Interactomics and targeted protein degradation for kinase substrate discovery

Reversible phosphorylation is one of the most important post translational modifications that has allowed us as a species to quickly adapt to changing molecular environments due to external stimulation. This process is only capable through the activity of kinases to carry out the targeting of specific substrates defined by their recognition motif allowing for selective phosphorylation and activation and inactivation of distinct pathways as well as other changes that permit cell survival. By being so important for the maintenance of the cells disruption often leads to worsening of the cells, leading to various diseases like cancer, immunological and neurodegenerative disorders. This is why the comprehension of kinases, and their substrates is so important for making progress in the medical care of the patients. For this exact purpose we chose Polo like kinase 2 (Plk2) as the target of study. Even though a great deal of knowledge exists about it, like the fact that it plays roles during the cell cycle, cell differentiation, ontogenesis, stress response, tumorigenesis, neurodegenerative diseases, and inflammation. The mechanisms at play i.e the specific interactions that permits its function in all these systems is not entirely known. As such, we employed the use CRISPR/Cas9 as the method of genetic modification for the endogenous tagging of Plk2 with the auxin inducible degron (AID) for the selective degradation of our target. After having observed through LC/MS the limitations of being a low abundant protein, we combined proximity labeling through Turbo-ID and targeted degradation as a method of accomplishing this task. We were successful in the identification of novel substrates of Plk2 in the cell cycle, and capable of study the interactome of Plk2 in asynchronous and G1/S arrested cells. Providing a sundry of new insight into Plk2 biology and the diversification of its interactions. AID technology in combination with proximity labeling provides the next steps in the ability of study for any type of protein. Capable of gathering information about the context of the protein in regard to its environment, the interactions, and what the absence of that protein would have for specific pathways, while minimizing off-target effects

Impact of the COVID-19 pandemic on HIV care in Guatemala

Objectives: To describe the impact of the coronavirus disease 2019 (COVID-19) pandemic on the diagnosis of human immunodeficiency virus (HIV) and deaths from opportunistic infections in Guatemala. Methods: A retrospective study was conducted to investigate the impact of the COVID-19 pandemic on people with HIV at a referral clinic (Clinica Familiar Luis Angel García, CFLAG), as well as the disruption of services at a diagnostic laboratory hub (DLH) which provides diagnosis for opportunistic infections to a network of 13 HIV healthcare facilities. Comparative analysis was undertaken using the months March-August from two different time periods: (i) pre-COVID-19 (2017-2019); and (ii) during the COVID-19 period (2020). Results: During the COVID-19 period, 7360 HIV tests were performed at Clinica Familiar Luis Angel García, compared with an average of 16,218 tests in the pre-COVID-19 period; a reduction of 54.7% [95% confidence interval (CI) 53.8-55.4%],Deaths from opportunistic infections at 90 days were 10.7% higher in 2020 compared with 2019 (27.3% vs 16.6%; P = 0.05). Clinical samples sent to the DLH for diagnosis of opportunistic infections decreased by 43.7% in 2020 (95% CI 41.0-46.2%). Conclusion: The COVID-19 pandemic is having a substantial impact on HIV care in Guatemala. Diagnostic services for HIV have been severely affected and deaths from opportunistic infections have increased. The lessons learnt must guide the introduction of strategies to reduce the impact of the pandemic.S

The Diagnostic Laboratory Hub: A New Health Care System Reveals the Incidence and Mortality of Tuberculosis, Histoplasmosis, and Cryptococcosis of PWH in Guatemala.

A Diagnostic Laboratory Hub (DLH) was set up in Guatemala to provide opportunistic infection (OI) diagnosis for people with HIV (PWH). Patients newly presenting for HIV, PWH not receiving antiretrovirals (ARVs) for >90 days but returned to care (Return/Restart), and PWH on ARVs with symptoms of OIs (ARV treatment) were prospectively included. Screening for tuberculosis, nontuberculous mycobacteria (NTM), histoplasmosis, and cryptococcosis was done. Samples were couriered to the DLH, and results were transmitted electronically. Demographic, diagnostic results, disease burden, treatment, and follow-up to 180 days were analyzed. In 2017, 1953 patients were included, 923 new HIV infections (an estimated 44% of all new HIV infections in Guatemala), 701 on ARV treatment, and 315 Return/Restart. Three hundred seventeen (16.2%) had an OI: 35.9% tuberculosis, 31.2% histoplasmosis, 18.6% cryptococcosis, 4.4% NTM, and 9.8% coinfections. Histoplasmosis was the most frequent AIDS-defining illness; 51.2% of new patients had <200 CD4 cells/mm3 with a 29.4% OI incidence; 14.3% of OIs in new HIV infections occurred with CD4 counts of 200-350 cells/mm3. OIs were the main risk factor for premature death for new HIV infections. At 180 days, patients with OIs and advanced HIV had 73-fold greater risk of death than those without advanced disease who were OI-free. The DLH OI screening approach provides adequate diagnostic services and obtains relevant data. We propose a CD4 screening threshold of <350 cells/mm3. Mortality remains high, and improved interventions are required, including expansion of the DLH and access to antifungal drugs, especially liposomal amphotericin B and flucytosine.Financial support. This work was supported by Global Action Fund for Fungal Infections and JYLAG, a charity Foundation based in Switzerland (E.A. received this funding under the proposal: “Minimising HIV deaths through rapid fungal diagnosis and better care in Guatemala”). Other contributions came from AIDS Health Foundation (AHF) Guatemala, Intrahealth International and Ministry of health in Guatemala (MSPAS).S

Assessment of oceanographic services for the monitoring of highly anthropised coastal lagoons: The Mar Menor case study

Ocean monitoring systems are designed for continuous monitoring to track their evolution and anticipate environmental issues. However, they are often based on IoT systems that offer little spatial coverage and are hard to maintain. Satellite remote sensing offers good geographical coverage but they also face several challenges to become a monitoring system. This paper introduces an easy-to-use software tool to crawl water-quality data from up to 6 satellite instruments from the ESA and NASA. Particularly, Chl-a data is deeply analyzed in terms of reliability and data coverage for a highly anthropised coastal lagoon (Mar Menor, Spain), where serious socio-environmental issues are happening. Our results show a good linear correlation between in situ data and SRS data, reaching values close to 0.9, and stating the relevance of organic matter inputs from ephemeral streams in Chl-a concentrations. Moreover, temporal granularity is increased from 5 to 1.5 days by combining SRS sources.Preprin

Fases finales del crecimiento y características metabólicas en bovinos de raza Wagyu y cruzados con Angus (Wangus) en un sistema de producción español

La raza wagyu (WY) se caracteriza por dar lugar a una carne con un alto grado de infiltración, de alta calidad (Shahrai et al. 2020) y de alto interés en el mercado mundial. Carne más engrasada que la tradicional en bovino también se origina en animales de raza Angus, aunque ambas razas presentan ritmos de crecimientos y calidad de la carne diferenciadas (Radunz et al. 2009; Shahrai et al. 2020). El cruce de la raza Angus con Wagyu (wangus, WN) es, por lo tanto, de interés productivo. Sin embargo, hay pocos estudios sobre las características productivas de animales WY y WN en sistemas de producción españoles. Por tanto, el objetivo de este estudio fue evaluar el crecimiento y las características metabólicas de WY y WN en la fase final del engorde

Crecimiento y características metabólicas diferenciadas de bovinos de razas con alto engrasamiento en un sistema de producción español

La producción de carne de vacuno con grasa altamente infiltrada tiene gran valor en el mercado mundial, debido a su excelente calidad y características organolépticas. Este tipo de carne puede obtenerse en su máxima calidad, de la raza wagyu (de origen japonés), de su cruce con la raza angus (wangus) o de la raza angus en pureza o en cruces con otras razas europeas (cruce comercial). La raza wagyu y sus cruces han sido poco estudiadas en sistemas de producción europeos y, por tanto, es difícil saber su comportamiento productivo y su eficiencia económica, comparada con razas europeas. El objetivo de este estudio fue comparar el crecimiento y las características metabólicas (diferenciadas en wagyu; Piao da C. et al., 2015) de animales wagyu (WY), wangus (WN) y cruces comerciales (CC) en las primeras etapas de su crecimiento

Variable thermal plasticity of leaf functional traits in Andean tropical montane forests

Tropical montane forests (TMFs) are biodiversity hotspots and provide vital ecosystem services, but they are disproportionately vulnerable to climate warming. In the Andes, cold-affiliated species from high elevations are being displaced at the hot end of their thermal distributions by warm-affiliated species migrating upwards from lower elevations, leading to compositional shifts. Leaf functional traits are strong indicators of plant performance and at the community level have been shown to vary along elevation gradients, reflecting plant adaptations to different environmental niches. However, the plastic response of such traits to relatively rapid temperature change in Andean TMF species remains unknown. We used three common garden plantations within a thermosequence in the Colombian Andes to investigate the warming and cooling responses of key leaf functional traits in eight cold- and warm-affiliated species with variable thermal niches. Cold-affiliated species shifted their foliar nutrient concentrations when exposed to warming, while all other traits did not significantly change; contrastingly, warm-affiliated species were able to adjust structural, nutrient and water-use efficiency traits from acquisitive to conservative strategies in response to cooling. Our findings suggest that cold-affiliated species will struggle to acclimate functional traits to warming, conferring warm-affiliated species a competitive advantage under climate change

Multi-Country Evaluation of the Sensitivity and Specificity of Two Commercially-Available NS1 ELISA Assays for Dengue Diagnosis

Dengue is the most important mosquito-borne viral disease of humans and an enormous public health burden in affected countries. Early, sensitive and specific diagnosis of dengue is needed for appropriate patient management as well as for early epidemic detection. Commercially available assays that detect the dengue virus protein NS1 in the plasma/serum of patients offer the possibility of early and rapid diagnosis. Here we evaluated two commercially available ELISA kits for NS1 detection (Pan-E Dengue Early ELISA and the Platelia™ Dengue NS1 Ag). Results were compared against a reference diagnosis in 1385 patients in 6 countries in Asia and the Americas. Collectively, this multi-country study suggests that the best performing NS1 assay (Platelia) had moderate sensitivity (median 64%, range 34–76%) and high specificity (100%) for the diagnosis of dengue. The combination of NS1 and IgM detection in samples collected in the first few days of fever increased the overall dengue diagnostic sensitivity