bluuebunny/crossref_metadata_2025 huggingface dataset:
Full-Length SSU rRNA Gene Sequencing Allows Species-Level Detection of Bacteria, Archaea, and Yeasts Present in Milk
Authors: Abellan-Schneyder Isabel , Siebert Annemarie , Hofmann Katharina , Wenning Mareike , Neuhaus Klaus
Published: June 2021

Full-length SSU rRNA gene sequencing allows species-level identification of the microorganisms present in milk samples. Here, we used bulk-tank raw milk samples of two German dairies and detected, using this method, a great diversity of bacteria, archaea, and yeasts within the samples. Moreover, the species-level classification was improved in comparison to short amplicon sequencing. Therefore, we anticipate that this approach might be useful for the detection of possible mastitis-causing species, as well as for the control of spoilage-associated microorganisms. In a proof of concept, we showed that we were able to identify several putative mastitis-causing or mastitis-associated species such as Streptococcusuberis, Streptococcusagalactiae, Streptococcusdysgalactiae, Escherichiacoli and Staphylococcusaureus, as well as several Candida species. Overall, the presented full-length approach for the sequencing of SSU rRNA is easy to conduct, able to be standardized, and allows the screening of microorganisms in labs with Illumina sequencing machines.
Morphological Features and Cold-Response Gene Expression in Mesophilic Bacillus cereus Group and Psychrotolerant Bacillus cereus Group under Low Temperature
Authors: Park Kyung-Min , Kim Hyun-Jung , Kim Min-Sun , Koo Minseon
Published: June 2021

At low temperatures, psychrotolerant B. cereus group strains exhibit a higher growth rate than mesophilic strains do. However, the different survival responses of the psychrotolerant strain (BCG34) and the mesophilic strain (BCGT) at low temperatures are unclear. We investigated the morphological and genomic features of BCGT and BCG34 to characterize their growth strategies at low temperatures. At low temperatures, morphological changes were observed only in BCGT. These morphological changes included the elongation of rod-shaped cells, whereas the cell shape in BCG34 was unchanged at the low temperature. A transcriptomic analysis revealed that both species exhibited different growth-related traits during low-temperature growth. The BCGT strain induces fatty acid biosynthesis, sulfur assimilation, and methionine and cysteine biosynthesis as a survival mechanism in cold systems. Increases in energy metabolism and fatty acid biosynthesis in the mesophilic B. cereus group strain might explain its ability to grow at low temperatures. Several pathways involved in carbohydrate mechanisms were downregulated to conserve the energy required for growth. Peptidoglycan biosynthesis was upregulated, implying that a change of gene expression in both RNA-Seq and RT-qPCR contributed to sustaining its growth and rod shape at low temperatures. These results improve our understanding of the growth response of the B. cereus group, including psychrotolerant B. cereus group strains, at low temperatures and provide information for improving bacterial inhibition strategies in the food industry.
Nanoscale Material Heterogeneity of Glowworm Capture Threads Revealed by AFM
Authors: Piorkowski Dakota , He Bo-Ching , Blamires Sean J. , Tso I-Min , Kane Deborah M.
Published: June 2021

Adhesive materials used by many arthropods for biological functions incorporate sticky substances and a supporting material that operate synergistically by exploiting substrate attachment and energy dissipation. While there has been much focus on the composition and properties of the sticky glues of these bio-composites, less attention has been given to the materials that support them. In particular, as these materials are primarily responsible for dissipation during adhesive pull-off, little is known of the structures that give rise to functionality, especially at the nano-scale. In this study we used tapping mode atomic force microscopy (TM-AFM) to analyze unstretched and stretched glowworm (Arachnocampa tasmaniensis) capture threads and revealed nano-scale features corresponding to variation in surface structure and elastic modulus near the surface of the silk. Phase images demonstrated a high resolution of viscoelastic variation and revealed mostly globular and elongated features in the material. Increased vertical orientation of 11–15 nm wide fibrillar features was observed in stretched threads. Fast Fourier transform analysis of phase images confirmed these results. Relative viscoelastic properties were also highly variable at inter- and intra-individual levels. Results of this study demonstrate the practical usefulness of TM-AFM, especially phase angle imaging, in investigating the nano-scale structures that give rise to macro-scale function of soft and highly heterogeneous materials of both natural and synthetic origins.
Zinc Biofortification in Food Crops Could Alleviate the Zinc Malnutrition in Human Health
Authors: Praharaj Subhashisa , Skalicky Milan , Maitra Sagar , Bhadra Preetha , Shankar Tanmoy , Brestic Marian , Hejnak Vaclav , Vachova Pavla , Hossain Akbar
Published: June 2021

Micronutrient malnutrition is a global health issue and needs immediate attention. Over two billion people across the globe suffer from micronutrient malnutrition. The widespread zinc (Zn) deficiency in soils, poor zinc intake by humans in their diet, low bioavailability, and health consequences has led the research community to think of an economic as well as sustainable strategy for the alleviation of zinc deficiency. Strategies like fortification and diet supplements, though effective, are not economical and most people in low-income countries cannot afford them, and they are the most vulnerable to Zn deficiency. In this regard, the biofortification of staple food crops with Zn has been considered a useful strategy. An agronomic biofortification approach that uses crop fertilization with Zn-based fertilizers at the appropriate time to ensure grain Zn enrichment has been found to be cost-effective, easy to practice, and efficient. Genetic biofortification, though time-consuming, is also highly effective. Moreover, a Zn-rich genotype once developed can also be used for many years without any recurring cost. Hence, both agronomic and genetic biofortification can be a very useful tool in alleviating Zn deficiency.
Intrinsic and Chemotherapeutic Stressors Modulate ABCC-Like Transport in Trypanosoma cruzi
Authors: da Costa Kelli Monteiro , Salustiano Eduardo J. , Valente Raphael do Carmo , Freire-de-Lima Leonardo , Mendonça-Previato Lucia , Previato José Osvaldo
Published: June 2021

Trypanosoma cruzi is the etiologic agent for Chagas disease, which affects 6–7 million people worldwide. The biological diversity of the parasite reflects on inefficiency of benznidazole, which is a first choice chemotherapy, on chronic patients. ABC transporters that extrude xenobiotics, metabolites, and mediators are overexpressed in resistant cells and contribute to chemotherapy failure. An ABCC-like transport was identified in the Y strain and extrudes thiol-conjugated compounds. As thiols represent a line of defense towards reactive species, we aimed to verify whether ABCC-like transport could participate in the regulation of responses to stressor stimuli. In order to achieve this, ABCC-like activity was measured by flow cytometry using fluorescent substrates. The present study reveals the participation of glutathione and ceramides on ABCC-like transport, which are both implicated in stress. Hemin modulated the ABCC-like efflux which suggests that this protein might be involved in cellular detoxification. Additionally, all strains evaluated exhibited ABCC-like activity, while no ABCB1-like activity was detected. Results suggest that ABCC-like efflux is not associated with natural resistance to benznidazole, since sensitive strains showed higher activity than the resistant ones. Although benznidazole is not a direct substrate, ABCC-like efflux increased after prolonged drug exposure and this indicates that the ABCC-like efflux mediated protection against cell stress depends on the glutathione biosynthesis pathway.
Advances in Enzyme and Ionic Liquid Immobilization for Enhanced in MOFs for Biodiesel Production
Authors: Shomal Reem , Ogubadejo Babatunde , Shittu Toyin , Mahmoud Eyas , Du Wei , Al-Zuhair Sulaiman
Published: June 2021

Biodiesel is a promising candidate for sustainable and renewable energy and extensive research is being conducted worldwide to optimize its production process. The employed catalyst is an important parameter in biodiesel production. Metal–organic frameworks (MOFs), which are a set of highly porous materials comprising coordinated bonds between metals and organic ligands, have recently been proposed as catalysts. MOFs exhibit high tunability, possess high crystallinity and surface area, and their order can vary from the atomic to the microscale level. However, their catalytic sites are confined inside their porous structure, limiting their accessibility for biodiesel production. Modification of MOF structure by immobilizing enzymes or ionic liquids (ILs) could be a solution to this challenge and can lead to better performance and provide catalytic systems with higher activities. This review compiles the recent advances in catalytic transesterification for biodiesel production using enzymes or ILs. The available literature clearly indicates that MOFs are the most suitable immobilization supports, leading to higher biodiesel production without affecting the catalytic activity while increasing the catalyst stability and reusability in several cycles.
Functional Domains and Evolutionary History of the PMEL and GPNMB Family Proteins
Authors: Chrystal Paul W. , Footz Tim , Hodges Elizabeth D. , Jensen Justin A. , Walter Michael A. , Allison W. Ted
Published: June 2021

The ancient paralogs premelanosome protein (PMEL) and glycoprotein nonmetastatic melanoma protein B (GPNMB) have independently emerged as intriguing disease loci in recent years. Both proteins possess common functional domains and variants that cause a shared spectrum of overlapping phenotypes and disease associations: melanin-based pigmentation, cancer, neurodegenerative disease and glaucoma. Surprisingly, these proteins have yet to be shown to physically or genetically interact within the same cellular pathway. This juxtaposition inspired us to compare and contrast this family across a breadth of species to better understand the divergent evolutionary trajectories of two related, but distinct, genes. In this study, we investigated the evolutionary history of PMEL and GPNMB in clade-representative species and identified TMEM130 as the most ancient paralog of the family. By curating the functional domains in each paralog, we identified many commonalities dating back to the emergence of the gene family in basal metazoans. PMEL and GPNMB have gained functional domains since their divergence from TMEM130, including the core amyloid fragment (CAF) that is critical for the amyloid potential of PMEL. Additionally, the PMEL gene has acquired the enigmatic repeat domain (RPT), composed of a variable number of imperfect tandem repeats; this domain acts in an accessory role to control amyloid formation. Our analyses revealed the vast variability in sequence, length and repeat number in homologous RPT domains between craniates, even within the same taxonomic class. We hope that these analyses inspire further investigation into a gene family that is remarkable from the evolutionary, pathological and cell biology perspectives.
Quantum Confined Stark Effect on the Linear and Nonlinear Optical Properties of SiGe/Si Semi Oblate and Prolate Quantum Dots Grown in Si Wetting Layer
Authors: Varsha  , Kria Mohamed , Hamdaoui Jawad El , Pérez Laura M. , Prasad Vinod , El-Yadri Mohamed , Laroze David , Feddi El Mustapha
Published: June 2021

We have studied the parallel and perpendicular electric field effects on the system of SiGe prolate and oblate quantum dots numerically, taking into account the wetting layer and quantum dot size effects. Using the effective-mass approximation in the two bands model, we computationally calculated the extensive variation of dipole matrix (DM) elements, bandgap and non-linear optical properties, including absorption coefficients, refractive index changes, second harmonic generation and third harmonic generation as a function of the electric field, wetting layer size and the size of the quantum dot. The redshift is observed for the non-linear optical properties with the increasing electric field and an increase in wetting layer thickness. The sensitivity to the electric field toward the shape of the quantum dot is also observed. This study is resourceful for all the researchers as it provides a pragmatic model by considering oblate and prolate shaped quantum dots by explaining the optical and electronic properties precisely, as a consequence of the confined stark shift and wetting layer.
FEBID 3D-Nanoprinting at Low Substrate Temperatures: Pushing the Speed While Keeping the Quality
Authors: Hinum-Wagner Jakob , Kuhness David , Kothleitner Gerald , Winkler Robert , Plank Harald
Published: June 2021

High-fidelity 3D printing of nanoscale objects is an increasing relevant but challenging task. Among the few fabrication techniques, focused electron beam induced deposition (FEBID) has demonstrated its high potential due to its direct-write character, nanoscale capabilities in 3D space and a very high design flexibility. A limitation, however, is the low fabrication speed, which often restricts 3D-FEBID for the fabrication of single objects. In this study, we approach that challenge by reducing the substrate temperatures with a homemade Peltier stage and investigate the effects on Pt based 3D deposits in a temperature range of 5–30 °C. The findings reveal a volume growth rate boost up to a factor of 5.6, while the shape fidelity in 3D space is maintained. From a materials point of view, the internal nanogranular composition is practically unaffected down to 10 °C, followed by a slight grain size increase for even lower temperatures. The study is complemented by a comprehensive discussion about the growth mechanism for a more general picture. The combined findings demonstrate that FEBID on low substrate temperatures is not only much faster, but practically free of drawbacks during high fidelity 3D nanofabrication.
The Contribution of Registered Dietitians in the Management of Hyperemesis Gravidarum in the United Kingdom
Authors: Maslin Kate , Billson Hazel A. , Dean Caitlin R. , Abayomi Julie
Published: June 2021

Hyperemesis Gravidarum (HG) is a condition at the extreme end of the pregnancy sickness spectrum, which can cause poor oral intake, malnutrition, dehydration and weight loss. The aim of this study is to explore the role of Registered Dietitians (RD) in the management of HG in the United Kingdom (UK). A survey was designed and distributed electronically to members of the British Dietetic Association. There were 45 respondents, 76% (n = 34) worked in secondary care hospitals, 11% (n = 5) were in maternal health specialist roles. The most commonly used referral criteria was the Malnutrition Universal Screening Tool (40%, n = 18), followed by second admission (36%, n = 16). However 36% (n = 16) reported no specific referral criteria. About 87% (n = 37) of respondents did not have specific clinical guidelines to follow. Oral nutrition supplements were used by 73% (n = 33) either ‘sometimes’ or ‘most of the time’. Enteral and parenteral nutrition were less commonly used. There was an inconsistent use of referral criteria to dietetic services and a lack of specific clinical guidelines and patient resources. Further training for all clinicians and earlier recognition of malnutrition, alongside investment in the role of dietitians were recommended to improve the nutritional care of those with HG.
Polyphenols and Ischemic Stroke: Insight into One of the Best Strategies for Prevention and Treatment
Authors: Pacifici Francesca , Rovella Valentina , Pastore Donatella , Bellia Alfonso , Abete Pasquale , Donadel Giulia , Santini Silvia , Beck Heinz , Ricordi Camillo , Daniele Nicola Di , Lauro Davide , Della-Morte David
Published: June 2021

Ischemic stroke (IS) is still among the leading causes of death and disability worldwide. The pathogenic mechanisms beyond its development are several and are complex and this is the main reason why a functional therapy is still missed. The beneficial effects of natural compounds against cardiovascular diseases and IS have been investigated for a long time. In this article, we reviewed the association between the most studied polyphenols and stroke protection in terms of prevention, effect on acute phase, and rehabilitation. We described experimental and epidemiological studies reporting the role of flavonols, phenolic acid, and stilbens on ischemic mechanisms leading to stroke. We analyzed the principal animal models used to evaluate the impact of these micronutrients to cerebral blood flow and to molecular pathways involved in oxidative stress and inflammation modulation, such as sirtuins. We reported the most significant clinical trials demonstrated as the persistent use of polyphenols is clinically relevant in terms of the reduction of vascular risk factors for IS, such as Atrial Fibrillation. Interestingly, different kinds of polyphenols provide brain protection by activating different pathways and mechanisms, like inducing antithrombotic effect, such as Honokiol. For this reason, we discussed an appropriate integrative use of them as a possible therapeutic alternative against stroke.
Potential Protective Protein Components of Cow’s Milk against Certain Tumor Entities
Authors: Leischner Christian , Egert Sarah , Burkard Markus , Venturelli Sascha
Published: June 2021

Milk and dairy products, especially from cow’s milk, play a major role in the daily human diet. It is therefore hardly surprising that the subject of milk is being extensively researched and that many effects of individual milk components have been characterized as a result. With the wealth of results available today, the influence of milk on the development of various types of cancer and, in particular, its often protective effects have been shown both in vitro and in vivo and in the evaluation of large-scale cohort and case-control studies. Various caseins, diverse whey proteins such as α-lactalbumin (α-LA), bovine α-lactalbumin made lethal to tumor cells (BAMLET), β-lactoglobulin (β-LG), or bovine serum albumin (BSA), and numerous milk fat components, such as conjugated linoleic acid (CLA), milk fat globule membrane (MFGM), or butyrate, as well as calcium and other protein components such as lactoferrin (Lf), lactoferricin (Lfcin), and casomorphines, show antitumor or cytotoxic effects on cells from different tumor entities. With regard to a balanced and health-promoting diet, milk consumption plays a major role in a global context. This work provides an overview of what is known about the antitumoral properties of proteins derived from cow’s milk and their modes of action.
Currents Nutritional Practices of Nutritionists in the Management of Type 2 Diabetes Patients at Public Health Centres in Padang, Indonesia
Authors: Puri Ice Yolanda , Mohd Yusof Barakatun-Nisak , Abu Zaid Zalina , Ismail Amin , Haron Hasnah , Lipoeto Nur Indrawaty
Published: June 2021

(1) Background: The interest in nutrition practices and education is slowly gaining traction among Indonesian nutritionists. However, there is a lack of local studies that evaluate nutritional practices, especially in the management of type 2 diabetes (T2DM). This cross-sectional study aimed to determine the nutritional practices among nutritionists and the adequacy of the current practices in the management of Type 2 Diabetes Mellitus (T2DM) patients at the Public Health Clinic in Padang (PHC), Indonesia. (2) Methods: An online survey form was distributed to all the nutritionists (n = 50) involved in the management of T2DM patients in their daily practices at the PHC. Socio-demographic characteristics, the current practice of T2DM, the need for DM nutrition education, and an evaluation questionnaire on the Indonesian Non-Communicable Diseases guideline and the Public Health Centre guideline were captured in the survey. (3) Result: A total of 48 completed survey forms were received, providing a response rate of 96% from the recruited nutritionists. One-third (37.5%) of the respondents counselled between one and ten patients per day. Nearly half (41.7%) conducted a monthly follow-up session for the patients at their respective PHC in the previous three months. Each nutritionist educated five to ten T2DM patients. The most common nutrition education topics delivered included appropriate menus (89.6%) as well as the etiology and symptoms of T2DM (85.5%). Almost all the nutritionists (93.8%) used leaflets and about 35.4% used poster education. Around 70.8% of counseling sessions lasted 30 min and two-thirds (66.7%) of the sessions included nutrition education. Based on the results, about half (52.1%) of them claimed that T2DM patients were reluctant to attend individual nutrition education. One-fifth of them (20.8%) claimed that it was because the T2DM patients were not interested in the tool kits and materials used. (4) Conclusions: T2DM patients are reluctant to attend individual nutrition education due to uninteresting tool kits and materials.
Diet, Physical Activity and Gestational Weight Gain Patterns among Pregnant Women Living with Obesity in the North East of England: The GLOWING Pilot Trial
Authors: Heslehurst Nicola , Flynn Angela C. , Ngongalah Lem , McParlin Catherine , Dalrymple Kathryn V. , Best Kate E. , Rankin Judith , McColl Elaine
Published: June 2021

Maternal diet, physical activity (PA) behaviours, and gestational weight gain (GWG) are important for optimum health of women and their babies. This secondary analysis of the GLOWING pilot cluster trial explored these among women living with obesity in high deprivation. Pregnant women completed food frequency, PA and psychosocial questionnaires. Weights were retrieved from medical records and measured during routine appointments with midwives. Descriptive and regression analyses were stratified by obesity class. A total of 163 women were recruited; 54.0% had class 1 obesity, 25.8% class 2, 20.2% class 3, and 76.1% lived in the two most deprived quintiles. Women had suboptimal dietary intake, particularly for oily fish, fruit and vegetables. PA was predominantly light intensity, from household, care and occupational activities. Most women gained weight outside of Institute of Medicine (IOM) guideline recommendations (87.8%); women in class 3 obesity were most likely to have inadequate GWG below IOM recommendations (58.3%, p < 0.01) and reduced odds of excessive GWG compared with class 1 (AOR 0.13, 95% 0.04–0.45). Deprived women with obesity have a double inequality as both increase pregnancy risks. This population requires support to meet guideline recommendations for diet, PA and GWG. Further research exploring obesity classes would inform policies and care to achieve the best pregnancy outcomes.
Evaluation of the Cholesterol-Lowering Mechanism of Enterococcus faecium Strain 132 and Lactobacillus paracasei Strain 201 in Hypercholesterolemia Rats
Authors: Yang Lingshuang , Xie Xinqiang , Li Ying , Wu Lei , Fan Congcong , Liang Tingting , Xi Yu , Yang Shuanghong , Li Haixin , Zhang Jumei , Ding Yu , Xue Liang , Chen Moutong , Wang Juan , Wu Qingping
Published: June 2021

Hypercholesterolemia can cause many diseases, but it can effectively regulated by Lactobacillus. This study aimed to evaluate the cholesterol-lowering mechanism of Enterococcus faecium strain 132 and Lactobacillusparacasei strain 201. These results showed that both the strains decreased serum total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), triglycerides (TG), liver TC and TG and increased fecal TC, TG and total bile acid (TBA) levels. Additionally, both strains also reduced glutamic-pyruvic transaminase (ALT), glutamic oxaloacetic transaminase (AST) and levels of tissue inflammation levels to improve the lipid profile, and they reduced fat accumulation partially by alleviating inflammatory responses. Furthermore, both strains regulated the expression of the CYP8B1, CYP7A1, SREBP-1, SCD1 and LDL-R gene to promote cholesterol metabolism and reduce TG accumulation. Interventions with both strains also altered the gut microbiota, and decreasing the abundance of Veillonellaceae, Erysipelotrichaceae and Prevotella. Furthermore, fecal acetic acid and propionic acid were increased by this intervention. Overall, the results suggested that E. faecium strain 132 and L. paracasei strain 201 can alleviate hypercholesterolemia in rats and might be applied as a new type of hypercholesterolemia agent in functional foods.
Epigenome-Wide Study Identified Methylation Sites Associated with the Risk of Obesity
Authors: Nikpay Majid , Ravati Sepehr , Dent Robert , McPherson Ruth
Published: June 2021

Here, we performed a genome-wide search for methylation sites that contribute to the risk of obesity. We integrated methylation quantitative trait locus (mQTL) data with BMI GWAS information through a SNP-based multiomics approach to identify genomic regions where mQTLs for a methylation site co-localize with obesity risk SNPs. We then tested whether the identified site contributed to BMI through Mendelian randomization. We identified multiple methylation sites causally contributing to the risk of obesity. We validated these findings through a replication stage. By integrating expression quantitative trait locus (eQTL) data, we noted that lower methylation at cg21178254 site upstream of CCNL1 contributes to obesity by increasing the expression of this gene. Higher methylation at cg02814054 increases the risk of obesity by lowering the expression of MAST3, whereas lower methylation at cg06028605 contributes to obesity by decreasing the expression of SLC5A11. Finally, we noted that rare variants within 2p23.3 impact obesity by making the cg01884057 site more susceptible to methylation, which consequently lowers the expression of POMC, ADCY3 and DNAJC27. In this study, we identify methylation sites associated with the risk of obesity and reveal the mechanism whereby a number of these sites exert their effects. This study provides a framework to perform an omics-wide association study for a phenotype and to understand the mechanism whereby a rare variant causes a disease.
Adherence to the Mediterranean Diet in a Portuguese Immigrant Community in the Central Valley of California
Authors: Couto Roberto M. , Frugé Andrew D. , Greene Michael W.
Published: June 2021

The Mediterranean Diet (MedDiet) is a healthy eating pattern associated with a better quality of life among older adults and reduced risk of non-communicable diseases. Little is known about the MedDiet in immigrant communities from countries in which the MedDiet is a settled cultural heritage. Thus, we examined MedDiet adherence and perceived knowledge, benefits, and barriers to the MedDiet in a Portuguese immigrant community in Turlock, California. A cross-sectional study was conducted with 208 participants in Turlock and Livermore, California, which was used as a reference population. Univariate, multivariable, and logistic regression models were used for data analysis. Compared to the Livermore group, the Turlock group was younger and less educated, but had a higher average MedDiet score and active adherence to a MedDiet (p < 0.001 for both). In the Turlock group, convenience, sensory appeal, and health were observed to be significant barriers to the MedDiet (p < 0.05), while health, weight loss, natural content, familiarity, price, sensory appeal, and mood were significant benefit factors (p < 0.05). In conclusion, participants in Turlock had greater MedDiet adherence despite lower education attainment. Furthermore, the perceived benefits of the MedDiet were key factors in MedDiet perception and adherence in a Portuguese immigrant community.
Antibiotic Susceptibility of Bartonella Grown in Different Culture Conditions
Authors: Gadila Shiva Kumar Goud , Embers Monica E.
Published: June 2021

Bartonellosis is caused by a Gram-negative intracellular bacterium with a zoonotic transmission. The disease, caused by any of several genospecies of Bartonella can range from a benign, self-limited condition to a highly morbid and life-threatening illness. The current standard of care antibiotics are generally effective in acute infection; these include azithromycin or erythromycin, doxycycline, gentamicin, rifampin, and ciprofloxacin. However, treatment of chronic infection remains problematic. We tested six different antibiotics for their ability to stop the growth of Bartonella sp. in the standard insect media and in an enrichment media. All antibiotics (ceftriaxone, doxycycline, gentamycin, azithromycin, ampicillin, and azlocillin) had minimum inhibitory concentrations (MICs) below 0.5 µg/mL in the BAPGM enrichment media but were ineffective at inhibiting growth when the standard insect media was used. Azlocillin was the most potent, with a MIC of 0.01 µg/mL. When Bartonella was tested under intracellular growth conditions, none of the antibiotics were efficacious singly. However, growth inhibition was observed when azlocillin and azithromycin were combined. These studies illustrate the impact of growth medium and intracellular environment on antibiotic susceptibility testing and indicate that azlocillin combined with azithromycin may be an effective drug combination for the treatment of Bartonellosis.
Serology- and Blood-PCR-Based Screening for Schistosomiasis in Pregnant Women in Madagascar—A Cross-Sectional Study and Test Comparison Approach
Authors: Hoffmann Tanja , Carsjens Imke , Rakotozandrindrainy Raphaël , Girmann Mirko , Randriamampionona Njary , Maïga-Ascofaré Oumou , Podbielski Andreas , Hahn Andreas , Frickmann Hagen , Schwarz Norbert Georg
Published: June 2021

This work was conducted as a cross sectional study to define the disease burden of schistosomiasis in pregnant Madagascan women and to evaluate serological and molecular diagnostic assays. A total of 1154 residual EDTA blood samples from pregnant Madagascan women were assessed. The nucleic acid extractions were subjected to in-house real-time PCRs specifically targeting S. mansoni complex, S. haematobium complex, and African Schistosoma spp. on genus level, while the EDTA plasma samples were analyzed using Schistosoma-specific IgG and IgM commercial ELISA and immunofluorescence assays. The analyses indicated an overall prevalence of schistosomiasis in Madagascan pregnant women of 40.4%, with only minor regional differences and differences between serology- and blood PCR-based surveillance. The S. mansoni specific real-time PCR showed superior sensitivity of 74% (specificity 80%) compared with the genus-specific real-time PCR (sensitivity 13%, specificity 100%) in blood. The laborious immunofluorescence (sensitivity IgM 49%, IgG 87%, specificity IgM 85%, IgG 96%) scored only slightly better than the automatable ELISA (sensitivity IgM 38%, IgG 88%, specificity IgM 78%, IgG 91%). Infections with S. mansoni were detected only. The high prevalence of schistosomiasis recorded here among pregnant women in Madagascar calls for actions in order to reduce the disease burden.
Experimental Evidence of Epizootic Epitheliotropic Disease Virus (Salmoid Herpesvirus-3, Alloherpesviridae) Transmission via Contaminated Fomites and Subsequent Prevention Using a Disinfectant
Authors: Purbayu Mochamad A. , Shavalier Megan A. , Faisal Mohamed , Loch Thomas P.
Published: June 2021

Epizootic epitheliotropic disease virus (EEDV) has caused considerable mortality in hatchery-reared lake trout Salvelinus namaycush in the Great Lakes Basin, and yet the routes of transmission and efficacious means of prevention remain poorly understood. To determine whether EEDV can be transmitted via contaminated fomites and clarify whether such transmission could be prevented via fomite disinfection, juvenile lake trout (n = 20 per treatment) were handled in nets previously soaked in an EEDV suspension (7.29 × 104–2.25 × 105 virus copies/mL of water) that were further immersed in either 1% Virkon® Aquatic (“disinfected” treatment, in triplicate) or in sample diluent (“EEDV-contaminated” treatment). Negative control nets were soaked in sterile sample diluent only. Characteristic gross signs of EED developed in the “EEDV-contaminated” treatment group, which was followed by 80% mortality, whereas no gross signs of disease and 0–5% mortality occurred in the negative control and “disinfected” treatment groups, respectively. EEDV was detected via qPCR in 90% of the “EEDV-contaminated” treatment fish, however, it was not detected in any fish within the negative control or “disinfected” treatment groups. Study findings not only demonstrate that EEDV can be readily transmitted via contaminated fomites, but importantly suggest that Virkon® Aquatic is an efficacious option for preventing EEDV contagion via the disinfection of hatchery tools, thereby highlighting a promising tool for improving lake trout hatchery biosecurity and minimizing EEDV-linked losses.
Periodontal Pathogens and Preterm Birth: Current Knowledge and Further Interventions
Authors: Terzic Milan , Aimagambetova Gulzhanat , Terzic Sanja , Radunovic Milena , Bapayeva Gauri , Laganà Antonio Simone
Published: June 2021

Preterm labor is defined as a birth before 37 weeks of gestation and occurs in 5–20% of pregnancies. Preterm labor, as multifactorial entity associated with a high risk of neonatal morbidity and mortality, is influenced by maternal, fetal and environmental factors. Microbiological studies suggest that infectious pathogens may account for 25–40% of preterm birth. Infections of different sites, like genital, urinary tract infections, and pneumonia, are linked to the preterm labor. The most recent epidemiological studies consistently report that maternal periodontal disease is associated with preterm delivery, as well as the association between the presence of pathogenic oral bacteria in the placenta and adverse pregnancy outcomes. On the other hand, some previously published papers found periodontal bacteria in placentas of term pregnancies. In spite of a huge research done on the topic, both experimental and clinical, there are many controversial opinions about the role of periodontal infections in preterm birth. Thus, this comprehensive review addresses this very important topic and evaluates novel strategies of preventive and therapeutic approaches.
Relative Age Effect in Physical Fitness during the Elementary School Years
Authors: Drenowatz Clemens , Ferrari Gerson , Greier Klaus , Hinterkörner Franz
Published: June 2021

Age-groups are commonly implemented in education and sports in order to provide fair and equal opportunities. Various studies, however, have shown a competitive advantage for early born children over their relatively younger peers, which is referred to as relative age effect. The present study examined differences in various components of physical fitness in Austrian elementary-school children. A total of 18,168 children (51% boys) between 6 and 11 years of age provided valid data on anthropometric characteristics and physical fitness. Specifically, children completed eight fitness tests that assessed cardiorespiratory endurance, muscular strength and power, speed, agility, flexibility and object control. Across age-specific quartiles, older children were significantly taller and heavier than their younger peers. Older children also displayed better performance for strength and power, speed, agility and object control, while differences in cardio-respiratory endurance were less pronounced. These results highlight the presence of a relative age effect during the elementary school years and emphasize the need to consider individual differences in the evaluation of children’s performance. As all children should be given equal opportunities to engage successfully in physical education and sports, physical education teachers and youth coaches need to be educated on the implications of a relative age effect.
Targeting the Copper Transport System to Improve Treatment Efficacies of Platinum-Containing Drugs in Cancer Chemotherapy
Authors: Kuo Macus Tien , Huang Yu-Fang , Chou Cheng-Yang , Chen Helen H. W.
Published: June 2021

The platinum (Pt)-containing antitumor drugs including cisplatin (cis-diamminedichloroplatinum II, cDDP), carboplatin, and oxaliplatin, have been the mainstay of cancer chemotherapy. These drugs are effective in treating many human malignancies. The major cell-killing target of Pt drugs is DNA. Recent findings underscored the important roles of Pt drug transport system in cancer therapy. While many mechanisms have been proposed for Pt-drug transport, the high-affinity copper transporter (hCtr1), Cu chaperone (Atox1), and Cu exporters (ATP7A and ATP7B) are also involved in cDDP transport, highlighting Cu homeostasis regulation in Pt-based cancer therapy. It was demonstrated that by reducing cellular Cu bioavailable levels by Cu chelators, hCtr1 is transcriptionally upregulated by transcription factor Sp1, which binds the promoters of Sp1 and hCtr1. In contrast, elevated Cu poisons Sp1, resulting in suppression of hCtr1 and Sp1, constituting the Cu-Sp1-hCtr1 mutually regulatory loop. Clinical investigations using copper chelator (trientine) in carboplatin treatment have been conducted for overcoming Pt drug resistance due in part to defective transport. While results are encouraging, future development may include targeting multiple steps in Cu transport system for improving the efficacies of Pt-based cancer chemotherapy. The focus of this review is to delineate the mechanistic interrelationships between Cu homeostasis regulation and antitumor efficacy of Pt drugs.
Acrylonitrile Derivatives against Trypanosoma cruzi: In Vitro Activity and Programmed Cell Death Study
Authors: Bethencourt-Estrella Carlos , Delgado-Hernández Samuel , López-Arencibia Atteneri , San Nicolás-Hernández Desirée , Sifaoui Ines , Tejedor David , García-Tellado Fernando , Lorenzo-Morales Jacob , Piñero José
Published: June 2021

The neglected infection known as Chagas disease, caused by the protozoan parasite Trypanosoma cruzi, results in more than 7000 deaths per year, with an increasing number of cases in non-endemic areas such as Europe or the United States. Moreover, with the current available therapy, only two compounds which are active against the acute phase of the disease are readily available. In addition, these therapeutic agents display multiple undesired side effects such as high toxicity, they are expensive, the treatment is lengthy and the resistant strain has emerged. Therefore, there is a need to find new compounds against Chagas disease which should be active against the parasite but also cause low toxicity to the patients. In the present work, the activity of novel acrylonitriles against Trypanosoma cruzi was evaluated as well as the analysis of the physiological events induced in the treated parasites related to the cell death process. Hence, the characteristic features of an apoptosis-like process such as chromatin condensation and mitochondrial membrane potential, among others, were studied. From the 32 compounds tested against the epimastigote stage of T. cruzi, 11 were selected based on their selectivity index to determine if these compounds were able to induce programmed cell death (PCD) in the treated parasites. Furthermore, acrylonitriles Q5, Q7, Q19, Q27 and Q29 were shown to trigger physiological events related in the PCD. Therefore, this study highlights the therapeutic potential of acrylonitriles as novel trypanocidal agents.
Trimethoxylated Halogenated Chalcones as Dual Inhibitors of MAO-B and BACE-1 for the Treatment of Neurodegenerative Disorders
Authors: Vishal Vishal Payyalot , Oh Jong Min , Khames Ahmed , Abdelgawad Mohamed A. , Nair Aathira Sujathan , Nath Lekshmi R. , Gambacorta Nicola , Ciriaco Fulvio , Nicolotti Orazio , Kim Hoon , Mathew Bijo
Published: June 2021

Six halogenated trimethoxy chalcone derivatives (CH1–CH6) were synthesized and spectrally characterized. The compounds were further evaluated for their inhibitory potential against monoamine oxidases (MAOs) and β-secretase (BACE-1). Six compounds inhibited MAO-B more effectively than MAO-A, and the 2′,3′,4′-methoxy moiety in CH4–CH6 was more effective for MAO-B inhibition than the 2′,4′,6′-methoxy moiety in CH1–CH3. Compound CH5 most potently inhibited MAO-B, with an IC50 value of 0.46 µM, followed by CH4 (IC50 = 0.84 µM). In 2′,3′,4′-methoxy derivatives (CH4-CH6), the order of inhibition was –Br in CH5 > -Cl in CH4 > -F in CH6 at the para-position in ring B of chalcone. CH4 and CH5 were selective for MAO-B, with selectivity index (SI) values of 15.1 and 31.3, respectively, over MAO-A. CH4 and CH5 moderately inhibited BACE-1 with IC50 values of 13.6 and 19.8 µM, respectively. When CH4 and CH5 were assessed for their cell viability studies on the normal African Green Monkey kidney cell line (VERO) using MTT assays, it was noted that both compounds were found to be safe, and only a slightly toxic effect was observed in concentrations above 200 µg/mL. CH4 and CH5 decreased reactive oxygen species (ROS) levels of VERO cells treated with H2O2, indicating both compounds retained protective effects on the cells by antioxidant activities. All compounds showed high blood brain barrier permeabilities analyzed by a parallel artificial membrane permeability assay (PAMPA). Molecular docking and ADME prediction of the lead compounds provided more insights into the rationale behind the binding and the CNS drug likeness. From non-test mutagenicity and cardiotoxicity studies, CH4 and CH5 were non-mutagenic and non-/weak-cardiotoxic. These results suggest that CH4 and CH5 could be considered candidates for the cure of neurological dysfunctions.
Thiolated-Polymer-Based Nanoparticles as an Avant-Garde Approach for Anticancer Therapies—Reviewing Thiomers from Chitosan and Hyaluronic Acid
Authors: Grosso Roberto , de-Paz M.-Violante
Published: June 2021

Thiomers (or thiolated polymers) have broken through as avant-garde approaches in anticancer therapy. Their distinguished reactivity and properties, closely linked to their final applications, justify the extensive research conducted on their preparation and use as smart drug-delivery systems (DDSs). Multiple studies have demonstrated that thiomer-rich nanoformulations can overcome major drawbacks found when administering diverse active pharmaceutical ingredients (APIs), especially in cancer therapy. This work focuses on providing a complete and concise review of the synthetic tools available to thiolate cationic and anionic polymers, in particular chitosan (CTS) and hyaluronic acid (HA), respectively, drawing attention to the most successful procedures. Their chemical reactivity and most relevant properties regarding their use in anticancer formulations are also discussed. In addition, a variety of NP formation procedures are outlined, as well as their use in cancer therapy, particularly for taxanes and siRNA. It is expected that the current work could clarify the main synthetic strategies available, with their scope and drawbacks, as well as provide some insight into thiomer chemistry. Therefore, this review can inspire new research strategies in the development of efficient formulations for the treatment of cancer.
Preliminary Investigation on Simvastatin-Loaded Polymeric Micelles in View of the Treatment of the Back of the Eye
Authors: Pescina Silvia , Sonvico Fabio , Clementino Adryana , Padula Cristina , Santi Patrizia , Nicoli Sara
Published: June 2021

There is increasing consensus in considering statins beneficial for age-related macular degeneration and in general, for immune and inflammatory mediated diseases affecting the posterior segment of the eye. However, all available data relate to oral administration, and safety and effectiveness of statins directly administered to the eye are not yet known, despite their ophthalmic administration could be beneficial. The aim was the development and the characterization of polymeric micelles based on TPGS or TPGS/poloxamer 407 to increase simvastatin solubility and stability and to enhance the delivery of the drug to the posterior segment of the eye via trans-scleral permeation. Simvastatin was chosen as a model statin and its active hydroxy acid metabolite was investigated as well. Results demonstrated that polymeric micelles increased simvastatin solubility at least 30-fold and particularly TPGS/poloxamer 407 mixed micelles, successfully stabilized simvastatin over time, preventing the hydrolysis when stored for 1 month at 4 °C. Furthermore, both TPGS (1.3 mPas) and mixed micelles (33.2 mPas) showed low viscosity, suitable for periocular administration. TPGS micelles resulted the best performing in delivery simvastatin either across conjunctiva or sclera in ex vivo porcine models. The data pave the way for a future viable ocular administration of statins.
Effects of p-Cresol on Oxidative Stress, Glutathione Depletion, and Necrosis in HepaRG Cells: Comparisons to Other Uremic Toxins and the Role of p-Cresol Glucuronide Formation
Authors: Zhu Sang , Rong Yan , Kiang Tony K. L.
Published: June 2021

The toxicological effects of p-cresol have primarily been attributed to its metabolism products; however, very little human data are available in the key organ (i.e., liver) responsible for the generation of these metabolites. Experiments were conducted in HepaRG cells utilizing the following markers of cellular toxicity: 2′-7′-dichlorofluorescein (DCF; oxidative stress) formation, total cellular glutathione (GSH) concentration, and lactate dehydrogenase (LDH; cellular necrosis) release. Concentrations of p-cresol, p-cresol sulfate, and p-cresol glucuronide were determined using validated assays. p-Cresol exposure resulted in concentration- and time-dependent changes in DCF (EC50 = 0.64 ± 0.37 mM at 24 h of exposure) formation, GSH (EC50 = 1.00 ± 0.07 mM) concentration, and LDH (EC50 = 0.85 ± 0.14 mM) release at toxicologically relevant conditions. p-Cresol was also relatively more toxic than 3-carboxy-4-methyl-5-propyl-2-furanpropanoic acid, indole-3-acetic acid, indoxyl sulfate, kynurenic acid, and hippuric acid on all markers. Although the exogenous administration of p-cresol sulfate and p-cresol glucuronide generated high intracellular concentrations of these metabolites, both metabolites were less toxic compared to p-cresol at equal-molar conditions. Moreover, p-cresol glucuronide was the predominant metabolite generated in situ from p-cresol exposure. Selective attenuation of glucuronidation (without affecting p-cresol sulfate formation, while increasing p-cresol accumulation) using independent chemical inhibitors (i.e., 0.75 mM l-borneol, 75 µM amentoflavone, or 100 µM diclofenac) consistently resulted in further increases in LDH release associated with p-cresol exposure (by 28.3 ± 5.3%, 30.0 ± 8.2% or 27.3 ± 6.8%, respectively, compared to p-cresol treatment). These novel data indicated that p-cresol was a relatively potent toxicant, and that glucuronidation was unlikely to be associated with the manifestation of its toxic effects in HepaRG cells.