Category: Disability & Diversity studies as a professional basis for diversity-aware education and training in medicine.

The Regulation of Nitrate Reductases in Response to Abiotic Stress in Arabidopsis TRANSLATE Journal: International Journ

On: By: Brent
The two homologous genes, NIA1 and NIA2, encode nitrate reductases in Arabidopsis, which govern the reduction of nitrate to nitrite. This step is the rate-limiting step of the nitrate assimilation and utilization. Therefore, the regulation of NIA1 and NIA2 is important for plant development and growth. Although they are similar in sequence and structure, their regulations are different. Genetic analysis uncovers that NIA1, rather than NIA2, plays a predominant role in adopting to ABA stress. Although both long-term stress conditions can cause an improvement in NIA1 levels, a decrease in NIA1 levels under short-term treatments seems to be necessary for plants to switch from the growth status into the adopting status. Interestingly, the downregulation of the NR is distinct under different stress conditions. Under ABA treatment, the NR proteins are degraded via a 26S-proteasome dependent manner, while the transcriptional regulation is the main manner to rapidly reduce the NIA1 levels under nitrogen deficiency and NaCl stress conditions. These results indicate that under stress conditions, the regulation of NIA1 is complex, and it plays a key role in regulating the balance between growth and adaptation.

Evaluation of the Simpson’s Method to Determine Left Ventricular Ejection Fraction Using the Transgastric Two-Chamber View

Introduction: Left ventricular chamber size and functional assessment by transesophageal echocardiography can be difficult if visualization is poor in the mid-esophageal views. However, the accuracy of using the Simpson’s method in the transgastric 2-chamber (TG2C) as an alternative approach has not been assessed.
Methods: The Simpson’s method was performed by 2 independent reviewers using midesophageal 2-chamber (ME2C) and TG2C views. Echocardiographic images were retrieved retrospectively for 49 adult cardiac surgical patients.
Results: Two-way random effects intraclass correlation coefficients demonstrated no significant interobserver variability. Linear mixed effects models showed no significant differences in ME2C and TG2C measurements with regard to EDV (P=.4407), ESV (P=.5113), or EF (P=.0610).Compared to the ME2C view, the TG2C view had better image quality of the LV walls (image quality score median [interquartile range]: 2.00 [.00] vs 1.00 [1.00]; P<.0001), but worse image quality of the mitral annulus (1.00 [1.00] vs 2.00 [.00]; P<.0001) and LV apex (.00 [1.00] vs 2.00 [1.00]; P<.0001).
Conclusions: This study suggests the Simpson’s method can be applied to the TG2C view as an alternative to the standard midesophageal method to estimate chamber volumes and EF.
Keywords: Simpson’s method; intraoperative assessment; left ventricular ejection fraction; monitoring; research; transesophageal echocardiography.

Intracellular MUC20 variant 2 maintains mitochondrial calcium homeostasis and enhances drug resistance in gastric cancer

Background: Signet ring cell carcinoma (SRCC) is a particular histologic variant of gastric cancer (GC). However, the critical factor related to the aggressive characteristics of SRCC has not been determined.
Methods: We collected surgically resected tissues from 360 GC patients in the Kumamoto University cohort and generated survival curves via the Kaplan-Meier method. In vitro, we identified the specific transcript variant of MUC20 in SRCC cells by direct sequencing and investigated the role of MUC20 in GC progression using GC cells with MUC20 silencing and forced expression. In vivo, we examined chemoresistance using MUC20 variant 2 (MUC20v2)-overexpressing non-SRCC cells to construct a xenograft mouse model.
Results: We analyzed a comprehensive GC cell line database to identify the specifically expressed genes in gastric SRCC. We focused on MUC20 and investigated its role in GC progression. Survival analysis revealed that GC patients with high MUC20 expression exhibited a poor prognosis and that MUC20 expression was significantly correlated with SRCC histological type. Moreover, we found that gastric SRCC cells specifically expressed MUC20v2, which was dominantly expressed in the cytoplasm. Silencing MUC20v2 caused cell death with characteristic morphological changes in gastric SRCC cells. To further determine the types of cell death, we examined apoptosis, pyroptosis and ferroptosis by detecting cleaved PARP, gasdermin E-N-terminal (GSDME-N), and lipid reactive oxygen species (ROS) levels, respectively. We found that apoptosis and pyroptosis occurred in MUC20-silenced gastric SRCC cells. In addition, MUC20v2-overexpressing GC cells exhibited chemoresistance to cisplatin (CDDP) and paclitaxel (PTX). RNA sequencing revealed that the pathways involved in intracellular calcium regulation were significantly upregulated in MUC20v2-overexpressing GC cells.
Notably, forced expression of MUC20v2 in the cytoplasm of GC cells led to the maintenance of mitochondrial calcium homeostasis and mitochondrial membrane potential (MMP), which promoted cell survival and chemoresistance by suppressing apoptosis and pyroptosis. Finally, we investigated the significance of MUC20v2 in a xenograft model treated with CDDP and showed that MUC20v2 overexpression caused chemoresistance by inhibiting cell death.
Conclusion: These findings highlight the novel functions of MUC20v2, which may confer cell survival and drug resistance in GC cells.
Significance: MUC20v2 protects GC cells from apoptosis and pyroptosis by maintaining mitochondrial calcium levels and mitochondrial membrane potential and subsequently induces drug resistance.
Keywords: Drug resistance; Intracellular mucin 20; Mitochondrial calcium homeostasis; Mitochondrial membrane potential; Signet ring cell carcinoma.

Antioxidant, Antimicrobial Activities and Characterization of Polyphenol-Enriched Extract of Egyptian Celery ( Apium graveolens L., Apiaceae) Aerial Parts via UPLC/ESI/TOF-MS

Medicinal plant extracts are increasingly considered a major source of innovative medications and healthcare products.
This study focused on preparing a polyphenol enriched water extract of Egyptian celery “Apium graveolens L., Apiaceae” aerial parts (TAE) in an endeavor to accentuate its antioxidant capacity as well as its antimicrobial activity. (TAE) of celery was partitioned against different organic solvents to yield dichloromethane (DCM), ethyl acetate (EAC), and butanol (BUOH) fractions. (TAE) and the organic fractions thereof besides the remaining mother liquor (ML) were all screened for their antioxidant capacity using various protocols viz. monitoring the reducing amplitudes for ferric ions (FRAP), and radical scavenging potentials of oxygen (ORAC), 2,2′-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS), 2,2-diphenyl-1-picrylhydrazyl (DPPH), and metal chelation assays.
The examination procedure revealed both (TAE) extract and (DCM) fraction, to pertain the highest antioxidant potentials, where the IC50 of the (TAE) using ABTS and metal chelation assays were ca. 34.52 ± 3.25 and 246.6 ± 5.78 µg/mL, respectively. The (DCM) fraction recorded effective results using the FRAP, ORAC, and DPPH assays ca. 233.47 ± 15.14 and 1076 ± 25.73 μM Trolox equivalents/mg sample and an IC50 474.4 ± 19.8 µg/mL, respectively.
Additionally, both (TAE) and (DCM) fraction exerted antimicrobial activities recording inhibition zones (mm) (13.4 ± 1.5) and (12.0 ± 1.0) against Staphylococcus aureus and (11.0 ± 1.2) and (10.0 ± 1.3) against Escherichia coli, respectively, with no anti-fungal activity. Minimum inhibitory concentration (MIC) of (TAE) and (DCM) fraction were 1250 and 2500 µg/mL, respectively. UPLC/ESI/TOF-MS unveiled the chemical profile of both (TAE) and (DCM) fraction to encompass a myriad of active polyphenolic constituents including phenylpropanoids, coumarins, apigenin, luteolin, and chrysoeriol conjugates.

Real world data on cardiometabolic diseases in U.S. adults during the SARS-CoV-2 pandemic: a decentralized registry study

Background: Pre-existing cardiometabolic comorbidities place SARS-CoV-2 positive patients at a greater risk for poorer clinical course and mortality than those without it. We aimed to analyze real-world registry data focused primarily on participants with cardiometabolic diseases (CMD), which were remotely obtained via a digital platform.
Methods: Participants were divided into two groups: CMD or no cardiometabolic disease (non-CMD). They were evaluated based on their medical history, current medications/supplements, COVID-19 status, demographics, and baseline characteristics. The frequency of medications/supplements for CMD were compared using relative risks and 95% confidence intervals. The WHO (Five) Well-Being Index (WHO-5) were collected monthly for 6 months to assess psychological well-being which included cheerfulness, calmness, vigor, rest, and engagement with daily activities of interest.
Results: The 791 enrollees represented 49 U.S. states.
The CMD group had significantly higher (p < 0.0001) BMI (mean + 3.04 kg/m2) and age (mean + 9.15 years) compared to non-CMD group. In the CMD group, participants who tested positive for COVID-19 had lower (p < 0.0001) well-being scores than those without COVID-19. For the 274 participants on CMD medications/supplements, there was no statistical difference in risk of COVID-19 contracture based on medication/supplement type; however, all six participants who were not being treated for CMD were COVID-19 positive (RR ~ 104). For 89 participants who were on treatment for diabetes or insulin resistance, there was a 90% reduced risk of COVID-19 incidence (p = 0.0187).
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Conclusion: The well-being score of the CMD group was dependent on whether they tested positive for COVID-19. Type of CMD treatment did not impact COVID-19 status, but absence of treatment significantly increased COVID-19 incidence. With respect to SARS-CoV-2, our analysis supports continued use of the statins, ACE-I, ARBs, and diabetes medications in CMD patients.
Trial registration: ClinicalTrials.gov Identifier: NCT04348942.
Keywords: Angiotensin; Cardiovascular; Diabetes; Insulin resistance; Metabolism; SARS-CoV-2.

Genome-Wide Characterization of SARS-CoV-2 Cytopathogenic Proteins in the Search of Antiviral Targets

On: By: Brent
Therapeutic inhibition of critical viral functions is important for curtailing coronavirus disease 2019 (COVID-19). We sought to identify antiviral targets through the genome-wide characterization of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) proteins that are crucial for viral pathogenesis and that cause harmful cytopathogenic effects.
All 29 viral proteins were tested in a fission yeast cell-based system using inducible gene expression. Twelve proteins, including eight nonstructural proteins (NSP1, NSP3, NSP4, NSP5, NSP6, NSP13, NSP14, and NSP15) and four accessory proteins (ORF3a, ORF6, ORF7a, and ORF7b), were identified that altered cellular proliferation and integrity and induced cell death. Cell death correlated with the activation of cellular oxidative stress. Of the 12 proteins, ORF3a was chosen for further study in mammalian cells because it plays an important role in viral pathogenesis and its activities are linked to lung tissue damage and a cytokine storm.
In human pulmonary and kidney epithelial cells, ORF3a induced cellular oxidative stress associated with apoptosis and necrosis and caused activation of proinflammatory response with production of the cytokines tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6), and IFN-β1, possibly through the activation of nuclear factor kappa B (NF-κB).
To further characterize the mechanism, we tested a natural ORF3a Beta variant, Q57H, and a mutant with deletion of the highly conserved residue, ΔG188. Compared with wild-type ORF3a, the ΔG188 variant yielded more robust activation of cellular oxidative stress, cell death, and innate immune response. Since cellular oxidative stress and inflammation contribute to cell death and tissue damage linked to the severity of COVID-19, our findings suggest that ORF3a is a promising, novel therapeutic target against COVID-19.
 IMPORTANCE The ongoing COVID-19 pandemic caused by SARS-CoV-2 has claimed over 5.5 million lives with more than 300 million people infected worldwide. While vaccines are effective, the emergence of new viral variants could jeopardize vaccine protection. Treatment of COVID-19 by antiviral drugs provides an alternative to battle against the disease. The goal of this study was to identify viral therapeutic targets that can be used in antiviral drug discovery. Utilizing a genome-wide functional analysis in a fission yeast cell-based system, we identified 12 viral candidates, including ORF3a, which cause cellular oxidative stress, inflammation, apoptosis, and necrosis that contribute to cytopathogenicity and COVID-19. Our findings indicate that antiviral agents targeting ORF3a could have a great impact on COVID-19.

Copolymers Containing 1-Methyl-2-phenyl-imidazole Moieties as Permanent Dipole Generating Units: Synthesis, Spectroscopic, Electrochemical, and Photovoltaic Properties

New donor-acceptor conjugated alternating or random copolymers containing 1-methyl-2-phenylbenzimidazole and benzothiadiazole (P1), diketopyrrolopyrrole (P4), or both acceptors (P2) are reported. The specific feature of these copolymers is the presence of a permanent dipole-bearing moiety (1-methyl-2-phenyl imidazole (MPI)) fused with the 1,4-phenylene ring of the polymer main chain.
For comparative reasons, polymers of the same main chain but deprived of the MPI group were prepared, namely, P5 with diketopyrrolopyrrole and P3 with both acceptors. The presence of the permanent dipole results in an increase of the optical band gap from 1.51 eV in P3 to 1.57 eV in P2 and from 1.49 eV in P5 to 1.55 eV in P4. It also has a measurable effect on the ionization potential (IP) and electrochemical band gap (EgCV), leading to their decrease from 5.00 and 1.83 eV in P3 to 4.92 and 1.79 eV in P2 as well as from 5.09 and 1.87 eV in P5 to 4.94 and 1.81 eV in P4.
 Moreover, the presence of permanent dipole lowers the exciton binding energy (Eb) from 0.32 eV in P3 to 0.22 eV in P2 and from 0.38 eV in P5 to 0.26 eV in P4. These dipole-induced changes in the polymer properties should be beneficial for photovoltaic applications. Bulk heterojunction solar cells fabricated from these polymers (with PC71BM acceptor) show low series resistance (rs), indicating good electrical transport properties. The measured power conversion efficiency (PCE) of 0.54% is limited by the unfavorable morphology of the active layer.

Preparation of Naringenin Nanosuspension and Its Antitussive and Expectorant Effects

Naringenin (NRG) is a natural flavonoid compound abundantly present in citrus fruits and has the potential to treat respiratory disorders. However, the clinical therapeutic effect of NRG is limited by its low bioavailability due to poor solubility.
To enhance the solubility, naringenin nanosuspensions (NRG-NSps) were prepared by applying tocopherol polyethylene glycol succinate (TPGS) as the nanocarrier via the media-milling method. The particle size, morphology, and drug-loading content of NRG-NSps were examined, and the stability was evaluated by detecting particle size changes in different physiological media. NRG-NSps exhibited a flaky appearance with a mean diameter of 216.9 nm, and the drug-loading content was 66.7%. NRG-NSps exhibited good storage stability and media stability.
NRG-NSps presented a sustainable release profile, and the cumulative drug-release rate approached approximately 95% within 7 d. NRG-NSps improved the antitussive effect significantly compared with the original NRG, the cough frequency was decreased from 22 to 15 times, and the cough incubation period was prolonged from 85.3 to 121.6 s. Besides, NRG-NSps also enhanced expectorant effects significantly, and phenol red secretion was increased from 1.02 to 1.45 μg/mL. These results indicate that NRG-NSps could enhance the bioavailability of NRG significantly and possess a potential clinical application.
Keywords: antitussive effect; bioavailability; expectorant effect; media-milling method; naringenin nanosuspension.

Distinct effects of volatile and intravenous anaesthetics on presynaptic calcium dynamics in mouse hippocampal GABAergic neurones

Background: General anaesthetics have marked effects on synaptic transmission, but their neuronal and circuit-level effects remain unclear. The volatile anaesthetic isoflurane differentially inhibits synaptic vesicle exocytosis in specific neuronal subtypes, but whether other common anaesthetics also have neurone-subtype-specific actions is unknown.
Methods: We used the genetically encoded fluorescent Ca2+ sensor GCaMP6f to compare the pharmacological effects of isoflurane, sevoflurane, propofol, and ketamine on presynaptic excitability in hippocampal glutamatergic neurones and in hippocampal parvalbumin-, somatostatin-, and vasoactive intestinal peptide-expressing (PV+, SST+, and VIP+, respectively) GABAergic interneurones.
Results: Isoflurane and sevoflurane depressed activity-driven presynaptic Ca2+ transients in a neurone-type-specific manner, with greater potency for inhibition of glutamate and SST+ compared with PV+ and VIP+ neurone presynaptic activation. In contrast, clinical concentrations of propofol (1 μM) or ketamine (15 μM) had no significant effects on presynaptic activation. Propofol potentiated evoked Ca2+ entry in PV+ interneurones but only at a supraclinical concentration (3 μM).
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Conclusions: Anaesthetic-agent-selective effects on presynaptic Ca2+ entry have functional implications for hippocampal circuit function during i.v. or volatile anaesthetic-mediated anaesthesia. Hippocampal interneurones have distinct subtype-specific sensitivities to volatile anaesthetic actions on presynaptic Ca2+, which are similar between isoflurane and sevoflurane.
Keywords: calcium; interneurone; isoflurane; ketamine; mechanisms of anaesthesia; presynaptic; propofol; sevoflurane.

Impact of DPP-4 inhibitors on plasma levels of BNP and NT-pro-BNP in type 2 diabetes mellitus

On: By: Brent
Background: Dipeptidyl peptidase-4 inhibitors (DPP-4i) decrease glucose levels by regulating incretin peptides in type 2 diabetes mellitus (T2DM). This study aimed to determine the modulatory effect of DPP-4i on brain natriuretic peptide (BNP) and N-terminal pro-brain natriuretic peptide (NT-pro-BNP) in patients with T2DM.
Methods: PubMed, Embase and the Cochrane Library were systematically searched to identify randomized controlled trials (RCTs) evaluating the impact of DPP-4i on BNP or NT-pro-BNP. A fixed- or random-effects model was used for quantitative analysis, according to the heterogeneity. Sensitivity analysis and publication bias were performed using standard methods.
Results: Nine trials with 3056 patients with T2DM were included. Meta-analysis revealed that DPP-4i did not significantly modulate the BNP (0.21 pg/mL, 95% CI – 2.36-2.79) or NT-pro-BNP level (- 7.34 pg/mL, 95% CI – 24.27-9.59). DPP-4i demonstrated no stronger effect on modulating BNP (5.17 pg/mL, 95% CI – 7.48-17.82) or NT-pro-BNP (- 9.95 pg/mL, 95% CI – 44.61-24.71) than active comparators. Pooled analysis was robust and stable after sensitivity analysis.
Conclusions: DPP-4i exhibits no significant effect on modulating BNP or NT-pro-BNP and shows no stronger effect than traditional antidiabetic agents in T2DM.
Keywords: BNP; Dipeptidyl peptidase-4 inhibitors; NT-pro-BNP; Randomized controlled trials; Type 2 diabetes mellitus.

Sociodemographic Determinants of Poles’ Attitudes towards the Forest during the COVID-19 Pandemic

Attitudes towards forest ecosystems have been changing together with human needs, which is amplified with society’s increasing need to spend recreation time in the forest. The phenomenon has been particularly visible during the COVID-19 pandemic. The aim of this study was to determine the attitude of Poles to forests during the COVID-19 pandemic. The research was based on (1) a sociodemographic background questionnaire that consisted of questions about the independent variables and (2) the LAS scale-an independently prepared tool for measuring attitudes towards the forest. In the survey, 1025 people participated (673 women). The age of the subjects was between 19 and 68.
The attitude towards the forest was analysed in three dimensions: Benefits, Involvement, and Fears. The Mann-Whitney U test and Kruskal-Wallis one-way analysis of variance by ranks were used for statistical analysis. Women and people with primary education expressed the most fears connected with going to the forest. Men and people living in the countryside and in small towns, as well as respondents who were professionally active and performing work connected with forests were the most involved in exploring the forest and working for its benefit. Concerning the forest, concerned women, people from the highest age group, respondents with university education, and white-collar workers notice the most benefits from recreational activities in the forest.

Relationship between Translational and Rotational Dynamics of Alkyltriethylammonium-Based Ionic Liquids

1H spin-lattice relaxation experiments have been performed for a series of ionic liquids including bis(trifluoromethanesulfonyl)imide anion and cations of a varying alkyl chain length: triethylhexylammonium, triethyloctylammonium, decyltriethylammonium, dodecyltriethylammonium, triethyltetradecylammonium, and hexadecyltriethylammonium. The relaxation studies were carried out in abroad frequency range covering three orders of magnitude, from 10 kHz to 10 MHz, versus temperature.
On the basis of a thorough, quantitative analysis of this reach data set, parameters characterizing the relative, cation-cation, translation diffusion (relative diffusion coefficients and translational correlation times), and rotational motion of the cation (rotational correlation times) were determined. Relationships between these quantities and their dependence on the alkyl chain length were discussed in comparison to analogous properties of molecular liquids. It was shown, among other findings, that the ratio between the translational and rotational correlation times is smaller than for molecular liquids and considerably dependent on temperature. Moreover, a comparison of relative and self-diffusion coefficients indicate correlated translational dynamics of the cations.

Incorporation of a FRET Pair into a Riboswitch RNA to Measure Mg 2+ Concentration and RNA Conformational Change in Cell

Riboswitches are natural biosensors that can regulate gene expression by sensing small molecules. Knowledge of the structural dynamics of riboswitches is crucial to elucidate their regulatory mechanism and develop RNA biosensors. In this work, we incorporated the fluorophore, Cy3, and its quencher, TQ3, into a full-length adenine riboswitch RNA and its isolated aptamer domain to monitor the dynamics of the RNAs in vitro and in cell. The adenine riboswitch was sensitive to Mg2+ concentrations and could be used as a biosensor to measure cellular Mg2+ concentrations. Additionally, the TQ3/Cy3-labeled adenine riboswitch yielded a Mg2+ concentration that was similar to that measured using a commercial assay kit.
Furthermore, the fluorescence response to the adenine of the TQ3/Cy3-labeled riboswitch RNA was applied to determine the proportions of multiple RNA conformational changes in cells. The strategy developed in this work can be used to probe the dynamics of other RNAs in cells and may facilitate the developments of RNA biosensors, drugs and engineering.

Circadian Control of REDOX Reactions in the Macrophage Inflammatory Response

Significance: Macrophages are immune sentinels located throughout the body that function in both the amplification and resolution of the inflammatory response. The circadian clock has emerged as a central regulator of macrophage inflammation. Reduction-oxidation (REDOX) reactions are central to both circadian clock and macrophage function. Recent Advances: Circadian regulation of metabolism controls the macrophage inflammatory response, whereby disruption of the clock causes dysfunctional inflammation. Altering metabolism and reactive oxygen/nitrogen species (RONS) production rescues the inflammatory phenotype of clock-disrupted macrophages.
Critical issues: The circadian clock possesses many layers of regulation. Understanding how REDOX reactions coordinate clock function is critical to uncover the full extent of circadian regulation of macrophage inflammation. We provide insights into how circadian regulation of REDOX affects macrophage pattern recognition receptor signaling, immunometabolism, phagocytosis, and inflammasome activation.
Future directions: Many diseases associated with aberrant macrophage derived inflammation exhibit time of day rhythms in disease symptoms and severity and are sensitive to circadian disruption. Macrophage function is highly dependent on REDOX reactions that signal through RONS. Future studies are needed to evaluate the extent of circadian control of macrophage inflammation, specifically in the context of REDOX signaling.

The health consequences of child marriage: a systematic review of the evidence

Background: Child marriage, defined as marriage before 18 years of age, is a violation of human rights and a marker of gender inequality. Growing attention to this issue on the global development agenda also reflects concerns that it may negatively impact health. We conducted a systematic review to synthesize existing research on the consequences of child marriage on health and to assess the risk of bias in this body of literature.
Methods and findings: We searched databases focused on biomedicine and global health for studies that estimated the effect of marrying before the age of 18 on any physical or mental health outcome or health behaviour. We identified 58 eligible articles, nearly all of which relied on cross-sectional data sources from sub-Saharan Africa or South Asia. The most studied health outcomes were indicators of fertility and fertility control, maternal health care, and intimate partner violence. All studies were at serious to critical risk of bias. Research consistently found that women who marry before the age of 18 begin having children at earlier ages and give birth to a larger number of children when compared to those who marry at 18 or later, but whether these outcomes were desired was not considered.
Across studies, women who married as children were also consistently less likely to give birth in health care facilities or with assistance from skilled providers. Studies also uniformly concluded that child marriage increases the likelihood of experiencing physical violence from an intimate partner. However, research in many other domains, including use of contraception, unwanted pregnancy, and sexual violence came to divergent conclusions and challenge some common narratives regarding child marriage.
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Conclusions: There are many reasons to be concerned about child marriage. However, evidence that child marriage causes the health outcomes described in this review is severely limited. There is more heterogeneity in the results of these studies than is often recognized. For these reasons, greater caution is warranted when discussing the potential impact of child marriage on health. We provide suggestions for avoiding common biases and improving the strength of the evidence on this subject.
Trial registration: The protocol of this systematic review was registered with PROSPERO (CRD42020182652) in May 2020.
Keywords: Child marriage; Social determinants of health; Systematic review.

Magnetic Immobilization and Growth of Nannochloropsis oceanica and Scenedasmus almeriensis

On: By: Brent
Microalgae are used in industrial and pharmaceutical applications. Their performance on biological applications may be improved by their immobilization. This study presents a way of cell immobilization using microalgae carrying magnetic properties. Nannochloropsis oceanica and Scenedasmus almeriensis cells were treated enzymatically (cellulase) and mechanically (glass beads), generating protoplasts as a means of incorporation of magnetic nanoparticles.
Scanning electron microscopy images verified the successful cell wall destruction for both of the examined microalgae cells. Subsequently, protoplasts were transformed with magnetic nanoparticles by a continuous electroporation method and then cultured on a magnetic surface. Regeneration of transformed protoplasts was optimized using various organic carbon and amino acid supplements. Both protoplast preparation methods demonstrated similar efficiency. Casamino acids, as source of amino acids, were the most efficient compound for N. oceanica protoplasts regeneration in enzymatic and mechanical treatment, while for S. almeriensis protoplasts regeneration, fructose, as source of organic carbon, was the most effective. Protoplasts transformation efficiency values with magnetic nanoparticles after enzymatic or mechanical treatments for N. oceanica and S. almeriensis were 17.8% and 10.7%, and 18.6% and 15.7%, respectively. Finally, selected magnetic cells were immobilized and grown on a vertical magnetic surface exposed to light and without any supplement.

Crassaminicella thermophila sp. nov., a moderately thermophilic bacterium isolated from a deep-sea hydrothermal vent chimney and emended description of the genus Crassaminicella

A novel moderately thermophilic, anaerobic, heterotrophic bacterium (strain SY095T) was isolated from a hydrothermal vent chimney located on the Southwest Indian Ridge at a depth of 2730 m. Cells were Gram-stain-positive, motile, straight to slightly curved rods forming terminal endospores. SY095T was grown at 45-60 °C (optimum 50-55 °C), pH 6.0-7.5 (optimum 7.0), and in a salinity of 1-4.5 % (w/v) NaCl (optimum 2.5 %). Substrates utilized by SY095T included fructose, glucose, maltose, N-acetyl glucosamine and tryptone. Casamino acid and amino acids (glutamate, glutamine, lysine, methionine, serine and histidine) were also utilized. The main end products from glucose fermentation were acetate, H2 and CO2. Elemental sulphur, sulphate, thiosulphate, sulphite, fumarate, nitrate, nitrite and Fe(III) were not used as terminal electron acceptors.
The predominant cellular fatty acids were C14 : 0 (60.5%) and C16 : 0 (7.6 %). The main polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, five unidentified phospholipids and two unidentified aminophospholipids. No respiratory quinones were detected. The chromosomal DNA G+C content was 30.8 mol%. The results of phylogenetic analysis of the 16S rRNA gene sequences indicated that SY095T was closely related to Crassaminicella profunda Ra1766HT (95.8 % 16S rRNA gene sequence identity). SY095T exhibited 78.1 % average nucleotide identity (ANI) to C. profunda Ra1766HT. The in silico DNA-DNA hybridization (DDH) value indicated that SY095T shared 22.7 % DNA relatedness with C. profunda Ra1766HT. On the basis of its phenotypic, genotypic and phylogenetic characteristics, SY095T is suggested to represent a novel species of the genus Crassaminicella, for which the name Crassaminicella thermophila sp. nov. is proposed. The type strain is SY095T (=JCM 34213=MCCC 1K04191). An emended description of the genus Crassaminicella is also proposed.

Desulfomarina profundi gen. nov., sp. nov., a novel mesophilic, hydrogen-oxidizing, sulphate-reducing chemolithoautotroph isolated from a deep-sea hydrothermal vent chimney

A novel mesophilic, strictly anaerobic, chemolithoautotrophic sulphate-reducing bacterium, designated strain KT2T, was isolated from a deep-sea hydrothermal vent chimney at the Suiyo Seamount in the Izu-Bonin Arc. Strain KT2T grew at 25-40 °C (optimum 35 °C) and pH 5.5-7.0 (optimum 6.6) in the presence of 25-45 g l-1 NaCl (optimum 30 g l-1). Growth occurred with molecular hydrogen as the electron donor and sulphate, thiosulphate, and sulphite as the electron acceptors. The isolate utilized CO2 as the sole carbon source for chemolithoautotrophic growth on H2. Glycerol, succinate, fumarate, malate, glutamate, or casamino acids could serve as an alternative electron donor in the presence of CO2. Malate, citrate, glutamate, and casamino acids were used as fermentative substrates for weak growth.
The G+C content of genomic DNA was 46.1 %. Phylogenetic analysis of the 16S rRNA gene sequence indicated that strain KT2T is a member of the family Desulfobulbaceae, showing a sequence similarity of 94.3 % with Desulforhopalus singaporensis. Phylogenomic analysis based on concatenated 156 single-copy marker genes confirmed the same topology as the 16S rRNA gene phylogeny. The ANI and AAI values between strain KT2T and related genera of the family Desulfobulbaceae were 65.6-68.6 % and 53.1-62.9 %. Based on the genomic, molecular, and physiological characteristics, strain KT2T represents a novel genus and species within the family Desulfobulbaceae, for which the name Desulfomarina profundi gen. nov., sp. nov. is proposed, with KT2T (=JCM 34118T = DSM 111364T) as the type strain.

Evaluation of immunochromatographic test of Shiga toxin 2e in enrichment cultures of swine edema disease clinical samples

To simplify the diagnosis of swine edema disease, overnight culture supernatants of swine clinical samples were assayed using immunochromatographic test strips we developed previously. Small-intestinal contents, mesenteric lymph nodes, and fecal samples were cultured in casamino acid-yeast extract broth overnight, after which supernatants were loaded onto immunochromatographic test strips to determine whether they could detect Shiga toxin 2e (Stx2e). Among 23 clinical samples in which PCR-identified stx2e-positive E. coli were isolated, samples from seven of ten small-intestinal contents, one of three mesenteric lymph nodes and six of ten fecal samples showed Stx2e-positive reactions in the protein-based immunochromatographic test. Additionally, one small-intestinal content sample, in which stx2e-positive E. coli were not isolated, showed an Stx2e-positive reaction.
Furthermore, the immunochromatographic test results of the samples were associated with the toxin concentration determined by sandwich ELISA and cytotoxicity assay results on Vero cells. The toxin concentration range of the samples with positive and negative reactions were 2.1-196.2 ng/ml and 0-12.8 ng/ml, respectively. The sensitivity and specificity of this immunochromatographic test strip calculated from all clinical samples analyzed in this study were 60.9% and 94.4%, respectively. Our immunochromatographic test strip has strong potential for simple and accurate diagnosis for edema disease by detecting toxin expression, complementing the PCR method.

Staphylococcus aureus Strain-Dependent Biofilm Formation in Bone-Like Environment

Staphylococcus aureus species is an important threat for hospital healthcare because of frequent colonization of indwelling medical devices such as bone and joint prostheses through biofilm formations, leading to therapeutic failure. Furthermore, bacteria within biofilm are less sensitive to the host immune system responses and to potential antibiotic treatments. We suggested that the periprosthetic bone environment is stressful for bacteria, influencing biofilm development. To provide insights into S. aureus biofilm properties of three strains [including one methicillin-resistant S. aureus (MRSA)] under this specific environment, we assessed several parameters related to bone conditions and expected to affect biofilm characteristics. We reported that the three strains harbored different behaviors in response to the lack of oxygen, casamino acids and glucose starvation, and high concentration of magnesium.

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Each strain presented different biofilm biomass and live adherent cells proportion, or matrix production and composition. However, the three strains shared common responses in a bone-like environment: a similar production of extracellular DNA and engagement of the SOS response. This study is a step toward a better understanding of periprosthetic joint infections and highlights targets, which could be common among S. aureus strains and for future antibiofilm strategies.
Keywords: MRSA; MSSA; biofilms; bone microenvironment; prosthetic joint infection.

Differentiation of canine adipose mesenchymal stem cells into insulin-producing cells: comparison of different culture medium compositions

On: By: Brent
The aim of this study was to differentiate canine adipose-derived mesenchymal stem cells (ADMSCs) into insulin-producing cells by using culture media with different compositions to determine the most efficient media. Stem cells isolated from the fat tissues close to the bitch uterus were distributed into 6 groups: (1) Dulbecco’s modified Eagle medium (DMEM)-high glucose (HG), β-mercaptoethanol, and nicotinamide; (2) DMEM–HG, β-mercaptoethanol, nicotinamide, and exendin-4; (3) DMEM–HG, β-mercaptoethanol, nicotinamide, exendin-4, B27, nonessential amino acids, and l-glutamine; (4) DMEM–HG, β-mercaptoethanol, and nicotinamide (for the initial 8-d period), and DMEM–HG, β-mercaptoethanol, nicotinamide, exendin-4, B27, nonessential amino acids, l-glutamine, and basic fibroblast growth factor (for the remaining 8-d period); (5) DMEM–HG and fetal bovine serum; and (6) DMEM-low glucose and fetal bovine serum (standard control group). Adipose-derived mesenchymal stem cells from groups 1 to 5 gradually became round in shape and gathered in clusters.
These changes differed between the groups. In group 3, the cell clusters were apparently more in numbers and gathered as bigger aggregates. Dithizone staining showed that groups 3 and 4 were similar in terms of the mean area of each aggregate stained for insulin. However, only in group 4, the number of insulin aggregates and the total area of aggregates stained were significantly bigger than in the other groups. The mRNA expression of PDX1, BETA2, MafA, and Insulin were also confirmed in all the groups. We conclude that by manipulating the composition of the culture medium it is possible to induce canine ADMSCs into insulin-producing cells, and the 2-staged protocol that was used promoted the best differentiation.

Development of a 3D Tissue-Engineered Skeletal Muscle and Bone Co-culture System.

In vitro 3D tissue-engineered (TE) structures have been shown to better represent in vivo tissue morphology and biochemical pathways than monolayer culture, and are less ethically questionable than animal models. However, to create systems with even greater relevance, multiple integrated tissue systems should be recreated in vitro. In the present study, the effects and conditions most suitable for the co-culture of TE skeletal muscle and bone are investigated. High-glucose Dulbecco’s modified Eagle medium (HG–DMEM) supplemented with 20% fetal bovine serum followed by HG–DMEM with 2% horse serum is found to enable proliferation of both C2C12 muscle precursor cells and TE85 human osteosarcoma cells, fusion of C2C12s into myotubes, as well as an upregulation of RUNX2/CBFa1 in TE85s.
Myotube formation is also evident within indirect contact monolayer cultures. Finally, in 3D co-cultures, TE85 collagen/hydroxyapatite constructs have significantly greater expression of RUNX2/CBFa1 and osteocalcin/BGLAP in the presence of collagen-based C2C12 skeletal muscle constructs; however, fusion within these constructs appears reduced. This work demonstrates the first report of the simultaneous co-culture and differentiation of 3D TE skeletal muscle and bone, and represents a significant step toward a full in vitro 3D musculoskeletal junction model.

Reduced cystathionine-γ-lyase (CSE) expression is involved in high glucose induced MMP14 expression in adipocytes and adipose tissues.

In the present study, we investigate the effect of reduced cystathionine-γ-lyase (CSE) expression in high glucose induced metalloproteinases14 (MMP14) expression in adipocytes and visceral adipose tissues. Diabetic mice were prepared by injections of STZ and the expression of CSE, MMP14 in visceral adipose tissues were determined. Adipocytes were differentiated from 3T3-L1 cells and treated with high glucose (HG), H2S slow-releasing compound GYY4137 or transfected with CSE siRNA. Then the expression of CSE, MMP14 were determined by western blotting. CSE knockout mice were generated by crossing CSE+/- heterozygous mice and given intraperitoneally (i.p.) injections of GYY4137, and then the expression of CSE and MMP14 in visceral adipose tissues were determined by quantitative real-time PCR and western blotting.
The following results were obtained from the study. In adipose tissues of diabetic mice, the mRNA and protein expression of MMP14 increased while the mRNA and protein expression of CSE decreased. In 3T3-L1 adipocytes, both HG DMEM and CSE siRNA transfection increased the mRNA and protein of MMP14. The addition of GYY4137 inhibited HG-induced upregulation of MMP14 expression. In CSE knockout mice, the mRNA and protein expression of MMP14 in adipose tissues increased, which could be inhibited by i.p. injections of GYY4137. In conclusion, high glucose increased the expression of MMP14 in adipocytes and visceral adipose tissues through inhibiting the expression of CSE.

Probucol promotes high glucose-induced proliferation and inhibits apoptosis by reducing reactive oxygen species generation in Müller cells.

To explore the protective effect of probucol on human retinal Müller cells cultured in high glucose.Primary Müller cells from human retinas were cultured in complete DMEM. Third-generation Müller cells were identified using glutamine synthetase (GS) antibody and randomly divided into three groups: normoglycemia (NG, 5.5 mmol/L); hyperglycemia (HG, 30 mmol/L); and hyperglycemia (30 mmol/L) with probucol (10 μmol/L; HGPB). After a 24-h intervention, cell proliferation, apoptosis, and cellular reactive oxygen species (ROS) were measured with a CCK-8 kit, flow cytometry, and DCFH-DA probe, respectively. Kelch-like ECH-associated protein 1 (Keap1), NF-E2-related factor 2 (Nrf2), and glutamate cysteine ligase catalytic subunit (GCLC) protein expression were detected by immunofluorescence staining.
For NG, HG, and HGPB, optical density (OD) values for cell proliferation were 0.98 ± 0.23, 0.58 ± 0.11, and 0.73 ± 0.11; apoptotic rates were 2.79 ± 0.52%, 7.70 ± 0.44%, and 4.00 ± 0.95%; and intracellular ROS were 20.89 ± 5.14, 55.17 ± 14.07, and 26.28 ± 4.73, respectively. Compared to NG, OD was markedly decreased (P < 0.01), apoptosis was increased (P < 0.001), and intracellular ROS level was significantly higher than in HG (P < 0.01). Compared to HG, OD was markedly increased (P < 0.01), apoptosis was meaningfully decreased (P < 0.01), and intracellular ROS level was significantly lower than in HGPB (P < 0.01). GS, Keap1, Nrf2, and GCLC had positive expression. Probucol could inhibit intracellular ROS generation, promote proliferation, and decrease apoptosis of human retinal Müller cells cultured in high glucose. This might also be associated with Keap1/Nrf2/ARE oxidative stress signaling pathway activation.

High Glucose-reduced Apoptosis in Human Breast Cancer Cells Is Mediated by Activation of NF-κB.

Tumor cells rely on glycolysis for their energy supply with the production of lactate even in normoxia condition, which is named aerobic glycolysis or Warburg effect. Therefore, high glucose (HG) concentration provides a favorable condition for increasing proliferation, angiogenesis and decreasing apoptosis, but its molecular mechanisms are still unknown. The objective of this study is to investigate HG condition on tumor cells behavior including proliferation, apoptosis, and an angiogenesis mediator. In this study, MCF-7 derived from human breast adenocarcinoma, were cultured in DMEM with two different concentrations of glucose for 48 h (5.5 mM as normal glucose (NG) condition and 25 mM as HG condition).
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We used Zingiber officinale extraction for the inhibition of NF-κB. Cell proliferation assay was done by direct counting, cell viability by MTT method, bcl-2 by Immunocytochemistry, apoptosis by Hoechst/PI double staining and vascular endothelium growth factor (VEGF) by ELISA.
Results showed that HG increased lactate production, significantly. HG increased cell proliferation, cell viability, VEGF secretion, and bcl-2 expression while it decreased apoptosis. However, when HG was combined with Zingiber officinale extraction, cell proliferation, cell viability, VEGF secretion and bcl-2 expression decreased and apoptosis increased significantly due to inhibition of NF-κB. Results revealed that HG increased cell proliferation, angiogenesis and decreased apoptosis due to activation of NF-κB pathway. Moreover, the probable mechanism of the activation of NF-κB in HG is increasing reactive oxygen species (ROS) in this condition that can activate NF-κB directly.

Microbe-derived antioxidants attenuate cobalt chloride-induced mitochondrial function, autophagy and BNIP3-dependent mitophagy pathways in BRL3A cells

On: By: Brent
Environmental excessive cobalt (Co) exposure increases risks of public health. This study aimed to evaluate the potential mechanism of microbe-derived antioxidants (MA) blend fermented by probiotics in attenuating cobalt chloride (CoCl2)-induced toxicology in buffalo rat liver (BRL3A) cells. Herein, results showed that some phenolic acids increased in MA compared with the samples before fermentation through UHPLC-QTOF-MS analysis. Also, the contents of essential and non-essential amino acids, their derivatives and minerals were rich in MA. The DPPH, O2–, OH– and ABTS+ scavenging ability of MA is comparable to those of vitamin C and better than mitoquinone mesylate (mitoQ).
In vitro cell experiments showed that CoCl2 treatment increased the percentage of apoptosis cells, lactate dehydrogenase and genes involved in glycolysis, increased ATP production and decreased mitochondrial membrane potential, increased genes involved in canonical autophagy process (including initiation, autophagosomes maturation and fusion with lysosome) and BNIP3-dependent mitophagy pathways in BRL3A cells, while MA attenuated CoCl2-induced reactive oxygen species (ROS) production, apoptosis, mitochondrial protein expression and dysfunction, and BNIP3-dependent mitophagy. Collectively, these results provide insights into the role of MA in reversing CoCl2-induced toxicology in BRL3A cells, providing the promising constituents for decreasing Co-induced toxicology in functional foods.

Cardioprotective effects of alantolactone on isoproterenol-induced cardiac injury and cobalt chloride-induced cardiomyocyte injury

Objectives: Alantolactone (AL) is a compound extracted from the roots of Inula Racemosa that has shown beneficial effects in cardiovascular disease. However, the cardioprotective mechanism of AL against hypoxic/ischemic (H/I) injury is still unclear. This research aimed to determine AL’s ability to protect the heart against isoproterenol (ISO)-induced MI injury in vivo and cobalt chloride (CoCl2) induced H/I injury in vitro.
Methods: Electrocardiography (ECG), lactate dehydrogenase (LDH), creatine kinase (CK), and cardiac troponin I (cTnI) assays in addition to histological analysis of the myocardium were used to investigate the effects of AL in vivo. Influences of AL on L-type Ca2+ current (ICa-L) in isolated rat myocytes were observed by the patch-clamp technique. Furthermore, cell viability, apoptosis, oxidative stress injury, mitochondrial membrane potential, and intracellular Ca2+ concentration were examined in vitro.
Results: The results indicated that AL treatment ameliorated the morphological and ECG changes associated with MI, and decreased levels of LDH, CK, and cTnI. Furthermore, pretreatment with AL elevated antioxidant enzyme activity and suppressed ROS production. AL prevented H/I-induced apoptosis, mitochondria damage, and calcium overload while reducing ICa-L in a concentration and time dependent fashion. The 50% inhibiting concentration (IC50) and maximal inhibitory effect (Emax) of AL were 17.29 μmol/L and 57.73 ± 1.05%, respectively.
Conclusion: AL attenuated MI-related injury by reducing oxidative stress, apoptosis, calcium overload, and mitochondria damage. These cardioprotective effects may be related to the direct inhibition of ICa-L.
Keywords: L-type calcium channel currents; alantolactone; calcium influx; cardio-protection; oxidative stress.

Tongxin formula protects H9c2 cardiomyocytes from cobalt chloride-induced hypoxic injury via inhibition of apoptosis

In this study, the effect of the Tongxin formula (TXF) on the apoptosis of H9c2 cardiomyocytes induced by cobalt chloride (CoCl2) was investigated, and the potential mechanism was explored. A hypoxic injury model of H9c2 cardiomyocytes was established using CoCl2. The cell viability was measured using a Cell Counting Kit-8 assay. The lactate dehydrogenase (LDH) release and caspase-3 activity were measured using spectrophotometry. The apoptosis was measured via Annexin V-FITC/PI staining and flow cytometry. The changes in the mitochondrial membrane potential were examined using immunofluorescence microscopy following the loading of JC-1 probes.
The expressions of apoptosis-related proteins and key proteins in the phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) pathway were examined via immunoblotting. The different TXF concentrations studied significantly improved the percentage of viability of cardiomyocytes with hypoxic injury, and the LDH release, apoptotic rate, caspase-3 activity, and levels of cleaved caspase-3 protein were reduced in the injured cells. Additionally, the TXF group had increased mitochondrial membrane potential, upregulated expression of Bcl-2 and p-Akt proteins, and significantly reduced expression of cleaved caspase-3 protein in the cells with hypoxic injury.
Moreover, in the TXF group, the treatment significantly reduced the BAX protein expression, but the difference was not statistically significant compared with the CoCl2 group. In this study, TXF regulated the expression of apoptosis-related proteins, inhibited apoptosis, increased the mitochondrial membrane potential, and alleviated damage to the mitochondrial membrane, thereby protecting the cardiomyocytes from hypoxic injury. The underlying mechanism could be related to activation of the PI3K/Akt signaling pathway and upregulation of the Bcl-2 protein.

The biological effect of cobalt chloride mimetic-hypoxia on nucleus pulposus cells and the comparability with physical hypoxia in vitro

Objective: Nucleus pulposus cells (NPCs) are cells e induce mimetic-hypoxia for NPCs and the comparison withxtracted from the intervertebral disc and are important for research into intervertebral disc degeneration (IVDD). NPCs live in an avascular and relatively hypoxic environment. Cobalt chloride (CoCl2) has been used in many cell studies to mimic hypoxia. The objective of this study was to explore the possibility of using CoCl2 to hypoxia (1% O2) in vitro.
 Materials and methods: Rat nucleus pulposus cells of Passage 3-5 were used in this research. Cell viability, rate of cell apoptosis, ROS (reactive oxygen species) generation, cell migration, extracellular pH and extracellular matrix metabolism were determined to compare the influence of hypoxia (1% O2) and CoCl2 on NPCs.
Results: We found that the effects of CoCl2 on NPCs was dose-dependent. At the proper concentration, CoCl2 could be used to elicit chemical hypoxia for nucleus pulposus cells in vitro and many biological effects, analogous to physical hypoxia (1% O2), could be achieved such as enhanced cell viability, decreased apoptosis and activated extracellular matrix metabolism. On the other hand, CoCl2 mimetic-hypoxia did not affect NPCs glycolysis and migration compared to physical hypoxia. In addition, high concentration of CoCl2 (>200 μM) is harmful to NPCs with high rates of apoptosis and ECM (extracellular matrix) degradation.
 Conclusions: It is feasible and convenient to use CoCl2 to induce chemical mimetic hypoxia for culturing NPCs on the premise of appropriate concentration. But in aspects of cell migration and glycolysis, CoCl2 could not achieve similar results with physical hypoxia. This study may provide a convenient method and enlightenment to induce mimetic-hypoxia for researchers studying NPCs and IVVD.
Keywords: Apoptosis; Cobalt chloride; Extracellular matrix synthesis; Hypoxia; Migration; Nucleus pulposus cells.

Synthesis and structure determination of racemic (Δ/Λ)-tris-(ethyl-enedi-amine)-cobalt(III) trichloride hemi(hexa-aqua-sodium chloride)

The synthesis and crystal structure of the title racemic compound, [Co(C2H8N2)3]Cl3.{[Na(H2O)6]Cl}0.5, are reported. The trivalent cobalt atom, which resides on a crystallographic threefold axis, is chelated by a single ethyl-ene di-amine (en) ligand and yields the tris-chelate [Co(en)3]3+ cation with distorted octa-hedral geometry after the application of crystal symmetry.
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The sodium cation (site symmetry ), has a single water mol-ecule bound to it in the asymmetric unit and yields a distorted, octa-hedrally coordinated hydrated [Na(H2O)6]+ cation after the application of symmetry. One of the chloride ions lies on a general position and the other has site symmetry. An extensive array of C-H⋯O, N-H⋯Cl and O-H⋯Cl hydrogen bonds exists between the ethyl-ene di-amine ligands, the water mol-ecules of hydration, and the anions present, thereby furnishing solid-state stability.
Keywords: crystal structure; enanti­omer; ethyl­ene di­amine; racemate.

Fenton-like reaction, glutathione reduction, and photothermal ablation-built-in hydrogels crosslinked by cupric sulfate for loco-regional cancer therapy

On: By: Brent
Fenton-like reaction-associated chemodynamic therapy (CDT) and hyperthermia-inducing photothermal therapy (PTT)-combined crosslinked hydrogel systems were developed for loco-regional cancer therapy. Cupric sulfate (Cu) has been employed to crosslink the catechol-functionalized hyaluronic acid (HC) polymer-based gel via metal-catechol coordination and covalent bonding of the catechol group (by pH adjustment). Cu can also be used as a hydroxyl radical-generating agent with endogenous H2O2 in cancer cells mediated by Fenton-like reaction and it can reduce intracellular glutathione (GSH) levels leading to the inhibition of reactive oxygen species (ROS) scavenging.
These two strategies can amplify the ROS-initiated CDT efficiency for combating cancer. The Cu-incorporated crosslinked hydrogel structure with pH modulation was appropriate for injectable gel formation via a single syringe. The incorporation of indocyanine green (ICG) into the hydrogel network and near-infrared (NIR) laser irradiation provided a temperature elevation sufficient for induction of hyperthermia in cancer therapy. It is expected that the designed HC/Cu/ICG hydrogel can be used safely and efficiently for local CDT and PTT of breast cancer.

The Role of Plasmapheresis in Treating Lethal Cupric Sulfate Poisoning.

The mortality rate of cupric sulfate is relatively high in contrast to that of other heavy metals. Cases of orally ingested cupric sulfate poisoning are very rare, with a reported half lethal dose of 10 g. Cupric sulfate poisoning leads to gastrointestinal corrosion, intravascular hemolysis, hemolytic anemia, methemoglobinemia and acute renal and hepatic impairment. Without proper and prompt treatment, multiple organ failure and death occur. Here, we present the first report that removal of the excessive intravascular copper ions by plasmapheresis was accompanied by complete recovery.

Preparation of cupric sulfate-based self-emulsifiable nanocomposites and their application to the photothermal therapy of colon adenocarcinoma.

Nanocomposites (NCs) of cupric sulfate monohydrate (CuSO4) were fabricated by hot-melt extrusion (HME) system equipped with twin screws. Micron-sized bulk powder of CuSO4 was dispersed in the mixture of surfactants (Span 80 and Tween 80) and hydrophilic polymer (polyethylene glycol (PEG) 6000) by HME process. Reduction of surface tension by surfactants and homogeneous dispersion in hydrophilic polymer along with HME technique were introduced to prepare CuSO4 NCs.
Dispersion of CuSO4 NCs exhibited approximately 204 nm hydrodynamic size, unimodal size distribution, and positive zeta potential values. Encapsulation of CuSO4 in CuSO4 NCs and the physicochemical interactions between CuSO4 and pharmaceutical excipients were investigated by solid-state studies. Of note, CuSO4 NCs group exhibited higher antiproliferation efficacies, compared with bulk CuSO4, in Caco-2 (human adenocarcinoma) cells at 75 and 100 μg/mL CuSO4 concentrations (p < 0.05). Also, near-infrared laser irradiation to CuSO4 NCs group elevated the antiproliferation efficacies, compared with non-irradiation group, in Caco-2 cells. After intravenous injection in mice, CuSO4 NCs did not show severe in vivo toxicities. Developed CuSO4 NCs can be one of promising candidates of photothermal therapeutic agents for colon cancers.

Ultralow Sample Volume Cupric Sulfate Oxidation Method for the Analysis of Dissolved Lignin.

A novel cupric sulfate (CuSO4) oxidation method was developed to enable the analysis of dissolved lignin phenols in small volumes of open ocean seawater (<200 mL for deep ocean) or river water (<17 mL) samples. Dissolved lignin phenols were isolated by automated reversed-phase (octadecyl) extraction from seawater before oxidation, whereas for freshwater samples, oxidation was performed without prior extraction. Optimized reaction conditions using alkaline CuSO4 at 150 °C effectively limited losses of lignin phenols and suppressed side reactions at 5-100 μg of sample organic carbon.
The method yielded up to ∼33% higher lignin phenol concentrations and 24-36% lower acid/aldehyde ratios of lignin phenols than existing CuO oxidation methods. The microscale design (200 μL reaction volume) resulted in extremely low blanks allowing accurate and precise quantification of lignin phenols. A comparison of silica-based octadecyl-bonded sorbents (C18) with copolymer-type sorbents (PPL) indicated that sorbent type affected concentrations and diagnostic ratios of dissolved lignin phenols. For robust intercomparability of measured lignin phenol concentrations and in consideration of method detection limits for quantification, a constant sample organic carbon content of ∼30 μg C and the addition of 150 μg ascorbic acid-C are recommended. Small sample volumes avoid time-consuming extraction steps in the field, and the simplified oxidation and sample processing procedures make the new method ideal for high-throughput analysis of dissolved lignin phenols in marine and freshwater environments.

Dioxins contamination in the feed additive (feed grade cupric sulfate) tied to chlorine industry.

The sources of polychlorinated dibenzo-p-dioxins/dibenzofurans (PCDD/Fs) found in animal feed additive (feed grade cupric sulfate, CuSO4) were investigated and traced back to the formation of chlorinated organic compounds in the chlor-alkali industry. PCDD/Fs could be transported through the supply chain: hydrochloric acid (HCl) by-produced during formation of chlorinated organic compounds in chlor-alkali industry → spent acid etching solution (acid-SES) generated in printed circuit board production → industrial cupric salt → CuSO4 in animal feed, and finally enter the food chain.
The concentration ranges in HCl and acid-SES were similar, of which the level in acid-SES was also consistent with that in various cupric salt products including CuSO4 based on Cu element content. PCDD/Fs also showed very similar congener profiles in all the sample types. This indicates a probable direct transport pathway of PCDD/Fs into the food chain, which may eventually be exposed to humans through consumption. To date this is the first study in China that systematically reports on the PCDD/Fs transport from industrial pollution sources to industrial processes and finally enters the human food chain.

Synergistic antifungal activity of sodium hypochlorite, hydrogen peroxide, and cupric sulfate against Penicillium digitatum.

Oxidizing compounds such as sodium hypochlorite (NaCIO) and hydrogen peroxide (H2O2) are widely used in food sanitization because of their antimicrobial effects. We applied these compounds and metals to analyze their antifungal activity against Penicillium digitatum, the causal agent of citrus green mold. The MICs were 300 ppm for NaClO and 300 mM for H2O2 when these compounds were individually applied for 2 min to conidia suspensions. To minimize the concentration of these compounds, we developed and standardized a sequential treatment for conidia that resulted in loss of viability on growth plates and loss of infectivity on lemons.
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The in vitro treatment consists of preincubation with 10 ppm of NaClO followed by incubation with 100 mM H2O2 and 6 mM CuSO4 (cupric sulfate). The combination of NaClO and H2O2 in the presence of CuSO4 produces a synergistic effect (fractional inhibitory concentration index of 0.36). The sequential treatment applied in situ on lemon peel 24 h after the fruit was inoculated with conidia produced a significant delay in the fungal infection. The in vitro treatment was effective on both imazalil-sensitive and imazalil-resistant strains of P. digitatum and Geotrichum candidum, the causal agent of citrus sour rot. However, this treatment inhibited 90% of mycelial growth for Penicillium italicum (citrus blue mold). These results indicate that sequential treatment may be useful for postharvest control of citrus fruit diseases.

Polyhydroxylated Cyclopentane β-Amino Acids Derived from d-Mannose and d-Galactose: Synthesis and Protocol for Incorporation into Peptides

On: By: Brent
A stereoselective synthesis of polyhydroxylated cyclopentane β-amino acids from hexoses is reported. The reaction sequence comprises, as key steps, ring-closing metathesis of a polysubstituted diene intermediate followed by the stereoselective aza-Michael functionalization of the resulting cyclopent-1-ene-1-carboxylic acid ester. Examples of synthesis of polysubstituted 2-aminocyclopentanecarboxylic acid derivatives starting from protected d-mannose and d-galactose are presented. A general protocol for the incorporation of these highly functionalized alicyclic β-amino acids into peptides is also reported.

Characterization of the low energy conformations and differential reactivities of D-glucose and D–mannose based oxepines

Carbohydrate-based oxepines are valuable intermediates for the synthesis of septanose carbohydrates. Here we report the characterization of the preferred conformations of D-glucose and D-mannose based oxepines 1 and 2 using computational chemistry and NMR spectroscopy. Monte Carlo conformational searches on 1 and 2 were performed, followed by DFT optimization and single-point energy calculations on the low energy conformations of each oxepine. Coupling constants were computed for all unique conformations at a B3LYP/6-31G(d,p)u+1s level of theory and weighted based on a Boltzmann distribution. These values were then compared to the experimental values collected using 3JH,H values collected from 1H NMR spectra. Information from the MC/DFT approach was then used in a least squares method that correlated DFT calculated and observed 3JH, H coupling constants.
The conformations of 1 and 2 are largely governed by a combination of the rigidifying enol ether element in combination with the reduction of unfavorable interactions. The vinylogous anomeric effect (VAE) emerged as a consequence, rather than a driver of conformations. Oxepine 1 showed greater reactivity in Ferrier rearrangement reactions relative to oxepine 2, in line with its greater %VAE.

ral D–mannose treatment suppresses experimental autoimmune encephalomyelitis via induction of regulatory T cells

D-mannose (D-m) is a glucose epimer found in natural products, especially fruits. In mouse models of diabetes and airway inflammation, D-m supplementation via drinking water attenuated pathology by modifying cellular energy metabolism, leading to the activation of latent transforming growth factor beta (TGF-β), which in turn induced T regulatory cells (Tregs). Given that Tregs are important in controlling neuroinflammation in experimental autoimmune encephalomyelitis (EAE) and likely in multiple sclerosis (MS), we hypothesized that D-m could also suppress EAE.
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We found that D-m delayed disease onset and reduced disease severity in two models of EAE. Importantly, D-m treatment prevented relapses in a relapsing-remitting model of EAE, which mimics the most common clinical manifestation of MS. EAE suppression was accompanied by increased frequency of CD4+FoxP3+ Tregs in the central nervous system, suggesting that EAE suppression resulted from Treg cell induction by D-m. These findings suggest that D-m has the potential to be a safe and low-cost complementary therapy for MS.

Comparison of Rat Primary Midbrain Neurons Cultured in DMEM/F12 and Neurobasal Mediums

On: By: Brent
Introduction: Midbrain dopaminergic neurons are involved in various brain functions, including motor behavior, reinforcement, motivation, learning, and cognition. Primary dopaminergic neurons and also several lines of these cells are extensively used in cell culture studies. Primary dopaminergic neurons prepared from rodents have been cultured in both DMEM/F12 and neurobasal mediums in several studies. However, there is no document reporting the comparison of these two mediums. So in this study, we evaluated the neurons and astroglial cells in primary midbrain neurons from rat embryos cultured in DMEM/F12 and neurobasal mediums.
Methods: Primary mesencephalon cells were prepared from the E14.5 rat embryo. Then they were seeded in two different mediums (Dulbecco’s Modified Eagle Medium/Nutrient Mixture F-12 [DMEM/F12] and neurobasal). On day 3 and day 5, half of the medium was replaced with a fresh medium. On day 7, β3-tubulin-, GFAP (Glial fibrillary acidic protein)- and Tyrosine Hydroxylase TH-positive cells were characterized as neurons, astrocytes, and dopaminergic neurons, respectively, using immunohistochemistry. Furthermore, the morphology of the cells in both mediums was observed under light microscopy on days 1, 3, and 5.
Results: The cells cultured in both mediums were similar under light microscopy regarding the cell number, but in a neurobasal medium, the cells have aggregated and formed clustering structures. Although GFAP-immunoreactive cells were lower in neurobasal compared to DMEM/F12, the number of β3-tubulin- and TH-positive cells in both cultures was the same.
Conclusion: This study’s findings demonstrated that primary midbrain cells from the E14.5 rat embryo could grow in both DMEM/F12 and neurobasal mediums. Therefore, considering the high price of a neurobasal medium, it can be replaced with DMEM/F12 for culturing primary dopaminergic neurons.
Keywords: B27-supplemented neurobasal; DMEM/F12 medium; Dopaminergic neurons; Rat mesencephalon cell culture.

KAv-1 is Better Suited to Chick Fibroblast Culture than DMEM or 199 Media

2021
Cultured cells are a useful resource for poultry scientists, since these cells allow scientists to evaluate biological responses to conditions such as infectious diseases in vitro while mimicking the whole-body response in birds. However avian cell culture requires an optimized basal medium, and there are currently relatively few options for this basal medium (medium 199 and KAv-1).
This means that there is still room for the development of an optimal basal medium for avian cell culture. Here we compare KAv-1 medium, Dulbecco’s modified Eagle medium (DMEM) and medium 199 during the culture of chick fibroblasts and determine that KAv-1 remains the optimal medium for these assays.
Our results show that DNA damage is reduced in fibroblasts cultured in the KAv-1 medium, when compared to both DMEM and Medium 199 and that these cells also display improved growth dynamics in KAv-1 medium when compared to both DMEM and medium 199. To the best of our knowledge, this is the first study to describe a comparative analysis of culture media for avian cells, which would provide useful information for poultry scientists.
Keywords: DNA damage; cell culture; cell growth; cellular senescence; chick fibroblasts; growth medium.

Injectable DMEM-induced phenylboronic acid-modified hyaluronic acid self-crosslinking hydrogel for potential applications in tissue repair

Most of traditional injectable hydrogels based on light curing or enzyme crosslinking are difficult to control the crosslinking time accurately and lack tissue adhesion, which leads to difficult clinical application and poor tissue repair effect.
In this study, a novel injectable DMEM (Dulbecco’s Modified Eagle’s Medium)-induced phenylboronic acid-modified hyaluronic acid self-crosslinking hydrogel was designed and prepared by combining the phenylboronic acid and a diol on hyaluronic acid as the main network, in which dynamically reversible phenylboronic acid esters imparted good self-healing properties and tissue adhesion properties to the hydrogels.
Cell medium that induced the formation of the hydrogel could simulate the pH of the physiological environment and provide uniform nutrients for the encapsulated cells. In addition, in vitro cell experiments indicated that the DMEM-induced phenylboronic acid-modified hyaluronic acid self-crosslinking hydrogel was capable of supporting cell loading and proliferation, thus being a promising candidate for tissue repair materials.

Type of culture medium determines properties of cultivated retinal endothelial cells: induction of substantial phenotypic conversion by standard DMEM

Contradictory behavior of microvascular retinal endothelial cells (REC) – a reliable in vitro model to study retinal diseases – have recently been reported which might result from cultivating the cells in standard DMEM not optimized for this cell type. Therefore, we studied DMEM’s effects on phenotype and behavior of immortalized bovine REC. Cells were cultivated in endothelial cell growth medium (ECGM) until a confluent monolayer was reached and then further kept for 1-4 days in ECGM, DMEM, or mixes thereof all supplemented with 5% fetal bovine serum, endothelial cell growth supplement, 90 μg/ml heparin, and 100 nM hydrocortisone.
Within hours of cultivation in DMEM, the cell index – measured to assess the cell layer’s barrier function – dropped to ~5% of the initial value and only slowly recovered, not only accompanied by stronger expression of HSP70 mRNA and secretion of interleukin-6, but also by lower expressions of tight junction proteins claudin-5, claudin-1 or of the marker of cell type conversion caveolin-1.
Altered subcellular localizations of EC-typic claudin-5, vascular endothelial cadherin and von Willebrand factor were also observed. Taken together, all experiments with (retinal) EC cultivated in common DMEM need to be interpreted very cautiously and should at least include phenotypic validation.
Keywords: Barrier dysfunction; Cell culture medium; Cell index measurements; Cell type conversion; DMEM; ECGM-MV; Endothelial cells; Paracellular flow; Phenotype; Retinal endothelial cells; Transcellular transport.

Assessing the protective effects of different surface coatings on NaYF 4:Yb 3+, Er 3+ upconverting nanoparticles in buffer and DMEM

We studied the dissolution behavior of β NaYF4:Yb(20%), Er(2%) UCNP of two different sizes in biologically relevant media i.e., water (neutral pH), phosphate buffered saline (PBS), and Dulbecco’s modified Eagle medium (DMEM) at different temperatures and particle concentrations. Special emphasis was dedicated to assess the influence of different surface functionalizations, particularly the potential of mesoporous and microporous silica shells of different thicknesses for UCNP stabilization and protection.
Dissolution was quantified electrochemically using a fluoride ion selective electrode (ISE) and by inductively coupled plasma optical emission spectrometry (ICP OES). In addition, dissolution was monitored fluorometrically. These experiments revealed that a thick microporous silica shell drastically decreased dissolution.
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Our results also underline the critical influence of the chemical composition of the aqueous environment on UCNP dissolution. In DMEM, we observed the formation of a layer of adsorbed molecules on the UCNP surface that protected the UCNP from dissolution and enhanced their fluorescence.Examination of this layer by X-ray photoelectron spectroscopy (XPS) and mass spectrometry (MS) suggested that mainly phenylalanine, lysine, and glucose are adsorbed from DMEM. These findings should be considered in the future for cellular toxicity studies with UCNP and other nanoparticles and the design of new biocompatible surface coatings.

Disability & Diversity studies as a professional basis for diversity-aware education and training in medicine.

Disability & Diversity studies as a professional basis for diversity-aware education and training in medicine.
On: By: Brent
The “Disability and Diversity Studies” (DDS) are research fields which, much like social work, take care of social inclusion and exclusion processes. Dimensions of incapacity and variety can result in disadvantages and inequalities in the person life and social coexistence of individuals. The DDS look at these inequalities and establish intersectional relationships between variety classes.
The idea of intersectionality opens up the view of the restriction of diversities, which might result in the intensification of inequalities and a number of discriminations: e. g., in the case of being a girl and member of an ethnic minority. The start line is subsequently not the distinction class per se, however the intersection of several classes [5]. This information of categorizing classification and exclusion in their intersectionality is key for the dissolution of social, societal, political and financial inequality.
The DDS bachelor’s program on the Carinthia University of Applied Sciences, which mixes Disability and Diversity Studies, focuses on these research areas and develops sensible solutions. In medical education and training, too, it’s essential that lecturers and college students, however also sufferers, recognise the advanced interrelationships of divergences in medical follow and the ensuing stigma that should be eliminated.
The DDS can serve as a basis for taking these interrelationships into consideration, for incorporating inventive approaches to solutions and a diversity-sensitive perspective into the doctor-patient relationship and medical remedy. For instance, the primary and so far solely World Report on Disability from 2011 famous a nonetheless present destructive infiltration of doctor-patient-interactions by means of stigmatization of persons with disabilities and deviations.
Misunderstandings, lack of awareness and improper presettings can endanger the remedy [32]. In order to create the framework circumstances for an acceptable consideration of variety and incapacity in this system, it’s essential to impart six core competencies to potential physicians [20]:
If potential, this could all the time be designed in the respective training courses of all well being care professions in a patient-centred method, throughout all occupational teams and beneath the premise “nothing for us with out us” [1]. This corresponds to the rules of Disability Studies.
Disability & Diversity studies as a professional basis for diversity-aware education and training in medicine.
Disability & Diversity studies as a professional basis for diversity-aware education and training in drugs.