Development and Evaluation of a New qPCR Assay for the Detection of Mycoplasma in Cell Cultures.

In recent years, cell culture has become an important tool not only in research laboratories, but also in diagnostic and biotechnological development laboratories. Mycoplasma contamination is present in up to 35% of cell cultures used in research and in cell therapies. This fact represents a significant problem since such contamination can cause disastrous effects on eukaryotic cells by altering their cellular parameters, which, in turn, can lead to unreliable experimental results. For this reason, it is mandatory to carry out continuous testing for the presence of Mycoplasma in cell culture and the development of appropriate methodologies for this purpose. An ideal detection methodology should be fast, sensitive, and reliable. In this study, we propose an alternative detection method based on real-time PCR in conjunction with a novel combination of primers and probes that have been improved to increase their efficiency. The new PCR method demonstrates 100% sensitivity and specificity results in the detection of common Mycoplasma species that contaminate cell cultures. Whilst 11 of 45 tested supernatants were positive for Mycoplasma (24.4%) using the new PCR method (corresponding to 5 of the 14 lines tested (35.71%)), only 10 of 45 supernatants showed positive results with the commercial Venor®GeM qEP and Plasmotest® kit. In addition, the new PCR method exhibits a high capacity to detect less-frequent Mycoplasma species, such as those related to the M. mycoides cluster. The use of an alternative Mycoplasma-detection method in cell culture labs can guarantee the detection of Mycoplasma contamination, especially in cases when dubious results are recorded.

Thin Film Composite Polyamide Reverse Osmosis Membrane Technology towards a Circular Economy.

It is estimated that Reverse Osmosis (RO) desalination will produce, by 2025, more than 2,000,000 end-of-life membranes annually worldwide. This review examines the implementation of circular economy principles in RO technology through a comprehensive analysis of the RO membrane life cycle (manufacturing, usage, and end-of-life management). Future RO design should incorporate a biobased composition (biopolymers, recycled materials, and green solvents), improve the durability of the membranes (fouling and chlorine resistance), and facilitate the recyclability of the modules. Moreover, proper membrane maintenance at the usage phase, attained through the implementation of feed pre-treatment, early fouling detection, and membrane cleaning methods can help extend the service time of RO elements. Currently, end-of-life membranes are dumped in landfills, which is contrary to the waste hierarchy. This review analyses up to now developed alternative valorisation routes of end-of-life RO membranes, including reuse, direct and indirect recycling, and energy recovery, placing a special focus on emerging indirect recycling strategies. Lastly, Life Cycle Assessment is presented as a holistic methodology to evaluate the environmental and economic burdens of membrane recycling strategies. According to the European Commission's objectives set through the Green Deal, future perspectives indicate that end-of-life membrane valorisation strategies will keep gaining increasing interest in the upcoming years.

Chlorophyll soft-sensor based on machine learning models for algal bloom predictions.

Harmful algal blooms (HABs) are a growing concern to public health and aquatic ecosystems. Long-term water monitoring conducted by hand poses several limitations to the proper implementation of water safety plans. This work combines automatic high-frequency monitoring (AFHM) systems with machine learning (ML) techniques to build a data-driven chlorophyll-a (Chl-a) soft-sensor. Massive data for water temperature, pH, electrical conductivity (EC) and system battery were taken for three years at intervals of 15 min from two different areas of As Conchas freshwater reservoir (NW Spain). We designed a set of soft-sensors based on compact and energy efficient ML algorithms to infer Chl-a fluorescence by using low-cost input variables and to be deployed on buoys with limited battery and hardware resources. Input and output aggregations were applied in ML models to increase their inference performance. A component capable of triggering a 10 [Formula: see text]g/L Chl-a alert was also developed. The results showed that Chl-a soft-sensors could be a rapid and inexpensive tool to support manual sampling in water bodies at risk.

Nitrate-Selective Anion Exchange Membranes Prepared using Discarded Reverse Osmosis Membranes as Support.

The present work shows a methodology for the preparation of membranes with a high affinity for nitrates. For this purpose, a polymeric mixture containing an anion exchange resin was extended on a recycled pressure filtration membrane used as mechanical support. Different ion exchange resins were tested. The influence in ion fractionation of (i) the type of ion exchange resin, (ii) the use of a recycled membrane as support and (iii) the operating current density during the separation process were studied. Results revealed that the employed anion exchange resin could tune up the transport numbers of the anions in the membrane and enhance the transport of nitrates over sulfates. The use of the recycled filtration membrane as support further increased the transport of nitrates in detriment of sulfates in nitrate-selective membranes. Moreover, it considerably improved the mechanical stability of the membranes. Lowering the operational current density also boosted ion fractionation. In addition, the use of recycled membranes as support in membrane preparation is presented as an alternative management route of discarded reverse osmosis membranes, coupling with the challenging management of waste generated by the desalination industry. These membranes could be used for nitrate recovery from wastewater or for nitrate separation from groundwater.

Influence of acid/base activation treatment in the performance of recycled electromembrane for fresh water production by electrodialysis.

In this study, an activation treatment for recycled anion exchange membranes is proposed. Following the circular economy approach, these membranes were prepared by using end-of-life reverse osmosis membranes as mechanical support. The end-of-life membrane was previously used and discarded by desalination plants after overcoming its lifespan. The activation treatment was based on the subsequent immersion of the membranes in diluted acid and alkali solutions. This treatment promoted the complete dissociation of the functional groups in the membrane, making them more reactive to the counter ions. The effects of acid and alkali concentrations and exposition times on the electrochemical properties were studied and the best combination was selected. In such a way, a decrease of 37% in membrane electrical resistance was achieved. The performance of activated and non-activated membranes in brackish water desalination by electrodialysis was compared. The results showed that the proposed activation treatment increased the flux of fresh water more than four-fold (from 1.2 to 4.9 L h-1·m-2), with a considerable reduction of energy consumption (from 5.2 to 3.0 kWh·m-3) and a great improvement in current efficiency (from 38% to 71%). In conclusion, this work shows a simple and low cost methodology for the improvement of the electrochemical properties of recycled electromembranes and thus, their performance in electrodialysis.

PEGylated ß-cyclodextrins: Click synthesis and in vitro biological insights.

We present three easily rationalized star-shaped PEGylated ß-cyclodextrin (ßCD) derivatives synthesized via conjugation of different molecular weight PEG chains (5000, 2000, and 550 Da) to the ßCD primary face by click chemistry (ßCD-PEG5000, ßCD-PEG2000, ßCD-PEG550 respectively). ßCDPEG systems are envisioned to further carry bioactive molecules, therefore, their interactions with biological interfaces must be determined at an early stage of development. Hence, the effect of ßCDPEGs chain length on cell viability was investigated. To this aim, three models were selected: Vero cells for their fibroblast-like features; HeLa cells that are commonly used for preliminary viability screening; and human peripheral monocytes which are macrophage precursors. Of the three pegylated derivatives, ßCD-PEG550 was the one that significantly affected HeLa cells and human monocytes viability. Despite the popularity of PEGylation approach, our results underscore the importance of careful and systematic PEGylated materials design for their future success in drug delivery systems.

Interferon alpha induction of metallothionein in rat liver is not linked to interleukin-1, interleukin-6, or tumor necrosis factor alpha.

Synthesis of metallothionein (MT) is induced by interferon-alpha (IFN-alpha) in vitro and in vivo. In addition, IFN-alpha promotes redistribution of zinc (Zn) from the plasma to the liver in mice. However, it is not clear if IFN-alpha induces hepatic MT synthesis directly or indirectly via liberation of other cytokines. In order to address this issue, we determined hepatic MT levels, Zn concentration in plasma, liver, and urine, and plasma levels interleukin-1 (IL-1), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNFalpha) in rats following intramuscular injection of human IFN-alpha (1.5 x 10(6) UI/m(2)). Animals were housed in metabolic cages and sacrificed at various times after IFN-alpha administration. Zn concentrations in serum, urine, and hepatic tissue were determined by atomic absorption spectrophotometry. MT protein was measured using the MT silver saturation method and expression of MT-1 and MT-2 mRNA was measured by RT-PCR. Plasma levels of rat IL-1, IL-6, and TNFalpha were determined using an ELISA method. Hepatic MT levels began to increase at 2 h following IFN-alpha administration and reached maximum levels at 12 h post-treatment. Induction of MT gene expression was confirmed by increases in MT-1 and MT-2 mRNA levels 6, 12, and 18 h after IFN-alpha administration. IFN-alpha treatment also resulted in biphasic increases in hepatic Zn, with levels peaking at 2 h, the time-point when MT levels are first increased, and again at 18 h. Concurrently, there were decreases in serum Zn levels at these time points, suggesting IFN-alpha induced movement of Zn from the blood to hepatic tissue. The decrease in serum Zn was not due to increased excretion since urinary Zn levels were unaffected following IFN-alpha treatment. IFN-alpha administration had no effect on plasma IL-1, IL-6, and TNFalpha levels. These results show that IFN-alpha promotes the increase of hepatic MT levels and plasma/liver redistribution directly, without IL-1, IL-6, or TNFalpha participation.

El valor del exámen oftalmoscópico en los hipertensos arteriales: una guia para el internista

Aunque data de 1949, para el lector interesado (y con tiempo) será valido por las extensas y detalladas descripciones semiológica-diagnósticas que contiene. Los signos oftalmoscópicos, los cuadros oftalmoscópicos de los hipertensos arteriales. Las clasificaciones de los hipertensos arteriales que tienen como base las comprobaciones oftalmoscópicas. Valor diagnóstico, pronostico y de indicación y control terapeútico del exámen oftalmoscópico en las enfermedades hipertensivas. El exámen del fondo del ojo por el internista

El valor del exámen oftalmoscópico en los hipertensos arteriales: una guia para el internista

Aunque data de 1949, para el lector interesado (y con tiempo) será valido por las extensas y detalladas descripciones semiológica-diagnósticas que contiene. Los signos oftalmoscópicos, los cuadros oftalmoscópicos de los hipertensos arteriales. Las clasificaciones de los hipertensos arteriales que tienen como base las comprobaciones oftalmoscópicas. Valor diagnóstico, pronostico y de indicación y control terapeútico del exámen oftalmoscópico en las enfermedades hipertensivas. El exámen del fondo del ojo por el internista