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The RS case is one of the latest rulings issued by the Court of Justice in the neglected Romanian rule of law saga. Together with other European counterparts, the Romanian Constitutional Court is openly questioning some basic postulates on which the European legal order is based. It joins a growing number of constitutional jurisdictions that are challenging the Luxembourgian conception of the principle of primacy and its interpretative authority. They use for this purpose a worrying articulation of national identity in terms that are incompatible with the effectiveness, uniformity and coherence of Union law. This article aims to address a series of judicial disagreements between the Romanian Constitutional Court and the CJEU, with particular attention to the RS case, in which it becomes evident the pernicious consequences of the tough crisis of the rule of law for the most elementary principles of the European legal system.

Abstract Filamentous fungi are critical in the transition to a more sustainable food system. While genetic modification of these organisms has promise for enhancing the nutritional value, sensory appeal, and scalability of fungal foods, genetic tools and demonstrated use cases for bioengineered food production by edible strains are lacking. Here, we develop a modular synthetic biology toolkit for Aspergillus oryzae, an edible fungus used in fermented foods, protein production, and meat alternatives. Our toolkit includes a CRISPR-Cas9 method for gene integration, neutral loci, and tunable promoters. We use these tools to elevate intracellular levels of the nutraceutical ergothioneine and the flavor-and color molecule heme in the edible biomass. The strain overproducing heme is red in color and is readily formulated into imitation meat patties with minimal processing. These findings highlight the promise of synthetic biology to enhance fungal foods and provide useful genetic tools for applications in food production and beyond.

Human activities directly impact estuaries, where the biota is exposed to sediment contamination. A contamination assessment was carried out in several estuaries in the Southern Iberian Peninsula. Sediment samples were analyzed for the presence of metals/metalloids, and bioaccumulation tests were conducted with clams (Ruditapes philippinarum). Huelva Estuary had the highest contamination levels, while the inner bay of Cádiz and the outer stations of the estuaries from Guadiana, Guadalquivir, Palmones, and Guadarranque were the lesser contaminated. All sampling points (except Huelva) had low contamination levels of As and Cd, but they had high concentrations of Cu. The elements Pb, Cd, Zn, Cu, As, and Hg displayed correlations between the concentrations in sediments and the biota. High bioaccumulation of Zn, Cu, and As was observed in Huelva and Barbate. Important insights into the sediment contamination in Southern Iberian Peninsula estuaries suggest greater management and conservation efforts in these critical ecosystems.

ABSTRACTTrichoderma species are known for their mycoparasitic activity against phytopathogenic fungi that cause significant economic losses in agriculture. During mycoparasitism, Trichoderma spp. recognize molecules produced by the host fungus and release secondary metabolites and hydrolytic enzymes to kill and degrade the host’s cell wall. Here, we explored the participation of the Trichoderma atroviride RNAi machinery in the interaction with six phytopathogenic fungi of economic importance. We determined that both Argonaute-3 and Dicer-2 play an essential role during mycoparasitism. Using an RNA-Seq approach, we identified that perception, detox, and cell wall degradation depend on the T. atroviride-RNAi when interacting with Alternaria alternata, Rhizoctonia solani AG2, and R. solani AG5. Furthermore, we constructed a gene co-expression network that provides evidence of two gene modules regulated by RNAi, which play crucial roles in essential processes during mycoparasitism. In addition, based on small RNA-seq, we conclude that siRNAs regulate amino acid and carbon metabolism and communication during the Trichoderma-host interaction. Interestingly, our data suggest that siRNAs might regulate allorecognition (het) and transport genes in a cross-species manner. Thus, these results reveal a fine-tuned regulation in T. atroviride dependent on siRNAs that is essential during the biocontrol of phytopathogenic fungi, showing a greater complexity of this process than previously established.IMPORTANCEThere is an increasing need for plant disease control without chemical pesticides to avoid environmental pollution and resistance, and the health risks associated with the application of pesticides are increasing. Employing Trichoderma species in agriculture to control fungal diseases is an alternative plant protection strategy that overcomes these issues without utilizing chemical fungicides. Therefore, understanding the biocontrol mechanisms used by Trichoderma species to antagonize other fungi is critical. Although there has been extensive research about the mechanisms involved in the mycoparasitic capability of Trichoderma species, there are still unsolved questions related to how Trichoderma regulates recognition, attack, and defense mechanisms during interaction with a fungal host. In this work, we report that the Argonaute and Dicer components of the RNAi machinery and the small RNAs they process are essential for gene regulation during mycoparasitism by Trichoderma atroviride.

Food production is one of the most environmentally damaging human activities. In the face of climate change, it is essential to rethink our dietary habits and explore potential alternative foods catering both towards human and planetary needs. Fungal mycelium might be an attractive alternative protein source due to its rapid growth on sustainable substrates as well as promising nutritional and organoleptic properties. The natural biodiversity of filamentous fungi is vast and represents an untapped reservoir for food innovation. However, fungi are known to produce bioactive compounds that may affect human health, both positively and negatively. To narrow the search for safe and culinarily attractive fungal species, mycelia of edible fruiting-body forming fungi provide a promising starting point. Here, we explore whether the culinary attractiveness and safety of the commonly eaten mushroom, Pleurotus ostreatus, can also be translated to its mycelium.Whole-genome sequencing and pan-genome analysis revealed a high degree of genetic variability within the genus Pleurotus, suggesting that gastronomic traits as well as food safety may differ between strains. A representative strain, P. ostreatus M2191, was further analyzed for the food safety, nutritional properties and culinary applicability of its mycelium. No regulated mycotoxins were detected in either the fruiting body nor the mycelium. Yet, P. ostreatus is known to produce four peptide toxins, Ostreatin, Ostreolysin and Pleurotoysin A/B. These were found to be lower in the mycelium compared to fruiting bodies, which are already considered safe for consumption. Instead, a number of secondary metabolites with potential health benefits were detected in the fungal mycelium. In silico analysis of the proteome suggested low allergenicity. In addition, the fruiting body and the mycelium showed similar nutritional value, which was dependent on the growth substrate.To highlight the culinary potential of mycelium, we created a dish served at the two-star restaurant the Alchemist in Copenhagen, Denmark. Sensory analysis of the mycelium dish by an untrained consumer panel indicated consumer liking and openness to fungal mycelia.Based on sustainability, safety, culinary potential, and consumer acceptance, our findings suggest that P. ostreatus mycelium has great potential for use as a novel food source.

Ancient Andean agricultural landscapes have been the subject of a large number of archaeological and agro-ecological studies, which generally refer to regions with favourable environmental conditions or, in the case of arid and semi-arid environments, those with irrigation facilities. The aim of this article is to present and analyse the pre-Hispanic rain-fed farming systems widely represented in two adjacent regions of Bolivia’s arid and cold southern Altiplano. The search for archaeological agricultural areas combined aerial analysis and field surveys. Agro-ecological characterisation was based on historical and ethnographic studies of the region’s present-day populations. Despite their geographical proximity, similar environmental conditions, and same agropastoral way of life, the typology of cultivated areas developed in the southern altiplano differs significantly. Within this same framework of adaptation and resilience, the sectorisation of agricultural systems observed in these two regions reveals a regional productive specialisation that favoured internal exchanges and exchanges with other regions. These differences are related to two models of non-centralised, low-inequality societies—one strongly based on cohesion and the other characterised by greater fragmentation and social conflict—underlining the limits of strict environmental determinism in shaping agricultural landscapes. These results provide new food for thought in the debate on the use and value of rain-fed agricultural practices and more broadly on the diversity of adaptations by human societies in extreme and unstable environmental contexts.

La existencia en Coctaca de grandes áreas agrícolas prehispánicas cautivó tempranamente la atención de los arqueólogos, siendo considerada como uno de los principales referentes de la agricultura nativa. Los nuevos estudios indican que, con el establecimiento de los inkas en la región, tuvo lugar en Coctaca una dinámica de neta intensificación de la producción agrícola en la que se readecuaron espacios de cultivo prexistentes y se crearon otros nuevos, en un área aproximada de 450 ha. Se resalta particularmente en las características del sistema agrícola el aprovechamiento pormenorizado de los recursos hídricos y la mitigación de las limitantes ambientales. Más allá de la agricultura, la colosal inversión volcada en este desarrollo productivo hace de Coctaca una de las mayores y más evidentes manifestaciones del poder del Estado inkaico en esta parte de los Andes.

BackgroundStroke is a leading cause of lifelong disability worldwide, partially driven by a reduced ability to use the upper limb in daily life causing increased dependence on caregivers. However, post-stroke functional impairments have only been investigated using limited clinical scores, during short-term longitudinal studies in relatively small patient cohorts. With the addition of technology-based assessments, we propose to complement clinical assessments with more sensitive and objective measures that could more holistically inform on upper limb impairment recovery after stroke, its impact on upper limb use in daily life, and on overall quality of life. This paper describes a pragmatic, longitudinal, observational study protocol aiming to gather a uniquely rich multimodal database to comprehensively describe the time course of upper limb recovery in a representative cohort of 400 Asian adults after stroke. Particularly, we will characterize the longitudinal relationship between upper limb recovery, common post-stroke impairments, functional independence and quality of life.MethodsParticipants with stroke will be tested at up to eight time points, from within a month to 3 years post-stroke, to capture the influence of transitioning from hospital to community settings. We will perform a battery of established clinical assessments to describe the factors most likely to influence upper limb recovery. Further, we will gather digital health biomarkers from robotic or wearable sensing technology-assisted assessments to sensitively characterize motor and somatosensory impairments and upper limb use in daily life. We will also use both quantitative and qualitative measures to understand health-related quality of life. Lastly, we will describe neurophysiological motor status using transcranial magnetic stimulation.StatisticsDescriptive analyses will be first performed to understand post-stroke upper limb impairments and recovery at various time points. The relationships between digital biomarkers and various domains will be explored to inform key aspects of upper limb recovery and its dynamics using correlation matrices. Multiple statistical models will be constructed to characterize the time course of upper limb recovery post-stroke. Subgroups of stroke survivors exhibiting distinct recovery profiles will be identified.ConclusionThis is the first study complementing clinical assessments with technology-assisted digital biomarkers to investigate upper limb sensorimotor recovery in Asian stroke survivors. Overall, this study will yield a multimodal data set that longitudinally characterizes post-stroke upper limb recovery in functional impairments, daily-life upper limb use, and health-related quality of life in a large cohort of Asian stroke survivors. This data set generates valuable information on post-stroke upper limb recovery and potentially allows researchers to identify different recovery profiles of subgroups of Asian stroke survivors. This enables the comparisons between the characteristics and recovery profiles of stroke survivors in different regions. Thus, this study lays out the basis to identify early predictors for upper limb recovery, inform clinical decision-making in Asian stroke survivors and establish tailored therapy programs.Clinical trial registrationClinicalTrials.gov, identifier: NCT05322837.

Using wearable technologies in the home setting is an emerging option for self-directed rehabilitation. A comprehensive review of its application as a treatment in home-based stroke rehabilitation is lacking. This review aimed to 1) map the interventions that have used wearable technologies in home-based physical rehabilitation for stroke, and 2) provide a synthesis of the effectiveness of wearable technologies as a treatment choice. Electronic databases of the Cochrane Library, MEDLINE, CINAHL, and Web of Science were systematically searched for work published from their inception to February 2022. This scoping review adopted Arksey and O’Malley’s framework in the study procedure. Two independent reviewers screened and selected the studies. Twenty-seven were selected in this review. These studies were summarized descriptively, and the level of evidence was assessed. This review identified that most research focused on improving the hemiparetic upper limb (UL) function and a lack of studies applying wearable technologies in home-based lower limb (LL) rehabilitation. Virtual reality (VR), stimulation-based training, robotic therapy, and activity trackers are the interventions identified that apply wearable technologies. Among the UL interventions, “strong” evidence was found to support stimulation-based training, “moderate” evidence for activity trackers, “limited” evidence for VR, and “inconsistent evidence” for robotic training. Due to the lack of studies, understanding the effects of LL wearable technologies remains “very limited.” With newer technologies like soft wearable robotics, research in this area will grow exponentially. Future research can focus on identifying components of LL rehabilitation that can be effectively addressed using wearable technologies.

Summary: Polycyclopropanated (POP) compounds show promise as fuels as their energy density can be greater than jet and rocket fuels in current use, but realizing their full potential requires significant development. This protocol guides the production of polycyclopropanated fatty acids in Streptomyces; POP production in another host remains to be demonstrated. This method can serve as a baseline for further development of POP as well as other polyketide products.For complete details on the use and execution of this protocol, please refer to Cruz-Morales et al. (2022).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Background The use of wearable technology offers a promising home-based self-directed option for upper limb training. Although product usability is a crucial aspect of users’ acceptance of a wearable device, usability studies in wearable devices are rare, with most studies focusing primarily on clinical validity. Objective This study aimed to explore the usability of a wristwatch device called “Smart reminder” for home-based upper limb telerehabilitation for persons with stroke. Methods Eleven stroke participants used the proposed wristwatch for at least two weeks and underwent a home-based telerehabilitation program. A mixed-methods design was used to explore the usability of the wristwatch. Quantitative data were collected through the System Usability Scale (SUS) questionnaire, and the participants’ rate of therapy compliance (gathered from the device) was reported descriptively. In addition, qualitative data were collected through semi-structured interviews with the participants and were analyzed using thematic analysis. Results The results demonstrated that the usability of the proposed wristwatch and telerehabilitation system was rated highly by the participants, with a high SUS mean score of 84.3 (12.3) and high therapy compliance rate (mean = 91%). Qualitatively, all participants reported positive experiences with the wristwatch and indicated keenness to use it again. Participants reported physical improvements and felt motivated to exercise after using the wristwatch. They found the device easy and convenient and appreciated the remote monitoring function. Meanwhile, they highlighted critical considerations for the design of the device and program, including technical support, a wearable design of the device, graded exercise content according to ability, and flexibility in exercise schedules. Finally, they suggested that an interim review with the therapist on their progress might help them continue using the wristwatch. Conclusions This study's results supported the proposed wearable device's usability and showed strong acceptance by the participants for using it as a home-based upper limb telerehabilitation intervention.

La identidad constitucional europea es una categoría jurídica emergente en el Derecho de la Unión Europea. A pesar del silencio que guardan los Tratados constitutivos, el Tribunal de Justicia ha proclamado por primera vez de manera explícita su existencia. En particular, declara que está constituida por los valores fundamentales a que se refiere el capital artículo 2 TUE. Su alumbramiento se enmarca en el contexto de la crisis del Estado de Derecho como una insoslayable respuesta judicial al discurso iliberal preponderante en Polonia y Hungría. Por todo ello, interesa profundizar en el significado y ámbito de aplicación de esta figura jurídica, así como también en el paralelismo que guarda con la cláusula de identidad nacional (artículo 4.2 TUE). Recibido: 28.11.2022 Aceptado: 13.12.2022

Shooting is practiced mostly by police or military, it has become an Olympic sport where the air pistol test stands out, which requires extreme mental concentration and precision in its movements to achieve effectiveness in the shot. In this work, the authors aim to identify the kinematic data of the arm-weapon complex in normal and excited state during the execution of the shot, considering variables such as heart rate, the effectiveness of the shot and the angles of the main joints (glecohumeral) that allow the execution of the technique. As part of the methodology, this research was implemented in the closed shooting range of the Escuela Superior Militar "Eloy Alfaro" and seven athletes that form the shooting teams of the Ecuadorian Army and Air Force were analyzed. The heart rate (in four phases), the effectiveness of the shot (in three opportunities) and the angle of the glenohumeral joint in resting and excited states were analyzed, where the pertinent comparisons were made. As results of the study, it is shown that the mean values of the heart rate were significantly different (p=0.001), being higher in the excited state. The effectiveness of the shot does not establish significant differences (p=0.620) between states, as well as the glenohumeral joint angles (p=0.209). It is concluded that there are no significant differences in the effectiveness of shooting in excitement and at rest, an indicator of good physical preparation, although there is less effectiveness of shooting in the excited state.

Abstract Background The psychological well-being of university students is an important factor in successfully coping with the demands of academic life. This study aimed to assess the impact of a peer-led intervention of mental health promotion combined with coping-strategy-based group workshops on mental health awareness and help-seeking behavior among university students in Hong Kong. Method A mixed-method concurrent design was used for this study. Quantitative data, based on one-group pretest-posttest design, were collected using Mental Health Knowledge Schedule Questionnaire to assess mental health awareness, and Attitude Towards Seeking Professional Help Questionnaire-Short Form to examine help-seeking behavior of university students from The Hong Kong Polytechnic University. Qualitative data were collected from written post-activity reflections and focus group discussions which were thematically analyzed. Results A total of 62 university students (mean age: 23.2 ± 5.1 years) were included in this study. Mental health awareness was significantly improved (p = 0.015, 95% Confidence Interval of − 2.670, − 0.297) after program implementation. Help-seeking behavior mean score increased from pretest to posttest, however, no significant difference was observed (p = 0.188, 95% CI = − 1.775, 0.355). Qualitative analysis revealed that the program helped participants learn about coping strategies to help themselves and others with mental health challenges. Conclusions The peer-led intervention provided a positive impact through increased mental health awareness and knowledge of coping strategies on self-help and helping others among university students. Further study could focus on the impact of the program when applied regularly throughout the entire academic year.

Articulando las informaciones brindadas por las fuentes documentales coloniales con datos arqueológicos y aquellos resultantes de pruebas de arqueología experimental, en este artículo nos centraremos en una determinada categoría de horno de reverbero, utilizado en el espacio surandino desde los primeros momentos de la Colonia, principalmente para la producción de plata. Testimonios de esta categoría de horno, compuesto de tres cuerpos (cámara de fuego, cámara de trabajo abovedada y chimenea), fueron identificados en distintos centros mineros-metalúrgicos estudiados en el sur de Bolivia (Potosí, Santa Isabel, Mina Santiago, Escoriani, entre otros). Contestando la caracterización generalizada de estos hornos como de tradición europea, en este trabajo proponemos que esta tecnología resultó de un desarrollo tecnológico local, mostrando algunas continuidades significativas con la metalurgia prehispánica, la cual posteriormente se difundió en Europa.

According to information provided by colonial documentary sources, a large number of ceremonial spaces linked to the ancient cults of the tutelary wak’a were identified in different regions of the central-southern Andean highlands (Bolivia). Placed on the tops of low hills and close to the settlements, they are distinguished by a variable number of stone walls arranged in a concentric manner. Although their chronology begins in pre-Hispanic times, the majority of these sites continued to be in use in the long term. Articulating new archaeological data with historical and ethnographic information, this paper presents and analyzes different aspects of these ceremonial spaces in the Andean highlands.

Hydroxyglutarate synthase (HglS) converts 2-oxoadipate to D-2- hydroxyglutarate during lysine catabolism in bacteria. Here the authors use structural and biochemical approaches to show that HglS acts via successive decarboxylation and intramolecular hydroxylation and that homologous enzymes catalyze the final step of lysine catabolism in plants.

The article analyzes how the Mexican subnational governments responded the COVID 19 outbreak during the first stage of the crisis. The response of two subnational governments whose territory is part of the largest metropolitan area in the country that has been the focus of the pandemic is presented. We ask about the coordination instruments to face COVID-19 between the federal policy and programs with the subnational governments and argue that, during the first stage of the management of the COVID-19 crisis the institutional and organizational design of the Mexican political-administrative system has had a relevant impact on the government’s response, as well in the coordination policies. The article demonstrates that in a crisis, coordination instruments do not depend on political parties but consist of a blend of weak and suitable articulation of policies of the subnational governments with the federal strategy, as well as the need to improve the instruments of collaboration between the various levels of government in the context of a healthcare system in transformation.

Objective. To evaluate and compare vaccination coverage among children aged 12-23 and 24-35 months living in localities with less than 100 000 inhabitants in Encuesta Nacional de Salud y Nutrición (Ensanut) 2012 and Ensanut 100k (2018). Materials and methods. Estimate of coverage with both surveys. Results. Between 2012 and 2018, according to proof and self-report, the coverage of the basic scheme was maintained in children aged 12-23 (51.6 vs. 60.2%) and 24-35 months (51.4 vs. 50.0%). Similarly, only with proof (53.9 vs. 51.3% and 52.8 vs. 44.2%). In children aged 24-35 months, the coverage of the reinforced basic scheme reinforcements with probative document and self-report (30.9 vs. 34.0%) and only with reinforcements (30.2 vs. 27.8%) was maintained. Coverage with second and third doses of hepatitis B in both age groups decreased; additionally, first dose of measlesmumps-rubella vaccine (SRP, in Spanish) and third dose of Pentavalent in children aged 24-35 months. Conclusions. Coverages were maintained by schemes, despite reductions in hepatitis B, pentavalent and SRP.

Objective. To compare the prevalence of acute respiratory infections (ARI) and acute diarrheal disease (ADD) among children younger than five years of age living in localities with less than 100 000 inhabitants in Encuesta Nacional de Salud y Nutrición (Ensanut) 2012 and Ensanut 100k (2018). In Ensanut 100k, we evaluate the associated factors. Materials and methods. Analysis of both surveys and of the Mexican Meteorological System. Results. The estimated prevalence of ARI was 45.1% in 2012 vs. 32.9% in 2018. The decrease was significant among medium and high-income households. There were no changes in trends for ADD. Among households with lower EC, ARI was associated with roofing material, temperature, and rainy precipitation while ADD was associated with lack of piped water. Conclusions. The estimated prevalence of ARI has decreased in medium and high income households. Some households and weather conditions are associated with ARI and ADD.

Abstract Background Determine the impact of poor oral health on the oral health-related quality of life (OHRQoL) in community-dwelling older adults. Methods Cross-sectional study of community-dwelling older adults in Mexico City. Sociodemographic characteristics were obtained and assessed their OHRQoL according to the Geriatric/General Oral Health Assessment Index (GOHAI). Clinical evaluation of their oral health: painful chewing, use of dentures, dry mouth, xerostomia, plaque, calculus, coronal and root caries, tooth loss and gingival bleeding. Finally, we determined the oral health of participants through Latent Class Analysis (LCA), excluding totally edentulous. The strength of association was determined (Odds Ratio [OR] and 95% confidence interval [95% CI]) through logical regression between the oral health categories (latent classes) and OHRoL in older adults, adjusted with the other variables included in the study: age, sex, marital status, living arrangements (lives alone), educational level, paid work status, comorbidity, cognitive deterioration, depression and use of medical and dental services in the previous 12 months. Results The mean (SD) GOHAI score for the 228 older adults to 46.5 (8.7), number of classes to characterize oral health through LCA was three (entropy 0.805). The GOHAI mean for Class 3 (57.0%), acceptable oral health was 50.1 (7.1); totally edentulous (9.6%), 47.9 (8.4); for Class 2 (16.7%), regular oral health, 43.8 (9.3); and for Class 1 (16.7%), poor oral health, 42.2 (9.7). Significant differences were observed among means (p

Objectives: To estimate hepatitis B virus (HBV) seroprevalence from natural infection or vaccination in 10–25-year-olds in Mexico, using the 2012 National Health and Nutrition Survey (ENSANUT). Methods: Randomly selected serum samples (1,581) from adolescents and young adults, representative of 38,924,584 Mexicans, were analyzed to detect hepatitis B surface antigen (HBsAg), hepatitis B surface antibody (anti-HBs) and hepatitis B core antibody (anti-HBc). Weighted HBV seroprevalence in the Mexican population and association with sociodemographic variables were calculated. Results: Overall weighted seroprevalence from natural infection (positive for anti-HBs and anti-HBc) was 0.23% (95% confidence interval [95% CI] 0.10–0.52). No HBsAg was detected, indicating no acute or chronic infection. Vaccine-derived immunity (positive ≥ 10.0 mIU/ml for anti-HBs and negative to anti-HBc) was 44.7% (95% CI: 40.2–49.4) overall; lower in persons aged 20–25 years (40.83%) than in persons aged 10–19 years (47.7%). Among the population analyzed, 54.2% (95% CI: 49.6–58.8) were seronegative to HBV (negative for all three markers) and no sociodemographic risk factors were identified. Conclusions: HBV seroprevalence from natural infection was low. Vaccination-induced immunity was higher among Mexican adolescents than young adults, possibly due to vaccination policies since 1999.

Breast cancer is one of the most frequent cancer types worldwide and the first cause of cancer-related deaths in women. Although significant therapeutic advances have been achieved with drugs such as tamoxifen and trastuzumab, breast cancer still caused 627,000 deaths in 2018. Since cancer is a multifactorial disease, it has become necessary to develop new molecular therapies that can target several relevant cellular processes at once. Ion channels are versatile regulators of several physiological- and pathophysiological-related mechanisms, including cancer-relevant processes such as tumor progression, apoptosis inhibition, proliferation, migration, invasion, and chemoresistance. Ion channels are the main regulators of cellular functions, conducting ions selectively through a pore-forming structure located in the plasma membrane, protein–protein interactions one of their main regulatory mechanisms. Among the different ion channel families, the Transient Receptor Potential (TRP) family stands out in the context of breast cancer since several members have been proposed as prognostic markers in this pathology. However, only a few approaches exist to block their specific activity during tumoral progress. In this article, we describe several TRP channels that have been involved in breast cancer progress with a particular focus on their binding partners that have also been described as drivers of breast cancer progression. Here, we propose disrupting these interactions as attractive and potential new therapeutic targets for treating this neoplastic disease.

Background: passive immunotherapy is a therapeutic alternative for patients with COVID-19. Equine polyclonal antibodies (EpAbs) could represent a source of scalable neutralizing antibodies against SARS-CoV-2. Methods: we conducted a double-blind, randomized, placebo-controlled trial to assess efficacy and safety of EpAbs (INM005) in hospitalized adult patients with moderate and severe COVID-19 pneumonia in 19 hospitals of Argentina. Primary endpoint was improvement in at least two categories in WHO ordinal clinical scale at day 28 or hospital discharge (ClinicalTrials.gov number NCT04494984). Findings: between August 1st and October 26th, 2020, a total of 245 patients were enrolled. Enrolled patients were assigned to receive two blinded doses of INM005 (n = 118) or placebo (n = 123). Median age was 54 years old, 65•1% were male and 61% had moderate disease at baseline. Median time from symptoms onset to study treatment was 6 days (interquartile range 5 to 8). No statistically significant difference was noted between study groups on primary endpoint (risk difference [95% IC]: 5•28% [-3•95; 14•50]; p = 0•15). Rate of improvement in at least two categories was statistically significantly higher for INM005 at days 14 and 21 of follow-up. Time to improvement in two ordinal categories or hospital discharge was 14•2 (± 0•7) days in the INM005 group and 16•3 (± 0•7) days in the placebo group, hazard ratio 1•31 (95% CI 1•0 to 1•74). Subgroup analyses showed a beneficial effect of INM005 over severe patients and in those with negative baseline antibodies. Overall mortality was 6•9% the INM005 group and 11•4% in the placebo group (risk difference [95% IC]: 0•57 [0•24 to 1•37]). Adverse events of special interest were mild or moderate; no anaphylaxis was reported. Interpretation: Albeit not having reached the primary endpoint, we found clinical improvement of hospitalized patients with SARS-CoV-2 pneumonia, particularly those with severe disease.

In 1908, after a short visit to the region, Eric Boman published the first detailed information about pre-Hispanic and Colonial Era mining and metallurgy in Cobres. Today, 110 years after the Swedish researcher's visit, recent studies allow for a better understanding of the different productive and technological dynamics that developed over the long run in this locality in the Salta highlands, based on the exploitation of the metalliferous minerals. Here, drawing on new archaeological data, we examine pre-Hispanic metal production in Cobres and analyze the discontinuities and transformations that occurred during the subsequent Colonial Period. Beyond certain particularities, the case of Cobres highlights the relevance of small Andean mining sites that were often left aside or overshadowed historically and in regional historiography, by the major productive and political centers.

In the 12,000 years preceding the Industrial Revolution, human activities led to significant changes in land cover, plant and animal distributions, surface hydrology, and biochemical cycles. Earth system models suggest that this anthropogenic land cover change influenced regional and global climate. However, the representation of past land use in earth system models is currently oversimplified. As a result, there are large uncertainties in the current understanding of the past and current state of the earth system. In order to improve representation of the variety and scale of impacts that past land use had on the earth system, a global effort is underway to aggregate and synthesize archaeological and historical evidence of land use systems. Here we present a simple, hierarchical classification of land use systems designed to be used with archaeological and historical data at a global scale and a schema of codes that identify land use practices common to a range of systems, both implemented in a geospatial database. The classification scheme and database resulted from an extensive process of consultation with researchers worldwide. Our scheme is designed to deliver consistent, empirically robust data for the improvement of land use models, while simultaneously allowing for a comparative, detailed mapping of land use relevant to the needs of historical scholars. To illustrate the benefits of the classification scheme and methods for mapping historical land use, we apply it to Mesopotamia and Arabia at 6 kya (c. 4000 BCE). The scheme will be used to describe land use by the Past Global Changes (PAGES) LandCover6k working group, an international project comprised of archaeologists, historians, geographers, paleoecologists, and modelers. Beyond this, the scheme has a wide utility for creating a common language between research and policy communities, linking archaeologists with climate modelers, biodiversity conservation workers and initiatives.

The scarcity or unpredictability of natural resources is a threat to cooperation within human societies. Exacerbated competition between individuals could affect social cohesion and collective action, generate conflicts over natural resources, and compromise their sustainable use. Yet, our in-depth archaeological study of the arid Andean highlands of Bolivia reveals the sustainable development of a complex agrarian society in a harsh environment marked, moreover, by a prolonged climatic degradation from the 13th to the 15th centuries. The 49 community settlements studied comprised independent family households that managed their own economic resources. A detailed study of the granary and housing structures of 549 of these households provided a strong quantitative data set for an analysis of Gini coefficients for grain storage capacity and housing area. This agro-pastoral society flourished with neither notable inequalities of wealth between villagers nor apparent long-lasting conflicts between villages. By sharing local knowledge, labor, and natural resources, this society succeeded both in limiting power and wealth concentration, and in sustainably producing food surpluses to be exchanged with neighboring populations. These results indicate a high degree of social cohesion and low levels of social and wealth inequality, similar to other well-established horticultural and agricultural societies around the world. We propose a conceptual model of low inequality in agrarian societies subject to extreme or unstable environments, where the sharing of knowledge, resources, and labor are the adaptive social responses to cope with the uncertainty in natural resources. The sustainability of the society is then guaranteed by a balance between collective action and family-based social organization.

Neurological and neuropsychiatric disorders are mediated by several pathophysiological mechanisms, including developmental and degenerative abnormalities caused primarily by disturbances in cell migration, structural plasticity of the synapse, and blood-vessel barrier function. In this context, critical pathways involved in the pathogenesis of these diseases are related to structural, scaffolding, and enzymatic activity-bearing proteins, which participate in Ca2+- and Ras Homologs (Rho) GTPases-mediated signaling. Rho GTPases are GDP/GTP binding proteins that regulate the cytoskeletal structure, cellular protrusion, and migration. These proteins cycle between GTP-bound (active) and GDP-bound (inactive) states due to their intrinsic GTPase activity and their dynamic regulation by GEFs, GAPs, and GDIs. One of the most important upstream inputs that modulate Rho GTPases activity is Ca2+ signaling, positioning ion channels as pivotal molecular entities for Rho GTPases regulation. Multiple non-selective cationic channels belonging to the Transient Receptor Potential (TRP) family participate in cytoskeletal-dependent processes through Ca2+-mediated modulation of Rho GTPases. Moreover, these ion channels have a role in several neuropathological events such as neuronal cell death, brain tumor progression and strokes. Although Rho GTPases-dependent pathways have been extensively studied, how they converge with TRP channels in the development or progression of neuropathologies is poorly understood. Herein, we review recent evidence and insights that link TRP channels activity to downstream Rho GTPase signaling or modulation. Moreover, using the TRIP database, we establish associations between possible mediators of Rho GTPase signaling with TRP ion channels. As such, we propose mechanisms that might explain the TRP-dependent modulation of Rho GTPases as possible pathways participating in the emergence or maintenance of neuropathological conditions.

Pseudomonas putida is a promising bacterial chassis for metabolic engineering given its ability to metabolize a wide array of carbon sources, especially aromatic compounds derived from lignin. However, this omnivorous metabolism can also be a hindrance when it can naturally metabolize products produced from engineered pathways. Herein we show that P. putida is able to use valerolactam as a sole carbon source, as well as degrade caprolactam. Lactams represent important nylon precursors, and are produced in quantities exceeding one million tons per year (Zhang et al., 2017). To better understand this metabolism we use a combination of Random Barcode Transposon Sequencing (RB-TnSeq) and shotgun proteomics to identify the oplBA locus as the likely responsible amide hydrolase that initiates valerolactam catabolism. Deletion of the oplBA genes prevented P. putida from growing on valerolactam, prevented the degradation of valerolactam in rich media, and dramatically reduced caprolactam degradation under the same conditions. Deletion of oplBA, as well as pathways that compete for precursors L-lysine or 5-aminovalerate, increased the titer of valerolactam from undetectable after 48 h of production to ~90 mg/L. This work may serve as a template to rapidly eliminate undesirable metabolism in non-model hosts in future metabolic engineering efforts.

Clavulanic acid is a bacterial specialized metabolite, which inhibits certain serine β-lactamases, enzymes that inactivate β-lactam antibiotics to confer resistance. Due to this activity, clavulanic acid is widely used in combination with penicillin and cephalosporin (β-lactam) antibiotics to treat infections caused by β-lactamase-producing bacteria. Clavulanic acid is industrially produced by fermenting Streptomyces clavuligerus, as large-scale chemical synthesis is not commercially feasible. Other than S. clavuligerus, Streptomyces jumonjinensis and Streptomyces katsurahamanus also produce clavulanic acid along with cephamycin C, but information regarding their genome sequences is not available. In addition, the Streptomyces contain many biosynthetic gene clusters thought to be “cryptic,” as the specialized metabolites produced by them are not known. Therefore, we sequenced the genomes of S. jumonjinensis and S. katsurahamanus, and examined their metabolomes using untargeted mass spectrometry along with S. clavuligerus for comparison. We analyzed the biosynthetic gene cluster content of the three species to correlate their biosynthetic capacities, by matching them with the specialized metabolites detected in the current study. It was recently reported that S. clavuligerus can produce the plant-associated metabolite naringenin, and we describe more examples of such specialized metabolites in extracts from the three Streptomyces species. Detailed comparisons of the biosynthetic gene clusters involved in clavulanic acid (and cephamycin C) production were also performed, and based on our analyses, we propose the core set of genes responsible for producing this medicinally important metabolite.

ABSTRACT Despite intensive study for 50 years, the biochemical and genetic links between lysine metabolism and central metabolism in Pseudomonas putida remain unresolved. To establish these biochemical links, we leveraged random barcode transposon sequencing (RB-TnSeq), a genome-wide assay measuring the fitness of thousands of genes in parallel, to identify multiple novel enzymes in both l- and d-lysine metabolism. We first describe three pathway enzymes that catabolize l-2-aminoadipate (l-2AA) to 2-ketoglutarate (2KG), connecting d-lysine to the TCA cycle. One of these enzymes, P. putida 5260 (PP_5260), contains a DUF1338 domain, representing a family with no previously described biological function. Our work also identified the recently described coenzyme A (CoA)-independent route of l-lysine degradation that results in metabolization to succinate. We expanded on previous findings by demonstrating that glutarate hydroxylase CsiD is promiscuous in its 2-oxoacid selectivity. Proteomics of selected pathway enzymes revealed that expression of catabolic genes is highly sensitive to the presence of particular pathway metabolites, implying intensive local and global regulation. This work demonstrated the utility of RB-TnSeq for discovering novel metabolic pathways in even well-studied bacteria, as well as its utility a powerful tool for validating previous research. IMPORTANCE P. putida lysine metabolism can produce multiple commodity chemicals, conferring great biotechnological value. Despite much research, the connection of lysine catabolism to central metabolism in P. putida remained undefined. Here, we used random barcode transposon sequencing to fill the gaps of lysine metabolism in P. putida. We describe a route of 2-oxoadipate (2OA) catabolism, which utilizes DUF1338-containing protein P. putida 5260 (PP_5260) in bacteria. Despite its prevalence in many domains of life, DUF1338-containing proteins have had no known biochemical function. We demonstrate that PP_5260 is a metalloenzyme which catalyzes an unusual route of decarboxylation of 2OA to d-2-hydroxyglutarate (d-2HG). Our screen also identified a recently described novel glutarate metabolic pathway. We validate previous results and expand the understanding of glutarate hydroxylase CsiD by showing that can it use either 2OA or 2KG as a cosubstrate. Our work demonstrated that biological novelty can be rapidly identified using unbiased experimental genetics and that RB-TnSeq can be used to rapidly validate previous results.

The natural grasslands that form part of the “Campos” of South America contain a large number of species at the paddock level (high α diversity), but little differentiation among paddocks (low β diversity). Thus, forage resources at the farm level have slightly different seasonal growth peaks, in part due to the low frequency of fertilization, the lack of differential grazing management and mowing of these plant communities. To stimulate diversification of these forage resources, it is possible to take advantage of differential responses of each type of plant community to changes in their use. Characterizing species by functional traits allows to orient the use of forage resources according their functional composition, thus increasing the diversity of vegetation types, which favors differentiation of growth peaks among paddocks. After an initial division between C 3 and C4 species, 63 Poaceae species were classified into eight groups or plant functional types (PFT) according to their preferred degree of soil fertility and use intensity. Based on the leaf dry matter content (LDMC) measured in experiments in Brazil, Uruguay and Argentina, we distinguished four PFT, two for C3 species and two for C4 species, with LDMC less than or equal to 300 mg/g. The species of these four PFT are adapted to fertile environments and intensive defoliation. Fertilizing and using more intensively vegetation dominated by species with these low LDMC can diversify the use value of paddocks, thus facilitating use of forage resources at the farm level. https://doi.org/10.25260/EA.19.29.2.0.727

This article examines the agrarian landscape in one part of the southern Andes (Quebrada of Humahuaca, Jujuy, Argentina). The region possesses extensive and well-preserved archaeological remains of agricultural systems, which stretch back to pre-Hispanic times. In this study, we employ an interdisciplinary approach in our analysis of the components that structure the agrarian landscape, especially those historical processes that intervened in its formation. The creation of a cartographic base, built from remote sensing and fieldwork data, allowed for the identification of four principal components of the landscape, each of which correspond to distinct phases or periods that mark the region’s history. Our study shows that, in contrast to what is observed in many other rural areas, the successive productive dynamics that developed in the area did not result in the destruction of previous productive structures. Rather, the agrarian landscape in the study area presents a multi-temporal agglutinating combination or composition, which transcends historical discontinuities in the productive matrix. This is owing to the partial reutilisation of previous structures in each period; however, religious and cultural factors play an important role. The agrarian landscape we studied is not only a passive result of human activity, but also a force influencing the productive and lifestyle decisions of the peasant populations that live there today. Our research amplifies the understanding of agrarian landscapes in the Andes and shows how past temporalities are articulated with the present through a dialectical process.

Cell migration is critical for several physiological and pathophysiological processes. It depends on the coordinated action of kinases, phosphatases, Rho-GTPases proteins, and Ca2+ signaling. Interestingly, ubiquitination events have emerged as regulatory elements of migration. Thus, the role of proteins involved in ubiquitination processes could be relevant to a complete understanding of pro-migratory mechanisms. KCTD5 is a member of Potassium Channel Tetramerization Domain (KCTD) proteins that have been proposed as a putative adaptor for Cullin3-E3 ubiquitin ligase and a novel regulatory protein of TRPM4 channels. Here, we study whether KCTD5 participates in cell migration-associated mechanisms, such as focal adhesion dynamics and cellular spreading. Our results show that KCTD5 CRISPR/Cas9- and shRNA-based depletion in B16-F10 cells promoted an increase in cell migration and cell spreading, and a decrease in the focal adhesion area, consistent with an increased focal adhesion disassembly rate. The expression of a dominant-negative mutant of Rho-GTPases Rac1 precluded the KCTD5 depletion-induced increase in cell spreading. Additionally, KCTD5 silencing decreased the serum-induced Ca2+ response, and the reversion of this with ionomycin abolished the KCTD5 knockdown-induced decrease in focal adhesion size. Together, these data suggest that KCTD5 acts as a regulator of cell migration by modulating cell spreading and focal adhesion dynamics through Rac1 activity and Ca2+ signaling, respectively.

Background: Transcranial direct-current stimulation (tDCS) facilitates cognitive improvement in healthy and pathological populations. It has been increasingly used in cases of mild cognitive impairment (MCI) and dementia. Our research question is: Can tDCS serve as a clinical intervention for improving the cognitive functions of persons with MCI (PwMCI) and dementia (PwD)?Objective: This systematic review evaluated the evidence to determine the efficacy of tDCS in improving cognitive outcomes in PwD and PwMCI.Methods: A systematic review was conducted of studies published up to November 2017 involving tDCS in cases of MCI and dementia. Studies were ranked according to the level of evidence (Oxford Center for Evidence-Based Medicine) and assessed for methodological quality (Risk of Bias Tool in the Cochrane Handbook for Systematic Reviews of Interventions). Data was extracted on all protocol variables to establish a reference framework for clinical interventions. Different modalities, tDCS alone or combined with cognitive training, compared with sham tDCS were examined in both short and long-term effects. Four randomized control trials (RCTs) with memory outcomes were pooled using the fixed-effect model for the meta-analysis.Results: Twelve studies with 195 PwD and four with 53 PwMCI met the inclusion criteria. Eleven articles were ranked as Level 1b. The results on the meta-analysis on pooled effects of memory indicated a statistically significant medium effect size of 0.39 (p = 0.04) for immediate effects. This improvement was not maintained in the long term 0.15 (p = 0.44).Conclusion: tDCS improves memory in PwD in the short term, it also seems to have a mild positive effect on memory and language in PwMCI. However, there is no conclusive advantage in coupling tDCS with cognitive training. More rigorous evidence is needed to establish whether tDCS can serve as an evidence-based intervention for both populations.

An Australian marine tunicate-derived fungus, Talaromyces sp. CMB-TU011 was subjected to a program of analytical microbioreactor (MATRIX) cultivations, supported by UHPLC-QTOF profiling, to reveal conditions for producing a new class of extensively N-methylated 11-12 residue linear peptides, talaropeptides A-D (2-5). The structures for 2-5, inclusive of absolute configurations, were determined by a combination of detailed spectroscopic and chemical (e.g., C3 and C18 Marfey's) analyses. We report on the biological properties of 2-5, including plasma stability, as well as antibacterial, antifungal and cell cytotoxicity. The talaropeptide mega non-ribosomal peptide synthetase (NRPS) is described, as second only in size to that for the fungus-derived immunosuppressant cyclosporine (an 11-residue extensively N-methylated cyclic peptide).

ABSTRACT The expansion of the genetic repertoire of an organism by gene duplication or horizontal gene transfer (HGT) can aid adaptation. Streptomyces bacteria are prolific producers of bioactive specialized metabolites that have adaptive functions in nature and have found extensive utility in human medicine. While the biosynthesis of these specialized metabolites is directed by dedicated biosynthetic gene clusters, little attention has been focused on how these organisms have evolved robustness in their genomes to facilitate the metabolic plasticity required to provide chemical precursors for biosynthesis during the complex metabolic transitions from vegetative growth to specialized metabolite production and sporulation. Here, we examine genetic redundancy in actinobacteria and show that specialized metabolite-producing bacterial families exhibit gene family expansion in primary metabolism. Focusing on a gene duplication event, we show that the two pyruvate kinases in the genome of Streptomyces coelicolor arose by an ancient duplication event and that each has evolved altered enzymatic kinetics, with Pyk1 having a 20-fold-higher kcat than Pyk2 (4,703 s−1 compared to 215 s−1, respectively), and yet both are constitutively expressed. The pyruvate kinase mutants were also found to be compromised in terms of fitness compared to wild-type Streptomyces. These data suggest that expanding gene families can help maintain cell functionality during metabolic perturbation such as nutrient limitation and/or specialized metabolite production. IMPORTANCE The rise of antimicrobial-resistant infections has prompted a resurgence in interest in understanding the production of specialized metabolites, such as antibiotics, by Streptomyces. The presence of multiple genes encoding the same enzymatic function is an aspect of Streptomyces biology that has received little attention; however, understanding how the metabolic expansion influences these organisms can help enhance production of clinically useful molecules. Here, we show that expanding the number of pyruvate kinases enables metabolic adaptation, increases strain fitness, and represents an excellent target for metabolic engineering of industrial specialized metabolite-producing bacteria and the activation of cryptic specialized metabolites.

Memory CD8+ T cells are ideal candidates for cancer immunotherapy because they can mediate long-term protection against tumors. However, the therapeutic potential of different in vitro-generated CD8+ T cell effector subsets to persist and become memory cells has not been fully characterized. Type 1 CD8+ T (Tc1) cells produce interferon-γ and are endowed with high cytotoxic capacity, whereas IL-17-producing CD8+ T (Tc17) cells are less cytotoxic but display enhanced self-renewal capacity. We sought to evaluate the functional properties of in vitro-generated Tc17 cells and elucidate their potential to become long lasting memory cells. Our results show that in vitro-generated Tc17 cells display a greater in vivo persistence and expansion in response to secondary antigen stimulation compared to Tc1 cells. When transferred into recipient mice, Tc17 cells persist in secondary lymphoid organs, present a recirculation behavior consistent with central memory T cells, and can shift to a Tc1 phenotype. Accordingly, Tc17 cells are endowed with a higher mitochondrial spare respiratory capacity than Tc1 cells and express higher levels of memory-related molecules than Tc1 cells. Together, these results demonstrate that in vitro-generated Tc17 cells acquire a central memory program and provide a lasting reservoir of Tc1 cells in vivo, thus supporting the use of Tc17 lymphocytes in the design of novel and more effective therapies.

Objective. The aim of this pilot study was to investigate whether the use of anodal transcranial direct current stimulation (tDCS) on the left dorsolateral prefrontal cortex could boost the effects of a cognitive stimulation (CS) programme using a tablet on five older adults with mild cognitive impairment (MCI). Method. A single-subject study of A-B-C-A design was used. After the baseline with the administration of CS (phase A), a sham treatment with CS was applied (B). Following the withdrawal of sham treatment, tDCS was introduced in combination with CS (C). Finally, phase A was replicated a second time. Results. tDCS had a significant effect on processing speed, selective attention, and planning ability tasks in terms of performance and completion time. Conclusion. tDCS appears to have a positive impact on some cognitive components in CS in persons with MCI. Further study on its long-term effects and generalization of power to daily activities is warranted.

Genotyping and georeferencing in tuberculosis (TB) have been used to characterize the distribution of the disease and occurrence of transmission within specific groups and communities.The objective of this study was to test the hypothesis that diabetes mellitus (DM) and pulmonary TB may occur in spatial and molecular aggregations.Retrospective cohort study of patients with pulmonary TB. The study area included 12 municipalities in the Sanitary Jurisdiction of Orizaba, Veracruz, México. Patients with acid-fast bacilli in sputum smears and/or Mycobacterium tuberculosis in sputum cultures were recruited from 1995 to 2010. Clinical (standardized questionnaire, physical examination, chest X-ray, blood glucose test and HIV test), microbiological, epidemiological, and molecular evaluations were carried out. Patients were considered "genotype-clustered" if two or more isolates from different patients were identified within 12 months of each other and had six or more IS6110 bands in an identical pattern, or < 6 bands with identical IS6110 RFLP patterns and spoligotype with the same spacer oligonucleotides. Residential and health care centers addresses were georeferenced. We used a Jeep hand GPS. The coordinates were transferred from the GPS files to ArcGIS using ArcMap 9.3. We evaluated global spatial aggregation of patients in IS6110-RFLP/ spoligotype clusters using global Moran´s I. Since global distribution was not random, we evaluated "hotspots" using Getis-Ord Gi* statistic. Using bivariate and multivariate analysis we analyzed sociodemographic, behavioral, clinic and bacteriological conditions associated with "hotspots". We used STATA® v13.1 for all statistical analysis.From 1995 to 2010, 1,370 patients >20 years were diagnosed with pulmonary TB; 33% had DM. The proportion of isolates that were genotyped was 80.7% (n = 1105), of which 31% (n = 342) were grouped in 91 genotype clusters with 2 to 23 patients each; 65.9% of total clusters were small (2 members) involving 35.08% of patients. Twenty three (22.7) percent of cases were classified as recent transmission. Moran`s I indicated that distribution of patients in IS6110-RFLP/spoligotype clusters was not random (Moran`s I = 0.035468, Z value = 7.0, p = 0.00). Local spatial analysis showed statistically significant spatial aggregation of patients in IS6110-RFLP/spoligotype clusters identifying "hotspots" and "coldspots". GI* statistic showed that the hotspot for spatial clustering was located in Camerino Z. Mendoza municipality; 14.6% (50/342) of patients in genotype clusters were located in a hotspot; of these, 60% (30/50) lived with DM. Using logistic regression the statistically significant variables associated with hotspots were: DM [adjusted Odds Ratio (aOR) 7.04, 95% Confidence interval (CI) 3.03-16.38] and attending the health center in Camerino Z. Mendoza (aOR18.04, 95% CI 7.35-44.28).The combination of molecular and epidemiological information with geospatial data allowed us to identify the concurrence of molecular clustering and spatial aggregation of patients with DM and TB. This information may be highly useful for TB control programs.

Streptomyces are Gram-positive bacteria that occupy diverse ecological niches including host-associations with animals and plants. Members of this genus are known for their overwhelming repertoire of natural products, which has been exploited for almost a century as a source of medicines and agrochemicals. Notwithstanding intense scientific and commercial interest in Streptomyces natural products, surprisingly little is known of the intra- and/or inter-species ecological roles played by these metabolites. In this report we describe the chemical structures, biological properties, and biosynthetic relationships between natural products produced by Streptomyces isolated from internal tissues of predatory Conus snails, collected from the Great Barrier Reef, Australia. Using chromatographic, spectroscopic and bioassays methodology, we demonstrate that Streptomyces isolated from five different Conus species produce identical chemical and antifungal profiles – comprising a suite of polycyclic tetramic acid macrolactams (PTMs). To investigate possible ecological (and evolutionary) relationships we used genome analyses to reveal a close taxonomic relationship with other sponge-derived and free-living PTM producing Streptomyces (i.e., Streptomyces albus). In-depth phylogenomic analysis of PTM biosynthetic gene clusters indicated PTM structure diversity was governed by a small repertoire of genetic elements, including discrete gene acquisition events involving dehydrogenases. Overall, our study shows a Streptomyces-Conus ecological relationship that is concomitant with specific PTM chemical profiles. We provide an evolutionary framework to explain this relationship, driven by anti-fungal properties that protect Conus snails from fungal pathogens.

Cancer is characterized by an uncontrolled cell proliferation rate even under low nutrient availability, which is sustained by a metabolic reprograming now recognized as a hallmark of cancer. Warburg was the first to establish the relationship between cancer and mitochondria; however, he interpreted enhanced aerobic glycolysis as mitochondrial dysfunction. Today it is accepted that many cancer cell types need fully functional mitochondria to maintain their homeostasis. Calcium (Ca2+)—a key regulator of several cellular processes—has proven to be essential for mitochondrial metabolism. Inositol 1,4,5-trisphosphate receptor (IP3R)-mediated Ca2+ transfer from the endoplasmic reticulum to the mitochondria through the mitochondrial calcium uniporter (MCU) proves to be essential for the maintenance of mitochondrial function and cellular energy balance. Both IP3R and MCU are overexpressed in several cancer cell types, and the inhibition of the Ca2+ communication between these two organelles causes proliferation arrest, migration decrease, and cell death through mechanisms that are not fully understood. In this review, we summarize and analyze the current findings in this area, emphasizing the critical role of Ca2+ and mitochondrial metabolism in cancer and its potential as a novel therapeutic target.

Dans les Andes, les populations locales contemporaines considèrent les chullpas, tombes abritant des momies, comme des vestiges d’une population antérieure à l’humanité actuelle, également appelée de ce nom. Le rapport de continuité et de filiation établi avec cette population du passé varie toutefois grandement selon les communautés. Certaines l’affirment tandis que d’autres la rejettent radicalement. Ces différences ont des effets sur le social et le politique ainsi que sur les conceptions des rapports au passé et à l’autochtonie. Dans plusieurs régions des Andes boliviennes, on comparera les modes de relation entretenus avec les chullpas, et les différents facteurs, récents ou plus anciens, à l’origine de ces variations. Dans le premier cas, des changements importants ont été induits par le développement du tourisme tandis que l’UNESCO, le gouvernement et diverses ONG ont favorisé un fort processus de patrimonialisation de la culture et du territoire. Dans le second, ces processus sont à ce jour inexistants, le rapport, ancien et conflictuel, entre les villages aymaras et celui de Chipaya (Uru) sont fondés sur une opposition devenue ontologique illustrée, notamment, par la façon dont sont définis et classés différents monuments et vestiges de la région.

Many studies have explored the relationship between diabetes mellitus (DM) and tuberculosis (TB) demonstrating increased risk of TB among patients with DM and poor prognosis of patients suffering from the association of DM/TB. Owing to a paucity of studies addressing this question, it remains unclear whether patients with DM and TB are more likely than TB patients without DM to be grouped into molecular clusters defined according to the genotype of the infecting Mycobacterium tuberculosis bacillus. That is, whether there is convincing molecular epidemiological evidence for TB transmission among DM patients. Objective: We performed a systematic review and meta-analysis to quantitatively evaluate the propensity for patients with DM and pulmonary TB (PTB) to cluster according to the genotype of the infecting M. tuberculosis bacillus.We conducted a systematic search in MEDLINE and LILACS from 1990 to June, 2016 with the following combinations of key words "tuberculosis AND transmission" OR "tuberculosis diabetes mellitus" OR "Mycobacterium tuberculosis molecular epidemiology" OR "RFLP-IS6110" OR "Spoligotyping" OR "MIRU-VNTR". Studies were included if they met the following criteria: (i) studies based on populations from defined geographical areas; (ii) use of genotyping by IS6110- restriction fragment length polymorphism (RFLP) analysis and spoligotyping or mycobacterial interspersed repetitive unit-variable number of tandem repeats (MIRU-VNTR) or other amplification methods to identify molecular clustering; (iii) genotyping and analysis of 50 or more cases of PTB; (iv) study duration of 11 months or more; (v) identification of quantitative risk factors for molecular clustering including DM; (vi) > 60% coverage of the study population; and (vii) patients with PTB confirmed bacteriologically. The exclusion criteria were: (i) Extrapulmonary TB; (ii) TB caused by nontuberculous mycobacteria; (iii) patients with PTB and HIV; (iv) pediatric PTB patients; (v) TB in closed environments (e.g. prisons, elderly homes, etc.); (vi) diabetes insipidus and (vii) outbreak reports. Hartung-Knapp-Sidik-Jonkman method was used to estimate the odds ratio (OR) of the association between DM with molecular clustering of cases with TB. In order to evaluate the degree of heterogeneity a statistical Q test was done. The publication bias was examined with Begg and Egger tests. Review Manager 5.3.5 CMA v.3 and Biostat and Software package R were used.Selection criteria were met by six articles which included 4076 patients with PTB of which 13% had DM. Twenty seven percent of the cases were clustered. The majority of cases (48%) were reported in a study in China with 31% clustering. The highest incidence of TB occurred in two studies from China. The global OR for molecular clustering was 0.84 (IC 95% 0.40-1.72). The heterogeneity between studies was moderate (I2 = 55%, p = 0.05), although there was no publication bias (Beggs test p = 0.353 and Eggers p = 0.429).There were very few studies meeting our selection criteria. The wide confidence interval indicates that there is not enough evidence to draw conclusions about the association. Clustering of patients with DM in TB transmission chains should be investigated in areas where both diseases are prevalent and focus on specific contexts.

Objetivo. Evaluar los efectos de diferentes tipos de alimentos sobre la productividad de Diaphanosoma sp., y Alona sp bajo condiciones de laboratorio. Materiales y métodos. Juveniles de Diaphanosoma sp., y Alona sp., fueron sembrados en acuarios de 20 L a una densidad de 6 org L-1 y mantenidos con aireación constante y fotoperíodo 12:12 (luz : oscuridad). Se alimentaron con Saccharomyces cereviseae y Chlorella vulgaris a concentración de 4 x 105 Cel/ml cada uno, en tres combinaciones: 1. C. vulgaris, 2. S. cereviseae, 3. C. vulgaris + S. cereviseae en proporciones iguales; de cada tratamiento se realizaron 3 réplicas. Con el propósito de determinar la densidad poblacional, cada 3 días y durante un período de 27 días, y se midió la concentración de individuos por ml. Resultados. Se obtuvieron mejores resultados en el tratamientos de microalgas + levadura, con valores de TCE=0.1323±0.07 y 0.1598±0.04 días-1; R=0.010 ±0.004 y 0.019±0.01 org ml-1 día-1, TD=5.2±0.5 y 4.3±0.38 días para Diaphanosoma sp., y Alona sp., respectivamente, pero sin presentar diferencias significativas (p>0.05). Conclusiones. La dieta mixta de levadura y microalgas produjo mayor eficiencia productiva en las dos especies estudiadas, constituyendo una alternativa de fuente de alimento vivo para la acuicultura.

Objetivo. Determinar la variación de la calidad, el contenido iónico y la osmolaridad del plasma seminal de Pseudoplatystoma metaense durante la estación reproductiva. Materiales y métodos. Se utilizaron machos sexualmente maduros durante un periodo reproductivo (marzo a agosto). La espermiación fue inducida con Extracto de Hipófisis de Carpa (4 mg/kg). Se evaluó el volumen (mL), movilidad masal (%), tiempo de activación (sg), espermatocrito (%); concentración espermática (106 sptz/µL) y viabilidad (%). También fue determinada la movilidad (%) y velocidad individual por medio de un Sistema de Análisis Espermático Asistido por Computador (CASA). La osmolaridad (mOsm/Kg) y la concentración de iones (Na+, Cl+, K+ y Mg2+) y glucosa fue determinada en plasma seminal obtenido por centrifugación del semen a 14.000 g. La concentración de iones y glucosa fue establecida a través de un sistema de reflectancia. Resultados. El semen de Pseudoplatystoma metaense presentó una movilidad masal mayor al 90%, con el menor valor de movilidad progresiva lineal rápida para el mes de abril (34.9±9.0%) y en general valores de espermatozoides inmóviles menores al 10%. El plasma seminal mostró una osmolaridad de 259.3± 3.5 mOsm/Kg, con una concentración promedio de glucosa e iones Na+, Cl+, K+ y Mg2+ de 142.7±2.8, 118±2.2, 4.8±0.4 y 0.33±0.0 mmol/L, respectivamente, durante la época reproductiva. Conclusiones. La calidad seminal de bagre rayado no presentó variaciones en la estación reproductiva mostrando una alta calidad para los procesos de fecundación.

Introducción: actualmente, las neoplasias malignas constituyen un grupo de enfermedades de gran importancia en la población anciana. Con la edad aumenta la incidencia de la mayoría de los tumores. En Cuba el 65 % de las personas que fallecen por cáncer son mayores de 60 años. Objetivos: caracterizar el comportamiento de la mortalidad por los tumores malignos más frecuentes en el adulto mayor del municipio Plaza de la Revolución durante el quinquenio 2004-2008. Métodos: estudio descriptivo en el que se tomaron todos los fallecidos por esta causa, y que tuvieran residencia en el municipio a partir de las bases de datos de mortalidad de la Dirección Nacional de Estadísticas. Se tuvieron en cuenta las variables: localización anatómica, sexo y edad. Resultados: la localización en próstata ocupó el primer lugar como causa de muerte, con una tasa de 254,1 por 100 000 hab, seguido por el pulmón (189,6). En hombres fueron más frecuentes próstata (254,1) y pulmón (234,8), y en mujeres pulmón (116,5) y mama (112,1). Según la edad, el grupo de 75 años y más fue el más afectado en ambos sexos. Conclusiones: los datos de este estudio confirman un importante crecimiento de la mortalidad en las principales localizaciones, lo que, junto al progresivo envejecimiento de la población, determinará un crecimiento del número anual de casos diagnosticados que generará, a su vez, un aumento de demanda de distintos servicios relacionados con la atención de los pacientes oncológicos en la tercera edad.Introduction: nowadays, the malignant neoplasias are a diseases group very significant in elderly persons. With advanced age increases the incidence of most tumors. In Cuba the 65 % of deceased persons due to cancer is aged over 65. Objectives: to characterize the behavior of mortality due to malignant tumors more frequent in elderly persons from the "Plaza de la Revolución" municipality during the five-year period 2004-2008. Methods: a descriptive study was conducted including all deceased persons due to this cause and residents of this municipality from the mortality database of National Management of Statistics. The following variables were into account: anatomical location, sex and age. Results: prostate location was in the first place as death cause with a rate of 254, 1 by 1000 000 inhabitants, followed by lungs (189,6). In men was predominance of prostate (254,1) and lungs (234,8) and in female predominance was of lungs (116,5) and breast (112,1). According to age, the group aged 75 and more was the more involved in both sexes. Conclusions: the data from present study confirm a significant increase of mortality in main locations what, together with progressive aging of population, will determine a growth in the yearly number of diagnosed cases, that will generate at the same time, an demand increase of different services related to care of third-age oncology patients.

OBJECTIVE. To compare the nosocomial infection (NI) rate obtained from a retrospective review of clinical charts with that from the routine nosocomial infection surveillance system in a community hospital. MATERIAL AND METHODS. Retrospective review of a randomized sample of clinical charts.Results were compared to standard surveillance using crude and adjusted analyses. RESULTS. A total of 440 discharges were reviewed, there were 27 episodes of NIs among 22 patients. Cumulated incidence was 6.13 NI per 100 discharges. Diarrhea, pneumonia and peritonitis were the most common infections. Predictors of NI by Cox regression analysis included pleural catheter (HR 16.38), entry through the emergency ward, hospitalization in the intensive care unit (HR 7.19), and placement of orotracheal tube (HR 5.54). CONCLUSIONS. Frequency of NIs in this community hospital was high and underestimated. We identified urgent needs in the areas of training and monitoring.OBJETIVO. Comparar la tasa de infecciones nosocomiales (IN) resultante de la revisión retrospectiva de expedientes clínicos con los resultados del sistema rutinario de vigilancia de IN de un hospital general. MATERIAL Y MÉTODOS. Revisión retrospectiva de una muestra seleccionada aleatoriamente de expedientes clínicos. Comparación con los resultados obtenidos por el sistema rutinario de vigilancia de IN. Análisis bivariado y multivariado de datos retrospectivos. RESULTADOS. De 440 egresos hubo 27 episodios de IN en 22 pacientes. La incidencia acumulada fue de 6.13 IN por 100 egresos. Las infecciones más frecuentes fueron diarrea, neumonía y peritonitis. Los predictores de IN fueron catéter pleural (HR 16.38), ingreso por urgencias y estancia en cuidados intensivos (HR 7.19), y colocación de tubo orotraqueal (HR 5.54). CONCLUSIONES. La frecuencia de IN fue elevada y subestimada por el sistema rutinario. Identificamos necesidades urgentes de monitoreo y entrenamiento en áreas específicas.