Cell Structural Proteins Research Articles

Mitigation of testicular damage induced by DMBA/TPA through Murraya koenigii (L.) Spreng.: Insights into antioxidant and anti-apoptotic mechanisms

Abstract BACKGROUND: 7,12-Dimethylbenz[a]anthracene (DMBA) is a known mutagen, teratogen, and toxicant besides being a significant factor responsible for skin carcinogenesis. During carcinogenesis, DMBA and its metabolites produce excessive free radicals and oxidative stress. This oxidative stress further targets various cell organelles. Consequences of the damage by DMBA also influence vital organs, such as the brain, liver, and testis, with complex reactions, such as an increase in mutation, modification in cell membranes, structural proteins, metabolic enzymes, and signaling proteins. Therefore, this study evaluated the possible role of hydroethanolic Murraya koenigii (L.) Spreng. leaves extract (HEMKLE) upon damages incurred in testis tissues against DMBA/12-O-tetradecanoyl phorbol-13-acetate (TPA) in skin tumor-bearing mice. METHODS: Forty male LACA mice were segregated into four groups: control, DMBA/TPA, HEMKLE, and HEMKLE + DMBA/TPA. Skin tumors were induced by DMBA (500 nmol/100 μL of acetone) and TPA (1.7 nmol/100 μL of acetone; applied topically). RESULTS: The protective response of HEMKLE (200 mg/kg body weight) to testicular damage during skin tumorigenesis was apparent by recovery of endogenous antioxidant enzymes, along with histoarchitecture in the HEMKLE + DMBA/TPA group contrasted with the DMBA/TPA group. Furthermore, in the HEMKLE + DMBA/TPA group, reduced messenger ribonucleic acid and protein expressions of proapoptotic genes (caspase-9 and caspase-3) and enhancement in Bcl-2 were observed, proposing the anti-apoptotic potential of HEMKLE in testicular tissues. CONCLUSION: HEMKLE could be used as a nutraceutical or antioxidant drug to protect the body from testicular insults.

Proteomic Analysis of Salivary Secretions from the Tea Green Leafhopper, Empoasca flavescens Fabrecius.

Saliva plays a crucial role in shaping the compatibility of piercing-sucking insects with their host plants. Understanding the complex composition of leafhopper saliva is important for developing effective and eco-friendly control strategies for the tea green leafhopper, Empoasca flavescens Fabrecius, a major piercing-sucking pest in Chinese tea plantations. This study explored the saliva proteins of tea green leafhopper adults using a custom collection device, consisting of two layers of Parafilm stretched over a sucrose diet. A total of 152 proteins were identified using liquid chromatography-tandem mass spectrometry (LC-MS/MS) following the filter-aided sample preparation (FASP). These proteins were categorized into six groups based on their functions, including enzymes, transport proteins, regulatory proteins, cell structure proteins, other proteins, and unknown proteins. Bioinformatics analyses predicted 16 secreted proteins, which were successfully cloned and transcriptionally analyzed across various tissues and developmental stages. Genes encoding putative salivary secretory proteins, including Efmucin1, EfOBP1, EfOBP2, EfOBP3, Efmucin2, low-density lipoprotein receptor-related protein (EfLRP), EFVg1, and EFVg2, exhibited high expressions in salivary gland (SG) tissues and feeding-associated expressions at different developmental stages. These findings shed light on the potential elicitors or effectors mediating the leafhopper feeding and defense responses in tea plants, providing insights into the coevolution of tea plants and leafhoppers. The study's conclusions open avenues for the development of innovative leafhopper control technologies that reduce the reliance on pesticides in the tea industry.

Utilizing Two Populations Derived from Tropical Maize for Genome-Wide Association Analysis of Banded Leaf and Sheath Blight Resistance

Banded leaf and sheath blight (BLSB) in maize is a soil-borne fungal disease caused by Rhizoctonia solani Kühn, resulting in significant yield losses. Investigating the genes responsible for regulating resistance to BLSB is crucial for yield enhancement. In this study, a multiparent maize population was developed, comprising two recombinant inbred line (RIL) populations totaling 442 F8RILs. The populations were generated by crossing two tropical inbred lines, CML444 and NK40-1, known for their BLSB resistance, as female parents, with the high-yielding but BLSB-susceptible inbred line Ye107 serving as the common male parent. Subsequently, we utilized 562,212 high-quality single nucleotide polymorphisms (SNPs) generated through genotyping-by-sequencing (GBS) for a comprehensive genome-wide association study (GWAS) aimed at identifying genes responsible for BLSB resistance. The objectives of this study were to (1) identify SNPs associated with BLSB resistance through genome-wide association analyses, (2) explore candidate genes regulating BLSB resistance in maize, and (3) investigate pathways involved in BLSB resistance and discover key candidate genes through Gene Ontology (GO) analysis. The GWAS analysis revealed nineteen SNPs significantly associated with BLSB that were consistently identified across four environments in the GWAS, with phenotypic variation explained (PVE) ranging from 2.48% to 11.71%. Screening a 40 kb region upstream and downstream of the significant SNPs revealed several potential candidate genes. By integrating information from maize GDB and the NCBI, we identified five novel candidate genes, namely, Zm00001d009723, Zm00001d009975, Zm00001d009566, Zm00001d009567, located on chromosome 8, and Zm00001d026376, on chromosome 10, related to BLSB resistance. These candidate genes exhibit association with various aspects, including maize cell membrane proteins and cell immune proteins, as well as connections to cell metabolism, transport, transcriptional regulation, and structural proteins. These proteins and biochemical processes play crucial roles in maize defense against BLSB. When Rhizoctonia solani invades maize plants, it induces the expression of genes encoding specific proteins and regulates corresponding metabolic pathways to thwart the invasion of this fungus. The present study significantly contributes to our understanding of the genetic basis of BLSB resistance in maize, offering valuable insights into novel candidate genes that could be instrumental in future breeding efforts to develop maize varieties with enhanced BLSB resistance.

Application of 4 × 44 Oligo Microarray to Transcriptomic Analysis of Immune Response in Rainbow Trout Infected with Aeromonas salmonicida

Rainbow trout, one of the most economically important aquaculture fish species worldwide, is affected by the pathogenic bacteria A. salmonicida, which causes furunculosis outbreaks, leading to huge economic losses. In this study, an oligonucleotide microarray was applied to identify transcriptional changes in the skin of rainbow trout individuals in response to a bacterial infection. Overall, 656 and 434 differentially expressed genes (DEGs) were identified at 2 and 6 days after a bacterial challenge (dpi), respectively. A comparison of moribund (2 dpi) and survivor fish (6 dpi) revealed 169 DEGs. Between these were many genes involved in immune response, including lysozymes, pattern recognition receptors (c-type lectins), antimicrobial peptides (cathelicidin and hepcidin), acute-phase proteins (serum amyloids and haptoglobin), complement cascade proteins (c3, c4, c6 and c7), interleukins (il11 and il1b) and chemokines (ccl19 and cxcl8). Alterations of leptin, eicosanoids and prostaglandins have been found, which suggest metabolic remodeling in conjunction with immune response. Further, the regulation of programmed cell death genes (caspase 8, bcl2 apoptosis regulator, nfkb inhibitor alpha and heme oxygenase) and structural proteins (collagens, myosins, keratins and metalloproteinases) was observed. This study provides, for the first time, a gene expression analysis of rainbow trout skin in response to A. salmonicida infection, revealing the complexity of defense strategies in response to furunculosis.

Insight into the antibacterial activity and mechanism of allicin and antioxidant of bamboo leaves against Shewanella putrefaciens

SummaryThe antibacterial activity and mechanism of allicin (All) and antioxidant of bamboo leaves (AOB) against Shewanella putrefaciens was studied. The minimum inhibitory concentration (MIC) and growth curve were used to evaluate the antibacterial activity of combining All and AOB against S. putrefaciens. The inactivation mechanism was reflected by the changes in Na+/K+‐ATPase activity, confocal laser scanning microscopy (CLSM), and the leakage of nucleic acid and protein. Scanning electron microscopy (SEM) was used to observe the morphology of bacterial cells and crystal violet biofilm reflected the ability to inhibit the formation of biofilm. Results showed that after being treated by All and AOB, S. putrefaciens was effectively inactivated, and the combined treatment had the strongest inactivation effect. The cell wall and cell membrane were destroyed by All and AOB, resulting in the release of cell contents. The results of Na+/K+‐ATPase activity demonstrated that the membrane permeability could be changed, and the cell structural proteins could be destroyed by All and AOB. SEM observation further proved the effects of All and AOB on bacterial morphology destructing. Therefore, the combination of All and AOB can be used as an efficient method for the preservation of aquatic products.

Bactericidal Efficacy and Mechanisms of Non-Electrolytic Slightly Acidic Hypochlorous Water on Pseudomonas fragi and Pseudomonas fluorescens.

Chilled pork is frequently contaminated with Pseudomonas fragi and Pseudomonas fluorescens. In this study, the bactericidal efficacy and mechanisms of non-electrolytic slightly acidic hypochlorous water (NE-SAHW) against two strains of these two species were evaluated. The results showed that the antibacterial efficacy of NE-SAHW was positively correlated with the concentration level of NE-SAHW and negatively correlated with the initial populations of the strains. The strains of small populations were completely inhibited when provided with each level of NE-SAHW. The killed cells of P. fragi were 0.94, 1.39, 4.02, and 5.60 log10 CFU/mL, respectively, and of P. fluorescens they were 1.21, 1.52, 4.14, and 5.74 log10 CFU/mL, respectively, when the initial populations of the strains were at high levels (about 7 log10 CFU/mL). Both strains were completely killed within 12 s with the available chlorine concentration (ACC) of 50 mg/L of NE-SAHW. Morphological changes in both cells were observed by using a Scanning Electron Microscope (SEM) and it was discovered that the cell membranes were damaged, which led to the leakage of the intracellular substances, including K+, nucleic acid, and protein. In terms of the Fourier Transform Infrared Spectroscopy (FTIR) results, NE-SAHW destroyed the structures of membrane proteins and cell structure proteins, and influenced the composition of polysaccharides. The bacteria were definitely dead after treatment by NE-SAHW compared to the control according to the results of flow cytometry. These results demonstrated the potential bactericidal property of NE-SAHW when applied to the meat and other food sterilization industries.

Detection of Novel Pathogenic Variants in Two Families with Recurrent Fetal Congenital Heart Defects.

Congenital heart disease (CHD) is the most common birth defect with strong genetic heterogeneity. To date, about 400 genes have been linked to CHD, including cell signaling molecules, transcription factors, and structural proteins that are important for heart development. Genetic analysis of CHD cases is crucial for clinical management and etiological analysis. Whole-exome sequencing (WES) was performed to identify the genetic variants in two independent CHD cases with DNA samples from fetuses and their parents, followed by the exclusion of aneuploidy and large copy number variations (CNVs). The WES results were verified by Sanger sequencing. In family A, a compound heterozygous variation in PLD1 gene consisting of c.1132dupA (p.I378fs) and c.1171C>T (p.R391C) was identified in the fetus. The two variants were inherited from the father (c.1132dupA) and the mother (c.1171C>T), respectively. In family B, a hemizygous variant ZIC3: c.861delG (p.G289Afs*119) was identified in the fetus, which was inherited from the heterozygous mother. We further confirmed that these variants PLD1: c.1132dupA and ZIC3: c.861delG were novel. The findings in our study identified novel variants to the mutation spectrum of CHD and provided reliable evidence for the recurrent risk and reproductive care options to the affected families. Our study also demonstrates that WES has considerable prospects of clinical application in prenatal diagnosis.

Epigenetics and stroke: role of DNA methylation and effect of aging on blood–brain barrier recovery

Incomplete recovery of blood–brain barrier (BBB) function contributes to stroke outcomes. How the BBB recovers after stroke remains largely unknown. Emerging evidence suggests that epigenetic factors play a significant role in regulating post-stroke BBB recovery. This study aimed to evaluate the epigenetic and transcriptional profile of cerebral microvessels after thromboembolic (TE) stroke to define potential causes of limited BBB recovery. RNA-sequencing and reduced representation bisulfite sequencing (RRBS) analyses were performed using microvessels isolated from young (6 months) and old (18 months) mice seven days poststroke compared to age-matched sham controls. DNA methylation profiling of poststroke brain microvessels revealed 11,287 differentially methylated regions (DMR) in old and 9818 DMR in young mice, corresponding to annotated genes. These DMR were enriched in genes encoding cell structural proteins (e.g., cell junction, and cell polarity, actin cytoskeleton, extracellular matrix), transporters and channels (e.g., potassium transmembrane transporter, organic anion and inorganic cation transporters, calcium ion transport), and proteins involved in endothelial cell processes (e.g., angiogenesis/vasculogenesis, cell signaling and transcription regulation). Integrated analysis of methylation and RNA sequencing identified changes in cell junctions (occludin), actin remodeling (ezrin) as well as signaling pathways like Rho GTPase (RhoA and Cdc42ep4). Aging as a hub of aberrant methylation affected BBB recovery processes by profound alterations (hypermethylation and repression) in structural protein expression (e.g., claudin-5) as well as activation of a set of genes involved in endothelial to mesenchymal transformation (e.g., Sox9, Snai1), repression of angiogenesis and epigenetic regulation. These findings revealed that DNA methylation plays an important role in regulating BBB repair after stroke, through regulating processes associated with BBB restoration and prevalently with processes enhancing BBB injury.

ITRAQ-mediated analysis of the relationship between proteomic changes and yak longissimus lumborum tenderness over the course of postmortem storage

To identify differentially expressed proteins associated with energy metabolism and tenderness during the postmortem aging of yak longissimus lumborum muscle samples, we collected tissue samples from yaks raised at different altitudes. At 12 h post-slaughter, we identified 290 differentially expressed proteins (DEPs) in these samples, whereas 436 such DEPs were detected after 72 h. Identified DEPs were clustered into four main functional categories: cell structural proteins, glycogen metabolic proteins, energy reserve metabolic proteins, and cellular polysaccharide metabolic proteins. Further bioinformatics analysis revealed that these proteins were associated with carbon metabolism, glycolysis, and the biosynthesis of amino acids. Our functional insights regarding these identified proteins contribute to a more detailed molecular understanding of the processes of energy metabolism in yak muscle tissue, and represent a valuable resource for future investigations.

Cathepsin L mediates glomerular endothelial cell injury by cleavaging complement C3 in trichloroethylene-sensitized mice

Objective: To observe the expressions of complement 3 (C3) and endothelial cell injury-associated proteins before and after cathepsin L (CTSL) blockade in renal injury of trichloroethylene (TCE) -sensitized mice. Methods: In June 2018, 41 SPF female BALB/c mice were divided respectively into blank control group (n=5) , vehicle control group (n=5) , TCE group (n=15) and TCE+CTSLi group (n=16) to establish trichloroethylene-sensitized mice model by pretreating the mice with intraperitoneal injection of CTSL inhibitor (CTSLi) and using TCE for the first and last challenge. According to the skin sensitization score, the mice were divided into positive group and negative group. 72 hours after the last challenge, the renal function indexes of the mice were detected, the pathological changes of mice kidneys were observed, and the glomerular C3 and endothelial cell damage-related proteins [vascular cell adhesion molecule 1 (VCAM-1) , tight junction protein 5 (Claudin-5) and Syndecan-1] expression levels were detected. Results: The sensitization rates of mice in TCE group and TCE+CTSLi group were 53.3% (8/15) and 50.0% (8/16) , respectively, and there was no significant difference between the two groups (P>0.05) . Compared with vehicle control group and the corresponding TCE negative group, the serum creatinine (CRE) and blood urea nitrogen (BUN) levels of mice in the TCE positive group was increased, while the TCE positive group were higher than the TCE+CTSLi positive group (P<0.05) . Pathological examination showed obvious vacuolar degeneration and cellular edema in the mice kidney of the TCE positive group. In the TCE+CTSLi positive group, the above pathological damage was significantly improved. Immunohistochemical results showed that the expression of glomerular C3 fragment and VCAM-1 in TCE positive group were significantly higher than that of the vehicle control and TCE negative group (P<0.05) , while TCE+CTSLi positive group was significantly lower than that of TCE positive group (P<0.05) . Western blot test results showed that the relative expression levels of Claudin-5 and Syndecan-1 protein in the mice glomeruli of TCE positive group were significantly lower than those in the vehicle control group and TCE negative group (P<0.05) . Compared with the TCE positive group, the Claudin-5 protein was increased in the kidney of the TCE+CTSLi positive group, but the difference was not statistically significant (P>0.05) , while the Syndecan-1 protein was significantly increased in the TCE+CTSLi positive group (P<0.05) . Conclusion: CTSL may mediate the glomerular structural damage by cutting complement C3, activating the complement system, damaging endothelial cell structural protein Syndecan-1 and overexpressing adhesion molecule VCAM-1 in TCE-sensitized mice. Inhibiting the expression of CTSL may be an effective way to protect the glomerular integrity of structure and function in pharmacology.

Gas plasma-spurred wound healing is accompanied by regulation of focal adhesion, matrix remodeling, and tissue oxygenation

In response to injury, efficient migration of skin cells to rapidly close the wound and restore barrier function requires a range of coordinated processes in cell spreading and migration. Gas plasma technology produces therapeutic reactive species that promote skin regeneration by driving proliferation and angiogenesis. However, the underlying molecular mechanisms regulating gas plasma-aided cell adhesion and matrix remodeling essential for wound closure remain elusive. Here, we combined in vitro analyses in primary dermal fibroblasts isolated from murine skin with in vivo studies in a murine wound model to demonstrate that gas plasma treatment changed phosphorylation of signaling molecules such as focal adhesion kinase and paxillin α in adhesion-associated complexes. In addition to cell spreading and migration, gas plasma exposure affected cell surface adhesion receptors (e.g., integrinα5β1, syndecan 4), structural proteins (e.g., vinculin, talin, actin), and transcription of genes associated with differentiation markers of fibroblasts-to-myofibroblasts and epithelial-to-mesenchymal transition, cellular protrusions, fibronectin fibrillogenesis, matrix metabolism, and matrix metalloproteinase activity. Finally, we documented that gas plasma exposure increased tissue oxygenation and skin perfusion during ROS-driven wound healing. Altogether, these results provide critical insights into the molecular machinery of gas plasma-assisted wound healing mechanisms.

Egg proteomic characterization of Seriola rivoliana in captivity

Continuous and sustained production of good quality eggs and larvae is required to have economically viable and ecologically sustainable rearing of Seriola rivoliana. Nonetheless, the complete production cycle of these species has been challenging to achieve due to high mortality during embryonic and larval stages. The objective of the project was to study the expression of the proteins involved in the embryonic process of almaco jack. Proteomic characterization of fertilized eggs was performed using two-dimensional electrophoresis and mass spectrometry. Different vitellogenin proteins A, B, C and Ab, β-actin, peroxiredoxin, superoxide dismutase 1, alpha subunit proteasome, and keratin II were identified to their functions related to embryonic development, energy metabolism, protein synthesis, cell structure, cytoskeleton, and antioxidant proteins with defense enzymatic activity.

Endocrinology and immunology of acne: Two sides of the same coin.

Current experimental research on acne pathophysiology has revealed a more complicated background than the classically reported four-factor aetiology. Cells of the pilosebaceous unit, which represent the template for the development of acne lesions, seem to be parallelly affected by endocrinological/metabolic factors as well as inflammatory/immunological ones that cooperate in sebocyte differentiation and lipogenesis. Indeed, the unique programme of sebocyte terminal differentiation and death, the so called holocrine secretion, is influenced by inflammatory and metabolic (lipid) signalling with common denominator the selective regulation of peroxisome proliferator-activated receptors. Autophagy provides substrates for energy generation and biosynthesis of new cell structure proteins contributing to the normally increased sebaceous gland metabolic functions, which are also regulated by extracellular calcium signalling, essential lipids and hormones. The ultimate differentiation product of human sebocytes, sebum, co-regulates the inflammatory sebocyte status. Sebum composition is controlled among others by Propionibacterium acnes and other bacteria, sexual hormones, neuropeptides, endogenous opioids and environmental agents, which may function as endocrine disruptors. Diet may also be an important source of substrates for the synthesis of pro-inflammatory and anti-inflammatory sebaceous lipids. Sebum changes might induce inflammation and initiate underlying immune mechanisms leading to acne lesions. Current new therapeutic efforts on acne concentrate on anti-inflammatory/immunologically active concepts, which are able to regulate sebaceous lipogenesis. At last, current molecular studies based on published molecular data sets confirmed the major role of inflammation in acne development.

Nano-scientific Application of Atomic Force Microscopy in Pathology: from Molecules to Tissues.

The advantages of atomic force microscopy (AFM) in biological research are its high imaging resolution, sensitivity, and ability to operate in physiological conditions. Over the past decades, rigorous studies have been performed to determine the potential applications of AFM techniques in disease diagnosis and prognosis. Many pathological conditions are accompanied by alterations in the morphology, adhesion properties, mechanical compliances, and molecular composition of cells and tissues. The accurate determination of such alterations can be utilized as a diagnostic and prognostic marker. Alteration in cell morphology represents changes in cell structure and membrane proteins induced by pathologic progression of diseases. Mechanical compliances are also modulated by the active rearrangements of cytoskeleton or extracellular matrix triggered by disease pathogenesis. In addition, adhesion is a critical step in the progression of many diseases including infectious and neurodegenerative diseases. Recent advances in AFM techniques have demonstrated their ability to obtain molecular composition as well as topographic information. The quantitative characterization of molecular alteration in biological specimens in terms of disease progression provides a new avenue to understand the underlying mechanisms of disease onset and progression. In this review, we have highlighted the application of diverse AFM techniques in pathological investigations.

Investigation of tear protein composition in healthy and dry eye subjects using trapped ion mobility spectrometry coupled with quadrupole time of flight

AbstractPurposeProteomic composition of tears is not yet fully characterized and has a great potential to be used as source for discovery of disease‐specific protein biomarkers. The present work is a preliminary evaluation of tear proteome using timsTOF Pro®, a new highly sensitive mass spectrometry technology.Material and MethodsThree tear samples from 3 healthy subjects (HS) and 2 from a dry eye (DE) patient were collected using plastic micropipettes and Schirmer’s strips respectively. After protein elution from the strips, all samples were treated under the same conditions. Proteins were reduced and alkylated then digested in solution with LysC and Trypsin prior to nanoLC‐MS/MS analysis (nanoElute and timsTOF Pro®, Bruker). Protein identification was performed with Xtandem.ResultsIn total, 1191 proteins were identified in both analytical groups with at least 1 specific peptide: 446 proteins in HS and 1034 proteins in the DE; 802 proteins were exclusively identified in DE (e.g. A2M, HPX, ALDH3A1, PKM, MYH14, ALDH3A1) while 157 were expressed only in controls (e.g. PLOD1, DNAJC3, GOLM1, RNASE4, SDF4, HYOU1). In DE cell structural associated proteins as well as proteins highly correlated with inflammatory and apoptotic processes were detected. Finally, among the proteins that are common to both groups 122 were upregulated in DE vs HS (e.g. ALB, TF, ENO1, ANXA1, GSTP1, S100A9) while 88 were downregulated (e.g. LTF, LCN1, LPO, CST4, LACRT, AZGP1). The upregulated proteins are associated with a blood‐lachrymal barrier failure, inflammatory &amp; immune responses while the downregulated ones sign a lachrymal gland dysfunction.ConclusionsDifferences in the protein expression between HS and DE arise due to the pathological conditions but also due to the different sampling methods used. This preliminary study could offer a roadmap for designing further studies for the in‐depth characterization of the tear protein profile expression under physiological and pathological conditions.

Behavioural and chronic toxicity of fullerene to Daphnia magna: Mechanisms revealed by transcriptomic analysis

Extensive application of fullerene nanoparticles (nC60) leads to potential environmental pollution. The acute toxic effects of nC60 have been largely investigated, but studies of behavioural and chronic toxicity at sublethal doses are still rare and the underlying molecular mechanisms remain unknown. The present study investigated behavioural and chronic toxicity of nC60 to Daphnia magna. The results showed that, in response to nC60 exposure, hopping, heartbeat frequencies and feeding ability of D. magna decreased significantly, displaying negative relationship with exposure time and dose. Chronic treatments with 0.1 mg/L or 1 mg/L nC60 for 21 days significantly reduced survival and reproduction of D. magna. These harmful effects suggested negative impacts of nC60 on aquatic ecosystems. Moreover, transcriptome analysis showed that the behavioural and chronic toxicity of nC60 to D. magna might be related to physiological functions such as cell structural repair, protein degradation, energy metabolism and reproduction. We found that nC60 accumulated in guts of D. magna, which should be responsible for the decrease of food ingestion and consequently inhibiting energy intake. Deficiency of energy not only affects behaviours but also declines reproduction in D. magna. Overall, this is the first study comprehensively considered the behavioral and chronic toxicity of nC60 to aquatic organism. The results should be helpful to better understand the ecological consequences of C60 released into water environments.

Tumour-treating fields (TTFields): Investigations on the mechanism of action by electromagnetic exposure of cells in telophase/cytokinesis

Tumour-treating fields (TTFields) use alternating electric fields which interfere with dividing cells, thereby reducing tumour growth. Previous reports suggest that electrical forces on cell structure proteins interfered with the chromosome separation during mitosis and induced apoptosis. In the present report we evaluate electromagnetic exposure of cells in telophase/cytokinesis in order to further analyse the mechanism of action on cells. We performed numerical electromagnetic simulations to analyse the field distribution in a cell during different mitotic phases. Based thereon, we developed an electric lumped element model of the mitotic cell. Both the electromagnetic simulation and the lumped element model predict a local increase of the specific absorption rate (SAR) as a measure of the electromagnetically induced power absorption density at the mitotic furrow which may help to explain the anti-proliferative effect. In accordance with other reports, cell culture experiments confirmed that TTFields reduce the proliferation of different glioma cell lines in a field strength- and frequency-dependent manner. Furthermore, we found an additional dependence on the commutation time of the electrical fields. The report gives new insights into TTFields’ anti-proliferative effect on tumours, which could help to improve future TTFields application systems.

The role of FHL2 in wound healing and inflammation.

The 4-and-a-half LIM domain protein 2 (FHL2) is a multifunctional adaptor protein that can interact with cell surface receptors, cytosolic adaptor and structural proteins, kinases, and nuclear transcription factors. It is involved in numerous functional activities, including the epithelial-mesenchymal transition, cell proliferation, apoptosis, adhesion, migration, structural stability, and gene expression. Despite this, FHL2-knockout (KO) mice are viable and fertile with no obvious abnormalities, rather suggesting a high capacity for fine-tuning adjustment and functional redundancy of FHL2. Indeed, challenging FHL2-KO cells or mice provided numerous evidences for the great functional significance of FHL2. In recent years, several reviews have been published describing the high capacity of FHL2 to bind diverse proteins as well as the versatile functions of FHL2, emphasizing in particular its role in cardiovascular diseases and carcinogenesis. Here, we view the function of FHL2 from a different perspective. We summarize the published data demonstrating the impact of FHL2 on wound healing and inflammation. FHL2 seems to be involved in numerous steps of these extremely complex and multidirectional but tightly regulated tissue remodeling processes, supporting tissue repair and coordinating inflammation. Deficiency of FHL2 not only slows down ongoing wound healing but also often turns it into a chronic condition.-Wixler, V. The role of FHL2 in wound healing and inflammation.

Mast cell chymase impairs bronchial epithelium integrity by degrading cell junction molecules of epithelial cells.

An increased degree of mast cell (MC) degranulation and damage to the epithelial lining are prominent features of bronchial asthma. In asthmatic airways, it seems likely that epithelial cells will be exposed to increased concentrations of proteases from MC, though their actions on the epithelium are still not very clear. Bronchial rings from human lung tissue or 16HBE cell monolayer were incubated with MC chymase in different doses or various inhibitors. The sections of paraffin-embedded tissue were haematoxylin-eosin stained and computerized by image analysis for epithelial damage-scale-evaluation; the cell viability, proliferation, adhesion and lactate dehydrogenase activity release were assayed; the expressions of gelatinases, cell junction molecules and structure proteins of 16HBE were examined. Mast cell chymase was found to provoke profound changes in the morphology of bronchi epithelial layer. Following incubation with chymase, there was 40% reduction in the length of epithelium that was intact, with detachment of columnar epithelial cells and basal cells. Chymase reduced epithelial cell proliferation and induced cell detachment, which were associated with the changes in secretion and activation of matrix metalloproteinase-2/9. In intact epithelial cell layers, immunocytochemistry study revealed that chymase reduced the expressions of occludin, claudin-4, ZO-1, E-cadherin, focal adhesion kinase and cytokeratin. Overall data of this study indicated that MC chymase can influence tissue remodelling, disrupt epithelial cell junctions, inhibit wound healing and impair the barrier function of epithelium, resulting in dysfunction of airway wall and ECM remodelling in pathogenesis of asthma. Mast cell chymase plays a key role in inducing the damage to bronchial epithelium in asthma.

Alteration of the colostrum whey proteome in mothers with gestational hypothyroidism.

BackgroundGestational hypothyroidism (G-HypoT) is one of the most common thyroid diseases in pregnant women. Human milk, which closely links the mother with infant, is an important factor to the infant health. Here, we analyzed the colostrum whey proteome of women with or without G-HypoT.Methods and resultsUsing high–mass accuracy and high-resolution liquid chromatography-tandem mass spectrometry (LC-MS/MS), 1055 proteins were identified. Tandem Mass Tags (TMT) analysis identified differentially expressed proteins between G-HypoT and non-G-HypoT mothers. Of 44 proteins identified, 15 proteins were significantly increased in G-HypoT colostrum whey, while 29 were significantly decreased. Analysis revealed that enzymes involved in carbohydrate metabolism, and that reflect the metabolic activities in breastfeeding women, including fructose-1, 6-bisphosphatase 1, phosphoglycerate mutase 1 were down-regulated. Cell structural proteins, biomarkers of mammary integrity development, including Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and actin were lower in G-HypoT colostrum whey. However, immune protein fragments like Ig gamma-3 chain C region increased in G-HypoT colostrum whey.ConclusionThese results implied that G-HypoT may changed human colostrum whey protein in composition level, decreasing levels of metabolic proteins and cell-structure proteins, while increasing levels of immune-related proteins, which may compromise or reflect mothers’ and infants’ health.