vitro. In AdMSCs-MCF-7 3D spheroids, a reduced proliferation of MCF-7 cells was observed in Int-α6+/ ColVIIhigh compared with Int-α6-/ ColVIIlow AdMSCs spheroids. In human tissue, high ColVII expression correlated to several positive prognostic markers. Staining for Cytokeratin-5 revealed that ColVIIhigh-expressing cells were predominantly myoepithelial cells. CONCLUSION: ColVII is associated with reduced proliferation of breast cancer cells in vitro. ColVII is strongly expressed in myoepithelial cells and in breast cancer tissue the high ColVII expression correlates with several well-known positive prognostic markers, highlighting its potential as a prognostic marker in breast cancer.

RE-START: Exploring the effectiveness of anti-calcitonin gene-related peptide resumption after discontinuation in migraine.

BACKGROUND AND PURPOSE: According to the latest European guidelines, discontinuation of monoclonal antibodies against calcitonin gene-related peptide (anti-CGRP MAb) may be considered after 12-18 months of treatment. However, some patients may worsen after discontinuation. In this study, we assessed the response following treatment resumption. METHODS: This was a prospective study conducted in 14 Headache Units in Spain. We included patients with response to anti-CGRP MAb with clinical worsening after withdrawal and resumption of treatment. Numbers of monthly migraine days (MMD) and monthly headache days (MHD) were obtained at four time points: before starting anti-CGRP MAb (T-baseline); last month of first treatment period (T-suspension); month of restart due to worsening (T-worsening); and 3 months after resumption (T-reintroduction). The response rate to resumption was calculated. Possible differences among periods were analysed according to MMD and MHD. RESULTS: A total of 360 patients, 82% women, with a median (interquartile range [IQR]) age at migraine onset of 18 (12) years. The median (IQR) MHD at T-baseline was 20 (13) and MMD was 5 (6); at T-suspension, the median (IQR) MHD was 5 (6) and MMD was 4 (5); at T-worsening, the median (IQR) MHD was 16 (13) and MMD was 12 (6); and at T-reintroduction, the median (IQR) MHD was 8 (8) and MHD was 5 (5). In the second period of treatment, a 50% response rate was achieved by 57.4% of patients in MHD and 65.8% in MMD. Multivariate models showed significant differences in MHD between the third month after reintroduction and last month before suspension of first treatment period (p < 0.001). CONCLUSION: The results suggest that anti-CGRP MAb therapy is effective after reintroduction. However, 3 months after resumption, one third of the sample reached the same improvement as after the first treatment period.

Lateral Evasive Maneuver with Shared Control Algorithm: A Simulator Study.

Shared control algorithms have emerged as a promising approach for enabling real-time driver automated system cooperation in automated vehicles. These algorithms allow human drivers to actively participate in the driving process while receiving continuous assistance from the automated system in specific scenarios. However, despite the theoretical benefits being analyzed in various works, further demonstrations of the effectiveness and user acceptance of these approaches in real-world scenarios are required due to the involvement of the human driver in the control loop. Given this perspective, this paper presents and analyzes the results of a simulator-based study conducted to evaluate a shared control algorithm for a critical lateral maneuver. The maneuver involves the automated system helping to avoid an oncoming motorcycle that enters the vehicle's lane. The study's goal is to assess the algorithm's performance, safety, and user acceptance within this specific scenario. For this purpose, objective measures, such as collision avoidance and lane departure prevention, as well as subjective measures related to the driver's sense of safety and comfort are studied. In addition, three levels of assistance (gentle, intermediate, and aggressive) are tested in two driver state conditions (focused and distracted). The findings have important implications for the development and execution of shared control algorithms, paving the way for their incorporation into actual vehicles.

Agonistic interactions and social behaviors in the Saharan Dorcas gazelle (Gazella dorcas neglecta): Using social network analysis to evaluate relationships and social structure in captive male groups.

Social interactions, including agonistic behavior, are very important for the management and welfare of individuals forming groups in captivity. One of the main concerns for the stability and durability of adult male groups is a noticeable level of intraspecific aggression. This study comprises a Social Network Analysis to illustrate social structure in different groups of captive Saharan Dorcas gazelles (Gazella dorcas neglecta). Our main objectives were to examine the relationship between agonistic, affiliative, and association networks and their reciprocity, assessing also whether the agonistic networks can conform to a linear dominance hierarchy. For these purposes, we recorded the behavior of 23 adult males organized in five herds, three composed only of Saharan Dorcas gazelle males and two mixed herds in which there were also Mhorr gazelle males (Nanger dama mohor). Observations were carried out during 295 h through scan sampling. We found no correlation between the affiliative and association networks in any group, although there was a significant correlation between the agonistic and association networks in mixed-species groups which was not present in single-species groups. Overall, there was no consistent reciprocity in either affiliative nor agonistic networks and none of the agonistic networks showed a linear structure. These results indicate that affiliative behavior in Saharan Dorcas gazelles offers distinctive and valuable information about the bonds between individuals, however, their dominance structure is far more complex than previously thought. As information provided by affiliative and proximity behaviors is different in this species, we suggest considering affiliative interactions to stablish affinity between individuals. Evaluating different social behaviors and not only agonistic interactions in later studies, is also recommended to develop a more accurately daily management in zoos that guarantee group stability and individuals' welfare, which will improve the conservation of captive populations.

Endophilin-B regulates autophagy during synapse development and neurodegeneration.

Synapses are critical for neuronal communication and brain function. To maintain neuronal homeostasis, synapses rely on autophagy. Autophagic alterations cause neurodegeneration and synaptic dysfunction is a feature in neurodegenerative diseases. In Parkinson's disease (PD), where the loss of synapses precedes dopaminergic neuron loss, various PD-causative proteins are involved in the regulation of autophagy. So far only a few factors regulating autophagy at the synapse have been identified and the molecular mechanisms underlying autophagy at the synapse is only partially understood. Here, we describe Endophilin-B (EndoB) as a novel player in the regulation of synaptic autophagy in health and disease. We demonstrate that EndoB is required for autophagosome biogenesis at the synapse, whereas the loss of EndoB blocks the autophagy induction promoted by the PD mutation LRRK2G2019S. We show that EndoB is required to prevent neuronal loss. Moreover, loss of EndoB in the Drosophila visual system leads to an increase in synaptic contacts between photoreceptor terminals and their post-synaptic synapses. These data confirm the role of autophagy in synaptic contact formation and neuronal survival.

Can Shared Control Improve Overtaking Performance? Combining Human and Automation Strengths for a Safer Maneuver.

The Shared Control (SC) cooperation scheme, where the driver and automated driving system control the vehicle together, has been gaining attention through the years as a promising option to improve road safety. As a result, advanced interaction methods can be investigated to enhance user experience, acceptance, and trust. Under this perspective, not only the development of algorithms and system applications are needed, but it is also essential to evaluate the system with real drivers, assess its impact on road safety, and understand how drivers accept and are willing to use this technology. In this sense, the contribution of this work is to conduct an experimental study to evaluate if a previously developed shared control system can improve overtaking performance on roads with oncoming traffic. The evaluation is performed in a Driver-in-the-Loop (DiL) simulator with 13 real drivers. The system based on SC is compared against a vehicle with conventional SAE-L2 functionalities. The evaluation includes both objective and subjective assessments. Results show that SC proved to be the best solution for assisting the driver during overtaking in terms of safety and acceptance. The SC's longer and smoother control transitions provide benefits to cooperative driving. The System Usability Scale (SUS) and the System Acceptance Scale (SAS) questionnaire show that the SC system was perceived as better in terms of usability, usefulness, and satisfaction.

Fusion pacing in patients with right bundle branch block who undergo cardiac resynchronization therapy.

PURPOSE: Patients with right bundle branch block (RBBB) are less likely to respond to cardiac resynchronization therapy (CRT). We aimed to assess whether patients with RBBB respond to CRT with biventricular fusion pacing. METHODS: Consecutive patients with RBBB at a single tertiary care center, who were implanted with a CRT device capable of biventricular fusion pacing using SyncAV programming, were assessed and compared to a historical cohort of CRT patients with RBBB. QRSd was measured and compared during intrinsic conduction, nominal CRT pacing and manual electrocardiogram-based optimized SyncAV programming. Left ventricular ejection fraction (LVEF) was also compared before and 6 months after CRT. RESULTS: We included 8 consecutive patients with RBBB (group 1) who were able to undergo SyncAV programming and 16 patients with RBBB (group 2) from a historical cohort. In group 1, compared to mean intrinsic conduction QRSd (155 ± 13 ms), mean nominally-paced QRSd was 156 ± 15 ms (ΔQRSd 1.3 ± 11.6; p = 0.77) and SyncAV-optimized paced QRSd was 135 ± 14 ms (ΔQRSd -20.0 ± 20.4; p = 0.03 and ΔQRSd -21.3 ± 16.3; p = 0.008; compared to intrinsic conduction and nominal pacing respectively). In group 2, mean QRSd with nominal pacing was 160 ± 24 ms (ΔQRSd 3.8 ± 33.4; p = 0.66 compared to intrinsic conduction). In group 1, baseline LVEF was 22.1 ± 11.5 and after 6 months of follow-up was 27.8 ± 8.6 (p = 0.047). In group 2, the baseline LVEF was 27.2 ± 10.6 and after 6 months of follow-up was 25.0 ± 10.0 (p = 0.45). CONCLUSIONS: CRT programed to allow biventricular fusion pacing significantly improved electrical synchrony and LVEF in patients with RBBB. Larger studies are required to confirm these findings.

Limb preference and personality in donkeys (Equus asinus).

Interhemispheric laterality has often been linked to different behavioural styles. This study investigates the link between limb preference and personality in donkeys. The sample consisted of 47 donkeys (Equus asinus), 30 males and 17 females. Limb preference was determined using observation of the leading limb in a motionless posture and personality was measured using the Donkey Temperament Questionnaire (French, J. M. (1993). Assessment of donkey temperament and the influence of home environment. Applied Animal Behaviour Science, 36(2), 249-257. doi:10.1016/0168-1591(93)90014-G) completed by the donkeys' keepers. A Principal Component Analysis obtained two components: Agreeableness and Extraversion. Age showed a positive relationship with Agreeableness, echoing trends in humans Donkeys did not show a population-level preference towards either side. Limb preference significantly predicted the trait difficult to handle: donkeys with a preference to keep the right foot forward when motionless were harder to handle. This study presents the first investigation into limb preference and personality in donkeys, although more research is needed to clarify whether there is a population-level limb preference bias in donkeys, and the relationship between limb preference and Agreeableness.

A Cost-Efficient 5G Non-Public Network Architectural Approach: Key Concepts and Enablers, Building Blocks and Potential Use Cases.

The provision of high data rate services to mobile users combined with improved quality of experience (i.e., zero latency multimedia content) drives technological evolution towards the design and implementation of fifth generation (5G) broadband wireless networks. To this end, a dynamic network design approach is adopted whereby network topology is configured according to service demands. In parallel, many private companies are interested in developing their own 5G networks, also referred to as non-public networks (NPNs), since this deployment is expected to leverage holistic production monitoring and support critical applications. In this context, this paper introduces a 5G NPN architectural approach, supporting among others various key enabling technologies, such as cell densification, disaggregated RAN with open interfaces, edge computing, and AI/ML-based network optimization. In the same framework, potential applications of our proposed approach in real world scenarios (e.g., support of mission critical services and computer vision analytics for emergencies) are described. Finally, scalability issues are also highlighted since a deployment framework of our architectural design in an additional real-world scenario related to Industry 4.0 (smart manufacturing) is also analyzed.

Epidemiology of hospitalized patients with COVID-19 in a tertiary care hospital.

INTRODUCTION: COVID-19, caused by the betacoronavirus SARS-CoV-2, has overwhelmed the world's health systems. OBJECTIVE: To describe the epidemiological characteristics of patients treated in a tertiary care hospital. METHODS: A retrospective cohort study of patients diagnosed with or suspected of having COVID-19 from March 23 to July 31, 2020 was conducted. RESULTS: 4,401 patients were hospitalized at Central Military Hospital, out of which 35 % were beneficiaries, 26 % civilians, 28 % active military personnel, and only 11 %, retired military personnel. Male gender predominated, both in hospitalized patients and in those who died, as well as the O+ group and absence of comorbidities; among the observed comorbidities, the main ones were overweight and diabetes. Hospitalized patients' median age was 49 years, while median age of those who died was 62 years; women older than 51 years had a higher risk of dying. Adjusted case fatality rate was 18.5 %; 50 % died within the first six days. CONCLUSIONS: In this study, the epidemiological characteristics and main comorbidities in Mexican patients with SARS-CoV-2 infection were identified.

INTRODUCCIÓN: COVID-19, causada por el betacoronavirus SARS-CoV-2, ha saturado los sistemas de salud del mundo. OBJETIVO: Describir las características epidemiológicas de los pacientes atendidos en un hospital de tercer nivel. MÉTODOS: Se realizó una cohorte retrospectiva de pacientes con diagnóstico o sospecha de COVID-19, del 23 de marzo al 31 de julio de 2020. RESULTADOS: En el Hospital Central Militar se hospitalizaron 4401 pacientes, 35 % derechohabientes, 26 % civiles, 28 % militares en activo y solo 11 %, militares retirados. Predominó el sexo masculino, tanto en los pacientes hospitalizados como en los que fallecieron, el grupo O+ y la ausencia de comorbilidades; entre las comorbilidades que se observaron, las principales fueron el sobrepeso y la diabetes. La mediana de edad de los pacientes hospitalizados fue de 49 años, mientras que 62 años fue la edad de quienes fallecieron; las mujeres mayores de 51 años tuvieron mayor riesgo de fallecer. La tasa de letalidad ajustada fue de 18.5 %; 50 % falleció durante los primeros seis días. CONCLUSIONES: En este estudio se lograron identificar las características epidemiológicas y se destacaron las principales comorbilidades en pacientes mexicanos con infección por SARS-CoV-2.

Loss of 5hmC identifies a new type of aberrant DNA hypermethylation in glioma.

Aberrant DNA hypermethylation is a hallmark of cancer although the underlying molecular mechanisms are still poorly understood. To study the possible role of 5-hydroxymethylcytosine (5hmC) in this process we analyzed the global and locus-specific genome-wide levels of 5hmC and 5-methylcytosine (5mC) in human primary samples from 12 non-tumoral brains and 53 gliomas. We found that the levels of 5hmC identified in non-tumoral samples were significantly reduced in gliomas. Strikingly, hypo-hydroxymethylation at 4627 (9.3%) CpG sites was associated with aberrant DNA hypermethylation and was strongly enriched in CpG island shores. The DNA regions containing these CpG sites were enriched in H3K4me2 and presented a different genuine chromatin signature to that characteristic of the genes classically aberrantly hypermethylated in cancer. As this 5mC gain is inversely correlated with loss of 5hmC and has not been identified with classical sodium bisulfite-based technologies, we conclude that our data identifies a novel 5hmC-dependent type of aberrant DNA hypermethylation in glioma.

Hormonal induction of spermiation in a Eurasian bufonid (Epidalea calamita).

BACKGROUND: Amphibian diversity is declining at an alarming rate due to habitat loss, invasive species, climate change and diseases. Captive assurance colonies have been established for some species at risk; however, many species do not breed well in captivity and the development of assisted reproductive technologies (ART) is critical to help sustain genetic diversity. To date, the majority of the work has been accomplished in species from the American continent and Australia, and there is a need to address similar breeding challenges in Eurasian and African species of amphibians. METHODS: The aim of this study was to develop a hormone protocol for stimulation of spermiation in Epidalea calamita as a model for Eurasian bufonids. Hence, the effect on sperm production and quality of three doses of chorionic gonadotropin hormone (5, 10 and 15 IU hCG/g) over time (1 to 24 h) was evaluated. In addition, cold storage (at 5 °C) of sperm for 24 and 48 h and three frequencies for hormonal treatment (weekly, biweekly and monthly) were examined. RESULTS: Hormone concentrations of 10 or 15 IU of hCG induced spermiation in 100% of males and produced sperm of comparable quality, while 5 IU hCG stimulated spermiation in only 40% of males. Total motility peaked between 1 to 4 h post-treatment with 10 IU hCG, whereas treatment with 15 IU hCG peaked between 2 to 6 h. After 24 h of cold storage total motility dropped by 20% and forward motility dropped by 10% for both the 10 and 15 IU treatments. Weekly hormone administration resulted in higher variation between trials in all motility parameters and a lower overall Total Motility and Forward Movement. Furthermore, the effect of exogenous hormone treatment overlapped between the last two trials in the weekly frequency. Sperm concentration was higher in the first trial for all frequencies but showed no differences among other trials. CONCLUSIONS: Overall, these results show that hormone concentration, time after treatment, frequency of hormone treatment and cold storage should be borne in mind when developing a hormone stimulation protocol for Eurasian amphibian species.

Natural genetic transformation of Vibrio parahaemolyticus via pVA1 plasmid acquisition as a potential mechanism causing AHPND.

Vibrio parahaemolyticus is the causative bacterium of acute hepatopancreatic necrosis disease (AHPND) in white shrimp Litopenaeus vannamei. This bacterium secretes protein toxins whose genes are encoded in an auto-transmissible plasmid called pVA1. The presence of this plasmid in V. parahaemolyticus is determinant for disease development. Its propagation is not only linked to bacterial colonisation capacity but also to horizontal gene transfer mechanisms. Nevertheless, the active uptake of plasmid, which is known as natural genetic transformation (NGT), has not yet been proposed as a possible acquisition mechanism of the pVA1 plasmid among Vibrio species. Previous studies suggest that some Vibrio species have the ability to undergo NGT in the presence of chitin. Therefore, the objective of this study was to evaluate the induction of NGT mediated by chitin in V. parahaemolyticus (ATCC-17802) through its ability to incorporate and express the pVA1 plasmid. The results showed that a reference strain that does not initially contain the plasmid can incorporate the plasmid under the appropriate transformation conditions, and cause mortality in white shrimp similar to that observed for pathogenic strains isolated from infectious outbreaks. Given the management and conditions of a shrimp farm with large amounts of chitinous exoskeletons, it is feasible that NGT could be a possible acquisition mechanism of plasmid pVA1 among Vibrio species, turning a non-causative strain of V. parahaemolyticus into a causative strain. With this study, we have expanded the knowledge of the pathogenesis process mediated by NGT and the understanding of the possible propagation mechanisms of emerging diseases in the aquaculture sector.

Cardiac resynchronization therapy reprogramming to improve electrical synchrony in patients with existing devices.

BACKGROUND: Optimal programming of cardiac resynchronization therapy (CRT) has not yet been fully elucidated. A novel algorithm (SyncAV) has been developed to improve electrical synchrony by fusion of the triple wavefronts: intrinsic, right ventricular (RV)-paced, and left ventricular (LV)-paced. METHODS: Consecutive patients at a single tertiary care center with a previously implanted CRT device with SyncAV algorithm (programmable negative AV hysteresis) were evaluated. QRS duration (QRSd) was measured during 1) intrinsic conduction, 2) existing CRT pacing as chronically programmed by treating physician, 3) using the device-based QuickOpt™ algorithm for optimization of AV and VV delays, and 4) ECG-based optimized SyncAV programming. The paced QRSd was assessed and compared to intrinsic conduction and between the different modes of programming. RESULTS: Of 64 consecutive, potentially eligible patients who underwent assessment, 34 patients who were able to undergo SyncAV programming were included. Mean intrinsic conduction QRSd was 163 ±â€¯24 ms. In comparison, the mean QRSd was 152 ±â€¯25 ms (-11.1 ±â€¯19.0) during existing CRT pacing, 160 ±â€¯25 ms (-4.1 ±â€¯25.2) using the QuickOpt™ algorithm and 138 ±â€¯23 (-24.9 ±â€¯17.2) using ECG-based optimized SyncAV programming. SyncAV optimization resulted in significant reductions in QRSd compared to existing CRT pacing (P = 0.02) and QuickOpt™ (P < 0.001). Of the 32% of patients who did not have QRS narrowing with existing CRT, 72% experienced QRS narrowing with SyncAV. CONCLUSION: ECG-based atrio-ventricular delay optimization using SyncAV significantly improved electrical synchrony in patients with a previously implanted CRT. Further studies are needed to assess the impact on long-term outcomes.

Production and characterization of a human lysosomal recombinant iduronate-2-sulfatase produced in Pichia pastoris.

Hunter syndrome (Mucopolysaccharidosis II, MPS II) is an X-linked lysosomal storage disease produced by the deficiency of the lysosomal enzyme iduronate-2-sulfatase (IDS). Currently, MPS II patients are mainly treated with enzyme replacement therapy (ERT) using recombinant enzymes produced in mammalian cells. As an alternative, several studies have shown the production of active and therapeutic forms of lysosomal proteins in microorganisms. In this paper, we report the production and characterization of a recombinant IDS produced in the yeast Pichia pastoris (prIDS). We evaluated the effect of culture conditions and gene sequence optimization on prIDS production. The results showed that the highest production of prIDS was obtained at oxygen-limited conditions using a codon-optimized IDS cDNA. The purified enzyme showed a final activity of 12.45 nmol mg-1 H-1 and an apparent molecular mass of about 90 kDa. The highest stability was achieved at pH 6.0, and prIDS also showed high stability in human serum. Noteworthy, the enzyme was taken up by culture cells in a dose-dependent manner through mannose receptors, which allowed the delivery of the enzyme to the lysosome. In summary, these results show the potential of Pichia pastoris as a host to produce an IDS intended for a MPS II ERT.

Knockdown of PTRF ameliorates adipocyte differentiation and functionality of human mesenchymal stem cells.

Healthy expansion of human adipose tissue requires mesenchymal stem cells (hMSC) able to proliferate and differentiate into mature adipocytes. Hence, characterization of those factors that coordinate hMSC-to-adipocyte transition is of paramount importance to modulate the adipose tissue expansion. It has been previously reported that the adipogenic program of hMSC can be disrupted by upregulating caveolar proteins, and polymerase I and transcript release factor (PTRF) is an integral component of caveolae, highly expressed in adipose tissue. Here, we hypothesized that the role of PTRF in adipocyte functionality might stem from an effect on hMSC. To test this hypothesis, we isolated hMSC from the subcutaneous fat depot. We found an upregulated expression of the PTRF associated with decreased adipogenic potential of hMSC, likely due to the existence of senescent adipocyte precursors. Employing short hairpin RNA-based constructs to stably reduce PTRF, we were able to restore insulin sensitivity and reduced basal lipolysis and leptin levels in human adipocytes with high levels of PTRF. Additionally, we pinpointed the detrimental effect caused by PTRF on the adipose tissue to the existence of senescent adipocyte precursors unable to proliferate and differentiate into adipocytes. This study provides evidence that impaired adipocyte functionality can be corrected, at least partially, by PTRF downregulation and warrants further in vivo research in patients with dysfunctional adipose tissue to prevent metabolic complications.

Human recombinant lysosomal enzymes produced in microorganisms.

Lysosomal storage diseases (LSDs) are caused by accumulation of partially degraded substrates within the lysosome, as a result of a function loss of a lysosomal protein. Recombinant lysosomal proteins are usually produced in mammalian cells, based on their capacity to carry out post-translational modifications similar to those observed in human native proteins. However, during the last years, a growing number of studies have shown the possibility to produce active forms of lysosomal proteins in other expression systems, such as plants and microorganisms. In this paper, we review the production and characterization of human lysosomal proteins, deficient in several LSDs, which have been produced in microorganisms. For this purpose, Escherichia coli, Saccharomyces cerevisiae, Pichia pastoris, Yarrowia lipolytica, and Ogataea minuta have been used as expression systems. The recombinant lysosomal proteins expressed in these hosts have shown similar substrate specificities, and temperature and pH stability profiles to those produced in mammalian cells. In addition, pre-clinical results have shown that recombinant lysosomal enzymes produced in microorganisms can be taken-up by cells and reduce the substrate accumulated within the lysosome. Recently, metabolic engineering in yeasts has allowed the production of lysosomal enzymes with tailored N-glycosylations, while progresses in E. coli N-glycosylations offer a potential platform to improve the production of these recombinant lysosomal enzymes. In summary, microorganisms represent convenient platform for the production of recombinant lysosomal proteins for biochemical and physicochemical characterization, as well as for the development of ERT for LSD.

Polymerase I and transcript release factor (PTRF) regulates adipocyte differentiation and determines adipose tissue expandability.

Impaired adipogenesis renders an adipose tissue unable to expand, leading to lipotoxicity and conditions such as diabetes and cardiovascular disease. While factors important for adipogenesis have been studied extensively, those that set the limits of adipose tissue expansion remain undetermined. Feeding a Western-type diet to apolipoprotein E2 knock-in mice, a model of metabolic syndrome, produced 3 groups of equally obese mice: mice with normal glucose tolerance, hyperinsulinemic yet glucose-tolerant mice, and prediabetic mice with impaired glucose tolerance and reduced circulating insulin. Using proteomics, we compared subcutaneous adipose tissues from mice in these groups and found that the expression of PTRF (polymerase I and transcript release factor) associated selectively with their glucose tolerance status. Lentiviral and pharmacologically overexpressed PTRF, whose function is critical for caveola formation, compromised adipocyte differentiation of cultured 3T3-L1cells. In human adipose tissue, PTRF mRNA levels positively correlated with markers of lipolysis and cellular senescence. Furthermore, a negative relationship between telomere length and PTRF mRNA levels was observed in human subcutaneous fat. PTRF is associated with limited adipose tissue expansion underpinning the key role of caveolae in adipocyte regulation. Furthermore, PTRF may be a suitable adipocyte marker for predicting pathological obesity and inform clinical management.

Understanding sociodemographic and sociocultural factors that characterize tobacco use and cessation during pregnancy among women in the Dominican Republic.

Tobacco use and exposure are serious public health problems that threaten to undermine improvements in maternal and child health, and add to already existing poor pregnancy outcomes in many low- and middle-income countries. The purpose of this study is to explore factors that characterize tobacco use and cessation during pregnancy among women in the Dominican Republic. This study was part of a larger trial and includes a sample of women who participated in baseline surveillance and community assessments (n = 613). Descriptive, bivariate, and multivariable analyses were conducted. Overall, 93.31 % (n = 572) of women experienced a past/current pregnancy and 22.44 % (n = 127) smoked during a past or current pregnancy. Among women who had smoked, 34.13 % (n = 43) stopped smoking due to a pregnancy, and 46.03 % (n = 58) were advised by a health care provider to quit smoking because of pregnancy. Women who were older, Catholic, and had a mother who used tobacco were three times more likely to smoke during a past or current pregnancy. Inability to read or write was also significantly associated with smoking during pregnancy. Women who were able to read and write and were from a tobacco growing community were three times more likely to quit smoking during pregnancy. This study provides a preliminary understanding of factors influencing tobacco use and cessation among pregnant women in the Dominican Republic. It also informs a critical area for public health research and intervention, indicating opportunities to engage the health care provider community in intervening with pregnant women and their families.

Cohort study of smoke-free homes in economically disadvantaged communities in the Dominican Republic.

OBJECTIVE: To analyze household smoking-ban prevalence over time and predictors among communities in the Dominican Republic, historically a significant tobacco-growing country with few tobacco control regulations. METHODS: Baseline (2004) and follow-up surveillance surveys (2006, 2007) (each n > 1 000 randomly selected households) conducted in six economically disadvantaged communities (three tobacco-growing and two each urban, peri-urban, and rural) assessed household members' demographics, health status, and household characteristics, including smoking restrictions. RESULTS: Between 2004 and 2007, household smoking-ban prevalence increased in all communities, with overall rates increasing from 23.9% (2004) to 45.3% (2007). Households with smokers adopted smoking bans at lower rates (6%-17%) versus those without smokers (which had an adoption rate of 35%-58%). Logistic regression models demonstrated that the associations between allowing smoking in households with no members who smoked and being located in a tobacco-growing community, being a Catholic household, and having a member with a cardiovascular problem were statistically significant. The association between having a child under age 5 or a member with a respiratory condition and prohibiting smoking in the home was not statistically significant. CONCLUSIONS: Prevalence of households banning smoking increased in all communities but remained well below rates in industrialized countries. For low- and middle-income countries or those in early stages of tobacco control, basic awareness-raising measures (including surveillance activities) may lead to statistically significant increases in household smoking-ban adoption, particularly among households with no smokers. An increase in household smoking-ban prevalence may result in changes in community norms that can lead to a further increase in the adoption of smoking bans. Having household members who smoke and being in a tobacco-growing community may mitigate the establishment of household bans. Increasing individuals' knowledge about the far-reaching health effects of secondhand smoke exposure on children and nonsmoking adults (healthy or unhealthy) may help overcome these obstacles.