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Background: With age, lungs undergo typical changes that lead to a deterioration of respiratory function. Our aim was to assess the role of age‐associated hyperphosphatemia in these changes. Methods: We used C57BL6 mice to study an ageing model in vivo and human lung fibroblasts were treated with a phosphate donor, beta‐glycerophosphate (BGP), to explore mechanisms involved. Respiratory function was registered with a double chamber plethysmograph. Lung structure was analysed by different staining, phosphate and cytokines levels by colorimeric kits, expression of fibrosis, inflammation and ET‐1 system by western blot or RT‐PCR. Results: Old mice showed hyperphosphatemia, along with lung fibrosis, loss of elastin, increased expression of pro‐inflammatory cytokines and impaired respiratory function. BGP induced inflammation and fibrosis in fibroblasts through the activation and binding of NFkB to the MCP‐1 or FN promoters. BGP increased ECE‐1 expression by inducing NFkB binding to the ECE‐1 promoter. QNZ, an NFkB inhibitor, blocked these effects. When ECE‐1 was inhibited with phosphoramidon, BGP‐induced inflammation and fibrosis were significantly reduced, suggesting a role for ET‐1 in BGP‐mediated effects.ET‐1 produced effects similar to those of BGP, which were also dependent on NFkB. To study the pathophysiological relevance of hyperphosphatemia in vivo, a low‐P diet was administered to a group of old animals, showing an improvement in fibrosis, inflammation and respiratory function compared to old mice on a standard diet. Conclusion: These results suggest that age‐related hyperphosphatemia induces inflammation, fibrosis, and impaired respiratory function in old mice; these effects appear to be mediated by ET‐1 and NFkB activation. [ABSTRACT FROM AUTHOR]

A subpopulation of NK cells with distinctive phenotype and function differentiates and expands specifically in response to infection by human cytomegalovirus (HCMV). A hallmark of these adaptive NK cells is their increased expression levels of the activating CD94/NKG2C receptor for HLA‐E, and lack of expression of its inhibitory homologue CD94/NKG2A. Their frequency is highly variable in HCMV+ individuals, and the basis for such differences is only partially understood. Here, we explore the possible influence of sequence polymorphism of the NKG2C (or KLRC2) gene on the expansion of NKG2C+NKG2A− NK cells in healthy HCMV‐seropositive donors. Our results show a significant association of greater proportions of adaptive NK cells with allele NKG2C*02. This is defined by two amino acid substitutions in comparison with the most prevalent allele, NKG2C*01, and associates with additional sequence polymorphisms in noncoding regions. Furthermore, we demonstrate consistently higher mRNA levels of NKG2C*02 in heterozygous individuals co‐expressing this allele in combination with NKG2C*01 or *03. This predominance is independent of polymorphisms in the promoter and 3′ UTRs and is appreciated also in HCMV‐seronegative donors. In summary, although additional factors are most likely implicated in the variable expansion of NKG2C+NKG2A− NK cells in response to HCMV, our results demonstrate that host immunogenetics, in particular NKG2C diversity, influences the magnitude of such response. [ABSTRACT FROM AUTHOR]

Motor imagery (MI) is the mental representation of a movement without its execution. It activates internal representations of the movement without external stimulus through different memory‐related processes. Although acute stress is frequent in the population and affects supraspinal structures essential for memory functionality, it is still unknown how that stress affects MI capacity and temporal congruence (TC) between execution and movement imagination. This study aimed to discover how acute stress may influence MI capacity and TC in the subscales of internal and external visual imagery and kinesthetic imagery. A double‐blind, randomized trial was conducted. Sixty‐two young, healthy subjects (mean age = 20.65 [2.54]; 39 females and 23 males) unfamiliar with the assessment and uses of MI were recruited. Participants were assigned by stratified randomization to the stress group or the control group. Stress was induced by the Maastricht Acute Stress Test (MAST), while the control group performed the MAST control protocol. MI capacity and TC were assessed before (t1) and after (t2) MAST stress or control using the Movement Imagery Questionnaire‐3 (MIQ‐3). Electrodermal activity and heart rate variability were further recorded as control variables to assess stress induction. Thirty subjects in the stress group and 26 subjects in the control group were analyzed. No significant group differences were observed when comparing MI capacity or TC in any subscales. These findings suggest that acute stress does not significantly affect MI capacity or TC in young, healthy, non‐experienced MI subjects. MI could thus be a relevant helpful technique in stressful situations. [ABSTRACT FROM AUTHOR]

Firm internationalization has been analyzed from different angles based on various theories. However, qualitative research on the stability of the factors that motivate and hinder internationalization across the export stages in emerging markets is limited. This study builds on the "stages" theories of internationalization, providing a detailed analysis of the perceived stability of stimuli and barriers during the exploration, entry, and expansion stages. We focus on a sample of Spanish wine firms exporting to China—a key wine market, with annual growth rates of over 25% between 2000 and 2020. The results suggest that the perception of most barriers is stage‐dependent, varying over time as commitment levels evolve. The main stimuli, on the other hand, remain constant across the different export stages. Moreover, we find that firm size, international experience, and access to local and international networks are key factors that influence the evolution of firms' internationalization in China, while lateral rigidity continues to constrain the expansion of smaller companies. These findings contribute to a deeper understanding of the opportunities and challenges of doing business in an emerging market, particularly in the institutional context in China, as well as their evolution over different stages of internationalization. Furthermore, they offer insights into the dynamics of the Chinese wine market and the implications for European exporters. [ABSTRACT FROM AUTHOR]

In recent years, therapies based on regenerative medicine have gained wide interest owing to their potential in organ transplantation and in the treatment of multiple diseases. These cell‐based therapies rely mainly on the therapeutic potential of stem cells as a result of their self‐renewal, differentiation, capacities and immunosuppressive properties. Adipose tissue contains a large number of mesenchymal stem cells compared to other tissues and is easily accessible through noninvasive techniques; hence the primary recovery of adipose‐derived mesenchymal stem cells (ASC) has become a critical step. Aqueous two‐phase systems (ATPS) have shown promise as a separation technique for mesenchymal stem cells (MSC) as a result of their scalability, cost‐effectiveness and compatibility. In this study, we investigated the partitioning behavior of the stromal vascular fraction cells, including ASC, in different ATPS compositions [polyethylene glycol, dextran (DEX), Ficoll and Ucon). Among the three ATPS tested, most of the ASC consistently migrates to the top phase as indicated by immunostaining studies for CD73, CD90 and CD105 stem cell markers. Nevertheless, the Ficoll–DEX system retrieves the greatest quantity of ASC recovered in the top phase (89% of total ASC), whereas the Ucon–DEX system effectively eliminates the most contaminating cells (38%). Our findings provide insights into the conditions under which ASC and contaminant cells concentrate in opposite phases in different ATPS, which can contribute to the development of efficient and scalable separation systems for MSC in regenerative medicine and tissue engineering applications. © 2024 Society of Chemical Industry (SCI). [ABSTRACT FROM AUTHOR]

Marine thraustochytrids produce metabolically important lipids such as the long‐chain omega‐3 polyunsaturated fatty acids, carotenoids, and sterols. The growth and lipid production in thraustochytrids depends on the composition of the culture medium that often contains yeast extract as a source of amino acids. This work discusses the effects of individual amino acids provided in the culture medium as the only source of nitrogen, on the production of biomass and lipids by the thraustochytrid Thraustochytrium sp. RT2316‐16. A reconstructed metabolic network based on the annotated genome of RT2316‐16 in combination with flux balance analysis was used to explain the observed growth and consumption of the nutrients. The culture kinetic parameters estimated from the experimental data were used to constrain the flux via the nutrient consumption rates and the specific growth rate of the triacylglycerol‐free biomass in the genome‐scale metabolic model (GEM) to predict the specific rate of ATP production for cell maintenance. A relationship was identified between the specific rate of ATP production for maintenance and the specific rate of glucose consumption. The GEM and the derived relationship for the production of ATP for maintenance were used in linear optimization problems, to successfully predict the specific growth rate of RT2316‐16 in different experimental conditions. [ABSTRACT FROM AUTHOR]

Background: Iron deficiency correction with ferric carboxymaltose improves symptoms and reduces rehospitalization in patients with reduced left ventricular ejection fraction. The mechanisms underlying these improvements are poorly understood. This study aimed to determine changes in left ventricular contractility after iron treatment as reflected in global longitudinal strain. Methods: Prospective single‐center study including 43 adults with reduced ejection fraction, non‐anemic iron deficiency, and functional class II‐III heart failure despite optimal medical treatment. Global longitudinal strain through speckle‐tracking echocardiography was measured at baseline and 4 weeks after ferric carboxymaltose. Results: A significant improvement in global longitudinal strain was detected (from −12.3% ± 4.0% at baseline to −15.6% ± 4.1%, p <.001); ferritin and transferrin saturation index had increased, but ejection fraction presented no significant changes (baseline 35.7% ± 4.6%, follow‐up 37.2% ± 6.6%, p =.073). Conclusions: In patients with heart failure and reduced ejection fraction, the correction of iron deficiency with ferric carboxymaltose is associated with an early improvement in global longitudinal strain, possibly suggesting a direct effect of iron correction on myocardial contractility. [ABSTRACT FROM AUTHOR]

Mesenchymal stem cells (MSCs) are recognized for their remarkable ability to differentiate into multiple cell types. They are also known to possess properties that can fight cancer, leading to attempts to modify MSCs for use in anticancer treatments. However, MSCs have also been found to participate in pathways that promote tumor growth. Many studies have been conducted to explore the potential of MSCs for clinical applications, but the results have been inconclusive, possibly due to the diverse nature of MSC populations. Furthermore, the conflicting roles of MSCs in inhibiting tumors and promoting tumor growth hinder their adaptation to anticancer therapies. Antitumorigenic and protumorigenic properties of MSCs in urological cancers such as bladder, prostate, and renal are not as well established, and data comparing them are still limited. MSCs hold significant promise as a vehicle for delivering anticancer agents and suicide genes to tumors. Presently, numerous studies have concentrated on the products derived from MSCs, such as extracellular vesicles (EVs), as a form of cell‐free therapy. This work aimed to review and discuss the current knowledge of MSCs and their EVs in urological cancer therapy. [ABSTRACT FROM AUTHOR]

Aims: This study aimed to assess, in patients with cardiogenic shock secondary to unprotected left main coronary artery‐related myocardial infarction (ULMCA‐related AMICS), the incidence and predictors of no recovery of left ventricular function during the admission. Methods and results: This was an observational study conducted at two tertiary care centres (2012–20). The main outcome measured was death or requirement for heart transplantation (HT) or left ventricular assist devices (LVAD) during the admission. A total of 70 patients were included. Percutaneous coronary intervention (PCI) was successful in 53/70 patients (75.7%). The combined endpoint of death or requirement of HT or LVAD during the admission occurred in 41/70 patients (58.6%). The highest incidence of the primary endpoint was observed among patients with profound shock and occluded left main coronary artery (LMCA) (20/23, 87%, P < 0.001). Although a successful PCI reduced the incidence of the event in the whole cohort (51.9% vs. 82.4% in failed PCI, P = 0.026), this association was not observed among this last group of complex patients (86.7% vs. 87.5% in failed PCI, P = 0.731). The predictive model included left ventricular ejection fraction, baseline ULMCA Thrombolysis In Myocardial Infarction flow, and severity of shock and showed an optimal ability for predicting death or requirements for HT or LVAD during the admission (area under the curve 0.865, P < 0.001). Conclusions: ULMCA‐related AMICS was associated with a high in‐hospital mortality or need for HT or LVAD. Prognosis was especially poor among patients with profound shock and baseline occluded LMCA, with a low probability of recovery regardless of successful PCI. [ABSTRACT FROM AUTHOR]

Background: The efficacy and safety of high versus medium doses of glucocorticoids for the treatment of patients with COVID‐19 has shown mixed outcomes in controlled trials and observational studies. We aimed to evaluate the effectiveness of methylprednisolone 250 mg bolus versus dexamethasone 6 mg in patients with severe COVID‐19. Methods: A randomised, open‐label, controlled trial was conducted between February and August 2021 at four hospitals in Spain. The trial was suspended after the first interim analysis since the investigators considered that continuing the trial would be futile. Patients were randomly assigned in a 1:1 ratio to receive dexamethasone 6 mg once daily for up to 10 days or methylprednisolone 250 mg once daily for 3 days. Results: Of the 128 randomised patients, 125 were analysed (mean age 60 ± 17 years; 82 males [66%]). Mortality at 28 days was 4.8% in the 250 mg methylprednisolone group versus 4.8% in the 6 mg dexamethasone group (absolute risk difference, 0.1% [95% CI, −8.8 to 9.1%]; p = 0.98). None of the secondary outcomes (admission to the intensive care unit, non‐invasive respiratory or high‐flow oxygen support, additional immunosuppressive drugs, or length of stay), or prespecified sensitivity analyses were statistically significant. Hyperglycaemia was more frequent in the methylprednisolone group at 27.0 versus 8.1% (absolute risk difference, −18.9% [95% CI, −31.8 to ‐ 5.6%]; p = 0.007). Conclusions: Among severe but not critical patients with COVID‐19, 250 mg/d for 3 days of methylprednisolone compared with 6 mg/d for 10 days of dexamethasone did not result in a decrease in mortality or intubation. [ABSTRACT FROM AUTHOR]

NK cells monitor altered molecular patterns in tumors and infected cells through an ample array of receptors. Two families of evolutionarily distant receptors have converged to enable human NK cells to sense levels of HLA class I ligands, frequently abnormal in altered cells. Whilst different forms of polymorphism are a hallmark of killer‐cell immunoglobulin‐like receptors and their classic HLA‐A, B, and C ligands, genetic diversity of killer‐cell lectin‐like receptors for the non‐classical HLA‐E (CD94/NKG2 heterodimers) is less conspicuous and has attracted less attention. A common pattern of diversification in both receptor families is evolution of pairs of inhibitory and activating homologs for a common ligand, the genes encoding activating receptors being more frequently affected by copy number variation (CNV). This is exemplified by the gene encoding the activating NKG2C subunit (KLRC2 or NKG2C), which marks an NK‐cell subpopulation that differentiates or expands in response to cytomegalovirus. We have studied NKG2C diversity in 240 South European individuals, using polymerase chain reaction and sequencing methods to assess both gene CNV and single‐nucleotide polymorphisms (SNPs) affecting its promoter, coding and 3′‐untranslated (3′UT) regions. Sequence analysis revealed eight common SNPs—one in the promoter, two in the coding sequence, and five in the 3′UT region. These SNPs associate strongly with each other, forming three conserved extended haplotypes (frequencies: 0.456, 0.221, and 0.117). Homo‐ and heterozygous combination of these, together with complete gene deletion (0.175) and additional haplotypes with frequencies lower than 0.015, generate a diversity of NKG2C genotypes of potential immunological importance. [ABSTRACT FROM AUTHOR]

Anti‐N‐methyl‐D‐aspartate receptor (anti‐NMDAR) encephalitis presents commonly with psychiatric symptoms. One cohort of these patients reported that antipsychotic administration led to neuroleptic intolerance (NI) in 19% of them, which was preventable by a prompt encephalitis diagnosis. To date, there is no clear description of the "neuroleptic intolerance" spectrum in general or during anti‐NMDAR encephalitis. We aimed to synthesize epidemiological and clinical characteristics of patients with NI and confirmed anti‐NMDAR encephalitis, the time to the encephalitis diagnosis, the disease course, outcomes at discharge, and associated factors. We systematically searched three databases, to include clinical cases, case series, and observational studies. Additionally, we reported one clinical case. Results were summarized using narrative synthesis and the quality of the included studies was assessed. We included 22 records representing 40 patients (28 females; mean age, 24.6). Overall, the evidence quality was low. Initially, most cases were admitted in psychiatric wards (70%) with purely psychiatric symptoms (37.5%). However, most of them developed subtle concomitant neurological symptoms. The mean time to anti‐NMDAR encephalitis diagnosis was 26.7 days, which was triggered by the NI in six patients. We found no association between clinical variables as delayed diagnosis, admission to psychiatric wards or the presence of malignancy with outcome variables as unfavorable outcomes at discharge, ICU, or mechanical ventilation requirement. A thorough neurological examination in young patients with new‐onset psychiatric symptoms could help emergency physicians, neurologists, and psychiatrists suspect anti‐NMDAR encephalitis earlier. Awareness of NI as a potential side effect during suspected or confirmed anti‐NMDAR encephalitis is encouraged. [ABSTRACT FROM AUTHOR]

Aqueous two‐phase systems (ATPS) have been demonstrated to be a simple and robust worldwide applied bioprocessing technology. From high value microscopic to macroscopic biological molecules obtained from a wide broad of matrices, ATPS have shown a high efficiency capacity for their concentration, enrichment, recovery, and purification. Particularly in Latin America, four relevant countries have paved the way and played important leading roles in this transformation history. This perspective article aims to present a critical analysis of the practical evolution of the main implementations of ATPS in Argentina, Brazil, Chile, and Mexico. Special attention is given to novel applications of this biphasic bioprocess that are being explored to accomplish visionary therapeutic tasks for the healthcare, biotechnology, and pharmaceutical sectors, as well as other environmental purposes. © 2021 Society of Chemical Industry (SCI). [ABSTRACT FROM AUTHOR]

Background: As the development of neuropathic symptoms contributes to pain severity and chronification after surgery, their early prediction is important to allow targeted treatment.Objectives: We longitudinally investigated trajectories of signs and symptoms in patients undergoing thoracotomy and assessed whether and at which time they were related to the development of neuropathic pain symptoms 6 months after surgery.Methods: Presurgical and 6 monthly postsurgical assessments included questionnaires for mental and physical well-being (e.g., depression/anxiety, pain catastrophizing, sleep quality, neuropathic pain symptoms), and quantitative sensory testing (QST).Results: QST trajectories indicated nerve impairment of the surgery site with predominant loss of function. Signs of recovery towards the end of the assessment period were observed for some tests. Unsupervised cluster analysis with NPSI scores 6 months after surgery as clustering variable identified one group with no/low levels of neuropathic symptoms and one with moderate levels. The two groups differed w.r.t. several signs and symptoms already at early time points. Notably, neuropathic pain anywhere in the body differed already preoperatively and sleep impairment differentiated the two groups at all time points. Regression analysis revealed three factors that seemed particularly suited to predicted 6 months NPSI scores, namely preoperative neuropathic pain symptoms, with contributions from sleep impairment 1 month after surgery and the presence of dynamic mechanical allodynia 3 months after surgery.Conclusions: Clinical routine should focus on the individual's physiological state, including pre-existing neuropathic pain and sleep quality to identify patients early who might be at risk to develop chronic post-surgical neuropathic pain.Significance: Development of neuropathies contributes to pain severity and pain chronification after surgery. Here we demonstrate trajectories of quantitative sensory tests (assessed at monthly intervals for 6 months after surgery) that reveal accurate time courses of gain/loss of nerve function following thoracotomy. Independent of the degree of neuropathic signs after surgery, the main predictors for post-surgical neuropathic pain are self-reported neuropathic pain before surgery and sleep quality shortly after surgery. [ABSTRACT FROM AUTHOR]

Aims: To evaluate echocardiographic and biomarker changes during chemotherapy, assess their ability to early detect and predict cardiotoxicity and to define the best time for their evaluation. Methods and results: Seventy‐two women with breast cancer (52 ± 9.8 years) treated with anthracyclines (26 also with trastuzumab), were evaluated for 14 months (6 echocardiograms/12 laboratory tests). We analysed: high‐sensitivity cardiac troponin T, NT‐proBNP, global longitudinal strain (GLS), left ventricle end‐systolic volume (LVESV), left ventricle end‐diastolic volume (LVEDV), and left ventricular ejection fraction (LVEF). Cardiotoxicity was defined as a reduction in LVEF>10% compared with baseline with LVEF<53%. High‐sensitivity troponin T levels rose gradually reaching a maximum peak at 96 ± 13 days after starting chemotherapy (P < 0.001) and 62.5% of patients presented increased values during treatment. NT‐proBNP augmented after each anthracycline cycle (mean pre‐cycle levels of 72 ± 68 pg/mL and post‐cycle levels of 260 ± 187 pg/mL; P < 0.0001). Cardiotoxicity was detected in 9.7% of patients (mean onset at 5.2 months). In the group with cardiotoxicity, the LVESV was higher compared with those without cardiotoxicity (40 mL vs. 29.5 mL; P = 0.045) at 1 month post‐anthracycline treatment and the decline in GLS was more pronounced (−17.6% vs. −21.4%; P = 0.03). Trastuzumab did not alter serum biomarkers, but it was associated with an increase in LVESV and LVEDV (P < 0.05). While baseline LVEF was an independent predictor of later cardiotoxicity (P = 0.039), LVESV and GLS resulted to be early detectors of cardiotoxicity [odds ratio = 1.12 (1.02–1.24), odds ratio = 0.66 (0.44–0.92), P < 0.05] at 1 month post‐anthracycline treatment. Neither high‐sensitivity troponin T nor NT‐proBNP was capable of predicting subsequent cardiotoxicity. Conclusions: One month after completion of anthracycline treatment is the optimal time to detect cardiotoxicity by means of imaging parameters (LVESV and GSL) and to determine maximal troponin rise. Baseline LVEF was a predictor of later cardiotoxicity. Trastuzumab therapy does not affect troponin values hence imaging techniques are recommended to detect trastuzumab‐induced cardiotoxicity. [ABSTRACT FROM AUTHOR]

This article develops from the revolt, or social outbreak that took place in October 2019 in Chile. With protests in all parts of continental Chile, these mobilisations have become the largest and longest mass mobilisations in the country's history, bringing together a significant number of anti‐neoliberal demands, which ended up overturning the legitimacy of the development model established by General Pinochet's dictatorship (1973–1990) and deepened during all post‐dictatorial governments (1990–2021). In this paper, the Freudian conceptions of masses are questioned, fundamentally embodied in Group Psychology and Analysis of the Ego, as well as in a subsidiary way in Totem and Taboo and On Narcissism. It is observed that the mobilisations in Chile lacked a leader, which challenges Freud's conclusions. In this way, it is postulated, making use of Rozitchner's work, that the mass is a contingent historical subject, whose conformation logics are open and influenced by its circumstances. Making use of Laclau's reception of nominalism, a reading of the October masses is offered as articulated by a signifier ('dignity') that serves as a symbolic support for the libidinal bond of its members. [ABSTRACT FROM AUTHOR]

Background: Hyperphosphatemia has been related to the development of sarcopenia in aging mice. We describe the intracellular mechanisms involved in the impairment of the myogenic differentiation promoted by hyperphosphatemia and analyse these mechanisms in the muscle from older mice. Methods: C2C12 cells were grown in 2% horse serum in order to promote myogenic differentiation, in the presence or absence of 10 mM beta‐glycerophosphate (BGP) for 7 days. Troponin T, paired box 7 (Pax‐7), myogenic factor 5 (Myf5), myogenic differentiation 1 (MyoD), myogenin (MyoG), myocyte enhancer factor 2 (MEF2C), P300/CBP‐associated factor (PCAF), histone deacetylase 1 (HDAC1), fibronectin, vimentin, and collagen I were analysed at 48, 72, and 168 h, by western blotting or by immunofluorescence staining visualized by confocal microscopy. Studies in mice were performed in 5‐ and 24‐month‐old C57BL6 mice. Three months before sacrifice, 21‐month‐old mice were fed with a standard diet or a low phosphate diet, containing 0.6% or 0.2% phosphate, respectively. Serum phosphate concentration was assessed by a colorimetric method and forelimb strength by a grip test. Fibrosis was observed in the tibialis anterior muscle by Sirius Red staining. In gastrocnemius muscle, MyoG, MEF2C, and fibronectin expressions were analysed by western blotting. Results: Cells differentiated in the presence of BGP showed near five times less expression of troponin T and kept higher levels of Pax‐7 than control cells indicating a reduced myogenic differentiation. BGP reduced Myf5 about 50% and diminished MyoD transcriptional activity by increasing the expression of HDAC1 and reducing the expression of PCAF. Consequently, BGP reduced to 50% the expression of MyoG and MEF2C. A significant increase in the expression of fibrosis markers as collagen I, vimentin, and fibronectin was found in cells treated with BGP. In mice, serum phosphate (17.24 ± 0.77 mg/dL young; 23.23 ± 0.81 mg/dL old; 19.09 ± 0.75 mg/dL old with low phosphate diet) correlates negatively (r = −0.515, P = 0.001) with the muscular strength (3.13 ± 0.07 gf/g young; 1.70 ± 0.12 gf/g old; 2.10 ± 0.09 gf/g old with low phosphate diet) and with the expression of MyoG (r = −0.535, P = 0.007) and positively with the expression of fibronectin (r = 0.503, P = 0.001) in gastrocnemius muscle. The tibialis anterior muscle from old mice showed muscular fibrosis. Older mice fed with a low phosphate diet showed improved muscular parameters relative to control mice of similar age. Conclusions: Hyperphosphatemia impairs myogenic differentiation, by inhibiting the transcriptional activity of MyoD, and enhances the expression of fibrotic genes in cultured myoblasts. Experiments carried out in older mice demonstrate a close relationship between age‐related hyperphosphatemia and the decrease in the expression of myogenic factors and the increase in factors related to muscle fibrosis. [ABSTRACT FROM AUTHOR]

Objective: Obesity-associated hypoandrogenemia is increasing in parallel to the obesity epidemic. The prevalence of hypoandrogenemia in nondiabetic young men with obesity is not known. This study aimed to evaluate the prevalence of hypoandrogenemia and associated risk factors in this population.Methods: This cross-sectional study included 266 nondiabetic men < 50 years of age with obesity who were referred from primary care. Total testosterone (high-performance liquid chromatography mass spectrometry), sex hormone-binding globulin, free testosterone (FT), luteinizing hormone (LH), high-sensitivity C-reactive protein, and homeostatic model assessment of insulin resistance were determined. Body composition and erectile function were also assessed. Hypoandrogenemia was defined as FT level < 70 pg/mL.Results: Subnormal FT concentrations were found in 25.6% of participants. Hypoandrogenemia prevalence was different along the BMI continuum, being > 75% in individuals with BMI ≥ 50 kg/m2 . A multivariate regression analysis indicated that increasing BMI (P < 0.001), age (P = 0.049), and reduced LH levels (P = 0.003) were independent risk factors for hypoandrogenemia.Conclusions: In a primary care-based cohort of nondiabetic young men with obesity, hypoandrogenemia was a very prevalent finding and was directly associated with adiposity. Obesity, age, and reduced LH levels were independent risk factors associated with hypoandrogenemia. Further prospective studies are needed to evaluate the long-term consequences of hypoandrogenemia in this population. [ABSTRACT FROM AUTHOR]

Background: During magnetic resonance–guided focused ultrasound for essential or parkinsonian tremor, adverse events (headache, nausea/vomiting, or anxiety) may alter the outcome of the procedure despite being mostly transient and mild. Objectives: Our aim was to analyze the relationship between demographic, procedural, and anesthetic characteristics with magnetic resonance/ultrasound‐related events. Methods: This was a retrospective study at the Clinica Universidad de Navarra of patients undergoing thalamotomy with magnetic resonance–guided focused ultrasound between September 2018 and October 2019. The anesthesia protocol included headache and nausea/vomiting prophylaxis and rescue therapy. Dexmedetomidine was used for anxiolysis in some patients after thorough multidisciplinary assessment. Results: A total of 123 patients were included. Headache was directly related to skull density ratio (P < 0.001) and skull thickness (P = 0.02). Patients with a skull density ratio less than 0.48 had 3 times the odds of experiencing moderate or severe headache (odds ratio [OR], 3.08; 95% confidence interval [CI], 1.21–7.82) and had a higher odds of aborting sonication due to pain. Sex was associated with increased nausea (P = 0.007). Women had 4 times the odds of nausea than men (OR, 4.4; 95% CI, 1.61–12.11). Dexmedetomidine did not reduce headache or nausea incidence. Patients who received dexmedetomidine had a higher number (P = 0.01) and total minutes of sonication (P = 0.01). Conclusions: Patients with lower skull density ratios and higher skull thicknesses could benefit from an aggressive analgesic prophylaxis. Women are more likely to experience nausea. Dexmedetomidine did not reduce headache and nausea, but increased the number and duration of sonications. Its exact effect on tremor is still unclear. [ABSTRACT FROM AUTHOR]

Purpose: To evaluate the intra and interobserver reproducibility of a new goniometric method for evaluating the isolated passive range of motion of the glenohumeral joint in an outpatient setting. Methods: This is a prospective observational study on healthy subjects. The Glenohumeral ROM Assessment with Scapular Pinch (GRASP) method is a new method for assessing the isolated range of motion (ROM) of the glenohumeral joint (GH) by a single examiner with a clinical goniometer. It measures the isolated glenohumeral passive abduction (GH-AB), passive external rotation (GH-ER) and internal rotation (GH-IR) with the arm at 45º of abduction. These three GH ROM parameters were measured in both shoulders of 30 healthy volunteers (15 males/15 females, mean age:41.6[SD = 10.3] years). The full shoulder passive abduction, passive external rotation and internal rotation 45º of abduction were measured by the same examiners with a goniometer for comparison. One examiner made two evaluations and a second examiner made a third one. The primary outcome was the intra- and interobserver reproducibility of the measurements assessed with intraclass correlation coefficients (ICC) and the Bland–Altman plot. Results: The intra-observer ICC for isolated glenohumeral ROM were: 0.84 ± 0.07 for GH-ABD, 0.63 ± 0.09 for GH-ER, and 0.61 ± 0.14 for GH-IR. The inter-observer ICC for isolated glenohumeral ROM were: 0.86 ± 0.06 for GH-ABD, 0.68 ± 0.12 for GH-ER, and 0.62 ± 0.14 for GH-IR. These results were similar to those obtained for full shoulder ROM assessment with a goniometer. Conclusion: The GRASP method is reproducible for quick assessment of isolated glenohumeral ROM. Level of evidence: III [ABSTRACT FROM AUTHOR]

European abandoned lands have been increasing in recent decades, and this is occurring in a climate change scenario in which the consequences of their interactions are still little known. We tested the hypothesis that the functionality of abandoned lands responds differently to year‐to‐year climate variations depending on the time since the cessation of agricultural activity. We constructed a chronosequence in Mediterranean abandoned vineyards at six different years of abandonment in 2016 (1, 3, 7, 11, 25, 40), in a current vineyard and in land not dedicated to vineyards. Soil total organic carbon (TOC), total nitrogen (TN) and available nutrients, enzyme activities relating to the main macronutrient cycles and plant‐community attributes were studied at 24 sites over three consecutive years. A total of 144 samplings were performed. TOC, TN, most enzyme activities and plant species density increased throughout the chronosequence, although interannual climate variability also had a significant influence. The extreme drought event increased the activity of enzymes related to the carbon (C) and nitrogen (N) cycle, as well as the stock of TOC after a subsequent decrease in TOC when environmental conditions became favourable. In contrast, plant species density and the cover of certain plant functional traits such as mycorrhizal symbiotic plants (Cistaceae) and N‐fixers (Fabaceae) decreased significantly. Our results also demonstrated that extreme drought events had a higher negative impact on lands that had been abandoned for longer. We therefore conclude that extreme drought events damage the functionality of Mediterranean abandoned agrosystems, thus affecting all its compartments and jeopardising its functionality. [ABSTRACT FROM AUTHOR]

Objective. To establish differentiated depressive subtypes using a latent class analysis (LCA), including clinical and functional indicators in a sample of depressed patients consulted in Chilean Primary Health Care. Methods. A LCA was performed on a sample of 297 depressed patients consulted in Chilean PHC. The Mini International Neuropsychiatric Interview, the Hamilton Depression Rating Scale, the Outcome Questionnaire -social role, and interpersonal subscales were as instruments. A regression analysis of the different subtypes with sociodemographic and adverse life experiences was performed. Results. In a sample characterized by 87.5% of women, two, three, and four latent class models were obtained. The three-class model likely represents the best clinical implications. In this model, the classes were labeled: "complex depression" (CD) (58% of the sample), "recurrent depression" (RD) (34%), and "single depression episode" (SD) (8%). Members of CD showed a higher probability of history of suicide attempts, interpersonal, and social dysfunction. Psychiatric comorbidities differentiated the RD from SD. According to a multinomial regression model, childhood trauma experiences, recent stressful life experiences, and intimate partner violence events were associated with the CD class (p < 0.01). Limitations. The vast majority of participants were females from Chile and the sample studied was not random. So, the results may not necessarily represent outpatient clinics. Conclusions. This study can provide additional evidence that depression, specifically in female gender, could be better understood as a complex heterogeneous disorder when clinical and functional indicators are studied. Furthermore, adverse life experiences starting in childhood could lead to a differentiated complex depressive subtype. [ABSTRACT FROM AUTHOR]

Production of biomass and lipids in batch cultures of the Antarctic thraustochytrid Oblongichytrium sp. RT2316‐13, is reported. The microorganism proved capable of producing nearly 67% docosahexaenoic acid (DHA) and 15% eicosapentaenoic acid (EPA) in its total lipid fraction. Biomass with a maximum total lipid content of 33.5% (wt/wt) could be produced at 15°C in batch culture using a medium containing glucose (20 g/L), yeast extract (10.5 g/L), and other minor components. A lower culture temperature (5°C) reduced biomass and lipid productivities compared to culture at 15°C, but enhanced the DHA and EPA content of the lipids by 6.4‐ and 3.3‐fold, respectively. Both a simple minimally structured mathematical model and a more complex genome‐scale metabolic model (GEM) allowed the fermentation profiles in batch cultures to be satisfactorily simulated, but the GEM provided much greater insight in the biochemical and physiological phenomena underlying the observed behavior. Unlike the simpler model, the GEM could be interrogated for the possible effects of various external factors such as oxygen supply, on the expected outcomes. In silico predictions of oxygen effects were consistent with literature observations for DHA producing thraustochytrids. [ABSTRACT FROM AUTHOR]

Purpose: To evaluate the intra and interobserver reproducibility of a new system that assesses the three-dimensional humero-scapulo-thoracic kinematics using wearable technology in an outpatient setting. To obtain normative data with the system for scapular angular motions in three planes. Methods: The SHoW Motion 3D kinematic tracking system is a motion analysis system that uses wireless wearable non-invasive inertial-magnetic sensors to assess the three-dimensional kinematics of the shoulder girdle. The sensors are placed over the skin in the sternum, scapular spine and arm to precisely define angular motions of the humerus and the scapula with three Degrees of Freedom (DOF) for each segment. The system was used to measure the scapular angular motions in three planes (upward/downward rotation, internal/external rotation and anterior/posterior tilt) during two shoulder full-range movements (flexion/extension and abduction/abduction) in both shoulders of 25 healthy volunteers (13 males and 12 females, mean age: 37 [standard deviation 11.1] years). In a first measuring session one examiner made two evaluations alternating with another examiner that made a third evaluation. In a second session, one week apart, the first examiner made a fourth evaluation. A mean curve was computed from the normalized data for each measurement to obtain normative data for scapular angular kinematics. Intra and inter-observer reproducibility was evaluated using Root Mean Square Error Estimation (RMSE) and Coefficients for Multiple Correlations (CMC). Results: Both shoulders of the 25 volunteers were evaluated four times. The two hundred resulting kinematic analyses were pooled to get normative values for relations between humeral elevation angles and the three angular movements of the scapula. The system showed at least very good (CMC > 0.90) intra and inter-observer reproducibility for scapular tilt and upward-downward rotations both in flexion and abduction. For scapular internal-external rotation the results were acceptable (CMC > 0.75) but not as good, especially for the abduction movement. RMSE calculations showed consistently good reproducibility with RSME< 4° for all three angles evaluated in flexion and abduction. Conclusion: The SHoW Motion 3D kinematic tracking system is a quick, reproducible and easy to use system for the assessment of scapular angular kinematics in healthy adults. The data obtained is similar to that obtained with other validated methods. Level of evidence: Level II. Clinical relevance: The presented system is portable, easy to use and fast. It also has good intra and inter-observer reproducibility, making it a good tool to assess objectively scapular dyskinesis in the clinical setting. The normative data obtained is consistent with previous information available. [ABSTRACT FROM AUTHOR]

In this work, we describe the use of Boron Dipyrromethene (BODIPY) as electron‐withdrawing group for activation of double bonds in asymmetric copper‐catalyzed [3+2] cycloaddition reactions with azomethine ylides. The reactions take place under smooth conditions and with high enantiomeric excess for a large number of different substituents, pointing out the high activation of the alkene by using a boron dipyrromethene as electron‐withdrawing group. Experimental, theoretical studies and comparison with other common electron‐withdrawing groups in asymmetric copper‐catalyzed [3+2] cycloadditions show the reasons of the different reactivity of the boron dipyrromethene derivatives, which can be exploited as a useful activating group in asymmetric catalysis. Additional experiments show that the so obtained pyrrolidines can be employed as biocompatible biosensors, which can be located in the endosomal compartments and do not present toxicity in three cell lines. [ABSTRACT FROM AUTHOR]

The novel NKG2C*03 allele encodes a hybrid of the NKG2C*01 and NKG2C*02 primary structures. [ABSTRACT FROM AUTHOR]

The aim was to evaluate the effect of size (long or chopped wheat straw), type (barley or wheat) and presence/absence (1 week) of straw supplementation on behaviour and welfare of fattening lambs. Sixteen lambs (65 days) were individually housed to be used in two simultaneous experiments (8 lambs each) for 5 weeks of fattening. After a habituation period, they were offered straw that differed either in size (Experiment 1) or type (Experiment 2), which was followed by a week without straw and then reallocation of straw. Maintenance behaviours, play and stereotypies were analysed using video recording. Physiological stress responses were measured using blood samples taken at the end of the week without straw and after the reallocation of the straw. A preference test was performed during the last week. Concentrate and roughage consumption, average daily gain and conversion index were estimated. Lambs provided with long straw spent more time eating forage, playing and performed fewer stereotypies (Experiment 1). Lambs given wheat straw increased their play behaviour compared with those given barley straw, without affecting stereotypies (Experiment 2). When the straw was removed, lambs performed more behaviours associated with stress; however, this was not reflected in blood stress indicators. Lambs clearly preferred long straw over chopped straw, but no clear preference was observed between wheat and barley straw. The amount of forage intake was consistent with the time spent eating forage, but the amount of concentrate intake did not vary with eating concentrate behaviour, since the amount of concentrate intake was higher when the straw was removed, especially in Experiment 2. Our results show that size more than type of straw supplement impact the behaviour of lambs. Long straw reduces the stereotypies and increases play. This may improve the welfare and performance of fattening lambs, adding ethical value to a highly appreciated product. [ABSTRACT FROM AUTHOR]

Familial hypercholesterolemia is an autosomal dominant disease of lipid metabolism caused by defects in the genes LDLR, APOB, and PCSK9. The prevalence of heterozygous familial hypercholesterolemia (HeFH) is estimated between 1/200 and 1/250. Early detection of patients with FH allows initiation of treatment, thus reducing the risk of coronary heart disease. In this study, we performed in vitro characterization of new LDLR variants found in our patients. Genetic analysis was performed by Next Generation Sequencing using a customized panel of 198 genes in DNA samples of 516 subjects with a clinical diagnosis of probable or definitive FH. All new LDLR variants found in our patients were functionally validated in CHO‐ldlA7 cells. The LDLR activity was measured by flow cytometry and LDLR expression was detected by immunofluorescence. Seven new variants at LDLR were tested: c.518 G>C;p.(Cys173Ser), c.[684 G>T;694 G>T];p.[Glu228Asp;Ala232Ser], c.926C>A;p.(Pro309His), c.1261A>G;p.(Ser421Gly), c.1594T>A;p.(Tyr532Asn), and c.2138delC;p.(Thr713Lysfs*17). We classified all variants as pathogenic except p.(Ser421Gly) and p.(Ala232Ser). The functional in vitro characterization of rare variants at the LDLR is a useful tool to classify the new variants. This approach allows us to confirm the genetic diagnosis of FH, avoiding the classification as "uncertain significant variants", and therefore, carry out cascade family screening. [ABSTRACT FROM AUTHOR]

We have previously shown that infectious pancreatic necrosis virus (IPNV) enters the embryo cell line CHSE‐214 by macropinocytosis. In this study, we have extended our investigation into SHK‐1 cells, a macrophage‐like cell line derived from the head kidney of Atlantic salmon, the most economically important host of IPNV. We show that IPNV infection stimulated fluid uptake in SHK‐1 cells above the constitutive macropinocytosis level. In addition, upon infection of SHK‐1 cells, IPNV produced several changes in actin dynamics, such as protrusions and ruffles, which are important features of macropinocytosis. We also observed that the Na+/H+ pump inhibitor EIPA blocked IPNV infection. On the other hand, IPNV entry was independent of clathrin, a possibility that could not be ruled out in CHSE 214 cells. In order to determine the possible role of accessory factors on the macropinocytic process, we tested several inhibitors that affect components of transduction pathways. While pharmacological intervention of PKI3, PAK‐1 and Rac1 did not affect IPNV infection, inhibition of Ras and Rho GTPases as well as Cdc42 resulted in a partial decrease in IPNV infection. Further studies will be required to determine the signalling pathway involved in the macropinocytosis‐mediated entry of IPNV into its target cells. [ABSTRACT FROM AUTHOR]

The novel C*07:37:01:02 was completely sequenced after haplo‐specific amplification from a European Caucasoid carrying B*07:02. [ABSTRACT FROM AUTHOR]

Summary: A metabolic network model of the green microalga Chlamydomonas reinhardtii was used to characterize photoautotrophic and heterotrophic (i.e. growth on stored compounds) growth under light and dark, respectively. The metabolic network comprised 2514 reactions distributed among nine intracellular compartments and the extracellular space. The metabolic network included all the key biochemical pathways for synthesis and metabolism of starch and triacylglycerols (TAGs). Under light and nitrogen limitation, the model simulated the accumulation of the energy‐rich compounds (TAGs and starch) in the cell. In the dark, the model could simulate cell growth and maintenance on stored compounds. The model‐predicted consumption rates of storage compounds (starch or TAGs) to enable growth in the dark, were found to be greater than the rates of synthesis under light. This implied utilization of the storage compounds for cell maintenance in the dark. Under constant illumination, the simulations of cell growth and intracellular starch content agreed closely with independent experimental data. In other simulations, compared with the case without photorespiration, light uptake rate increased 1.04‐fold when the ratio of the rates of oxygenation and carboxylation (Rubisco) was 0.1. Although extensive experimental work exists on culture and physiology of microalgae, it does not allow quantitative predictions of the influence of dark metabolism on the productivity of metabolites to be made. This limitation is overcome using the present model. A metabolic network model of Chlamydomonas reinhardtii is shown to simulate growth and synthesis of energy‐rich compounds (triacylglycerols and starch) under light. The same model also simulates dark growth and maintenance through consumption of the stored energy‐rich compounds. Significance Statement: A metabolic network model of Chlamydomonas reinhardtii is shown to simulate growth and synthesis of energy‐rich compounds (triacylglycerols and starch) under light. The same model also simulates dark growth and maintenance through consumption of the stored energy‐rich compounds. [ABSTRACT FROM AUTHOR]

Abstract: BACKGROUND: The bioavailability of therapeutic proteins is improved through PEGylation. This chemical modification involves the production of isomers with different numbers and sites of attached PEG chains, which are difficult to separate efficiently. Their purification with chromatography requires an understanding of the operation and the evaluation of different operational conditions. The General Rate Model (GRM) was applied for modelling the linear salt gradient elution of mono‐PEGylated and native lysozyme in Heparin Affinity Chromatography (HAC) considering mass balance equations for proteins in the bulk‐fluid phase, in the particle phase and the kinetic adsorption. RESULTS: The model was able to simulate the individual proteins and the separation of these in a PEGylation reaction using as proof‐of‐concept a mono‐PEGylated and native lysozyme mixture under changes of operational parameters such as the gradient length (5, 13, 25 column volumes) and flow (0.8 and 1.2 mL min‐1) with a relative error in retention times of less than 6% and correlation coefficients greater than 0.78. CONCLUSION: Simulation of the elution curves of PEGylated lysozyme in HAC was performed in this work and the diverse information generated by the model is explained through the physicochemical protein properties. This simulation represents a tool for optimization, prediction and future scale‐up of PEGylated proteins purification, which would reduce the investment in time and resources to test several operating conditions. © 2017 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Abstract: The first genome scale model (GSM) for Streptomyces leeuwenhoekii C34 was developed to study the biosynthesis pathways of specialized metabolites and to find metabolic engineering targets for enhancing their production. The model, iVR1007, consists of 1,722 reactions, 1,463 metabolites, and 1,007 genes, it includes the biosynthesis pathways of chaxamycins, chaxalactins, desferrioxamines, ectoine, and other specialized metabolites. iVR1007 was validated using experimental information of growth on 166 different sources of carbon, nitrogen and phosphorous, showing an 83.7% accuracy. The model was used to predict metabolic engineering targets for enhancing the biosynthesis of chaxamycins and chaxalactins. Gene knockouts, such as sle03600 (L‐homoserine O‐acetyltransferase), and sle39090 (trehalose‐phosphate synthase), that enhance the production of the specialized metabolites by increasing the pool of precursors were identified. Using the algorithm of flux scanning based on enforced objective flux (FSEOF) implemented in python, 35 and 25 over‐expression targets for increasing the production of chaxamycin A and chaxalactin A, respectively, that were not directly associated with their biosynthesis routes were identified. Nineteen over‐expression targets that were common to the two specialized metabolites studied, like the over‐expression of the acetyl carboxylase complex (sle47660 (accA) and any of the following genes: sle44630 (accA_1) or sle39830 (accA_2) or sle27560 (bccA) or sle59710) were identified. The predicted knockouts and over‐expression targets will be used to perform metabolic engineering of S. leeuwenhoekii C34 and obtain overproducer strains. [ABSTRACT FROM AUTHOR]

BACKGROUND The efficient, controlled and robust purification of conjugates from PEGylation has a growing demand in the biopharmaceutical's market. In general, the yield and purity reached through the conventional chromatographic modes are not particularly high or efficient. Affinity chromatography has so far scarcely been explored. The present work introduces the purification of mono-PEGylated lysozyme from a PEGylation reaction by heparin affinity chromatography (HAC) for the first time in a single step. Response surface methodology (RSM), particularly a Box-Behnken design (BBD) was employed to optimize the separation. RESULTS Protein adsorption of PEGylated and native lysozyme on Heparin Sepharose 6 Fast Flow resin was described by Langmuir isotherms, showing a relatively low affinity for the PEGylated proteins. From the experimental design, optimal elution conditions in a linear gradient of sodium chloride (NaCl) for the three response variables (yield, purity and productivity) were: gradient length of 13 column volumes (CVs), flow at 0.8 mL min−1 and protein load of 1 mg mL−1. Based on this optimization, a step gradient procedure was designed that achieved the purification of mono-PEGylated lysozyme with approximately 100% yield and purity in comparison with 92.7% and 99.7% with the linear gradient. Productivity was c. 0.048 ± 0.001 mg mL−1 min−1 using 0.05 mol L−1 NaCl for its elution. CONCLUSIONS Mono-PEGylated lysozyme was completely separated from a PEGylation mixture with high yield and purity using HAC for first time. Applying response surface methodology (RSM), adequate conditions for more than one requirement were found as well as optimal conditions for a linear gradient of NaCl. Based on this optimization a step gradient procedure was designed that achieved the purification of mono-PEGylated lysozyme in one step with advantages respect to time, resolution, yield and purity compared with other chromatographic modes such as hydrophobic interaction chromatography (HIC) and cation exchange chromatography (CEX). © 2017 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

BACKGROUND Gene therapy is a potent alternative for long-lasting inhibition of alcohol consumption. This study compares the purification of a recombinant adenoviral vector serotype 5 ( rAdV5) for use in gene therapy against alcoholism using two anion-exchange methods. RESULTS Two anion-exchange chromatography methods using fast protein liquid chromatography were compared using a packed-bed column (Q-Sepharose™ XL) and two monolithic columns ( CIM™ QA-1 and CIM™ DEAE-1). An improved and reproducible separation of recombinant adenovirus type 5 from cell lysate contaminants was achieved using the two strong anion-exchange columns in a two-step gradient chromatography. Higher adenovirus yields were achieved using the CIM QA-1 tube monolithic column at sample volumes of both 1 and 10 mL compared with the Q-Sepharose XL column. At higher flow rates, the CIM QA-1 tube monolithic column achieved better separation of the target fraction. Process recovery was improved from 28% using the Q-Sepharose XL column to 34% with the CIM QA-1 tube monolithic column quantified as vector genome. Analysis by SDS-PAGE demonstrated a purity of 70% for purified adenovirus using the CIM QA-1 tube monolithic column. CONCLUSION This study indicated that the use of a CIM QA-1 tube monolithic column is a better alternative than Q-Sepharose XL, and CIM DEAE-1 tube monolithic columns for the primary purification process of rAdV5 carrying the human aldehyde dehydrogenase-2 antisense gene. This purification strategy has been used as a basis to scale-up a GLP process for the production of material at the National Research Council of Canada to be used in preclinical trials of this gene therapy against alcoholism. © 2017 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

ABSTRACT In this work a biotechnological multiproduct batch plant that manufactures four different recombinant proteins for human application is described in some detail. This batch plant design is then optimized with regards to the size of equipment using a mixed-integer linear programming (MILP) formulation recently developed by us in order to find a hypothetical new biotechnological batch plant based on the information of real known processes for the production of the four recombinant protein products. The real plant was divided for practical purposes into two sub-processes or facilities: a fermentation facility and a purification facility. Knowing the specific steps conforming the downstream processing of each product, size, and time factors were computed and used as parameters to solve the aforementioned MILP reformulation. New constraints were included to permit the selection of some equipment-such as centrifuges and membrane filters-in a discrete set of sizes. For equipment that can be built according to customer needs-such as reactors-the original formulation was retained. Computational results show the ability of this optimization methodology to deal with real data giving reliable solutions for a multi-product batch plant composed of 44 unit operations in a relatively small amount of time showing that in the case studied it is possible to save up to a 66% of the capital investment in equipment given the cost data used. Biotechnol. Bioeng. 2017;114: 1252-1263. © 2017 Wiley Periodicals, Inc. [ABSTRACT FROM AUTHOR]

Biomining is defined as biotechnology for metal recovery from minerals, and is promoted by the concerted effort of a consortium of acidophile prokaryotes, comprised of members of the Bacteria and Archaea domains. Ferroplasma acidiphilum and Leptospirillum ferriphilum are the dominant species in extremely acid environments and have great use in bioleaching applications; however, the role of each species in this consortia is still a subject of research. The hypothesis of this work is that F. acidiphilum uses the organic matter secreted by L. ferriphilum for growth, maintaining low levels of organic compounds in the culture medium, preventing their toxic effects on L. ferriphilum. To test this hypothesis, a characterization of Ferroplasma acidiphilum strain BRL-115 was made with the objective of determining its optimal growth conditions. Subsequently, under the optimal conditions, L. ferriphilum and F. acidiphilum were tested growing in each other's supernatant, in order to define if there was exchange of metabolites between the species. With these results, a mixed culture in batch cyclic operation was performed to obtain main specific growth rates, which were used to evaluate a mixed metabolic model previously developed by our group. It was observed that F. acidiphilum, strain BRL-115 is a chemomixotrophic organism, and its growth is maximized with yeast extract at a concentration of 0.04% wt/vol. From the experiments of L. ferriphilum growing on F. acidiphilum supernatant and vice versa, it was observed that in both cases cell growth is favorably affected by the presence of the filtered medium of the other microorganism, proving a synergistic interaction between these species. Specific growth rates were obtained in cyclic batch operation of the mixed culture and were used as input data for a Flux Balance Analysis of the mixed metabolic model, obtaining a reasonable behavior of the metabolic fluxes and the system as a whole, therefore consolidating the model previously developed. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1390-1396, 2016 [ABSTRACT FROM AUTHOR]

Full and effective participation of indigenous peoples and local communities, and high accuracy estimates are two current requirements for the purposes of monitoring forests at international level. We produced two land cover maps, both of which were based on digital image processing (decision trees) using Rapideye imagery, and a land cover participatory map, for indigenous territories of eastern Panama. Accuracy of the three maps was evaluated using field data. Classification that was based on participatory mapping gave best overall accuracy of 83.7% (κ = 0.783), followed by the decision tree that included textural variables (DT2: overall accuracy of 79.9%, κ = 0.757). We have demonstrated for the first time that local knowledge can improve land cover classification and facilitate the identification of forest degradation. The plea of the UNFCC for the full and effective participation of local and indigenous people could, therefore, improve the accuracy of monitoring. [ABSTRACT FROM AUTHOR]

The oxidation process of sulfide minerals in natural environments is achieved by microbial communities from the Archaea and Bacteria domains. A metabolic reconstruction of two dominant species, Leptospirillum ferriphilum and Ferroplasma acidiphilum, which are always found together as a mixed culture in this natural environments, was made. The metabolic model, composed of 152 internal reactions and 29 transport reactions, describes the main interactions between these species, assuming that both use ferrous iron as energy source, and F. acidiphilum takes advantage of the organic compounds secreted by L. ferriphilum for chemomixotrophic growth. A first metabolic model for a mixed culture used in bacterial leaching is proposed in this article, which pretends to represent the characteristics of the mixed culture in a simplified manner. It was evaluated with experimental data through flux balance analysis (FBA) using as objective function the maximization of biomass. The growth yields on ferrous iron obtained for each microorganism are consistent with experimental data, and the flux distribution obtained allows understanding of the metabolic capabilities of both microorganisms growing together in a bioleaching process. The model was used to simulate the growth of F. acidiphilum on different substrates, to determine in silico which compounds maximize cell growth, and which are essential. Knockout simulations were carried out for L. ferriphilum and F. acidiphilum metabolic models, predicting key enzymes of central metabolism. The results of this analysis are consistent with experimental data from literature, showing a robust behavior of the metabolic model. © 2014 American Institute of Chemical Engineers Biotechnol. Prog., 31:307-315, 2015 [ABSTRACT FROM AUTHOR]

Two different approaches to highly symmetric macrocycles by reversible Nicholas reactions have been developed. The first sequence uses a bis[Co2(CO)6] complex derived from a double propargylic alcohol supporting two natural product moieties. The reactions with BF3 ·OEt2 and different benzene dimethanols yielded either (1:1) or (2:2) adducts, depending essentially on the nature of the tether joining both Co clusters. Alternatively, the Nicholas reactions of Co-complexed propargyl alcohol templates with one steroid and one monoterpene fragment as well as one aromatic terminus containing an alcohol moiety yielded the corresponding macrocycles derived from self-dimerization. Both routes to macrocycles are complementary and efficiently produce sophisticated natural-product-containing structures. [ABSTRACT FROM AUTHOR]