Dry weight model, capacitance and metabolic data as indicators of fungal biomass growth in solid state fermentation

Developing improved industrial bioprocesses has been a driver for the growing research attention to solid state fermentation, in particular involving filamentous fungi. Accurate description of fungal growth in these systems is crucial and certainly needed to enable optimal deployment of subsequent engineering work. This manuscript proposes a model based on total dry weight measurement to describe biomass growth for Aspergillus awamori on wheat grains in two systems: Petri dishes and a 1L packed bed bioreactor. The proposed dry weight model can be used not only for identifying growth phases of the fungus but also to calculate key growth parameters such as specific growth rate and maximum biomass concentration. The use of techniques based on capacitance measurements and on metabolic data were also used in order to estimate fungal growth and to validate the proposed model

Psychosocial stress and prematurity: impact on maternal and neonatal health

Trabajo fin de grado en EnfermeríaIntroducción: El estrés durante el embarazo puede aumentar el riesgo de parto prematuro que, a su vez, puede incrementar el estrés materno postparto y, con ello, afectar a la salud de la madre y del hijo. Estas repercusiones evidencian la necesidad de llevar a cabo intervenciones encaminadas a reducir el estrés materno, tanto previas a la concepción como durante la gestación y el periodo postparto. Objetivo: Analizar el estrés materno gestacional y postparto relacionado con la prematuridad y discutir las intervenciones que ayudan a su reducción. Comparar, en un estudio experimental, el nivel de estrés en madres de prematuros y a término. Metodología: Para la revisión bibliográfica se realizó una búsqueda de artículos en bases de datos científicas. El estudio experimental consta de dos cuestionarios (PSS-10 y LOTR-6) que fueron rellenados por madres lactantes en el postparto. Resultados/discusión: El estrés se debe detectar precozmente durante la gestación. Tras el parto, la reducción del estrés se puede realizar mediante intervenciones educativas, de apoyo psicoemocional e intervenciones que implican a los padres como cuidadores activos. En el estudio experimental, se observa una tendencia a un nivel mayor de estrés en las madres de prematuros en comparación a las madres a término. Conclusiones: Durante la gestación, lo más efectivo es realizar una evaluación integral. Tras el parto, la intervención que muestra mejores resultados es la descripción de la UCIN y del comportamiento y las características del prematuro. Los datos experimentales indican una tendencia a experimentar mayor estrés en las madres de prematuros.Introduction: Stress during pregnancy can increase the risk of premature delivery which, in turn, can increase maternal post-partum stress and affect the health of both mother and child. These implications highlight the need of interventions to reduce maternal stress, before conception, during pregnancy and the postpartum period. Aim: To analyze maternal gestational and postpartum stress related to prematurity and discuss interventions that help reducing it. To compare, in an experimental study, the level of stress in mothers of preterm and term infants. Methodology: For the bibliographic review, a search of articles in scientific databases was carried out. The experimental study consists of two questionnaires (PSS-10 and LOTR-6) that were filled out by postpartum nursing mothers. Results/discussion: Stress should be detected early during pregnancy. After delivery, stress reduction can be achieved through educational interventions, psycho-emotional support and interventions which involve parents as active caregivers. In the experimental study, there is a trend towards a higher level of stress is observed in mothers of preterm infants compared to full-term mothers. Conclusions: During pregnancy, a comprehensive evaluation is most effective. After birth, the intervention that shows the best results is the description of the NICU and the behaviour and characteristics of the premature infant. Experimental data indicate a tendency to experience greater stress in mothers of preterm infants

What do we know about carbon monoxide poisoning and cardiac compromise?

Carbon monoxide (CO) poisoning is one of the most common types of poisoning and the leading cause of death by poisoning worldwide. Cardiac injury caused by CO poisoning has been little described despite being a predictor of poor prognosis. We present the case of a healthy 24-year-old woman, admitted to our emergency room due to an episode of lipothymia without loss of consciousness. She reported holocranial headache for the previous two weeks associated with nausea and vomiting. Laboratory tests revealed blood gas analysis: pH 7.392, pCO2 32 mmHg, pO2 101 mmHg, lactate 3.5 mmol/l, HCO3 20.8 mmol/l; COHb 29.2%; serial troponin I 1.21→5.25→6.13→3.65 μg/l; myoglobin 1378→964→352 μg/l; and NT-proBNP 1330 pg/l. The electrocardiogram showed sinus rhythm, heart rate 110 bpm, and ST-segment depression of 2 mm in V4 and 1 mm in V5. Transthoracic echocardiography revealed a left ventricle with normal wall motion and preserved ejection fraction. Given the clinical and epidemiological context, myocardial and central nervous system ischemia due to prolonged CO exposure was assumed and normobaric oxygen therapy was immediately started. In view of evidence of injury to two major organ systems the indication for hyperbaric oxygen therapy was discussed with a specialist colleague, who suggested maintaining conservative treatment with oxygen therapy and in-hospital monitoring for 72 h. The patient was discharged on the third day and was still asymptomatic at 400 days of follow-up. Besides symptoms and signs of central nervous system dysfunction, myocardial damage should also always be considered in the context of CO poisoning. Hyperbaric therapy is still controversial and the lack of objective data highlights the need for new randomized studies.publishersversionpublishe

SMP-CONTAINING PROTEINS AT MEMBRANE CONTACT SITES: SUBCELLULAR LOCALIZATION AND CHARACTERIZATION.

Membrane contact sites (MCS) are microdomains where two membranes of two different organelles are in close apposition, but they do not fuse. MCS are essential for non-vesicular transport of lipids. This lipid transport is mediated by several families of proteins which all of them contain a lipid transport domain, as the synaptotagmin-like mitochondrial lipid-binding (SMP) domain. The most studied SMP protein is Arabidopsis SYT1 which is known to be involved in tolerance to multiple abiotic stresses. Later studies in other SMP proteins of the same family have shown that SYT1 and homologous such as SYT3 or SYT5 gave similar results. However, little information is available about the role other SMP proteins in plants. We have studied the occurrence of additional SMP proteins in A. thaliana and S. lycopersicum. In order to identify these proteins, SMP sequences from human and yeast were used to identify their remote orthologues in A. thaliana and S. lycopersicum, allowing the identification of several putative encoding SMP domains. We have found that some of the identified proteins are exclusive of plants as they do not have direct orthologs in yeast nor human. Transient expression in N. benthamiana leaves followed by confocal microscopy was used to study the subcellular localization of these proteins. Our results show that some of these proteins are localized at ER-Golgi contact sites and two other proteins at ER-Chloroplast sites. Finally, to determine whether these proteins are involved in abiotic stress tolerance, we have analysed the root growth and seed germination rates of Arabidopsis mutants for these genes under different conditions. Some of these mutants have shown different germination rates in media supplemented with NaCl and different rates of expanded cotyledons in media supplemented with ABA. These results suggest that some these proteins may be implicated in abiotic stress signalling through an ABA-dependent pathway.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech. This work is supported by grants from: Ministerio de Ciencia, Innovación y Universidades (grant PGC2018-098789-B-I00), UMA-FEDER (grant UMA18-FEDERJA-154) and Ministerio de Ciencia e Innovación (BIO2017-82609-R)

Pt(IV)-based nanoscale coordination polymers : antitumor activity, cellular uptake and interactions with nuclear DNA

Cisplatin has been for many years the gold standard chemotherapeutic drug for the treatment of a wide range of solid tumors, even though its use is commonly associated with serious side effects including non-selective toxicity, myelosuppression or development of cisplatin resistance, among others complications. Over the last decade, a number of nanoparticle formulations were developed to reduce its side effects and improve the selectivity and efficacy of this drug. In this study, we have developed a novel nanoparticle platform based on nanoscale coordination polymer named (Zn-Pt(IV)-NCPs) which contains a Pt(IV) prodrug, Zn and the linker ligand 1,4-Bis(imidazol-1-ylmethyl)benzene (bix). The main objective has been to gain insights into the mechanism of action of this nanostructured material in comparison with cisplatin and the free Pt(IV) prodrug in order to establish a correlation between nanostructuration and therapeutic activity. Zn-Pt(IV)-NCPs nanoparticles displayed an average size close to 200 nm as determined by DLS, a good stability in physiologic environments, and a controlled drug release of Pt. In vitro studies demonstrated that Pt(IV)-NCPs showed an enhanced cytotoxic effect against cell culture of cervical cancer, neuroblastoma and human adenocarcinoma cells in comparison with free Pt(IV) prodrug. Although no difference in cell uptake of Pt was observed for any of the three cell lines assayed, a higher amount of Pt bound to the DNA was found in the cells treated with the nanostructured Pt(IV) prodrug. These studies suggest that the nanostructuration of the prodrug facilitate its activation and induce a change in the mechanism of action related to an increased interaction with the DNA as corroborated by the studies of direct interaction of the Pt(IV) prodrug, nanostructured or not, with DNA

Localization and characterization of SMP-containing proteins in Membrane Contact Sites

Membrane contact sites (MCS) are discrete regions where two membranes from different organelles are closely apposed (10-30 nm). In those regions, non-vesicular transfer of lipids takes place to ensure proper organelle functioning. Arabidopsis SYT1 is one of the best characterized MCS protein, and it plays a relevant role in tolerance to abiotic stresses. SYT1 is a SMP (synaptotagmin-like mitocondrial lipid binding domain) containing protein localized at ER-PM contact sites. Recent studies suggest that this protein transfer glycerolipids between these two membranes. However, little is known about other SMP-containing proteins in plants, as their localization or their role in abiotic stress. We have focused on studying the rest of SMP-containing proteins in Arabidopsis thaliana and Solanum lycopersicum. To identify them, human E-Syt1 sequence was used to find the remote orthologues in plants. An interesting highlight of those results was that some SMP-containing proteins are exclusive from plants, there are no orthologues in human nor yeast. The subsequent step was the study of their subcellular location, that was carried out in Nicotiana benthamiana by transient expression of the SMP-containing proteins from Arabidopsis and Solanum, followed by confocal microscopy imaging. We have found that those proteins locate in different MCS across the cell: SYT6, NTMC2T6 and Tex2 localise in ER-Golgi contact sites, NTMC2T5 in ER-Chloroplast contact sites, and we have also confirmed that Solanum CLB1 and SYT5 localized at ER-PM contact sites as their Arabidopsis counterparts. Additionally, we have analysed the root growth, seed germination rates and fully expanded cotyledons of Arabidopsis mutants for these genes in media supplemented with salt or ABA, and our results suggest that some of these proteins might be implicated in abiotic stress signalling through an ABA pathway.This work is supported by grants from: Ministerio de Ciencia, Innovación y Universidades (grant PGC2018-098789-B-I00), UMA-FEDER (grant UMA18-FEDERJA-154) and Ministerio de Ciencia e Innovación (BIO2017-82609-R), and meeting assistance was granted by Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Diacylglycerol transport by Arabidopsis Synaptotagmin 1 at ERplasma membrane contact sites under abiotic stress.

Bulk lipid transport between membranes within cells involves vesicles, however membrane contact sites have recently been discovered as mediators of non-vesicular lipid transfer. ER-PM contact sites are conserved structures defined as regions of the endoplasmic reticulum (ER) that tightly associate with the plasma membrane (PM). Our recent data suggest that the constitutively expressed Arabidopsis Synaptotagmin 1 (SYT1) and the cold-induced homolog AtSYT3 are proteins located in these ER-PM contact sites that are essential for the tolerance various abiotic stresses. Arabidopsis SYTs proteins are integral membrane proteins that contain multiple Ca2+-binding C2 domains and a synaptotagmin-like mitochondrial lipid-binding protein (SMP) domain that contains a hydrophobic groove. In mammals, several SMP proteins are responsible for the inter-organelle transport of glycerophospholipids. Our experiments have demonstrated that there is a recruitment of AtSYT1 and AtSTYT3 to ER-PM contact sites under stress conditions and it requires phosphatidylinositol 4- phosphate, PI(4)P in the PM, in opposition to the recruitment of PI(4,5)P2 in mammals. Moreover, our recent high-resolution lipidome analysis suggest that saturated diacylglycerols (DAGs) are the lipids that AtSYT1 is transferring between the PM and ER. Additionally, we have identified AtDGK2 (diacylglycerol kinase 2) as a key interactor of AtSYT1. Generally, in response to a stress stimulus, a phospholipase C (PLC), hydrolyses PIP2 after the elevation of cytosolic Ca2+, generating DAGs which immediately can be converted to phosphatidic acid (PA) by DGKs.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech. The authors acknowledge the support by the Plan Propio from University of Malaga, Campus de Excelencia Internacional de Andalucía and by the Redes of Excelencia (BIO2014-56153-REDT) and BIO2017-82609-R, RYC-2016-21172 & PGC2018-098789 of the Ministerio de Economía, Industria y Competitividad

IDENTIFICATION OF SYT1 INTERACTORS CONNECTS CALCIUM SIGNALING, ENDOPLASMIC RETICULUM BENDING, PLASMODESMATA AND MEMBRANE CONTACT SITES

Synaptotagmin1 (SYT1) is an Arabidopsis thaliana protein essential for tolerance to several abiotic stresses (Schapire et al., 2008; Pérez-Sancho et al., 2015; Ruiz-Lopez et al., 2020). SYT1 forms endoplasmic reticulum-plasma membrane contact sites (ER-PM CS), microdomains conserved across eukaryotes where protein tethers maintain the membranes of the ER and the PM in close apposition (∼30 nm) without fusing. The short distance between membranes facilitates processes such as ion and lipid transport (Pérez-Sancho et al., 2016). For example, SYT1 transports diacylglycerol (DAG) from the PM to the ER during abiotic stress to assure PM integrity (Ruiz-Lopez et al., 2020). Usually, protein complexes form the core of contact sites. In particular, SYT1 forms dimers with SYT1, SYT3, SYT5 and CLB1; all members of the Arabidopsis SYT family and EPCS tethers (Lee et al., 2020; Ruiz-Lopez et al., 2020). We found that SYT1 interacts with proteins involved in different cellular processes by non-targeted proteomic approaches (IP-MS and TAP-tag). Thus, SYT1 interacts with reticulons (RTN), ER-resident proteins responsible for ER curvature, which is crucial for ER morphology and lipid transport at ER-PM CS (Collado et al., 2019). Additionally, SYT1 interacts with ECAs, Ca2+-ATPases located at the ER membrane. SYT1 also interacts with sterol methyltransferases (SMTs), key enzymes in the route of sitosterol and stigmasterol biosynthesis. The homeostasis of the sitosterol and stigmasterol is crucial for the tolerance to wound, heat and bacterial stress presumably by affecting PM fluidity. MCTPs (multiple C2 domains and transmembrane region proteins), plasmodesmata-exclusive proteins are also SYT1 interactors (Brault et al., 2019). We are now investigating the role of SYT1 in these processes using biochemical, genetic and cellular biology approaches

Melatonin affects the dynamic steady-state equilibrium of estrogen sulfates in human umbilical vein endothelial cells by regulating the balance between estrogen sulfatase and sulfotransferase

Melatonin is known to reduce the growth of endocrine-responsive breast cancers by interacting with estrogen signaling pathways. Estrogens play an important role in breast cancer, but also in various types of tissues, including vascular tissue. Estrogen sulfatase (STS) converts inactive estrogen sulfates into active estrogens, whereas estrogen sulfotransferase (EST) sulfonates estrogens to estrogen sulfates. Therefore, STS and EST are considered to be involved in the regulation of local estrogen levels in hormone‑dependent tumors and in non-pathologic tissues, such as those of the vascular system. Estrogens have a major impact on the vasculature, influencing vascular function, the expression of adhesion proteins, angiogenesis and the inflammatory state. In this study, we investigated the status of STS and EST in human umbilical vein endothelial cells (HUVECs) and the modulatory effects of melatonin. Both STS and EST were highly expressed in the HUVECs. The enzymatic activity correlated with the expression levels in these cells. Our findings also demonstrated that melatonin, at physiological concentrations, modulated the synthesis and transformation of biologically active estrogens in HUVECs through the inhibition of STS activity and expression, and the stimulation of EST activity and expression. Since melatonin decreased the STS levels and increased the EST levels, it modified the dynamic steady‑state equilibrium of estrogen sulfates by increasing the inactive estrogen levels and decreasing the active estrogen levels. Therefore, melatonin may modulate the known different biological actions of estrogens in endothelial cells, as well as in estrogen‑dependent tumors and non-pathologic tissues

Melatonin enhancement of the radiosensitivity of human breast cancer cells is associated with the modulation of proteins involved in estrogen biosynthesis

Enhancing the radiosensitivity of cancer cells is one of the most important tasks in clinical radiobiology. Endocrine therapy and radiotherapy are two cancer treatmentmodalities which are often given together in patients with locally-advanced breast cancer and positive hormone-receptor status. Oncostatic actions of melatonin are relevant on estrogen-dependent mammary tumors. In the present study, we wanted to evaluate the effects of the combination of ionizing radiation and melatonin on proteins involved in estrogen biosynthesis in breast cancer cells.We demonstrated a role of melatonin in mediating the sensitization of human breast cancer cells to the ionizing radiation by decreasing around 50% the activity and expression of proteins involved in the synthesis of estrogens in these cells. Thus, melatonin pretreatment before radiation reduces the amount of active estrogens at cancer cell level. Melatonin 1 nM induced a 2-fold change in p53 expression as compared to radiation alone. The regulatory action of melatonin on p53 could be a link between melatonin and its modulatory action on the sensitivity of breast cancer cells to ionizing radiation. These findings may have implications for designing clinical trials using melatonin and radiotherapy