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Biotechnology Progress 2023Upstream advances have led to increased mAb titers above 5 g/L in 14-day fed-batch cultures. This is accompanied by higher cell densities and process-related impurities...
Upstream advances have led to increased mAb titers above 5 g/L in 14-day fed-batch cultures. This is accompanied by higher cell densities and process-related impurities such as DNA and Host Cell Protein (HCP), which have caused challenges for downstream operations. Depth filtration remains a popular choice for harvesting CHO cell culture, and there is interest in utilizing these to remove process-related impurities at the harvest stage. Operation of the harvest stage has also been shown to affect the performance of the Protein A chromatography step. In addition, manufacturers are looking to move away from natural materials such as cellulose and Diatomaceous Earth (DE) for better filter consistency and security of supply. Therefore, there is an increased need for further understanding and knowledge of depth filtration. This study investigates the effect of depth filter material and loading on the Protein A resin lifetime with an industrially relevant high cell density feed material (40 million cells/ml). It focuses on the retention of process-related impurities such as DNA and HCP through breakthrough studies and a novel confocal microscopy method for imaging foulant in-situ. An increase in loading of the primary-synthetic filter by a third, led to earlier DNA breakthrough in the secondary filter, with DNA concentration at a throughput of 50 L/m being more than double. Confocal imaging of the depth filters showed that the foulant was pushed forward into the filter structure with higher loading. The additional two layers in the primary-synthetic filter led to better pressure profiles in both primary and secondary filters but did not help to retain HCP or DNA. Reduced filtrate clarity, as measured by OD600, was 1.6 fold lower in the final filtrate where a synthetic filter train was used. This was also associated with precipitation in the Protein A column feed. Confocal imaging of resin after 100 cycles showed that DNA build-up around the outside of the bead was associated with synthetic filter trains, leading to potential mass transfer problems.
Topics: Cricetinae; Animals; Cricetulus; Batch Cell Culture Techniques; CHO Cells; Filtration; Cell Count; Staphylococcal Protein A
PubMed: 36775837
DOI: 10.1002/btpr.3329 -
Environment International Feb 2023Desert dust is increasingly recognized as a major air pollutant affecting respiratory health. Since desert dust exposure cannot be regulated, the hazardousness of its...
Desert dust is increasingly recognized as a major air pollutant affecting respiratory health. Since desert dust exposure cannot be regulated, the hazardousness of its components must be understood to enable health risk mitigation strategies. Saharan dust (SD) comprises about half of the global desert dust and contains quartz, a toxic mineral dust that is known to cause severe lung diseases via oxidative stress and activation of the NLRP3 inflammasome-interleukin-1β pathway. We aimed to assess the physicochemical and microbial characteristics of SD responsible for toxic effects. Also, we studied the oxidative and pro-inflammatory potential of SD in alveolar epithelial cells and the activation of the NLRP3 inflammasome in macrophage-like cells in comparison to quartz dusts and synthetic amorphous silica (SAS). Characterization revealed that SD contained Fe, Al, trace metals, sulfate, diatomaceous earth, and endotoxin and had the capacity to generate hydroxyl radicals. We exposed A549 lung epithelial cells and wild-type and NLRP3 THP-1 macrophage-like cells to SD, three well-investigated quartz dusts, and SAS. SD induced oxidative stress in A549 cells after 24 h more potently than the quartz dusts. The quartz dusts and SAS upregulated interleukin 8 expression after 4 h and 24 h while SD only caused a transient upregulation. SD, the quartz dusts, and SAS induced interleukin-1β release from wild-type THP-1 cells>20-fold stronger than from NLRP3 THP-1 cells. Interleukin-1β release was lower for SD, in which microbial components including endotoxin were heat-destructed. In conclusion, microbial components in SD are pivotal for its toxicity. In the epithelium, the effects of SD contrasted with crystalline and amorphous silica in terms of potency and persistence. In macrophages, the strong involvement of the NLRP3 inflammasome emphasizes the acute and chronic health risks associated with desert dust exposure.
Topics: Cytokines; Dust; Endotoxins; Inflammasomes; Interleukin-1beta; NLR Family, Pyrin Domain-Containing 3 Protein; Oxidative Stress; Quartz; Silicon Dioxide; Humans; A549 Cells
PubMed: 36680803
DOI: 10.1016/j.envint.2023.107732 -
Scientific Reports Jan 2023In this study, injection moulding was applied to produce biocomposites consisting of polylactide (PLA) and amorphous diatomaceous earth used as a filler at different...
In this study, injection moulding was applied to produce biocomposites consisting of polylactide (PLA) and amorphous diatomaceous earth used as a filler at different concentrations. Natural wax and synthetic wax were added to improve processing properties, comparing the resulting biocomposites. The use of natural beeswax makes the composite environmentally friendly. The prepared composites contained 2.5, 5, 10 and 15% w/w filler. The test samples have been injection moulded. Rheological, mechanical, surface and other properties were assessed for the fabricated composites. The testing has shown that the use of wax additives has a significant influence on the mechanical properties (tensile strength, flexural strength, impact strength) and the hydrophilicity/hydrophobicity of composite surfaces. The addition of natural wax, especially at lower concentration, has a positive effect on the rheological properties of composites (melt flow rate, MFR), flexural modulus and impact strength. Different composite parameters are modified by different wax types so both natural and synthetic waxes, can be used interchangeably, depending on the required final material characteristics.
PubMed: 36670202
DOI: 10.1038/s41598-023-28435-0 -
Environmental Science and Pollution... Mar 2023The current study evaluates the efficacy of the entomopathogenic fungus Beauveria bassiana (Balsamo-Crivelli) Vuillemin (Hypocreales: Cordycipitaceae), diatomaceous...
The current study evaluates the efficacy of the entomopathogenic fungus Beauveria bassiana (Balsamo-Crivelli) Vuillemin (Hypocreales: Cordycipitaceae), diatomaceous earth (DE) (Protect-It), and the oxadiazine indoxacarb, at single or combined applications on wheat kernels, for the management of the rusty grain beetle, Cryptolestes ferrugineus (Stephens) (Coleoptera: Laemophloeidae), the red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae), the khapra beetle, Trogoderma granarium Everts (Coleoptera: Dermestidae), and the lesser grain borer, Rhyzopertha dominica (F.) (Coleoptera: Bostrychidae). The study was conducted between November 2020 and August 2021 in Faisalabad under a complete randomized block design. The combination of DE + indoxacarb was the most efficient as it caused higher overall mortalities ranging between 59.34 and 100%, and lower overall progeny production ranging between 8.35 and 33.70 individuals per vial, than all other treatments. Beauveria bassiana alone exhibited the lowest mortality rates ranging between 22.33 and 47.76%, and the highest offspring emergence, ranging between 51.33 and 78.55 individuals per vial. Similar pattern was observed when persistence bioassays were conducted. For a period of 120 days, the DE + indoxacarb was the most powerful combination against all tested species, providing overall mortality rates between 17.06 and 63.80%. The overall progeny production was lower for the insect individuals exposed on wheat treated with the DE + indoxacarb combination, ranging between 13.66 and 52.23 individuals per vial, and higher for those exposed to B. bassiana alone, ranging between 44.03 and 107.67 individuals per vial, for the entire duration of storage. However, the efficacy of all treatments decreased gradually during the course of storage. The findings of the current study indicate that the combinations of entomopathogenic fungi, DE, and indoxacarb can be used for the prolonged protection of stored wheat from the tested noxious insect species of stored products. Further research, which will include other inert dusts in combination with entomopathogenic fungi and indoxacarb, may provide additional knowledge towards an effective management of noxious species occurring in storages.
Topics: Animals; Beauveria; Coleoptera; Diatomaceous Earth; Hypocreales; Insecta; Insecticides; Triticum
PubMed: 36640233
DOI: 10.1007/s11356-022-25075-1 -
Frontiers in Veterinary Science 2022Water is one of the primary vectors for African swine fever virus (ASFV) transmission among swine herds. However, the low concentrations of ASFV in water represent a...
Water is one of the primary vectors for African swine fever virus (ASFV) transmission among swine herds. However, the low concentrations of ASFV in water represent a challenge for the detection of the virus by conventional PCR methods, and enrichment of the virus would increase the test sensitivity. In this study, aiming to enrich ASFV in water quickly and efficiently, a rapid and efficient water-borne virus enrichment system (MDEF, modified diatomaceous earth by ferric hydroxide colloid) was used to enrich ASFV in water. After enrichment by MDEF, conventional real-time PCR (qPCR) was used for ASFV detection. ASFV were inactivated and diluted in 10 L of water, of which 4 mL were collected after 60 min treatment using the MDEF system. Two thousand five hundred times reduction of the sample volume was achieved after enrichment. A high adsorption rate of about 99.99 (±0.01)% and a high recovery rate of 64.01 (±10.20)% to 179.65 (±25.53)% was achieved by using 1g modified diatomaceous earth for 10 L ASFV contaminated water. The limit of qPCR detection of ASFV decreased to 1 × 10 GU ml (genomic units per milliliter) from 1 × 10 GU ml after concentrating the spiked water from 10 L to 4 ml. Preliminary application of MDEF allowed successful detection of African swine fever virus (ASFV), porcine circovirus type 2 (PCV2), and pseudorabies virus (PRV) in sewage. Thus, the combination of modified diatomaceous earth and real-time PCR is a promising strategy for the detection of viruses in water.
PubMed: 36619955
DOI: 10.3389/fvets.2022.1045190 -
Materials (Basel, Switzerland) Dec 2022Portland cement porous concrete (PCPC) has received immense interest recently due to its environmental aids. Its porous structure helps to reduce the water runoff amount...
Portland cement porous concrete (PCPC) has received immense interest recently due to its environmental aids. Its porous structure helps to reduce the water runoff amount while improving the recharge of groundwater. Earlier studies have concentrated on illustrating and knowing the functional as well as structural properties of PCPC. However, very few studies are available on PCPC in combination with natural silica sources as supplementary cementitious materials (SCMs). Most SCMs are by-products of industrial manufacturing processes and cause some environmental concerns, but with their pozzolanic effect, they could be utilized as partial substitute materials for ordinary Portland cement (OPC) to enhance the strength as well as durability performance. The aim of this study is to evaluate the effects of diatomaceous earth (DE) as a supplementary cementitious material for partial substitution of OPC for Portland cement porous concrete application. Compression strength, split tensile strength, and flexural strength tests were performed to determine the effect of partial replacement. To investigate the impact of test variables, basic tests, including void content and water permeability, were also performed. Compared to the control concrete, the results show that a 15% replacement of cement with DE significantly increased the compressive strength (by 53%) while also providing adequate porosity and better water permeability. Statistical analysis (ANOVA) and regression analysis showed that there is a significant (p < 0.05) growth within the physical characteristics of concrete upon the replacement of cement by 15% DE. Collectively, the replacement of cement with DE could not only improve the concrete strength but also reduce the consumption of cement, thereby lessening the cost of construction as well as indirectly reducing the carbon footprint.
PubMed: 36614394
DOI: 10.3390/ma16010048 -
Sensors and Actuators. B, Chemical Mar 2023Rapid, sensitive, and specific detection of the severe acute respiratory syndrome coronavirus (SARS-CoV)- 2 during early infection is pivotal in controlling the spread...
Rapid, sensitive, and specific detection of the severe acute respiratory syndrome coronavirus (SARS-CoV)- 2 during early infection is pivotal in controlling the spread and pathological progression of Coronavirus Disease 2019 (COVID-19). Thus, highly accurate, affordable, and scalable point-of-care (POC) diagnostic technologies are necessary. Herein, we developed a rapid and efficient self-directed molecular diagnostic (SdMDx) system for SARS-CoV-2. This system combines the sample preparation step, including virus enrichment and extraction processes, which involve dimethyl suberimidate dihydrochloride and diatomaceous earth functionalized with 3-aminopropyl(diethoxy)methylsilane, and the detection step using loop-mediated isothermal amplification-lateral flow assay (LAMP-LFA). Using the SdMDx system, SARS-CoV-2 could be detected within 47 min by hand without the need for any larger instruments. The SdMDx system enabled detection as low as 0.05 PFU in the culture fluid of SARS-CoV-2-infected VeroE6 cells. We validated the accuracy of the SdMDx system on 38 clinical nasopharyngeal specimens. The clinical utility of the SdMDx system for targeting the S gene of SARS-CoV-2 showed 94.4% sensitivity and 100% specificity. This system is more sensitive than antigen and antibody assays, and it minimizes the use of complicated processes and reduces contamination risks. Accordingly, we demonstrated that the SdMDx system enables a rapid, accurate, simple, efficient, and inexpensive detection of SARS-CoV-2 at home, in emergency facilities, and in low-resource sites as a pre-screening platform and POC testing through self-operation and self-diagnosis.
PubMed: 36570722
DOI: 10.1016/j.snb.2022.133193 -
Polymers Dec 2022In the present study, tests were conducted on high-filled composite samples on a polylactide matrix, modified with diatomaceous earth, three types of silanes, and...
In the present study, tests were conducted on high-filled composite samples on a polylactide matrix, modified with diatomaceous earth, three types of silanes, and natural and synthetic wax. The obtained samples were characterized in terms of the effect of modifications on mechanical properties (tensile strength, flexural strength, and impact resistance) or processing properties, e.g., melt flow rate (MFR). The study showed that the modification had a favorable effect on the processing properties of the composites, associated with up to an eight-fold increase in flow rate index compared with the reference sample, especially for samples treated with methyltrimethoxysilane (MTMOS), and up to a ten-fold increase under low shear-rate flow conditions. The effect of the addition of waxes of different origins (synthetic and natural) was also determined, and it was shown that beeswax tended to reduce the flow rate of the composites regardless of the silane used. The addition of synthetic wax to composites increased the tendency to agglomerate diatomaceous earth, while natural wax had a positive effect on filler dispersion.
PubMed: 36559878
DOI: 10.3390/polym14245511 -
Toxins Dec 2022Polydopamine (PDA) is a synthetic eumelanin polymer mimicking the biopolymer secreted by mussels to attach to surfaces with a high binding strength. It exhibits unique...
Improving the In Vitro Removal of Indoxyl Sulfate and p-Cresyl Sulfate by Coating Diatomaceous Earth (DE) and Poly-vinyl-pyrrolidone-co-styrene (PVP-co-S) with Polydopamine.
Polydopamine (PDA) is a synthetic eumelanin polymer mimicking the biopolymer secreted by mussels to attach to surfaces with a high binding strength. It exhibits unique adhesive properties and has recently attracted considerable interest as a multifunctional thin film coating. In this study, we demonstrate that a PDA coating on silica- and polymer-based materials improves the entrapment and retention of uremic toxins produced in specific diseases. The low-cost natural nanotextured fossil diatomaceous earth (DE), an abundant source of mesoporous silica, and polyvinylpyrrolidone-co-Styrene (PVP-co-S), a commercial absorbent comprising polymeric particles, were easily coated with a PDA layer by oxidative polymerization of dopamine at mild basic aqueous conditions. An in-depth chemical-physical investigation of both the resulting PDA-coated materials was performed by SEM, AFM, UV-visible, Raman spectroscopy and spectroscopic ellipsometry. Finally, the obtained hybrid systems were successfully tested for the removal of two uremic toxins (indoxyl sulfate and p-cresyl sulfate) directly from patients' sera.
Topics: Humans; Indican; Povidone; Diatomaceous Earth; Sulfates; Uremic Toxins; Polymers; Silicon Dioxide; Polyvinyl Chloride; Styrenes
PubMed: 36548761
DOI: 10.3390/toxins14120864 -
Toxicology of diatomaceous earth, phyto oils and their admixed emulsions against adults of (Herbst).Toxicology Reports 2022(Herbst), one of the invasive stored pest, is resistant to the most of synthetic insecticides used against this it. Moreover, synthetic insecticides are a major threat...
(Herbst), one of the invasive stored pest, is resistant to the most of synthetic insecticides used against this it. Moreover, synthetic insecticides are a major threat to human health, the environment, and the ecosystem. The development of some smart tools is needed to minimize the use of hazardous chemicals. As an alternative, nano-insecticides are on the horizon. Emulsions are expressed as sustain release of insecticidal components to achieve maximum efficacy and low residual toxicity. In this study, some essential oils ( (DC.) Stapf, L., L., and L.), diatomaceous earth (DE), and their nano-emulsions are evaluated against . Essential oils and DE were tested at four (60 ppm, 30 ppm, 15 ppm, 7.5 ppm) different concentrations with and without emulsions, and data was recorded after 6, 12, 24, 48, and 72 h of exposure respectively. The mortality observed in essential oils of , , and without emulsion after 72 h of treatment at the highest concentration was 98%, 95%, and 85%, respectively. While, at the highest concentration the lowest mortalities were observed with DE and i.e. 65%. Insecticidal activity of emulsion essential oils increased to 100%, 98%, 90%, and 68.3% for , , , and , respectively. The results support that these admixed emulsions could be used as an alternative to synthetic insecticides in conventional formulations.
PubMed: 36518385
DOI: 10.1016/j.toxrep.2022.05.011