bluuebunny/crossref_metadata_2025 huggingface dataset:
Alkaline Dehydration of Human Urine Collected in Source-Separated Sanitation Systems Using Magnesium Oxide
Authors: Simha Prithvi , Friedrich Christopher , Randall Dyllon Garth , Vinnerås Björn
Published: January 2021

Fresh human urine, after it is alkalized to prevent the enzymatic hydrolysis of urea, can be dehydrated to reduce its volume and to produce a solid fertilizer. In this study, we investigated the suitability of MgO to alkalize and dehydrate urine. We selected MgO due to its low solubility (<2 g·L−1) and relatively high saturation pH (9.9 ± 0.2) in urine. Using a laboratory-scale setup, we dehydrated urine added to pure MgO and MgO mixed with co-substrates (biochar, wheat bran, or calcium hydroxide) at a temperature of 50°C. We found that, dehydrating urine added to a mixture of MgO (25% w/w), biochar, and wheat bran resulted in a mass reduction of >90% and N recovery of 80%, and yielded products with high concentrations of macronutrients (7.8% N, 0.7% P and 3.9% K). By modeling the chemical speciation in urine, we also showed that ammonia stripping rather than urea hydrolysis limited the N recovery, since the urine used in our study was partially hydrolyzed. To maximize the recovery of N during alkaline urine dehydration using MgO, we recommend treating fresh/un-hydrolysed urine a temperature <40°C, tailoring the drying substrate to capture NH4+ as struvite, and using co-substrates to limit the molecular diffusion of ammonia. Treating fresh urine by alkaline dehydration requires only 3.6 kg MgO cap−1y−1 and a cost of US$ 1.1 cap−1y−1. Therefore, the use of sparingly soluble alkaline compounds like MgO in urine-diverting sanitation systems holds much promise.
Forest Soil Colloids Enhance Delivery of Phosphorus Into a Diffusive Gradient in Thin Films (DGT) Sink
Authors: Konrad Alexander , Billiy Benjamin , Regenbogen Philipp , Bol Roland , Lang Friederike , Klumpp Erwin , Siemens Jan
Published: January 2021

Phosphorus (P) is preferentially bound to colloids in soil. On the one hand, colloids may facilitate soil P leaching leading to a decrease of plant available P, but on the other hand they can carry P to plant roots, thus supporting the P uptake of plants. We tested the magnitude and the kinetics of P delivery by colloids into a P sink mimicking plant roots using the Diffusive Gradients in Thin-Films (DGT) technique. Colloids were extracted with water from three forest soils differing in parent material using a method based on dispersion and sedimentation. Freeze-dried colloids, the respective bulk soil, and the colloid-free extraction residue were sterilized and mixed with quartz sand and silt to an equal P basis. The mixtures were wetted and the diffusive fluxes of P into the DGTs were measured under sterile, water unsaturated conditions. The colloids extracted from a P-poor sandy podzolic soil were highly enriched in iron and organic matter compared to the bulk soil and delivered more P at a higher rate into the sink compared to bulk soil and the colloid-free soil extraction residue. However, colloidal P delivery into the sink was smaller than P release and transport from the bulk soil developed on dolomite rock, and with no difference for a soil with intermediate phosphorus-stocks developed from gneiss. Our results provide evidence that both the mobility of colloids and their P binding strength control their contribution to the plant available P-pool of soils. Overall, our findings highlight the relevance of colloids for P delivery to plant roots.
Efficacy and Safety of Eculizumab in the Treatment of Transplant-Associated Thrombotic Microangiopathy: A Systematic Review and Meta-Analysis
Authors: Zhang Rui , Zhou Meng , Qi Jiaqian , Miao Wenjing , Zhang Ziyan , Wu Depei , Han Yue
Published: January 2021

Background

Transplant-associated thrombotic microangiopathy (TA-TMA) is a dangerous and life-threatening complication in patients undergoing hematopoietic stem cell transplantation (HSCT). Eculizumab has been used in the treatment of TA-TMA, and several studies have confirmed the benefit of Eculizumab in patients with TA-TMA. However, the results remain controversial. We conducted a systematic review and meta-analysis to evaluate the efficacy and safety of Eculizumab for TA-TMA.

Materials and Methods

We searched PubMed and Embase for studies on the efficacy and safety of Eculizumab in TA-TMA patients. Efficacy outcomes consisted of overall response rate (ORR), complete response rate (CRR), and survival rate at the last follow-up (SR). Safety outcomes were adverse events (AEs), including infection, sepsis, impaired liver function, infusion reactions, and death.

Results

A total of 116 patients from six studies were subjected to meta-analysis. The pooled estimates of ORR, CRR, and SR for TA-TMA patients were 71% (95% CI: 58–82%), 32% (95% CI: 11–56%), and 52% (95% CI: 40–65%), respectively. Only one patient presented with a severe rash, and infection was the most common AEs. The main causes of death were infection and GvHD.

Conclusion

Current evidence suggests that Eculizumab improves SR and ORR in patients with TA-TMA and that Eculizumab is well tolerated. However, the number of studies is limited, and the findings are based mainly on data from observational studies. Higher quality randomized controlled trials and more extensive prospective cohort studies are needed.
Evaluation of Proteoforms of the Transmembrane Chemokines CXCL16 and CX3CL1, Their Receptors, and Their Processing Metalloproteinases ADAM10 and ADAM17 in Proliferative Diabetic Retinopathy
Authors: Abu El-Asrar Ahmed M. , Nawaz Mohd Imtiaz , Ahmad Ajmal , De Zutter Alexandra , Siddiquei Mohammad Mairaj , Blanter Marfa , Allegaert Eef , Gikandi Priscilla W. , De Hertogh Gert , Van Damme Jo , Opdenakker Ghislain , Struyf Sofie
Published: January 2021

The transmembrane chemokine pathways CXCL16/CXCR6 and CX3CL1/CX3CR1 are strongly implicated in inflammation and angiogenesis. We investigated the involvement of these chemokine pathways and their processing metalloproteinases ADAM10 and ADAM17 in the pathophysiology of proliferative diabetic retinopathy (PDR). Vitreous samples from 32 PDR and 24 non-diabetic patients, epiretinal membranes from 18 patients with PDR, rat retinas, human retinal Müller glial cells and human retinal microvascular endothelial cells (HRMECs) were studied by enzyme-linked immunosorbent assay, immunohistochemistry and Western blot analysis. In vitro angiogenesis assays were performed and the adherence of leukocytes to CXCL16-stimulated HRMECs was assessed. CXCL16, CX3CL1, ADAM10, ADAM17 and vascular endothelial growth factor (VEGF) levels were significantly increased in vitreous samples from PDR patients. The levels of CXCL16 were 417-fold higher than those of CX3CL1 in PDR vitreous samples. Significant positive correlations were found between the levels of VEGF and the levels of CXCL16, CX3CL1, ADAM10 and ADAM17. Significant positive correlations were detected between the numbers of blood vessels expressing CD31, reflecting the angiogenic activity of PDR epiretinal membranes, and the numbers of blood vessels and stromal cells expressing CXCL16, CXCR6, ADAM10 and ADAM17. CXCL16 induced upregulation of phospho-ERK1/2, p65 subunit of NF-κB and VEGF in cultured Müller cells and tumor necrosis factor-α induced upregulation of soluble CXCL16 and ADAM17 in Müller cells. Treatment of HRMECs with CXCL16 resulted in increased expression of intercellular adhesion molecule-1 (ICAM-1) and increased leukocyte adhesion to HRMECs. CXCL16 induced HRMEC proliferation, formation of sprouts from HRMEC spheroids and phosphorylation of ERK1/2. Intravitreal administration of CXCL16 in normal rats induced significant upregulation of the p65 subunit of NF-κB, VEGF and ICAM-1 in the retina. Our findings suggest that the chemokine axis CXCL16/CXCR6 and the processing metalloproteinases ADAM10 and ADAM17 might serve a role in the initiation and progression of PDR.
Integrated Analysis of circRNA-miRNA-mRNA Regulatory Networks in the Intestine of Sebastes schlegelii Following Edwardsiella tarda Challenge
Authors: Cao Min , Yan Xu , Su Baofeng , Yang Ning , Fu Qiang , Xue Ting , Song Lin , Li Qi , Li Chao
Published: January 2021

Sebastes schlegelii, an important aquaculture species, has been widely cultured in East Asian countries. With the increase in the cultivation scale, various diseases have become major threats to the industry. Evidence has shown that non-coding RNAs (ncRNAs) have remarkable functions in the interactions between pathogens and their hosts. However, little is known about the mechanisms of circular RNAs (circRNAs) and coding RNAs in the process of preventing pathogen infection in the intestine in teleosts. In this study, we aimed to uncover the global landscape of mRNAs, circRNAs, and microRNAs (miRNAs) in response to Edwardsiella tarda infection at different time points (0, 2, 6, 12, and 24 h) and to construct regulatory networks for exploring the immune regulatory mechanism in the intestine of S. schlegelii. In total, 1,794 mRNAs, 87 circRNAs, and 79 miRNAs were differentially expressed. The differentially expressed RNAs were quantitatively validated using qRT-PCR. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis showed that most of the differentially expressed mRNA genes and the target genes of ncRNAs were related to immune signaling pathways, such as the NF-κB signal pathway, pathogen recognition receptors related to signaling pathways (Toll-like receptors and Nod-like receptors), and the chemokine signaling pathway. Based on these differentially expressed genes, 624 circRNA-miRNA pairs and 2,694 miRNA-mRNA pairs were predicted using the miRanda software. Integrated analyses generated 25 circRNA-miRNA-mRNA interaction networks. In a novel_circ_0004195/novel-530/IκB interaction network, novel_530 was upregulated, while its two targets, novel_circ_0004195 and IκB, were downregulated after E. tarda infection. In addition, two circRNA-miRNA-mRNA networks related to apoptosis (novel_circ_0003210/novel_152/apoptosis-stimulating of p53 protein 1) and interleukin (novel_circ_0001907/novel_127/interleukin-1 receptor type 2) were also identified in our study. We thus speculated that the downstream NF-κB signaling pathway, p53 signaling pathway, and apoptosis pathway might play vital roles in the immune response in the intestine of S. schlegelii. This study revealed a landscape of RNAs in the intestine of S. schlegelii during E. tarda infection and provided clues for further study on the immune mechanisms and signaling networks based on the circRNA-miRNA-mRNA axis in S. schlegelii.
GBP1 Facilitates Indoleamine 2,3-Dioxygenase Extracellular Secretion to Promote the Malignant Progression of Lung Cancer
Authors: Meng Yinnan , Wang Wei , Chen Meng , Chen Kuifei , Xia Xinhang , Zhou Suna , Yang Haihua
Published: January 2021

IDO1-mediated immune escape can lead to the malignant progression of tumors. However, the precise mechanism of IDO1 remains unclear. This study showed that IDO1 can bind to GBP1 and increase the extracellular secretion of IDO1 with the assistance of GBP1, thereby promoting the malignant proliferation and metastasis of lung cancer. In vitro study showed that the high expression levels of IDO1 and GBP1 in lung cancer cells promoted cell invasion and migration. In vivo study revealed that knock-down of IDO1 and GBP1 inhibited tumor growth and metastasis. In addition, Astragaloside IV reduces the extracellular secretion of IDO1 by blocking the interaction of IDO1 and GBP1, thereby reducing T cell exhaustion and inhibiting tumor progression. These results suggest that blocking the extracellular secretion of IDO1 may prevent T cell exhaustion and thereby enhance the effect of PD-1 inhibitors on cancer treatment.
Spatio-Temporal Variations in Community Size Structure of Arctic Protist Plankton in the Fram Strait
Authors: Lampe Vanessa , Nöthig Eva-Maria , Schartau Markus
Published: January 2021

The Arctic Ocean is subject to severe environmental changes, including the massive decline in sea ice due to continuous warming in many regions. Along with these changes, the Arctic Ocean's ecosystem is affected on various scales. The pelagic microbial food web of the Arctic is of particular interest, because it determines mass transfer to higher trophic levels. In this regard, variations in the size structure of the microbial community reflect changes in size-dependent bottom-up and top-down processes. Here we present analyses of microscopic data that resolve details on composition and cell size of unicellular plankton, based on samples collected between 2016 and 2018 in the Fram Strait. Using the Kernel Density Estimation method, we derived continuous size spectra (from 1 μm to ≈ 200 μm Equivalent Spherical Diameter, ESD) of cell abundance and biovolume. Specific size intervals (3–4, 8–10, 25–40, and 70–100 μm ESD) indicate size-selective predation as well as omnivory. In-between size ranges include loopholes with elevated cell abundance. By considering remote sensing data we could discriminate between polar Arctic- and Atlantic water within the Fram Strait and could relate our size spectra to the seasonal change in chlorophyll-a concentration. Our size spectra disclose the decline in total biovolume from summer to autumn. In October the phytoplankton biovolume size-spectra reveal a clear relative shift toward larger cell sizes (> 30 μm). Our analysis highlights details in size spectra that may help refining allometric relationships and predator-prey dependencies for size-based plankton ecosystem model applications.
How Fishery Collapses: The Case of Lepidopus caudatus (Pisces: Trichiuridae) in the Strait of Sicily (Central Mediterranean)
Authors: Falsone Fabio , Scannella Danilo , Geraci Michele Luca , Gancitano Vita , Vitale Sergio , Fiorentino Fabio
Published: January 2021

The silver scabbardfish Lepidopus caudatus is a mesopelagic species living on the shelf and slope down to 600 m in temperate seas all around the world. In the Mediterranean, the species is caught mainly by longlines with a marked seasonality. In the early 90s in the Strait of Sicily (Central Mediterranean Sea), a new fishery targeting L. caudatus was developed. This fishery uses an ad hoc pelagic trawl gear called “spatolara.” Vessels using spatolara have increased from 1 in 1993 to 10 in 2007 with a growth of catches of up to 1,200 tons in 2011. Development of this fishery was not regulated by any specific management measures and, due to the progressive reduction of catch to 169 tons, only one vessel was active in 2018. The availability of catch and biomass indices from trawl survey since the beginning of trawling exploitation allowed providing the first assessment of the state of L. caudatus stock in the Central Mediterranean (GFCM Geographical Sub-Area 16) by using data-limited methods. Catch-Maximum Sustainable Yield (CMSY) and Bayesian State Space Schaefer model (BSM) were fitted to landings and abundance indices (2004–2018). The Abundance-Maximum Sustainable Yield model (AMSY) was also applied to survey data from 1994 (1 year after the start of the spatolara fishery) to 2018 to further corroborate the results. BSM prediction of biomass levels was just above 50% of BMSY, whereas AMSY estimated the current stock levels below 50% of BMSY. The BSM was used for forecasting B/BMSY and catches under different fishing scenarios. Although current exploitation was very close to FMSY, more than a decade would be needed to rebuild the stock to biomass levels producing MSY. A faster rebuilding could be achieved by fishing at least 80% of FMSY, with minimal loss in yield over the next 5–8 years. Following the development of a new fishery since the beginning, the study provides a further example of how unregulated exploitation leads to a heavy overfished state of stock and collapse of fishing activities.
Mechanism Analysis of the Influence of Delay Period on Mechanical Properties of Reactive Powder Concrete
Authors: Wang Xiaohui , Zhao Qingxin , He Xiaojun , Zhang Shuang
Published: January 2021

In order to analyze the influence mechanism of delay period on the mechanical properties of reactive powder concrete (RPC), the compressive strength of RPC with delay periods of 18, 24, and 30 h was tested at the age of 7, 28, and 90 days, respectively. The results show that compared with the RPC with delay period of 18 h, the compressive strength of the RPC with delay periods of 24 and 30 h increases by 3.2 and 4.2%, respectively, and the long-term strength reduction ratio decreases by 22.8 and 71.9%, respectively. The constitutive model curves of RPC under different delay period show that the initial elastic modulus E increases with the delay period and the strength and rigidity of RPC increase with the extension of delay period. According to the non-evaporation water quantity test, it could be speculated that the quantities of hydration products of the RPC with delay periods of 24 and 30 h slightly increase compared with the RPC with delay period of 18 h. X-ray diffraction (XRD) analysis show that the delay periods of 24 and 30 h consume more 3CaO·SiO2 (C3S) and 2CaO·SiO2 (C2S) compared with delay period of 18 h. Seen from the scanning electron microscope (SEM) image, the structures of the three groups of samples are relatively dense and have no significant difference. Through energy dispersive X-ray spectroscopy (EDS) analysis, the calcium-silicon ratios of hydration products of the RPC with delay periods of 18, 24, and 30 h are 1.81, 1.56, and 1.54, respectively. The existence of C-S-H gel and Ca(OH)2 in hydration products is confirmed by thermogravimetric-differential scanning calorimetry (DSC-TG) analysis. An appropriate delay period (30 h in this paper) generates more hydration products, then improves the compactness of the internal structure and reduces the calcium-silicon ratio of hydration products, and it is conducive to the growth of RPC compressive strength and the stability of long-term compressive strength.
Idiopathic Pulmonary Fibrosis in Elderly Patients: Analysis of the INSIGHTS-IPF Observational Study
Authors: Leuschner Gabriela , Klotsche Jens , Kreuter Michael , Prasse Antje , Wirtz Hubert , Pittrow David , Frankenberger Marion , Behr Jürgen , Kneidinger Nikolaus ,   The INSIGHTS-IPF Registry Group
Published: January 2021

Background: An association between idiopathic pulmonary fibrosis (IPF) and advancing age is suspected since IPF occurs primarily in patients over 60 years of age. Though, little is known about the disease in the elderly. The aim of this study was to characterize elderly IPF patients using data from the longitudinal, German-wide INSIGHTS-IPF registry.

Methods: Patients were grouped into elderly (≥75 years) and nonelderly IPF (<75 years) at the time of enrollment into the study. Baseline clinical characteristics, comorbidities, health related quality of life (HRQoL), medical therapy and survival were compared between age groups. Effects of antifibrotic therapy on forced vital capacity (FVC) were analyzed over 24 months.

Results: Of 1,009 patients, 350 (34.7%) were ≥75 years old. Elderly IPF patients compared to younger patients had a higher number of comorbidities (3.6 ± 2.5 vs. 2.8 ± 2.3; p < 0.001). The mean ± SD EQ-5D score (0.64 ± 0.21 vs. 0.69 ± 0.21; p = 0.005), and the overall WHO-5 score (13.1 ± 5.9 vs. 14.3 ± 6.0; p = 0.015) were significantly lower while the UCSD-SOBQ (52.6 ± 31.2 vs. 45.5 ± 31.2; p = 0.030) was significantly higher in elderly patients, indicating a more impaired HRQoL and more breathlessness. At baseline, 55.4% of elderly and 56.8% of nonelderly patients with IPF were treated with antifibrotic therapy (p = 0.687). For FVC decline after initiation of antifibrotic therapy, there was neither a significant difference between age groups at the different time points over 24 months (beta: 0.41; 95%-CI: −0.98 to 1.81; p = 0.563) nor over the whole course of time (beta: −0.05; 95%-CI: −0.20 to 0.09; p = 0.478). All-cause mortality was higher in elderly patients (49.1 vs. 37.9%; HR 1.65; 95%-CI 1.36–2.00; p < 0.001). Antifibrotic therapy was associated with improved survival in IPF patients, independent from age (<75 years: beta 0.76; 95%-CI: 0.59–0.99; p = 0.049; ≥75 years: beta 0.71; 95%-CI: 0.51–0.98; p = 0.043).

Conclusion: In real life, a significant proportion of IPF patients are ≥75 years old, characterized by higher number of comorbidities and global reduced HRQoL. However, the effect of an antifibrotic therapy was similar between age groups and associated with a survival benefit emphasizing the importance for an early antifibrotic therapy in IPF, independent from age.
Cerebral Blood Flow Deviations in Critically Ill Patients: Potential Insult Contributing to Ischemic and Hyperemic Injury
Authors: Slessarev Marat , Mahmoud Ossama , McIntyre Christopher W. , Ellis Christopher G.
Published: January 2021

Background: Ischemic and hyperemic injury have emerged as biologic mechanisms that contribute to cognitive impairment in critically ill patients. Spontaneous deviations in cerebral blood flow (CBF) beyond ischemic and hyperemic thresholds may represent an insult that contributes to this brain injury, especially if they accumulate over time and coincide with impaired autoregulation.

Methods: We used transcranial Doppler to measure the proportion of time that CBF velocity (CBFv) deviated beyond previously reported ischemic and hyperemic thresholds in a cohort of critically ill patients with respiratory failure and/or shock within 48 h of ICU admission. We also assessed whether these CBFv deviations were more common during periods of impaired dynamic autoregulation, and whether they are explained by concurrent variations in mean arterial pressure (MAP) and end-tidal PCO2 (PetCO2).

Results: We enrolled 12 consecutive patients (three females) who were monitored for a mean duration of 462.6 ± 39.8 min. Across patients, CBFv deviated by more than 20–30% from its baseline for 17–24% of the analysis time. These CBFv deviations occurred equally during periods of preserved and impaired autoregulation, while concurrent variations in MAP and PetCO2 explained only 13–21% of these CBFv deviations.

Conclusion: CBFv deviations beyond ischemic and hyperemic thresholds are common in critically ill patients with respiratory failure or shock. These deviations occur irrespective of the state of dynamic autoregulation and are not explained by changes in MAP and CO2. Future studies should explore mechanisms responsible for these CBFv deviations and establish whether their cumulative burden predicts poor neurocognitive outcomes.
Clinical Characteristics of Itch in Renal Transplant Recipients
Authors: Krajewski Piotr K. , Olczyk Piotr , Krajewska Magdalena , Krajewski Wojciech , Szepietowski Jacek C.
Published: January 2021

Background:Chronic itch is the most common symptom in dermatology. End-stage renal disease-associated chronic itch (ESRDCI) is a common burden affecting up to 35% of patients treated with hemodialysis. Kidney transplant (KTx) is believed to be the best renal replacement therapy leading to the elimination of ESRDCI. The study was undertaken to characterize and assess the prevalence of itch among patients after renal transplantation.

Methods:Between October 2019 and January 2020, we analyzed the data of 197 patients comprising 121 males (61.4%) and 76 females (38.6%) and aged 54.5 ± 13.6 years. The data collection was performed with a specially designed questionnaire. Level of itch after renal transplantation was assessed with the use of a Numeral Rating Scale, a Visual Rating Scale, and 4-Item Itch Questionnaire. Moreover, the previous 3 days of itching were evaluated.

Results:The patients suffered from chronic renal disease for 20.2 ± 12.3 years, with a mean time of pre-transplant dialysis of 2.6 ± 2.4 years and a mean time after the KTx of 8.0 ± 6.5 years. The itch was present in 38.6% of the patients during the hemodialysis, and in 73.7% of cases, the itch ceased completely after the successful transplantation. Moreover, only 2.63% of the cases had no improvement. Nevertheless, the itch was reported in 42 (21.3%) renal transplant recipients (RTR), and in 22 (52.4%) cases, the itch appeared after transplantation. The majority of patients suffering from itch were women (54.8%). Itch in the last 3 days was reported in 21 patients. The itch's severity was assessed with a numerical rating scale (NRS), with the worst severity measured at 6 ± 2.2 points indicating moderate itch. In most cases (57.1%), itch affected multiple body areas. Extremities (50%) and the back (50%) were among the most frequently affected areas. The sensation had a mostly short duration and was most frequently reported in the evening. Only eight patients suffered for the whole day. Hot water was the most frequently reported (16.7%) alleviating factor, whilst sweat was responsible for itch aggravation in 35.9% of cases.

Conclusion:Our analysis on representative patients' population indicates that itch after KTx is an important problem. Moreover, it is worth noting that more than half of the RTR did not suffer from itch during dialysis.
Electrically Evoked Itch in Human Subjects
Authors: Solinski Hans Jürgen , Rukwied Roman
Published: January 2021

Administration of chemicals (pruritogens) into the skin evokes itch based on signal transduction mechanisms that generate action potentials mainly in mechanically sensitive and insensitive primary afferent C-fibers (pruriceptors). These signals from peripheral neurons are processed in spinal and supra-spinal centers of the central nervous system and finally generate the sensation of itch. Compared to chemical stimulation, electrical activation of pruriceptors would allow for better temporal control and thereby a more direct functional assessment of their activation. Here, we review the electrical stimulation paradigms which were used to evoke itch in humans in the past. We further evaluate recent attempts to explore electrically induced itch in atopic dermatitis patients. Possible mechanisms underlying successful pruritus generation in chronic itch patients by transdermal slowly depolarizing electrical stimulation are discussed.
“The Good, the Bad and the Double-Sword” Effects of Microplastics and Their Organic Additives in Marine Bacteria
Authors: Fernández-Juárez Víctor , López-Alforja Xabier , Frank-Comas Aida , Echeveste Pedro , Bennasar-Figueras Antoni , Ramis-Munar Guillem , Gomila Rosa María , Agawin Nona S. R.
Published: January 2021

Little is known about the direct effects of microplastics (MPs) and their organic additives on marine bacteria, considering their role in the nutrient cycles, e.g., N-cycles through the N2-fixation, or in the microbial food web. To fill this gap of knowledge, we exposed marine bacteria, specifically diazotrophs, to pure MPs which differ in physical properties (e.g., density, hydrophobicity, and/or size), namely, polyethylene, polypropylene, polyvinyl chloride and polystyrene, and to their most abundant associated organic additives (e.g., fluoranthene, 1,2,5,6,9,10-hexabromocyclododecane and dioctyl-phthalate). Growth, protein overproduction, direct physical interactions between MPs and bacteria, phosphorus acquisition mechanisms and/or N2-fixation rates were evaluated. Cyanobacteria were positively affected by environmental and high concentrations of MPs, as opposed to heterotrophic strains, that were only positively affected with high concentrations of ~120 μm-size MPs (detecting the overproduction of proteins related to plastic degradation and C-transport), and negatively affected by 1 μm-size PS beads. Generally, the organic additives had a deleterious effect in both autotrophic and heterotrophic bacteria and the magnitude of the effect is suggested to be dependent on bacterial size. Our results show species-specific responses of the autotrophic and heterotrophic bacteria tested and the responses (beneficial: the “good,” deleterious: the “bad” and/or both: the “double-sword”) were dependent on the type and concentration of MPs and additives. This suggests the need to determine the threshold levels of MPs and additives concentrations starting from which significant effects can be observed for key microbial populations in marine systems, and these data are necessary for effective environmental quality control management.
Persistence of a Stx-Encoding Bacteriophage in Minced Meat Investigated by Application of an Improved DNA Extraction Method and Digital Droplet PCR
Authors: Spilsberg B. , Sekse C. , Urdahl Anne M. , Nesse Live L. , Johannessen Gro S.
Published: January 2021

Shiga toxin-producing Escherichia coli (STEC) are important food-borne pathogens with Shiga toxins as the main virulence factor. Shiga toxins are encoded on Shiga toxin-encoding bacteriophages (Stx phages). Stx phages may exist as free bacteriophages in the environment or in foods or as prophages integrated into the host genome. From a food safety perspective, it is important to have knowledge on the survival and persistence of Stx phages in food products since these may integrate into the bacterial hosts through transduction if conditions are right. Here, we present the results from a study investigating the survival of a Stx phage in minced meat from beef stored at a suboptimal temperature (8°C) for food storage along with modifications and optimizations of the methods applied. Minced meat from beef was inoculated with known levels of a labeled Stx phage prior to storage. Phage filtrates were used for plaque assays and DNA extraction, followed by real-time PCR and digital droplet PCR (ddPCR). The results from the pilot study suggested that the initial DNA extraction protocol was not optimal, and several modifications were tested before a final protocol was defined. The final DNA extraction protocol comprised ultra-centrifugation of the entire phage filtrate for concentrating phages and two times phenol–chloroform extraction. The protocol was used for two spiking experiments. The DNA extraction protocol resulted in flexibility in the amount of DNA available for use in PCR analyses, ultimately increasing the sensitivity of the method used for quantification of phages in a sample. All three quantification methods employed (i.e., plaque assays, real-time PCR, and ddPCR) showed similar trends in the development of the phages during storage, where ddPCR has the benefit of giving absolute quantification of DNA copies in a simple experimental setup. The results indicate that the Stx phages persist and remain infective for at least 20 days under the storage conditions used in the present study. Stx phages in foods might represent a potential risk for humans. Although it can be speculated that transduction may take place at 8°C with subsequent forming of STEC, it can be expected to be a rare event. However, such an event may possibly take place under more optimal conditions, such as an increase in storage temperature of foods or in the gastrointestinal tract of humans.
Pursuing Advances in DNA Sequencing Technology to Solve a Complex Genomic Jigsaw Puzzle: The Agglutinin-Like Sequence (ALS) Genes of Candida tropicalis
Authors: Oh Soon-Hwan , Isenhower Allyson , Rodriguez-Bobadilla Rubi , Smith Brooke , Jones Jillian , Hubka Vit , Fields Christopher , Hernandez Alvaro , Hoyer Lois L.
Published: January 2021

The agglutinin-like sequence (ALS) gene family encodes cell-surface adhesins that interact with host and abiotic surfaces, promoting colonization by opportunistic fungal pathogens such asCandida tropicalis. Studies of Als protein contribution toC. tropicalisadhesion would benefit from an accurate catalog ofALSgene sequences as well as insight into relative gene expression levels. Even in the genomics era, this information has been elusive: genome assemblies are often broken withinALSgenes because of their extensive regions of highly conserved, repeated DNA sequences and because there are many similarALSgenes at different chromosomal locations. Here, we describe the benefit of long-read DNA sequencing technology to facilitate characterization ofC. tropicalis ALSloci. ThirteenALSloci inC. tropicalisstrain MYA-3404 were deduced from a genome assembly constructed from Illumina MiSeq and Oxford Nanopore MinION data. Although the MinION data were valuable, PCR amplification and Sanger sequencing ofALSloci were still required to complete and verify the gene sequences. Each predicted Als protein featured an N-terminal binding domain, a central domain of tandemly repeated sequences, and a C-terminal domain rich in Ser and Thr. The presence of a secretory signal peptide and consensus sequence for addition of a glycosylphosphatidylinositol (GPI) anchor was consistent with predicted protein localization to the cell surface. TaqMan assays were designed to recognize eachALSgene, as well as both alleles at the divergentCtrALS3882locus.C. tropicaliscells grown in five differentin vitroconditions showed differential expression of variousALSgenes. To place theC. tropicalisdata into a larger context, TaqMan assays were also designed and validated for analysis ofALSgene expression inCandida albicansandCandida dubliniensis. These comparisons identified the subset of highly expressedC. tropicalis ALSgenes that were predicted to encode proteins with the most abundant cell-surface presence, prioritizing them for subsequent functional analysis. Data presented here provide a solid foundation for future experimentation to deduceALSfamily contributions toC. tropicalisadhesion and pathogenesis.
Tracking Major Sources of Water Contamination Using Machine Learning
Authors: Wu Jianyong , Song Conghe , Dubinsky Eric A. , Stewart Jill R.
Published: January 2021

Current microbial source tracking techniques that rely on grab samples analyzed by individual endpoint assays are inadequate to explain microbial sources across space and time. Modeling and predicting host sources of microbial contamination could add a useful tool for watershed management. In this study, we tested and evaluated machine learning models to predict the major sources of microbial contamination in a watershed. We examined the relationship between microbial sources, land cover, weather, and hydrologic variables in a watershed in Northern California, United States. Six models, including K-nearest neighbors (KNN), Naïve Bayes, Support vector machine (SVM), simple neural network (NN), Random Forest, and XGBoost, were built to predict major microbial sources using land cover, weather and hydrologic variables. The results showed that these models successfully predicted microbial sources classified into two categories (human and non-human), with the average accuracy ranging from 69% (Naïve Bayes) to 88% (XGBoost). The area under curve (AUC) of the receiver operating characteristic (ROC) illustrated XGBoost had the best performance (average AUC = 0.88), followed by Random Forest (average AUC = 0.84), and KNN (average AUC = 0.74). The importance index obtained from Random Forest indicated that precipitation and temperature were the two most important factors to predict the dominant microbial source. These results suggest that machine learning models, particularly XGBoost, can predict the dominant sources of microbial contamination based on the relationship of microbial contaminants with daily weather and land cover, providing a powerful tool to understand microbial sources in water.
Antimicrobial Resistance in Bacteria Isolated From Cats and Dogs From the Iberian Peninsula
Authors: Li Yanli , Fernández Rubén , Durán Inma , Molina-López Rafael A. , Darwich Laila
Published: January 2021

Pet animals are assumed to be potential reservoirs in transferring antimicrobial resistance (AMR) to humans due to the extensively applied broad-spectrum antimicrobial agents and their close contact with humans. In this study, microbiological data and antimicrobial susceptibility results of dog (n= 5,086) and cat (n= 789) clinical samples from a private Laboratory of Diagnosis in Barcelona were analyzed. Samples came from different counties of the Iberian Peninsula during 2016–2018. In dogs, clinical samples were most commonly from otitis, and in cats from wounds, respiratory tract infections and conjunctivitis. In both pet groups,Staphylococcusspp. (31% in dogs vs 30% in cats),Streptococcusspp. (19% vs 17%),Pseudomonasspp. (16% vs 10%),Escherichia coli(8% vs 5.6%), andEnterococcusspp. (5.5% vs 6.8%) were shown as the most predominant bacteria. However, higher frequencies ofP. aeruginosa,P. canis, andS. pseudintermediuswere found in dogs, whileS. aureusandP. multocidawere more prevalent in cats. The antimicrobial susceptibility testing demonstrated thatEnterococcusspp. andPseudomonasspp. presented the highest levels of AMR in both dogs and cats. Within the Enterobacteriaceae,E. colishowed low levels of AMR compared toKlebsiella,Proteus, orEnterobacterspp. Respiratory tract infections caused byK. pneumoniaepresented higher AMR in cats. By contrast,Pasteurellaisolates from the respiratory tract were highly sensitive to all the antimicrobials in cats and dogs. Data from this study could be used to guide empirical antimicrobial selection in companion animal veterinary practices in the Iberian Peninsula.
Interacting Abiotic Factors Affect Growth and Aflatoxin B1 Production Profiles of Aspergillus flavus Strains on Pistachio-Based Matrices and Pistachio Nuts
Authors: Baazeem Alaa , Garcia-Cela Esther , Medina Angel , Magan Naresh
Published: January 2021

Pistachio nuts are an economically important commodity produced by many countries. They can be colonized by mycotoxigenic fungi, especiallyAspergillus flavus, resulting in contamination withaflatoxins (AFs), especially aflatoxin B1(AFB1), a Class 1a carcinogen. The objectives were to examine the effect of interactions between the two key abiotic factors, temperature and water activity (aw) on (a)in vitrogrowth and AFB1production by four strains ofA. flavusisolated from pistachio nuts, on a milled pistachio nut medium modified ionically (NaCl) and non-ionically (glycerol) in the range 20–35°C and 0.995–0.85 aw, (b) colonization of layers of raw pistachio nuts stored at different interacting temperature x awconditions and on relative AFB1production and (c) develop models to produce contour maps of the optimal and marginal boundary conditions for growth and AFB1production by up to 4 strains of this species. On pistachio nut-based media, optimum growth of four strains ofA. flavuswas at 0.98–0.95 awand 30–35°C. Optimum AFB1production was at 30–35°C and 0.98 aw. No significant differences in growth was found on ionic and non-ionically modified media. Colonization of layers of raw pistachio nuts was slower and contamination with AFB1significantly less than inin vitrostudies. Contour maps based on the pooled data for up to four strains (in vitro,in situ) showed the optimum and marginal conditions for growth and AFB1production. These data can be used to identify those conditions which represent a high, intermediate or low risk of colonization and AFB1contamination in the pistachio nut processing chain. These results are discussed in the context of the development of appropriate intervention strategies to minimize AFB1contamination of this economically important commodity.
Drug-Resistant Temporal Lobe Epilepsy Alters the Expression and Functional Coupling to Gαi/o Proteins of CB1 and CB2 Receptors in the Microvasculature of the Human Brain
Authors: Nuñez-Lumbreras María de los Ángeles , Castañeda-Cabral José Luis , Valle-Dorado María Guadalupe , Sánchez-Valle Vicente , Orozco-Suárez Sandra , Guevara-Guzmán Rosalinda , Martínez-Juárez Iris , Alonso-Vanegas Mario , Walter Fruzsina , Deli Maria A. , Carmona-Cruz Francia , Rocha Luisa
Published: January 2021

Cannabinoid receptors 1 and 2 (CB1 and CB2, respectively) play an important role in maintaining the integrity of the blood–brain barrier (BBB). On the other hand, BBB dysfunction is a common feature in drug-resistant epilepsy. The focus of the present study was to characterize protein expression levels and Gαi/o protein-induced activation by CB1 and CB2 receptors in the microvascular endothelial cells (MECs) isolated from the brain of patients with drug-resistant mesial temporal lobe epilepsy (DR-MTLE). MECs were isolated from the hippocampus and temporal neocortex of 12 patients with DR-MTLE and 12 non-epileptic autopsies. Immunofluorescence experiments were carried out to determine the localization of CB1 and CB2 receptors in the different cell elements of MECs. Protein expression levels of CB1 and CB2 receptors were determined by Western blot experiments. [35S]-GTPγS binding assay was used to evaluate the Gαi/o protein activation induced by specific agonists. Immunofluorescent double-labeling showed that CB1 and CB2 receptors colocalize with tight junction proteins (claudin-5, occludin, and zonula occludens-1), glial fibrillary acidic protein and platelet-derived growth factor receptor-β. These results support that CB1 and CB2 receptors are expressed in the human isolated microvessels fragments consisting of MECs, astrocyte end feet, and pericytes. The hippocampal microvasculature of patients with DR-MTLE presented lower protein expression of CB1 and CB2 receptors (66 and 43%, respectively; p < 0.001). However, its Gαi/o protein activation was with high efficiency (CB1, 251%, p < 0.0008; CB2, 255%, p < 0.0001). Microvasculature of temporal neocortex presented protein overexpression of CB1 and CB2 receptors (35 and 41%, respectively; p < 0.01). Their coupled Gαi/o protein activation was with higher efficiency for CB1 receptors (103%, p < 0.006), but lower potency (p < 0.004) for CB2 receptors. The present study revealed opposite changes in the protein expression of CB1 and CB2 receptors when hippocampus (diminished expression of CB1 and CB2) and temporal neocortex (increased expression of CB1 and CB2) were compared. However, the exposure to specific CB1 and CB2 agonists results in high efficiency for activation of coupled Gαi/o proteins in the brain microvasculature of patients with DR-MTLE. CB1 and CB2 receptors with high efficiency could represent a therapeutic target to maintain the integrity of the BBB in patients with DR-MTLE.
A Hypomorphic Dars1D367Y Model Recapitulates Key Aspects of the Leukodystrophy HBSL
Authors: Fröhlich Dominik , Mendes Marisa I. , Kueh Andrew J. , Bongers Andre , Herold Marco J. , Salomons Gajja S. , Housley Gary D. , Klugmann Matthias
Published: January 2021

Hypomyelination with brain stem and spinal cord involvement and leg spasticity (HBSL) is a leukodystrophy caused by missense mutations of the aspartyl-tRNA synthetase-encoding gene DARS1. The clinical picture includes the regression of acquired motor milestones, spasticity, ataxia, seizures, nystagmus, and intellectual disabilities. Morphologically, HBSL is characterized by a distinct pattern of hypomyelination in the central nervous system including the anterior brainstem, the cerebellar peduncles and the supratentorial white matter as well as the dorsal columns and the lateral corticospinal tracts of the spinal cord. Adequate HBSL animal models are lacking. Dars1 knockout mice are embryonic lethal precluding examination of the etiology. To address this, we introduced the HBSL-causing Dars1D367Y point mutation into the mouse genome. Surprisingly, mice carrying this mutation homozygously were phenotypically normal. As hypomorphic mutations are more severe in trans to a deletion, we crossed Dars1D367Y/D367Y mice with Dars1-null carriers. The resulting Dars1D367Y/− offspring displayed a strong developmental delay compared to control Dars1D367Y/+ littermates, starting during embryogenesis. Only a small fraction of Dars1D367Y/− mice were born, and half of these mice died with hydrocephalus during the first 3 weeks of life. Of the few Dars1D367Y/− mice that were born at term, 25% displayed microphthalmia. Throughout postnatal life, Dars1D367Y/− mice remained smaller and lighter than their Dars1D367Y/+ littermates. Despite this early developmental deficit, once they made it through early adolescence Dars1D367Y/− mice were phenotypically inconspicuous for most of their adult life, until they developed late onset motor deficits as well as vacuolization and demyelination of the spinal cord white matter. Expression levels of the major myelin proteins were reduced in Dars1D367Y/− mice compared to controls. Taken together, Dars1D367Y/− mice model aspects of the clinical picture of the corresponding missense mutation in HBSL. This model will enable studies of late onset deficits, which is precluded in Dars1 knockout mice, and can be leveraged to test potential HBSL therapeutics including DARS1 gene replacement therapy.
Clinical Phenotyping and Biomarkers in Spinal and Bulbar Muscular Atrophy
Authors: Millere Elina , Rots Dmitrijs , Glazere Ieva , Taurina Gita , Kurjane Natalja , Priedite Viktorija , Gailite Linda , Blennow Kaj , Zetterberg Henrik , Kenina Viktorija
Published: January 2021

Background: Spinal and bulbar muscular atrophy (SBMA) or Kennedy disease [OMIM: 313200] is a rare X-linked neuromuscular disease. Patients commonly present with muscle cramps, tremors, leg weakness, dysarthria and dysphagia.

Methods: We deeply phenotyped and evaluated the possible extent of affected systems in all patients with SBMA in Latvia (n = 5). In addition, neurophysiological studies and blood analyses were used to perform a molecular diagnosis and evaluate biochemical values. We analyzed neurofilament light (NfL) as a possible biomarker.

Results: Neurological examination revealed typical SBMA clinical manifestations; all patients had small or large nerve fiber neuropathy. Three of five patients had increased neurofilament light levels.

Conclusion: The study confirms the systemic involvement in patients suffering from SBMA. Increased NfL concentration was associated with either peripheral neuropathy or decreased body mass index. The complex phenotype of the disease should be kept in mind, as it could help to diagnose patients with SBMA.
The Biosafety and Risk Management in Preparation and Processing of Cerebrospinal Fluid and Other Neurological Specimens With Potential Coronavirus Infection
Authors: Chen Chien-Chin , Chiang Pei-Chun , Chen Tsung-Hsien
Published: January 2021

The recent outbreak of coronavirus disease 2019 (COVID-19), caused by Severe Acute Respiratory Syndrome Coronavirus 2, has become a global threat. Due to neurological manifestations presented throughout the coronavirus disease process, the potential involvement of COVID-19 in central nervous system has attracted considerable attention. Notably, the neurologic system could be widely affected, with various complications such as acute cerebrovascular events, encephalitis, Guillain-Barré syndrome, and acute necrotizing hemorrhagic encephalopathy. However, the risk assessment of exposure to potential biohazards in the context of the COVID-19 pandemic has not been clearly clarified regarding the sampling, preparation, and processing neurological specimens. Further risk managements and implantations are seldom discussed either. This article aims to provide current recommendations and evidence-based reviews on biosafety issues of preparation and processing of cerebrospinal fluid and neurological specimens with potential coronavirus infection from the bedside to the laboratory.