Published by MDPI AG
Online ISSN: 2227-9059
Schematic view of the 10-slot microwave antenna with an impedance π-matching network. The conducting material, Teflon, air, and dielectric are represented by black, green, light blue, and light brown, respectively. The width of the slot is 0.6 mm with a spacing of 0.8 mm between adjacent slots.
Three-dimensional simulation models corresponding to two liver tumors (triangulated surfaces) labeled as (a) 1.07 and (b) 1.03 in the 3D-IRCADb-01 database that contains several sets of CT scans of the patients [51].
Iso-contours consisting of totally ablated regions (solid light brown surface) and tumors (triangulated surface) (a) 1.07 [51] and (b) 1.03 [51] exposed to a frequency of 2.45 GHz, an input power of 15 W and 17 W for various ablation times.
(a) Dependence of the ablation time on the input power for tumor 1.03 [51] (blue squares) and tumor 1.07 [51] (red circles). (b) Temperature as a function of the ablation time for various input power values calculated in the center of the heating zone.
Simulation techniques are powerful tools for determining the optimal conditions necessary for microwave ablation to be efficient and safe for treating liver tumors. Owing to the complexity and computational resource consumption, most of the existing numerical models are two-dimensional axisymmetric models that emulate actual three-dimensional cancers and the surrounding tissue, which is often far from reality. Different tumor shapes and sizes require different input powers and ablation times to ensure the preservation of healthy tissues that can be determined only by the full three-dimensional simulations. This study aimed to tailor microwave ablation therapeutic conditions for complete tumor ablation with an adequate safety margin, while avoiding injury to the surrounding healthy tissue. Three-dimensional simulations were performed for a multi-slot microwave antenna immersed in two tumors obtained from the 3D-IRCADb-01 liver tumors database. The temperature dependence of the dielectric and thermal properties of healthy and tumoral liver tissues, blood perfusion, and water content are crucial for calculating the correct ablation time and, thereby, the correct ablation process. The developed three-dimensional simulation model may help practitioners in planning patient-individual procedures by determining the optimal input power and duration of the ablation process for the actual shape of the tumor. With proper input power, necrotic tissue is placed mainly in the tumor, and only a small amount of surrounding tissue is damaged.
Increased astrogliosis after kindling is ameliorated by ACT-03 treatment. (A-F) In nonkindled control animals, vimentin immunoreactivity (IR) was not observed in the parenchyma Increased astrogliosis after kindling is ameliorated by ACT-03 treatment. (A-F) In nonkindled control animals, vimentin immunoreactivity (IR) was not observed in the parenchyma (A,D), while vimentin-positive astrocytes with coarse processes and a reactive morphology (arrowheads) were observed in the hippocampus of kindled, vehicle-treated animals (B,E). In kindled animals treated with ACT-03, vimentin-positive cells were sparsely present and displayed less of a reactive morphology (C,F). (G-J) Quantification shows a larger vimentin-positive area in the dorsal and ventral dentate gyrus (DG) and CA1 area of kindled animals compared to non-kindled animals (p < 0.001). The vimentin-positive area in kindled, ACT-03-treated animals was smaller compared to kindled, vehicle-treated animals in the DG and CA1 of the dorsal hippocampus (p < 0.05). Scale bar in A for A-F; 100 µm; hematoxylin counterstained; ml, molecular later; gcl, granular cell layer; so, stratum oriens; pcl, pyramidal cell layer; sr, stratum radiatum. Bar graphs display mean + SEM, with individual values as dots; *, p < 0.05; ***, p < 0.001; ****, p < 0.0001.
Increased microgliosis after kindling is ameliorated by ACT-03 treatment. (A-F) Iba1-positive microglia with a resting morphology (arrows) were observed in the hippocampus of nonkindled animals (A,D), where Iba1-positive microglia in kindled, vehicle-treated animals have thick and coarse processes (arrowheads). In kindled, ACT-03-treated animals, Iba1-positive microglia with both resting and active morphologies were observed (C,F). (G-J) Quantification shows a higher number of Iba1-positive microglia in kindled versus non-kindled animals in multiple regions (p < 0.05). Iba1-positive cells was lower in the DG of the dorsal and ventral hippocampus of ACT-03-treated animals versus vehicle-treated animals. Scale bar in A for A-F; 100µm; hematoxylin counterstained; ml, molecular later; gcl, granular cell layer; so, stratum oriens; pcl, pyramidal cell layer; sr, stratum radiatum. Bar graphs display mean + SEM, with individual values as dots; *, p < 0.05; **, p < 0.01; ***, p < 0.001.
ACT-03 treatment rescues loss of barrier integrity in vitro. (A) Schematic representation o the in vitro blood-brain barrier (BBB) model containing human fetal astrocytes and human brai endothelial cells (hCMEC/D3) plated on a Transwell ® insert with 0.4µ m-sized pores. (B) Immuno fluorescent staining of the membrane shows GFAP-positive astrocytes (red) under a monolayer o ACT-03 treatment rescues loss of barrier integrity in vitro. (A) Schematic representation of the in vitro blood-brain barrier (BBB) model containing human fetal astrocytes and human brain endothelial cells (hCMEC/D3) plated on a Transwell ® insert with 0.4µm-sized pores. (B) Immunofluorescent staining of the membrane shows GFAP-positive astrocytes (red) under a monolayer of CD31-positive endothelial cells (green) of. (C) Scheme of experimental layout showing the stimulation of the in vitro BBB model with phorbol myristate acetate (PMA) and ionomycin in the presence of either vehicle or ACT-03 for 3 h or 6 h. Subsequently, medium was exchanged for Hank's balanced salt solution (HBSS) and 20 nM of Lucifer Yellow (LY) was added to the upper compartment of the insert. After 1 h of incubation, LY absorbance of the lower compartment of the Transwell ® was measured in triplicates using spectrophotometry. (D) Higher barrier permeability indicated by apparent permeability (P app ) was observed after 6 h of stimulation (p < 0.05). Addition of ACT-03 reduced barrier permeability compared to stimulation alone (p < 0.01), returning values to control level. (E) MTT assays of human fetal astrocytes and (F) endothelial cells show cell viability after PMA + iono stimulation. Monocultures of human fetal astrocytes were performed on n = 4 donors in 4 experimental replicates, endothelial cell cultures were performed with 6 experimental replicates. The co-culture in vitro BBB model was performed using n = 3 astrocytes donors with experimental triplicates. Scale bar: 12.5 µm. Bar graphs display mean + SEM, with individual values as dots; *, p < 0.05; **, p < 0.01; ***, p < 0.001. Within treatment comparisons indicated with red/green lines. Between treatment comparisons indicated with hooked black lines.
Primers used for quantitative real-time PCR.
Matrix metalloproteinases (MMPs) are endopeptidases responsible for the cleavage of intra- and extracellular proteins. Several brain MMPs have been implicated in neurological disorders including epilepsy. We recently showed that the novel gelatinase inhibitor ACT-03 has disease-modifying effects in models of epilepsy. Here, we studied its effects on neuroinflammation and blood–brain barrier (BBB) integrity. Using the rapid kindling rat model of epilepsy, we examined whether ACT-03 affected astro- and microgliosis in the brain using immunohistochemistry. Cellular and molecular alterations were further studied in vitro using human fetal astrocyte and brain endothelial cell (hCMEC/D3) cultures, with a focus on neuroinflammatory markers as well as on barrier permeability using an endothelial and astrocyte co-culture model. We observed less astro- and microgliosis in the brains of kindled animals treated with ACT-03 compared to control vehicle-treated animals. In vitro, ACT-03 treatment attenuated stimulation-induced mRNA expression of several pro-inflammatory factors in human fetal astrocytes and brain endothelial cells, as well as a loss of barrier integrity in endothelial and astrocyte co-cultures. Since ACT-03 has disease-modifying effects in epilepsy models, possibly via limiting gliosis, inflammation, and barrier integrity loss, it is of interest to further evaluate its effects in a clinical trial.
G. mellonella representing the different levels of melanization and cuticle discoloration.
Immunomodulation induced by the metal-tdda-phen complexes in combination with gentamicin (CN) (2 μg and 2 μg/larvae, and 4 μg and 4 μg/larvae) in G. mellonella after 2, 6, and 24 h post-injection. * indicate significant differences in relation to the PBS injected control (p < 0.05).
Survival (%) of G. mellonella inoculated with P. aeruginosa strains, ATCC 27853 (A-D), PAO1 (E-H), CF1 (I-L), CF2 (M-P), CF3 (Q-T, and treated with 2-10 μg/larvae of Mn-tdda-phen (A,E,I,M,Q), Cu-tdda-phen (B,F,J,N,R), Ag-tdda-phen (C,G,K,O,S), and gentamicin (D,H,L,T) over 96 h.
Drug-resistant Pseudomonas aeruginosa is rapidly developing resulting in a serious global threat. Immunocompromised patients are specifically at risk, especially those with cystic fibrosis (CF). Novel metal complexes incorporating 1,10-phenanthroline (phen) ligands have previously demonstrated antibacterial and anti-biofilm effects against resistant P. aeruginosa from CF patients in vitro. Herein, we present the in vivo efficacy of {[Cu(3,6,9-tdda)(phen)2]·3H2O·EtOH}n (Cu-tdda-phen), {[Mn(3,6,9-tdda)(phen)2]·3H2O·EtOH}n (Mn-tdda-phen) and [Ag2(3,6,9-tdda)(phen)4]·EtOH (Ag-tdda-phen) (tddaH2 = 3,6,9-trioxaundecanedioic acid). Individual treatments of these metal-tdda-phen complexes and in combination with the established antibiotic gentamicin were evaluated in vivo in larvae of Galleria mellonella infected with clinical isolates and laboratory strains of P. aeruginosa. G. mellonella were able to tolerate all test complexes up to 10 µg/larva. In addition, the immune response was affected by stimulation of immune cells (hemocytes) and genes that encode for immune-related peptides, specifically transferrin and inducible metallo-proteinase inhibitor. The amalgamation of metal-tdda-phen complexes and gentamicin further intensified this response at lower concentrations, clearing a P. aeruginosa infection that were previously resistant to gentamicin alone. Therefore this work highlights the anti-pseudomonal capabilities of metal-tdda-phen complexes alone and combined with gentamicin in an in vivo model.
The natural plant dietary polyphenols 1,2,3,4,6-O-Pentagalloylglucose (PGG) and proanthocyanidin (PAC) have potent antioxidant activity and a variety of pharmacological activities, including antiviral activity. In this study, we examined the inhibitory effect of PGG and PAC on SARS-CoV-2 virus infection, and elucidated its mode of action. PGG and PAC have dose-dependent inhibitory activity against SARS-CoV-2 infection in Vero cells. PGG has a lower IC50 (15.02 ± 0.75 μM) than PAC (25.90 ± 0.81 μM), suggesting that PGG has better inhibitory activity against SARS-CoV-2 than PAC. The PGG and PAC inhibit similar Mpro activities in a protease activity assay, with IC50 values of 25–26 μM. The effects of PGG and PAC on the activity of the other essential SARS-CoV-2 viral protein, RdRp, were analyzed using a cell-based activity assay system. The activity of RdRp is inhibited by PGG and PAC, and PGG has a lower IC50 (5.098 ± 1.089 μM) than PAC (21.022 ± 1.202 μM), which is consistent with their inhibitory capacity of SARS-CoV-2 infection. PGG and PAC also inhibit infection by SARS-CoV and MERS-CoV. These data indicate that PGG and PAC may be candidate broad-spectrum anticoronaviral therapeutic agents, simultaneously targeting the Mpro and RdRp proteins of SARS-CoV-2.
Overheating can affect solubility or lipophilicity, among other properties, of some anticancer drugs. These temperature-dependent changes can improve efficiency and selectivity of the drugs, since they may affect their bioavailability, diffusion through cell membrane or activity. One recent approach to create thermosensitive molecules is the incorporation of fluorine atoms in the chemical structure, since fluor can tune some chemical properties such as binding affinity. Herein we report the anticancer effect of gold derivatives with phosphanes derived from 1,3,5-triaza-7-phosphaadamantane (PTA) with long hydrocarbon chains and the homologous fluorinated chains. Besides, we analysed the influence of temperature in the cytotoxic effect. The studied gold(I) complexes with phosphanes derived from PTA showed antiproliferative effect on human colon carcinoma cells (Caco-2/TC7 cell line), probably by inhibiting cellular TrxR causing a dysfunction in the intracellular redox state. In addition, the cell cycle was altered by the activation of p53, and the complexes produce apoptosis through mitochondrial depolarization and the consequent activation of caspase-3. Furthermore, the results suggest that this cytotoxic effect is enhanced by hyperthermia and the presence of polyfluorinated chains.
The antimicrobial activity of KHQ 711, 712, 713, 714, 715, 716, 717 and 718. P. gingivalis cultures were treated with (A) KHQ 711 (6.25 and 3.125 µg/mL), (B) KHQ 712 (3.125 and 1.56 µg/mL), (C) KHQ 713 (1.56 and 0.78 µg/mL), (D) KHQ 714 (12.5 and 6.25 µg/mL), (E) KHQ 715 (3.125 and 1.56 µg/mL), (F) KHQ 716 (3.125 and 1.56 µg/mL), (G) KHQ 717 (3.125 and 1.56 µg/mL) and (H) KHQ 718 (3.125 and 1.56 µg/mL) at 37 • C for 72 h and check the growth at every 6 h.
Strains used in this study.
Minimum inhibitory concentration (MIC) values (µg/mL) against oral bacteria.
Inhibition zone diameters of KHQ 711, 712, 713, 714, 715, 716, 717 and 718 against P. gingivalis.
Background: Dental caries is considered to be a preventable disease, and various antimicrobial agents have been developed for the prevention of dental disease. However, many bacteria show resistance to existing agents. Methods/principal findings: In this study, four known 1,4-naphthoquinones and newly synthesized 10 pyrimidinone-fused 1,4-naphthoquinones, i.e. KHQ 701, 702, 711, 712, 713, 714, 715, 716, 717 and 718, were evaluated for antimicrobial activity against Enterococcus faecalis, Enterococcus faecium, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus mutans, Streptococcus sobrinus, Porphyromonas gingivalis, Actinomyces viscosus and Fusobacterium nucleatum. Pyrimidinone-fused 1,4-naphthoquinones were synthesized in good yields through a series of chemical reactions from a commercially available 1,4-dihydroxynaphthoic acid. MIC values of KHQ 711, 712, 713, 714, 715, 716, 717 and 718 were 6.25-50 μg/mL against E. faecalis (CCARM 5511), 6.25-25 μg/mL against E. faecium (KACC11954) and S. aureus (CCARM 3506), 1.56-25 μg/mL against S. epidermidis (KACC 13234), 3.125-100 μg/mL against S. mutans (KACC16833), 1.56-100 μg/mL against S. sobrinus (KCTC5809) and P. gingivalis (KCTC 5352), 3.125-50 μg/mL against A. viscosus (KCTC 9146) and 3.125-12.5 μg/mL against F. nucleatum (KCTC 2640) with a broth microdilution assay. A disk diffusion assay with KHQ derivatives also exhibited strong susceptibility with inhibition zones of 0.96 to 1.2 cm in size against P. gingivalis. Among the 10 compounds evaluated, KHQ 711, 712, 713, 715, 716 and 717 demonstrated strong antimicrobial activities against the 9 types of pathogenic oral bacteria. A pyrimidin-4-one moiety comprising a phenyl group at the C2 position and a benzyl group at the N3 position appears to be essential for physiological activity. Conclusion/significance: Pyrimidinone-fused 1,4-naphthoquinones synthesized from simple starting compounds and four known 1,4-naphthoquinones were synthesized and showed strong antibacterial activity to the 9 common oral bacteria. These results suggest that these derivatives should be prospective for the treatment of dental diseases caused by oral bacteria, including drug-resistant strains.
Electrostatic affinity between the RBD and ACE2. (a) Structural representation of the RBD (wild type, wt: blue; B.1.1.7: orange) in complex with ACE2 (grey). RBD residues with atoms within a maximum distance of 4 Å from ACE2 are shown as spheres with radii and colors according to their electrostatic linear interaction energy to ACE2. (b) View of the interacting interface of the RBD with ACE2. Residue color and atom size represent their electrostatic linear interaction energy to ACE2. (c) Quantification of electrostatic linear interaction energy for all residues within 4 Å distance of ACE2 (n = 4; two-way ANOVA; statistical significance assumed for * p < 0.05; full list of results in Table S4).
Van der Waals linear interaction energy between the RBD and ACE2. (a) Structural representation of the RBD (wild type, wt: blue; B.1.1.7: orange) in complex with ACE2. RBD residues
The B.1.1.7 variant of the SARS-CoV-2 virus shows enhanced infectiousness over the wild type virus, leading to increasing patient numbers in affected areas. Amino acid exchanges within the SARS-CoV-2 spike protein variant of B.1.1.7 affect inter-monomeric contact sites within the trimer (A570D and D614G) as well as the ACE2-receptor interface region (N501Y), which comprises the receptor-binding domain (RBD) of the spike protein. However, the molecular consequences of mutations within B.1.1.7 on spike protein dynamics and stability or ACE2 binding are largely unknown. Here, molecular dynamics simulations comparing SARS-CoV-2 wild type with the B.1.1.7 variant revealed inter-trimeric contact rearrangements, altering the structural flexibility within the spike protein trimer. Furthermore, we found increased flexibility in direct spatial proximity of the fusion peptide due to salt bridge rearrangements induced by the D614G mutation in B.1.1.7. This study also implies a reduced binding affinity for B.1.1.7 with ACE2, as the N501Y mutation restructures the RBD–ACE2 interface, significantly decreasing the linear interaction energy between the RBD and ACE2. Our results demonstrate how mutations found within B.1.1.7 enlarge the flexibility around the fusion peptide and change the RBD–ACE2 interface. We anticipate our findings to be starting points for in depth biochemical and cell biological analyses of B.1.1.7.
Enzymes are key proteins performing the basic functional activities in cells. In humans, enzymes can be also responsible for diseases, and the molecular mechanisms underlying the genotype to phenotype relationship are under investigation for diagnosis and medical care. Here, we focus on highlighting enzymes that are active in different metabolic pathways and become relevant hubs in protein interaction networks. We perform a statistics to derive our present knowledge on human metabolic pathways (the Kyoto Encyclopaedia of Genes and Genomes (KEGG)), and we found that activity aldehyde dehydrogenase (NAD(+)), described by Enzyme Commission number EC, and activity acetyl-CoA C-acetyltransferase (EC are the ones most frequently involved. By associating functional activities (EC numbers) to enzyme proteins, we found the proteins most frequently involved in metabolic pathways. With our analysis, we found that these proteins are endowed with the highest numbers of interaction partners when compared to all the enzymes in the pathways and with the highest numbers of predicted interaction sites. As specific enzyme protein test cases, we focus on Alpha-Aminoadipic Semialdehyde Dehydrogenase (ALDH7A1, EC and Acetyl-CoA acetyltransferase, cytosolic and mitochondrial (gene products of ACAT2 and ACAT1, respectively; EC With computational approaches we show that it is possible, by starting from the enzyme structure, to highlight clues of their multiple roles in different pathways and of putative mechanisms promoting the association of genes to disease.
1 H NMR of cholesterol methyl (0.68 ppm) from lipid extracts of normal and tumorous brain tissues (N = 3) on days nine (A) and 12 (B) and their cholesterol content (C) as quantified using AUCs of methyl shown in (A,B). (D) The choline methyls from the phosphatidylcholines (~3.35 ppm) and alkyl hydrogens (~1.5 ppm) can be identified in the 1 H spectra. The AUCs between 3.28 to 3.38 ppm and 1.45 to 1.55 ppm were used to estimate the relative amounts of phosphatidylcholine (E) and total lipid (F). (* p < 0.05).
The chemical exchange saturation transfer (CEST) signal at −1.6 ppm is attributed to the choline methyl on phosphatidylcholines and results from the relayed nuclear Overhauser effect (rNOE), that is, rNOE(−1.6). The formation of rNOE(−1.6) involving the cholesterol hydroxyl is shown in liposome models. We aimed to confirm the correlation between cholesterol content and rNOE(−1.6) in cell cultures, tissues, and animals. C57BL/6 mice (N = 9) bearing the C6 glioma tumor were imaged in a 7 T MRI scanner, and their rNOE(−1.6) images were cross-validated through cholesterol staining with filipin. Cholesterol quantification was obtained using an 18.8-T NMR spectrometer from the lipid extracts of the brain tissues from another group of mice (N = 3). The cholesterol content in the cultured cells was manipulated using methyl-β-cyclodextrin and a complex of cholesterol and methyl-β-cyclodextrin. The rNOE(−1.6) of the cell homogenates and their cholesterol levels were measured using a 9.4-T NMR spectrometer. The rNOE(−1.6) signal is hypointense in the C6 tumors of mice, which matches the filipin staining results, suggesting that their tumor region is cholesterol deficient. The tissue extracts also indicate less cholesterol and phosphatidylcholine contents in tumors than in normal brain tissues. The amplitude of rNOE(−1.6) is positively correlated with the cholesterol concentration in the cholesterol-manipulated cell cultures. Our results indicate that the cholesterol dependence of rNOE(−1.6) occurs in cell cultures and solid tumors of C6 glioma. Furthermore, when the concentration of phosphatidylcholine is carefully considered, rNOE(−1.6) can be developed as a cholesterol-weighted imaging technique.
Genotype frequencies of miRNA gene polymorphisms in control subjects and recurrent pregnancy loss patients.
Adjusted odds ratios for risk of recurrent pregnancy loss associated with miRNA polymorphisms combined with clinical factors.
This study investigated the genetic association between recurrent pregnancy loss (RPL) and microRNA (miRNA) polymorphisms in miR-10aA>T, miR-30cA>G, miR-181aT>C, and miR-499bA>G in Korean women. Blood samples were collected from 381 RPL patients and 281 control participants, and genotyping of miR-10aA>T, miR-30cA>G, miR-181aT>C, and miR-499bA>G was carried out by TaqMan miRNA RT-Real Time polymerase chain reaction (PCR). Four polymorphisms were identified, including miR-10aA>T, miR-30cA>G, miR-181aT>C, and miR-499bA>G. MiR-10a dominant model (AA vs. AT + TT) and miR-499bGG genotypes were associated with increased RPL risk (adjusted odds ratio [AOR] = 1.520, 95% confidence interval [CI] = 1.038–2.227, p = 0.032; AOR = 2.956, 95% CI = 1.168–7.482, p = 0.022, respectively). Additionally, both miR-499 dominant (AA vs. AG + GG) and recessive (AA + AG vs. GG) models were significantly associated with increased RPL risk (AOR = 1.465, 95% CI = 1.062–2.020, p = 0.020; AOR = 2.677, 95% CI = 1.066–6.725, p = 0.036, respectively). We further propose that miR-10aA>T, miR-30cA>G, and miR-499bA>G polymorphisms effects could contribute to RPL and should be considered during RPL patient evaluation.
Boron neutron capture therapy (BNCT) is based on the ability of the boron-10 (10B) isotope to capture epithermal neutrons, as a result of which the isotope becomes unstable and decays into kinetically active elements that destroy cells where the nuclear reaction has occurred. The boron-carrying compounds—L-para-boronophenylalanine (BPA) and sodium mercaptoundecahydro-closo-dodecaborate (BSH)—have low toxicity and, today, are the only representatives of such compounds approved for clinical trials. For the effectiveness and safety of BNCT, a low boron content in normal tissues and substantially higher content in tumor tissue are required. This study evaluated the boron concentration in intracranial grafts of human glioma U87MG cells and normal tissues of the brain and other organs of mice at 1, 2.5 and 5 h after administration of the boron-carrying compounds. A detailed statistical analysis of the boron biodistribution dynamics was performed to find a ‘window of opportunity’ for BNCT. The data demonstrate variations in boron accumulation in different tissues depending on the compound used, as well as significant inter-animal variation. The protocol of administration of BPA and BSH compounds used did not allow achieving the parameters necessary for the successful course of BNCT in a glioma orthotopic xenograft mouse model.
Percentage of participants in each hearing-loss severity, in the five WHO grades [18]. (A) Males; (B) females.
Background: Accurate data on the prevalence of hearing impairment and severity across age and gender are paramount to formulate hearing health policies. Here, we sought to analyze audiometric data from a large group of age-diverse people in Japan, which has not been previously described in detail. Methods: We analyzed retrospective hearing threshold data of 23,860 participants (10-99 years; left-right hearing threshold difference <15 dB; air-bone gap ≤10 dB) at 500, 1000, 2000, and 4000 Hz, and then classified them for hearing impairment severity according to the WHO Classification. Findings: There was a significant gender difference in median hearing thresholds, starting in 20-year-olds up to early 80-year-olds. Twenty-five percent of men in their late 50s had some level of HI, ~50% in their late 60s, and ~75% in their late 70s. For women, 25% had some level of HI in their early 60s, ~50% in their early 70s, and ~75% in their late 70s. For participants in their early 80s, 50% of either gender had moderate or more severe HI. Interpretation: Our results, derived from a large number of participants, provide basic information about the prevalence of hearing loss by age decade. Since people can expect to live longer than those in previous generations, our detailed data can inform national social systems responsible for hearing screening in making decisions about hearing-aid qualification, which may reduce barriers to older people's independence, productivity, and quality of life.
Altered postnatal microRNA expression profile in children descending from PTB pregnancies. Combined screening of 13 selected microRNAs with the best sensitivity (miR-1-3p, miR-16-5p, miR-17-5p, miR-20a-5p, miR-20b-5p, miR-26a-5p, miR-29a-3p, miR-103a-3p, miR-126-3p, miR-143-3p, miR-181a-5p, miR-195-5p, and miR-499a-5p) reveals 85.71% children with a potential cardiovascular risk at 10.0% FPR.
Clinical outcomes of preterm-and term-born children.
(1) Background: Preterm-born children have an increased cardiovascular risk with the first clinical manifestation during childhood and/or adolescence. (2) Methods: The occurrence of overweight/obesity, prehypertension/hypertension, valve problems or heart defects, and postnatal microRNA expression profiles were examined in preterm-born children at the age of 3 to 11 years descending from preterm prelabor rupture of membranes (PPROM) and spontaneous preterm birth (PTB) pregnancies. The whole peripheral blood gene expression of 29 selected microRNAs associated with cardiovascular diseases was the subject of our interest. (3) Results: Nearly one-third of preterm-born children (32.43%) had valve problems and/or heart defects. The occurrence of systolic and diastolic prehypertension/hypertension was also inconsiderable in a group of preterm-born children (27.03% and 18.92%). The vast majority of children descending from either PPROM (85.45%) or PTB pregnancies (85.71%) had also significantly altered microRNA expression profiles at 90.0% specificity. (4) Conclusions: Postnatal microRNA expression profiles were significantly influenced by antenatal and early postnatal factors (gestational age at delivery, birth weight of newborns, and condition of newborns at the moment of birth). These findings may contribute to the explanation of increased cardiovascular risk in preterm-born children. These findings strongly support the belief that preterm-born children should be dispensarized for a long time to have access to specialized medical care.
Swimmers plot with time to new biochemical recurrence (BCR) in Subset 1 (blue line) and Subset 2 (orange line) after r-RT. The red points represent patients with a PSA reduction of more than 50% three months after r-RT.
Kaplan-Meier curves of biochemical recurrence-free survival (b-RFS) after 68 Ga-labeled PSMA ligand PET-directed radiotherapy of prostate cancer: (A) overall population; (B) Subset 1 and Subset 2.
(1) Purpose: To investigate the role of 68Ga-PSMA-11 PET/CT in guiding retreatment stereotactic body radiation therapy (SBRT) in prostate cancer (PCa) patients in biochemical recurrence (BCR) after salvage radiotherapy (S-RT). (2) Methods: We retrospectively evaluated PCa patients previously treated with S-RT on the prostate bed and with proven serum prostate antigen (PSA) failure after S-RT. In all patients (pts), 68Ga-PSMA-11 PET/CT was positive in the prostate bed only and guided retreatment SBRT. All retreatments were performed by applying the same radiotherapy protocol (median dose of 18 Gy/3 fractions; IQR 18–21 Gy). The median follow-up was 27 months (range 4–35 months). (3) Results: 38 consecutive patients were considered in this analysis. The overall median PSA level before RT was 1.10 ng/mL (IQR 0.82–2.59). PSA decreased at 3 and 6 months after treatment, with a median value of 0.60 ng/mL (IQR 0.31–0.96; p < 0.001) and 0.51 ng/mL (IQR 0.29–1.17; p < 0.001), respectively. Overall, biochemical recurrence-free survival (b-RFS) was 15.0 months (95% CI 13–23). Grade-1 toxicity was reported in 31.6% of patients (12/38). (4) Conclusion: These results confirm that 68Ga-PSMA-11-PET/CT is able to identify the site of recurrence in patients who have failed S-RT, thus supporting the use of metastases-directed radiotherapy as a safe and effective treatment.
Scatterplot showing the correlation between baseline PSA (ug/L) levels and whole-body PSMA-TV (WBPSMATV).
Comparison of primary tumor PSMA-TV in WSA (A) and BSA (B) for two patients with GG 1 disease in maximum projection image, PET only and fused PET/CT. (A) corresponds to a 61-yearold WSA with PSA 73.31 ug/L. SUVmax-3.42, TL-PSMA-0.90, PSMA-TV of 0.29. (B) corresponds to a 60-year-old BSA with PSA of 22 ug/L. SUVmax-6.85, TL-PSMA-17.3, PSMA-TV-4.89.
Comparison of whole-body (WB) PSMA-TV in WSA (A) and BSA (B) for two patients with GG 2 disease. Left: maximum intensity projection image; middle column: PET and fused PET/CT of prostate; right column: PET and fused PET/CT metastases. (A) 58 year-old WSA with PSA 59 ug/L, with pelvic nodal metastases (red arrow). Bilateral ureteric tracer excretion seen, WBPSMA-TV-45. (B) 57-year-old BSA with PSA of 30 ug/L demonstrating pelvic skeletal metastases, WBPSMA-TV-58.
Patient characteristics.
Logistic regression of variables that predict for metastases.
Prostate adenocarcinoma (PCa) is a leading cause of mortality. Black males with high-risk PCa have a poorer prognosis compared to white males. Patients with International Society of Urological Pathology (ISUP) Grade Group (GG) 1 and 2 PCa have little potential for metastases post radical prostatectomy. 68Gallium prostate specific membrane antigen (68Ga-PSMA) PET/CT imaging for metastatic PCa is superior to conventional imaging in staging high-risk PCa. No strong evidence is available to support imaging low-risk patients. We aimed to evaluate the value of 68Ga-PSMA PET/CT in black and white South African (BSA and WSA) males with GG1 and 2 PCa at initial staging. We evaluated 25 WSA and 123 BSA males. The image findings were correlated with prostate specific antigen (PSA). PSA levels significantly correlated with both primary tumor and whole-body PSMA-tumor volume (PSMA-TV) and were higher in BSA males. No differences were noted in the occurrence of metastases; however, PSA, seminal vesicle invasion and black race predicted metastases. Our findings suggest higher PSMA expression and tumor burden in BSA with histologically low-risk PCa, and future research with immunohistochemistry evaluation will be essential to confirm these findings.
In vitro and in vivo characterization of NHance ® -mutated ARGX-111 for FcRn-mediated transcytosis. (a) NHance mutations promote transcytosis across MDCK II cells overexpressing human FcRn. Transwell assay showing apical-to-basolateral transport of Fc-engineered variants of ARGX-111. Measured antibody concentrations in the basolateral compartment are shown in ng/mL; (b) Tissue distribution of ARGX-111 in 32TG human FcRn transgenic mice. Tissue/blood ratio in mice (N = 3) analyzed 168 h after administration of 89 Zr-labelled ARGX-111 or 53E2 in the presence of hIgG or not. Bars represent mean ±SEM; * p = 0.0174; ** p = 0.0005.
Mean duration of study and best response. Each bar represents one patient and the length of the bar the duration of treatment from until C1D1 end of treatment (EOT). The doses and the administration interval are indicated by the color code. The best response as per investigator review is also indicated on the right for each patient. PR, partial response; SD, stable disease; PD, progressive disease; NE, not evaluable according to RECIST 1.1.
Patient demographics and baseline characteristics.
Summary of treatment-emergent adverse events (TEAEs) in ≥20% of all patients (dose escalation plus safety expansion) by preferred term (PT).
Pharmacokinetics of ARGX-111 at cycle 1 after the first ARGX-111 administration (actual time points).
Dysregulation of MET signaling has been implicated in tumorigenesis and metastasis. ARGX-111 combines complete blockade of this pathway with enhanced tumor cell killing and was investigated in 24 patients with MET-positive advanced cancers in a phase 1b study at four dose levels (0.3–10 mg/kg). ARGX-111 was well tolerated up to 3 mg/kg (MTD). Anti-tumor activity was observed in nearly half of the patients (46%) with a mean duration of treatment of 12 weeks. NHance® mutations in the Fc of ARGX-111 increased affinity for the neonatal Fc receptor (FcRn) at acidic pH, stimulating transcytosis across FcRn-expressing cells and radiolabeled ARGX-111 accumulated in lymphoid tissues, bone and liver, organs expressing FcRn at high levels in a biodistribution study using human FcRn transgenic mice. In line with this, we observed, in a patient with MET-amplified (>10 copies) gastric cancer, diminished metabolic activity in multiple metastatic lesions in lymphoid and bone tissues by 18F-FDG-PET/CT after two infusions with 0.3 mg/kg ARGX-111. When escalated to 1 mg/kg, a partial response was reached. Furthermore, decreased numbers of CTC (75%) possibly by the enhanced tumor cell killing witnessed the modes of action of the drug, warranting further clinical investigation of ARGX-111.
Chemical structures and radiolabeling positions for commonly used dopaminergic PET radiotracers. The predominate radiotracer for imaging of dopamine synthesis has remained[ 18 F]FDOPA, but syntheses of the carbon-11 labeled radiotracers β-[ 11 C]-L-DOPA and 6-[ 11 C]methylm-tyrosine were also accomplished. The β-[ 11 C]-L-DOPA forms [ 11 C]dopamine after AADC-mediated decarbozylation, and provides human brain images and uptake data much like 6-[ 18 F]fluorodopa. A study comparing β-[ 11 C]-L-DOPA, 6-[ 18 F]fluoroDOPA and
Representative radiotracers used for in vivo PET imaging of the neurochemistry of dopamine in the human brain. Tyrosine Hydroxylase-No In Vivo Radiotracers Available
The applications of positron emission tomography (PET) imaging to study brain biochemistry, and in particular the aspects of dopamine neurotransmission, have grown significantly over the 40 years since the first successful in vivo imaging studies in humans. In vivo PET imaging of dopaminergic functions of the central nervous system (CNS) including dopamine synthesis, vesicular storage, synaptic release and receptor binding, and reuptake processes, are now routinely used for studies in neurology, psychiatry, drug abuse and addiction, and drug development. Underlying these advances in PET imaging has been the development of the unique radiotracers labeled with positron-emitting radionuclides such as carbon-11 and fluorine-18. This review focuses on a selection of the more accepted and utilized PET radiotracers currently available, with a look at their past, present and future.
Adiponectin is an adipokine that mediates cellular cholesterol efflux and plays important roles in neuroinflammatory processes. In this study, we undertook positron emission tomography (PET) with the translocator protein (TSPO) ligand [11C]PK11195 and measured serum adiponectin levels in groups of treatment-naïve young adult patients with major depressive disorder (MDD) and matched healthy controls. Thirty treatment-naïve MDD patients (median age: 24 years) and twenty-three healthy controls underwent [11C]PK11195 PET. We quantified TSPO availability in brain as the [11C]PK11195 binding potential (BPND) using a reference tissue model in conjunction with the supervised cluster analysis (SVCA4) algorithm. Age, sex distribution, body mass index, and serum adiponectin levels did not differ between the groups. Between-group analysis using a region-of-interest approach showed significantly higher [11C]PK11195 BPND in the left anterior and right posterior cingulate cortices in MDD patients than in controls. Serum adiponectin levels had significant negative correlations with [11C]PK11195 BPND in the bilateral hippocampus in MDD patients, but significant positive correlations in the bilateral hippocampus in the control group. Our results indicate significantly higher TSPO binding in the anterior and posterior cingulate cortices in treatment-naïve young MDD patients, suggesting microglial activation in these limbic regions, which are involved in cognitive and emotional processing. The opposite correlations between [11C]PK11195 BPND in the hippocampus with serum adiponectin levels in MDD and control groups suggest that microglial activation in the hippocampus may respond differentially to adiponectin signaling in MDD and healthy subjects, possibly with respect to microglial phenotype.
Ranking order of MSLN immunostaining in cancers. Both the frequency of positive cases (blue dots) and the frequency of strongly positive cases (orange dots) are shown. Fifty-six additional tumor entities without any MSLN positive cases are not shown due to space restrictions.
MSLN immunostaining and tumor phenotype.
MSLN immunostaining and HPV status.
Mesothelin (MSLN) represents an attractive molecule for targeted cancer therapies. To identify tumors that might benefit from such therapies, tissue microarrays including 15,050 tumors from 122 different tumor types and 76 healthy organs were analyzed for MSLN expression by immunohistochemistry. Sixty-six (54%) tumor types showed at least occasional weak staining, including 50 (41%) tumor types with at least one strongly positive sample. Highest prevalence of MSLN positivity had ovarian carcinomas (serous 97%, clear cell 83%, endometrioid 77%, mucinous 71%, carcinosarcoma 65%), pancreatic adenocarcinoma (ductal 75%, ampullary 81%), endometrial carcinomas (clear cell 71%, serous 57%, carcinosarcoma 50%, endometrioid 45%), malignant mesothelioma (69%), and adenocarcinoma of the lung (55%). MSLN was rare in cancers of the breast (7% of 1138), kidney (7% of 807), thyroid gland (1% of 638), soft tissues (0.3% of 931), and prostate (0 of 481). High expression was linked to advanced pathological tumor (pT) stage (p < 0.0001) and metastasis (p < 0.0001) in 1619 colorectal adenocarcinomas, but unrelated to parameters of malignancy in 1072 breast-, 386 ovarian-, 174 lung-, 757 kidney-, 171 endometrial-, 373 gastric-, and 925 bladder carcinomas. In summary, numerous important cancer types with high-level MSLN expression might benefit from future anti-MSLN therapies, but MSLN’s prognostic relevance appears to be limited.
Differences in intestinal microbiota composition and diversity in normal renal function controls and advanced CKD patients receiving AST-120 or not. (A) Top 10 genera among groups. The relative abundances are expressed as percentage. "1" denotes 100%. (B) α-diversity (Chao 1) and (C) β-diversity (Bray-Curtis similarity index) of intestinal microbiota among groups. The boxplot reveals the median, the 25th, and the 75th percentile; * p < 0.001. (D) Nonmetric multidimensional scaling (NMDS) ordination based on weighted UniFrac of gut microbiomes. Significant sample-to-sample dissimilarities by analysis of similarity (ANOSIM, p < 0.001) test for determining differences in microbial composition among three groups. (E) Microbial taxa that best characterize each group were defined by applying linear discriminant analysis of effect size (LEfSe) on OTU tables.
Changes in circulating metabolites concentrations associated with AST-120 treatment in advanced CKD patients. The box-plot reveals the median, the 25th, and the 75th percentile. Wilcoxon rank sum test was used to detect differences among groups. * p < 0.05. CKD, chronic kidney disease; IS, indoxyl sulfate; pCS, p cresyl-sulfate.
Baseline characteristics of study population (n = 56).
Background: Animal studies have demonstrated that an oral absorbent AST-120 modulates gut environment. However, this phenomenon remains unclear in humans. This study aimed to assess the effects of AST-120 on the gut microbiota, related functional capability and metabolomic profiling in advanced chronic kidney diseases (CKD) patients. Methods: Eight advanced CKD patients with AST-120 (CKD+AST), 24 CKD patients (CKD), and 24 non-CKD controls were enrolled. We analyzed 16S rRNA pyrosequencing of feces and serum metabolomics profiling. Results: The CKD+AST group exhibited dispersed microbial community structure (β-diversity, p < 0.001) compared to other groups. The relative abundances of at least 16 genera were significantly different amongst the three groups. Increases of fatty acids-producing bacteria (Clostridium_sensu_stricto_1, Ruminococcus_2, Eubacterium_nodatum and Phascolarctobacterium) associated with elevated serum acetic acid and octanoic acid levels were found in CKD+AST group. Analysis of microbial gene function indicated that pathway modules relevant to metabolisms of lipids, amino acids and carbohydrates were differentially enriched between CKD+AST and CKD groups. Specifically, enrichments of gene markers of the biosynthesis of fatty acids were noted in the CKD+AST group. Conclusion: Advanced CKD patients exhibited significant gut dysbiosis. AST-120 can partially restore the gut microbiota and intervenes in a possible signature of short- and medium-chain fatty acids metabolism.
The flowchart indicating each time-point of experiment and behavioral test. Eight-week-old male C57BL/6J mice (or NLRP3 knockout mice) received sham surgery or 2-step 5/6 nephrectomy (NX) 2 weeks before AST-120 treatment. At week 0, mice were divided and treated with or without 10% AST-120. Blood was drawn from the tail vein at week 4 and week 8. Y maze tests were performed on week 4 and week 8. Morris water maze (MWM) tests were performed start from week 8. Mice were sacrificed at around week 10 and brain tissues were harvested for uremic toxin quantification, Western blotting and staining. NX: nephrectomy; WT: wild type; each interval of chart is a week.
The 5/6 nephrectomy-induced chronic kidney disease (CKD) mouse models show impaired working memory. The serum levels of BUN (panel A), creatinine (panel A), and indoxyl sulfate (IS, panel B) of the 5/6 nephrectomy mice were significantly higher than those of sham-control mice at 4 and 8 weeks. However, p-cresol sulfate (PCS) levels significantly increased at week 4 but dropped back at week 8 (panel B). These findings suggest that the 5/6 nephrectomy procedure could successfully establish CKD animal models. The Y-maze behavioral test was performed at 4 and 8 weeks after the 5/6 nephrectomy procedure to examine working memory, which is also referred to as hippocampal function (panel C). The Y-maze test revealed significantly decreased cognitive function compared with the sham control group at 8 weeks after 5/6 nephrectomy surgery. Wn (walking number): number of times the animal passes different arms within 5 min; Cn (cycle number): number of times the animal goes through the three arms (n = 8, * p ≤ 0.1; ** p ≤ 0.01; *** p ≤ 0.001).
AST-120 administration decreased serum and brain levels of indoxyl sulfate (IS) and p-cresol sulfate (PCS) in 5/6 nephrectomy-induced CKD animals. Blood samples were obtained at 4 weeks and 8 weeks of experimental mice for highperformance liquid chromatography (HPLC) analysis. HPLC revealed significantly decreased serum IS and PCS levels in AST-120 treated CKD mice at 4 weeks, as well as non-significantly lower IS and PCS levels at 8 weeks compared with those in CKD mice not treated with AST-120 (panel A,B). Moreover, AST-120-treated CKD mice showed significantly decreased IS levels in the frontal lobe and hippocampus (panel C,D); however, the PCS levels in the frontal lobe and hippocampus showed a non-significant decrease compared with those in CKD mice not treated with AST-120 (panel E,F) (* p ≤ 0.1; ** p ≤ 0.01; *** p ≤ 0.00; each group contained 4-6 animals).
There were increased NLRP3 inflammasomes in hippocampal microglia and astrocytes of CKD mice. Bright light microscopy of hematoxylin stain demonstrated the anatomy of hippocampus with hematoxylin and eosin stain. The following images were in CA3 region of the hippocampus (panel A). The demonstration of Immunohistochemistry staining against Iba-1, GFAP, and NLRP3 was used to examine the inflammatory response in the CA3 of hippocampus in CKD mice, which demonstrated colocalization of NLRP3 foci (green, panel A,B) with Iba-1 (red, panel B) and GFAP (red, panel C). Additionally, the quantitative results of images demonstrated a considerable increase in NLRP3 inflammasomes in
Chronic kidney disease (CKD) is characterized by the progressive loss of renal function; moreover, CKD progression commonly leads to multiple comorbidities, including neurological dysfunction and immune disorders. CKD-triggered neuroinflammation significantly contributes to cognitive impairment. This study aimed to investigate the contribution of uremic toxins to cognitive impairment. Serum creatinine, blood urea nitrogen (BUN), indoxyl sulfate (IS), and p-cresol sulfate (PCS) levels were measured using an enzyme-linked immunosorbent assay and high-performance liquid chromatography. The creatinine, BUN, IS, and PCS levels were increased from 4 weeks after 5/6-nephrectomy in mice, which suggested that 5/6-nephrectomy could yield a CKD animal model. Further, CKD mice showed significantly increased brain and serum indoxyl sulfate levels. Immunohistochemistry analysis revealed hippocampal inflammation and NLRP3-inflammasomes in astrocytes. Further, the Y-maze and Morris water maze tests revealed learning and memory defects in CKD mice. AST-120, which is also an IS absorbent, effectively reduced serum and hippocampal IS levels as well as reversed the cognitive impairment in CKD mice. Additionally, NLRP3-knockout mice that underwent 5/6-nephrectomy showed no change in cognitive function. These findings suggested that IS is an important uremic toxin that induces NLRP3 inflammasome-mediated not only in microglia, but it also occurred in astrocytic inflammation, which subsequently causes cognitive impairment.
Background: MicroRNAs (miRNAs) have been proposed as biomarkers in hepatocellular carcinoma (HCC). We aim at evaluating miR-21 and miR-122 in HCC patients treated with drug-eluting beads transarterial chemoembolization (DEB-TACE) as prognostic biomarkers and investigating their correlation with hypoxia inducible factor-1α (HIF-1α) serum levels. Methods: In this retrospective study, 12 healthy subjects, 28 cirrhotics, and 54 HCC patients (tested before and four weeks after DEB-TACE) were included. Whole blood miR-21 and miR-122 levels were measured by quantitative real time (qRT)-PCR, while serum HIF-1α was assessed by an enzyme-linked immunosorbent assay (ELISA) test. Results: The highest level of miR-21 was found in cirrhotics, while HCC patients had the highest level of miR-122 (which was even higher in "viral" HCC, p = 0.006). miR-21 ratio (after/before DEB-TACE) and miR-122 below their respective cut-offs identified patients with longer progression-free survival (p = 0.0002 and p = 0.02, respectively). The combined assessment of alpha-fetoprotein and miR-21 ratio, both independent prognostic predictors, identified early progressors among patients with complete or partial radiological response. miR-21 levels positively correlated with HIF-1α before (p = 0.045) and after DEB-TACE (p = 0.035). Conclusions: miR-21 ratio and miR-122 are useful prognostic markers after DEB-TACE. miR-21 correlates with HIF-1α and probably has a role in modulating angiogenesis in HCC.
miR-124-3p regulated the expression of MRP4. (A) Interaction diagrams between miR-124-3p and ABCC4 3′-UTR by computational prediction. (B) Luciferase reporter results showed that miR-124-3p could bind directly with ABCC4 3′ stands for negative control. (C) mRNA levels of ABCC4 was similar in miR-124-3p mimic transfected cells and control cell lines. (D) MRP4 protein expression was reduced in SMMC-7721 and Huh7 cells transfected with 100 nM miR-124-3p mimics for 72 h, while treatment with the miR-124-3p inhibitors had contrasting effects. Results are expressed as mean ± standard error of the mean (SEM) of at least three independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001, compared with control group. ## p < 0.01, ### p < 0.001, compared with NC inhibitor group. (E) RT-qPCR results showed downregulation of miR-124-3p in HCC tissues compared to the adjacent noncancerous tissues. U6 was used to normalize the miRNA expression, n = 19. (F) Significant downregulation of miR-124-3p was observed in HCC cell lines compared to that in the human primary hepatocytes. Differences between two experimental groups were analyzed by the Student's t test. * p <0.05, ** p <0.01, *** p <0.001.
The proposed molecular mechanism of how circRNA regulates the expression of MRP4. MRP4 is upregulated in HCC. miR-124-3p and miR-4524-5p reduced the expression of MRP4 at the protein but not mRNA level. circHIPK3, as a competitive endogenous RNA, binds with miR-124-3p and miR-4524-5p to decrease free miRNAs, which leads to MRP4 upregulation in HCC.
Multidrug resistance-associated protein 4 (MRP4), a member of the adenosine triphosphate (ATP) binding cassette transporter family, pumps various molecules out of the cell and is involved in cell communication and drug distribution. Several studies have reported the role of miRNAs in downregulating the expression of MRP4. However, regulation of MRP4 by circular RNA (circRNA) is yet to be elucidated. In this study, MRP4 was significantly upregulated in hepatocellular carcinoma (HCC) tissues compared to the adjacent noncancerous tissues. Computational prediction, luciferase reporter assay and miRNA transfection were used to investigate the interaction between miRNAs and MRP4. miR-124-3p and miR-4524-5p reduced the expression of MRP4 at the protein but not mRNA level. Circular RNA in vivo precipitation and luciferase reporter assays demonstrated that circHIPK3, as a competitive endogenous RNA, binds with miR-124-3p and miR-4524-5p. Further, knockdown of circHIPK3 resulted in downregulation of MRP4 protein, whereas cotransfection of circHIPK3-siRNA and miR-124-3p or miR-4524-5p inhibitors restored its expression. In conclusion, we report that miR-4524-5p downregulates the expression of MRP4 and circHIPK3 regulates MRP4 expression by sponging miR-124-3p and miR-4524-5p for the first time. Our results may provide novel insights into the prevention of MRP4-related proliferation and multiple drug resistance in HCC.
The secretome from mSOD1 MNs transfected with anti-miR-124 abolishes the upregulation of miR-124 in the spinal cord (SC) of ALS mice after 3 weeks of intrathecal injection. Expression of miRNA-124 in the lumbar SC of (A) SOD1-G93A (mSOD1) mice injected at 12 weeks of age with the vehicle (basal media of NSC-34 motor neurons (MNs)) in comparison with the respective wildtype (WT) animals, and (B) mSOD1 mice injected with the secretome derived from anti-miR-124-treated mSOD1 MNs (mSOD1 + sec) in comparison with those injected only with the vehicle. The results were obtained at 15 weeks of age and were normalized to SNORD110. Data are expressed as fold change vs. (A) WT + vehicle and (B) mSOD1 + vehicle (mean ± SEM) from at least 5 animals per group. ** p < 0.01 vs. WT + vehicle; # p < 0.05 vs. mSOD1 + vehicle, unpaired and parametric t-test (with Welch's correction when needed).
Motor performance, muscular strength, spontaneous activity, and corticospinal function in ALS mice are improved after 2 weeks of intrathecal injection of the secretome from anti-miR-124-treated mSOD1 motor neurons (MNs). Representative illustrations of (A) footprint test, (B) hanging wire test, (C) cylinder test, and (D) clasping/grasping reflexes test. Measurement of the (E) stride length (in centimeters, cm), (F) time holding onto the cage grid (in seconds, s), (G) number of times the mice reared up against the cylinder, and percentage (%) of animals showing the (H) clasping and (I) grasping reflexes in wild-type (WT)/SOD1-G93A (mSOD1) mice injected with the vehicle (basal media of NSC-34 MNs) and mSOD1 mice injected with the secretome derived from mSOD1 MNs modulated with anti-miR-124 (mSOD1 + sec). Data are expressed as mean ± SEM for (E-G) and percentage (%) for (H,I) from at least 5 animals per group. **** p < 0.0001, ** p < 0.01 and * p < 0.05 vs. WT + vehicle; #### p < 0.0001 and ## p < 0.01 vs. mSOD1 + vehicle. One-way ANOVA followed
Intrathecal injection of anti-miR-124-treated ALS MN-derived secretome in 12-week-old mSOD1 mice prevents loss of muscle fiber area and the deregulation of genes that direct synaptic proteins at 3 weeks after treatment. (A) Representative images of transversal sections of gastrocnemius muscle from the wild-type (WT) and SOD1-G93A (mSOD1) mice injected with the vehicle (basal media of NSC-34 motor neurons (MNs)), as well as mSOD1 mice injected with the secretome derived from anti-miR-124-treated mSOD1 MNs (mSOD1 + sec), stained with hematoxylin-eosin. Scale bars: 50 µm. (B) Respective average muscle fiber area (in µm). (C) Gene expression of synaptophysin, PSD-95, and NeuN from mSOD1 mice injected with the vehicle in comparison with the respective WT mice, and (D) from mSOD1 mice injected with the secretome in comparison with those injected with the vehicle. Results are mean (±SEM) for (B) and expressed as fold change vs.
Intrathecal injection of the secretome from anti-miR-124-modulated mSOD1 motor neurons (MNs) in the lumbar spinal cord of mSOD1 mice at 12 weeks of age prevents age-associated neurodegeneration after 3 weeks of its administration. (A) Representative images of the ventral horn of the lumbar section grey matter stained by Fluoro-Jade B fluorescence (square) from 15-week-old
Neuronal demise and deficits in synaptic signalling, axonal transport, CX3CL1-CX3CR1 axis, and myelination in the lumbar spinal cord (SC) of 15-week-old mSOD1 mice are prevented by the intrathecal injection of the secretome from modulated mSOD1 motor neurons (MNs) in 12-weekold animals. (A) Gene expression of neuronal-related NeuN, synaptophysin, PSD-95, dynein, kinesin, and CX3CL1; (B) protein expression of NeuN; and (C) myelin-associated genes (MBP, PLP, and GPR17) in the SC of 15-week-old SOD1-G93A (mSOD1) mice that were treated with the vehicle
Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease with short life expectancy and no effective therapy. We previously identified upregulated miR-124 in NSC-34-motor neurons (MNs) expressing human SOD1-G93A (mSOD1) and established its implication in mSOD1 MN degeneration and glial cell activation. When anti-miR-124-treated mSOD1 MN (preconditioned) secretome was incubated in spinal cord organotypic cultures from symptomatic mSOD1 mice, the dysregulated homeostatic balance was circumvented. To decipher the therapeutic potential of such preconditioned secretome, we intrathecally injected it in mSOD1 mice at the early stage of the disease (12-week-old). Preconditioned secretome prevented motor impairment and was effective in counteracting muscle atrophy, glial reactivity/dysfunction, and the neurodegeneration of the symptomatic mSOD1 mice. Deficits in corticospinal function and gait abnormalities were precluded, and the loss of gastrocnemius muscle fiber area was avoided. At the molecular level, the preconditioned secretome enhanced NeuN mRNA/protein expression levels and the PSD-95/TREM2/IL-10/arginase 1/MBP/PLP genes, thus avoiding the neuronal/glial cell dysregulation that characterizes ALS mice. It also prevented upregulated GFAP/Cx43/S100B/vimentin and inflammatory-associated miRNAs, specifically miR-146a/miR-155/miR-21, which are displayed by symptomatic animals. Collectively, our study highlights the intrathecal administration of the secretome from anti-miR-124-treated mSOD1 MNs as a therapeutic strategy for halting/delaying disease progression in an ALS mouse model.
Severe acute respiratory syndrome coronovirus-2 (SARS-CoV-2) is the cause of the coronavirus disease 2019 (COVID-19) pandemic. Vaccination is considered the core approach to containing the pandemic. There is currently insufficient evidence on the efficacy of these vaccines in immunosuppressed inflammatory bowel disease (IBD) patients. The aim of this study was to investigate the humoral response in immunosuppressed IBD patients after COVID-19 mRNA vaccination. In this prospective study, IgG antibody levels (AB) against the SARS-CoV-2 receptor-binding domain (spike-protein) were quantitatively determined. For assessing the potential neutralizing capacity, a SARS-CoV-2 surrogate neutralization test (sVNT) was employed in IBD patients (n = 95) and healthy controls (n = 38). Sera were examined prior to the first/second vaccination and 3/6 months after second vaccination. Patients showed lower sVNT (%) and IgG-S (AU/mL) AB both before the second vaccination (sVNT p < 0.001; AB p < 0.001) and 3 (sVNT p = 0.002; AB p = 0.001) and 6 months (sVNT p = 0.062; AB p = 0.061) after the second vaccination. Although seroconversion rates (sVNT, IgG-S) did not differ between the two groups 3 months after second vaccination, a significant difference was seen 6 months after second vaccination (sVNT p = 0.045). Before and three months after the second vaccination, patients treated with anti-tumor necrosis factor (TNF) agents showed significantly lower AB than healthy subjects. In conclusion, an early booster shot vaccination should be discussed for IBD patients on anti-TNF therapy.
Effect of SZV 1287 on proton-induced TRPV1 receptor activation on cultured primary sensory neurons. (a) The percentage of proton-responsive cells at pH 5.3 or pH 5.3 + 10 μM CZP is presented on control plates and on SZV 1287-pretreated (0.1, 1, and 10 μM) plates. Ca 2+ -responses are presented in % of total number of examined neurons, * p < 0.05 vs. control; one-way ANOVA (treatment factor F = 3.45, p = 0.017) followed by Dunnett's multiple comparison test. (b) The change in the fluorescence ratio (R = F340/F380) demonstrating the activation of individual cells is presented on the control, SZV 1287-pretreated (0.1, 1 and 10 μM) and CZP-treated (10 μM) plates. Each column represents mean + SEM of n = 65-97 cells/group, ** p < 0.01, *** p < 0.001 vs. the control; one-way ANOVA (treatment factor F = 8.73, p < 0.0001) followed by Dunnett's multiple comparison test. Abbreviation: CZP, capsazepine.
Effects of SZV 1287 on general locomotor activity and deep body temperature in mice. (a) General locomotor activity and (b) abdominal temperature in response to a single administration of enterosolvent capsules; (c) colonic temperature in response to single i.p. administration of 20 and 50
SZV 1287 (3-(4,5-diphenyl-1,3-oxazol-2-yl)propanal oxime) is a novel multi-target candidate under preclinical development for neuropathic pain. It inhibits amine oxidase copper containing 3, transient receptor potential ankyrin 1 and vanilloid 1 (TRPV1) receptors. Mainly under acidic conditions, it is transformed to the cyclooxygenase inhibitor oxaprozin, which is ineffective for neuropathy. Therefore, an enterosolvent capsule is suggested for oral formulation, which we investigated for nociception, basic kinetics, and thermoregulatory safety in mice. The antihyperalgesic effect of SZV 1287 (10, 20, 50, and 200 mg/kg, p.o.) was determined in partial sciatic nerve ligation-induced traumatic neuropathy by aesthesiometry, brain and plasma concentrations by HPLC, and deep body temperature by thermometry. Its effect on proton-induced TRPV1 activation involved in thermoregulation was assessed by microfluorimetry in cultured trigeminal neurons. The three higher SZV 1287 doses significantly, but not dose-dependently, reduced neuropathic hyperalgesia by 50% of its maximal effect. It was quickly absorbed; plasma concentration was stable for 2 h, and it entered into the brain. Although SZV 1287 significantly decreased the proton-induced TRPV1-mediated calcium-influx potentially leading to hyperthermia, it did not alter deep body temperature. Oral SZV 1287 inhibited neuropathic hyperalgesia and, despite TRPV1 antagonistic action and brain penetration, it did not influence thermoregulation, which makes it a promising analgesic candidate.
The changes in lung ventilation defect (a), gas-blood exchange time (b), brain white matter volume (c), and brain gray matter volume (d) among four patients from April 14 to December 18, after discharge, corresponding to patients 4, 5, 8, and 9 in Table 1.
Scheme 1. The SARS-CoV-2 virus affects the nervous and respiratory systems. Sketch illustrating brain 1 H MRI (top) and hyperpolarized 129 Xe gas lung MRI (bottom). The top schematic illustrates the segmentation and normalization of brain 1 H MRI. The bottom schematic shows two generations of lung respiratory airways of the Weibel lung model, with the main morphometric parameters including internal radius (r), external radius (R), and depth of alveolar sleeve (h). The diagram of the gas-blood exchange region of the alveoli is shown in the lower illustration. The total septal thickness, xenon gas exchange time constant, and hematocrit are assigned as d, T, and Hct, respectively.
Although the lungs are the primary organ involved, increasing evidence supports the neuroinvasive potential of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). This study investigates the potential relationship between coronavirus disease (COVID-19)-related deterioration of brain structure and the degree of damage to lung function. Nine COVID-19 patients were recruited in critical condition from Jin Yin-tan Hospital (Wuhan, China) who had been discharged between 4 February and 27 February 2020. The demographic, clinical, treatment, and laboratory data were extracted from the electronic medical records. All patients underwent chest CT imaging, 129Xe gas lung MRI, and 1H brain MRI. Four of the patients were followed up for 8 months. After nearly 12 months of recovery, we found no significant difference in lung ventilation defect percentage (VDP) between the COVID-19 group and the healthy group (3.8 ± 2.1% versus 3.7 ± 2.2%) using 129Xe MRI, and several lung-function-related parameters—such as gas–blood exchange time (T)—showed improvement (42.2 ms versus 32.5 ms). Combined with 1H brain MRI, we found that the change in gray matter volume (GMV) was strongly related to the degree of pulmonary function recovery—the greater the increase in GMV, the higher degree of pulmonary function damage.
Stevia rebaudiana (Asteraceae), commonly known as candyleaf, sweetleaf, or sugarleaf, is a branched bushy shrub whose leaves are used as a natural sweetener owing to the high content of sweet diterpenes. As part of our ongoing work to identify structurally novel and bioactive natural products, phytochemical investigation of the ethanolic extract of S. rebaudiana leaves led to the isolation of one new labdane-type diterpene, 6-O-acetyl-(12R)-epiblumdane (1), and nine known terpenoids, including six diterpenes (2–6 and 10), two monoterpenes (7 and 8), and one triterpene (9). The structure of the new compound 1 was elucidated via analysis of one- and two-dimensional nuclear magnetic resonance (NMR) spectroscopic data and high-resolution electrospray ionization mass spectrometry data, and its absolute configuration was established using electronic circular dichroism (ECD) calculations and gauge-including atomic orbital NMR chemical shift calculations, followed by DP4 + probability analysis. The isolated compounds 1–10 were evaluated for their effects on glucose-stimulated insulin secretion in the INS-1 rat pancreatic β-cell line. The new compound 1, 6-O-acetyl-(12R)-epiblumdane, stimulated glucose-stimulated insulin secretion in INS-1 pancreatic β-cells without inducing cytotoxicity. Thus, 6-O-acetyl-(12R)-epiblumdane (1), an active compound derived from S. rebaudiana leaves, can be used as a potential therapeutic agent to prevent type 2 diabetes.
Study design. GP = general practitioner; OGTT = oral glucose tolerance test; clin. chemistry = clinical chemistry (HbA1c, triglycerides and HDL, LDL, and total cholesterol). Anthropometry includes body height (only at baseline), body weight, waist circumference, and fat percentage.
Baseline characteristics by treatment group.
Remission data * for the intervention and usual care groups after the intervention (week 13) and at the one-and two-year follow-ups (week 52 and 104), expressed as the number and percentage of participants with T2DM at baseline **.
Changes in oral glucose tolerance test response from baseline to 13 weeks (end of interven- tion) for the main type 2 diabetes subtypes.
A type 2 diabetes mellitus (T2DM) subtyping method that determines the T2DM phenotype based on an extended oral glucose tolerance test is proposed. It assigns participants to one of seven subtypes according to their β-cell function and the presence of hepatic and/or muscle insulin resistance. The effectiveness of this subtyping approach and subsequent personalized lifestyle treatment in ameliorating T2DM was assessed in a primary care setting. Sixty participants, newly diagnosed with (pre)diabetes type 2 and not taking diabetes medication, completed the intervention. Retrospectively collected data of 60 people with T2DM from usual care were used as controls. Bodyweight (p < 0.01) and HbA1c (p < 0.01) were significantly reduced after 13 weeks in the intervention group, but not in the usual care group. The intervention group achieved 75.0% diabetes remission after 13 weeks (fasting glucose ≤ 6.9 mmol/L and HbA1c < 6.5% (48 mmol/mol)); for the usual care group, this was 22.0%. Lasting (two years) remission was especially achieved in subgroups with isolated hepatic insulin resistance. Our study shows that a personalized diagnosis and lifestyle intervention for T2DM in a primary care setting may be more effective in improving T2DM-related parameters than usual care, with long-term effects seen especially in subgroups with hepatic insulin resistance.
The characteristics of the studied women with PCOS and the control group (CG).
Correlations between potassium levels and arachidonic acid (AA) derivatives.
Potassium helps to maintain the water–electrolyte and acid–base balance. There is little research on the relationship between plasma fatty acids (FAs), inflammatory mediators and red blood cell potassium levels in women with polycystic ovary syndrome (PCOS). This study included 38 Caucasian women with PCOS. Potassium in the erythrocytes was determined by inductively coupled atomic plasma emission spectrometry. The FAs were analysed with gas chromatography, and liquid chromatography was used to separate the eicosanoids. The relationships between the potassium content and the amounts of fatty acids, as well as potassium and arachidonic acid (AAs) derivatives, were analysed. Significant negative correlations were found with, among others, pentadecanoic acid, palmitic acid, stearic acid and arachidic acid, whereas a positive correlation was found with neuronic acid. Positive correlations were observed with 9, 13 HODE (derivatives synthetized from linolenic acid) and 5 oxo ETE and 5 HETE (from 5 LOX pathway). Saturated fatty acids reduce the influx of potassium into the cell by destabilizing the pH of the cytosol, and thus exacerbating the inflammatory response through the activation of the AA cascade. Therefore, improving the flow of potassium inside the cell is important in the treatment of patients.
(A) Dentin fine microstructure: (a) CTR, (b) 3H, (c) 6H, (d) 12H, (e) 24H, (f) 36H, (g) 48 H, (h) 192H; (B) roughness surface of dentin fine microstructure; (C) dentin nanostructure: (a) CTR, (b) 3H, (c) 6H, (d) 12H, (e) 24H, (f) 36H, (g) 48 H, (h) 192H; (D) characteristics of dentin nanostructure; (E) HAP crystallite diameter. For (B,D), values are expressed in mean ± SD. a p < 0.05 compared to CTR group; b p < 0.05 compared to IRD3 group.
This study aimed to evaluate, in vitro, the effects of I-131 on enamel and dentin in healthy human incisive permanent maxillary teeth. Our in vitro model analogue with the in vivo conditions of differentiated thyroid carcinoma patients treated with I-131, consisted in a solution of I-131 dissolved in artificial saliva. A total of 48 teeth were divided into eight groups (n = 6): control, irradiation groups at 3, 6, 12, 24, 36, 48, and 192 h, respectively. At the end of radiation exposure, radioiodine activity of specimens was assessed. Fine microstructure, nanostructure, surface roughness, and hidroxyapatite (HAP) crystallite diameter were investigated by atomic force microscopy (AFM) to both enamel and dentin structures. There is a constant increase of radioactivity in dental structures at 3, 6, 12, 24 h, due to progressive retention and I-131 migration, with a maximum at 36 h. Enamel showed notable alterations, which was correlated with the increase of the treatment time. A relevant visible distance between the HAP prisms was observed after 24 h. The surface suffered a loss in its compact structure. I-131 acts in the same way on HAP crystallites in dentin as in those in enamel. It was noticed that their morpho-dimensional changes occurred only after 12 h of treatment. Radioiodine-131 determines degradation of enamel and dentin by starting from the alteration of the crystalline network of HAP prisms, transforming them from compact materials into an agglomeration of rocky submicron structures.
The androgen receptor (AR) plays a critical role in prostate cancer (PCa) development and metastasis. Thus, blocking AR activity and its downstream signaling constitutes a major strategy for PCa treatment. Here, we report on the potent anti-PCa activity of a small-molecule imidazoacridinone, C-1311. In AR-positive PCa cells, C-1311 was found to inhibit the transcriptional activity of AR, uncovering a novel mechanism that may be relevant for its anticancer effect. Mechanistically, C-1311 decreased the AR binding to the prostate-specific antigen (PSA) promoter, reduced the PSA protein level, and, as shown by transcriptome sequencing, downregulated numerous AR target genes. Importantly, AR-negative PCa cells were also sensitive to C-1311, suggesting a promising efficacy in the androgen-independent PCa sub-type. Irrespective of AR status, C-1311 induced DNA damage, arrested cell cycle progression, and induced apoptosis. RNA sequencing indicated significant differences in the transcriptional response to C-1311 between the PCa cells. Gene ontology analysis showed that in AR-dependent PCa cells, C-1311 mainly affected the DNA damage response pathways. In contrast, in AR-independent PCa cells, C-1311 targeted the cellular metabolism and inhibited the genes regulating glycolysis and gluconeogenesis. Together, these results indicate that C-1311 warrants further development for the treatment of PCa.
MiR-138-5p expression in the undamaged spinal cord: (A) Representative image of miR-138-5p expression (pink) in a transverse section of the thoracic spinal cord. (B-D) Co-staining of GFAP (astrocytes, (B), NeuN (neurons, (C) and APC (oligodendrocytes, (D) cell markers (green) MiR-138-5p expression in the undamaged spinal cord: (A) Representative image of miR-138-5p expression (pink) in a transverse section of the thoracic spinal cord. (B-D) Co-staining of GFAP (astrocytes, (B), NeuN (neurons, (C) and APC (oligodendrocytes, (D) cell markers (green) with miR-138-5p probe (red) in spinal cord sections. (E) Map of the Rexed laminae in the T9 spinal segment detailing the mean percentage of miR-138-5p positive neurons present in each lamina of the naive spinal cord (7 sections from 2 individuals).
MiR-138-5p reduces the expression of Caspase 3, Caspase 7, and BAK: (A) Gene expression data from RT-qPCR analysis of C6 cultures transiently transfected with miR-138-5p or the negative control (Neg Ctr) mimics. The dot plot represents the average of ΔCt from six independent experiments ± CI. Comparison between expression values was carried out through a paired t-test with ∆Ct data (n = 3). (B) Analyses of protein expression of C6 cultures transiently transfected with miR-138-5p or Neg Ctr mimics and analyzed using immunoblot and densitometry. The images represent the results of two independent replicates. The bar graph underneath shows the densitometry data of each apoptotic protein following transfection with miR-138-5p relative to their respective values after transfection with Neg Ctr, and comparisons between the two were carried out through a paired t-test. The bars represent the average ± SD of, at least, four independent experiments. Complete blot images are available at OSF (, accessed on 24 May 2022). * and ** indicate p < 0.05 and p < 0.01, respectively, relative to its corresponding negative control.
The central nervous system microRNA miR-138-5p has attracted much attention in cancer research because it inhibits pro-apoptotic genes including CASP3. We hypothesize that miR-138-5p downregulation after SCI leads to overexpression of pro-apoptotic genes, sensitizing neural cells to noxious stimuli. This study aimed to identify miR-138-5p targets among pro-apoptotic genes overexpressed following SCI and to confirm that miR-138-5p modulates cell death in neural cells. Gene expression and histological analyses revealed that the drop in miR-138-5p expression after SCI is due to the massive loss of neurons and oligodendrocytes and its downregulation in neurons. Computational analyses identified 176 potential targets of miR-138-5p becoming dysregulated after SCI, including apoptotic proteins CASP-3 and CASP-7, and BAK. Reporter, RT-qPCR, and immunoblot assays in neural cell cultures confirmed that miR-138-5p targets their 3′UTRs, reduces their expression and the enzymatic activity of CASP-3 and CASP-7, and protects cells from apoptotic stimuli. Subsequent RT-qPCR and histological analyses in a rat model of SCI revealed that miR-138-5p downregulation correlates with the overexpression of its pro-apoptotic targets. Our results suggest that the downregulation of miR-138-5p after SCI may have deleterious effects on neural cells, particularly on spinal neurons.
The Cancer Genome Atlas Program (TCGA) database shows inverse correlation between BRIC5 and miR-138 in their expression. (A) BIRC5 expression is up-regulated, while (B) miR-138 expression is down-regulated in most types of glioma compared to that of normal tissues. (C) Direct comparison between BIRC5 and miR-138 clearly reveals inverse correlation in all types of glioma compared to normal tissues. All error bars indicates standard deviations, and Student ttest was used to determine the significance in difference between the two groups. * p < 0.05, ** p < 0.01, *** p < 0.001.
Glioblastoma (GBM) is one of the most deadly cancers and poorly responses to chemotherapies, such as temozolomide (TMZ). Dysregulation of intrinsic signaling pathways in cancer cells are often resulted by dysregulated tumor suppressive microRNAs (miRNAs). Previously, we found miR-138 as one of tumor suppressive miRNAs that were significantly down-regulated in GBM. In this study, we demonstrated that ectopic over-expression of miR-138 sensitizes GBM cells to the treatment of TMZ and increased apoptotic cell death. Mechanistically, miR-138 directly repressed the expression of Survivin, an anti-apoptotic protein, to enhance caspase-induced apoptosis upon TMZ treatment. Using an intracranial GBM xenograft mice model, we also showed that combination of miR-138 with TMZ increases survival rates of the mice compared to the control mice treated with TMZ alone. This study provides strong preclinical evidence of the therapeutic benefit from restoration of miR-138 to sensitize the GBM tumor to conventional chemotherapy.
A. Candidate target genes of miR-139-5p. B. Candidate target genes of miR-139-3p. A. Candidate target genes of miR-139-5p
We previously found that both the guide and passenger strands of the miR-139 duplex (miR-139-5p and miR-139-3p, respectively) were downregulated in cancer tissues. Analysis of TCGA datasets revealed that low expression of miR-139-5p (p < 0.0001) and miR-139-3p (p < 0.0001) was closely associated with 5-year survival rates of patients with renal cell carcinoma (RCC). Ectopic expression assays showed that miR-139-5p and miR-139-3p acted as tumor-suppressive miRNAs in RCC cells. Here, 19 and 22 genes were identified as putative targets of miR-139-5p and miR-139-3p in RCC cells, respectively. Among these genes, high expression of PLXDC1, TET3, PXN, ARHGEF19, ELK1, DCBLD1, IKBKB, and CSF1 significantly predicted shorter survival in RCC patients according to TCGA analyses (p < 0.05). Importantly, the expression levels of four of these genes, PXN, ARHGEF19, ELK1, and IKBKB, were independent prognostic factors for patient survival (p < 0.05). We focused on PXN (paxillin) and investigated its potential oncogenic role in RCC cells. PXN knockdown significantly inhibited cancer cell migration and invasion, possibly by regulating epithelial–mesenchymal transition. Involvement of the miR-139-3p passenger strand in RCC molecular pathogenesis is a new concept. Analyses of tumor-suppressive-miRNA-mediated molecular networks provide important insights into the molecular pathogenesis of RCC.
Representative anatomical T2-weighted (rapid acquisition with relaxation enhancement (RARE)) images in sagittal (A) and axial (B) orientations, with oblique spectroscopy slice geometry shown for a subcutaneous tumor (t) in a rat and for a thermal lactate (lac) phantom for transmit power calibration. To measure within the target tumor, with the scanner reference frequency set to that of [1-13 C]pyruvate, the slice geometry is prescribed twice: once on the target tumor for [1-13 C]pyruvate (yellow-and-red outlines) and again offset by the chemical shift displacement of [1-13 C]lactate (dashed yellow outlines). This produces excitations that, at the location of the tumor, alternate between [1-13 C]pyruvate and [1-13 C]lactate frequencies. An additional slice may also be prescribed (dashed blue outlines) to target lactate in the phantom (solid blue outlines). Carbopol ® 980 gel (g, bright contrast) was placed around the tumor to improve its B0 field uniformity. The locations of the 13 C surface receive coils (c1 and c2, light blue outlines), muscle (m), and a point-resolved spectroscopy (PRESS) voxel (white box surrounding t) for shimming on proton signal within the tumor are also marked. Magnitude spectra (C) from [1-13 C]lactate in the phantom (blue) and from hyperpolarized [1-13 C]lactate in the tumor (yellow) after injection, exhibiting a frequency shift between the two sources of lactate signal. In cases where this shift is large, the placement of a slice prescribed to measure within the phantom may benefit from additional spatial offset to compensate for that frequency shift in order to excite the 13 C in the phantom.
Hyperpolarized 13C nuclear magnetic resonance spectroscopy can characterize in vivo tissue metabolism, including preclinical models of cancer and inflammatory disease. Broad bandwidth radiofrequency excitation is often paired with free induction decay readout for spectral separation, but quantification of low-signal downstream metabolites using this method can be impeded by spectral peak overlap or when frequency separation of the detected peaks exceeds the excitation bandwidth. In this work, alternating frequency narrow bandwidth (250 Hz) slice-selective excitation was used for 13C spectroscopy at 7 T in a subcutaneous xenograft rat model of human pancreatic cancer (PSN1) to improve quantification while measuring the dynamics of injected hyperpolarized [1-13C]lactate and its metabolite [1-13C]pyruvate. This method does not require sophisticated pulse sequences or specialized radiofrequency and gradient pulses, but rather uses nominally spatially offset slices to produce alternating frequency excitation with simpler slice-selective radiofrequency pulses. Additionally, point-resolved spectroscopy was used to calibrate the 13C frequency from the thermal proton signal in the target region. This excitation scheme isolates the small [1-13C]pyruvate peak from the similar-magnitude tail of the much larger injected [1-13C]lactate peak, facilitates quantification of the [1-13C]pyruvate signal, simplifies data processing, and could be employed for other substrates and preclinical models.
Ex vivo 13 C metabolic profiling after 13 C glucose or 13 C glutamine feeding (fluxomic experiments) in response to BRAF and/or MEK inhibition. Representative spectra of lactate C1 issued from the 13 C glucose metabolism in the control group (blue) and in the BRAFi/MEKi treated group (orange) (A). Quantification of the 13 C detectable metabolites in YUMM1.7 xenografts, represented by the area under the curve corrected for internal standard (TSP) and tumor mass, arbitrary units (a.u.), two-way anova, Holm-Sidak multiple comparisons test. Sample size n = 3 for Ctrl and BRAFi and n = 5 for MEKi and BRAFi/MEKi (Combo) (B). Representative spectra of glutamate issued from the 13 C glutamine metabolism in the control group (blue) and in the BRAFi/MEKi treated group Ex vivo 13 C metabolic profiling after 13 C glucose or 13 C glutamine feeding (fluxomic experiments) in response to BRAF and/or MEK inhibition. Representative spectra of lactate C1 issued from the 13 C glucose metabolism in the control group (blue) and in the BRAFi/MEKi treated group (orange) (A). Quantification of the 13 C detectable metabolites in YUMM1.7 xenografts, represented by the area under the curve corrected for internal standard (TSP) and tumor mass, arbitrary units (a.u.), two-way anova, Holm-Sidak multiple comparisons test. Sample size n = 3 for Ctrl and BRAF i and n = 5 for MEK i and BRAFi/MEK i (Combo) (B). Representative spectra of glutamate issued from the 13 C glutamine metabolism in the control group (blue) and in the BRAFi/MEKi treated group (orange) (C), and 13 C glutamate quantification (a.u.), one-way anova, Holm-Sidak multiple comparisons test n = 5/grp (D).
A vast majority of BRAF V600E mutated melanoma patients will develop resistance to combined BRAF/MEK inhibition after initial clinical response. Resistance to targeted therapy is described to be accompanied by specific metabolic changes in melanoma. The aim of this work was to evaluate metabolic imaging using 13C-MRS (Magnetic Resonance Spectroscopy) as a marker of response to BRAF/MEK inhibition in a syngeneic melanoma model. Tumor growth was significantly delayed in mice bearing YUMM1.7 melanoma xenografts treated with the BRAF inhibitor vemurafenib, and/or with the MEK inhibitor trametinib, in comparison with the control group. 13C-MRS was performed in vivo after injection of hyperpolarized (HP) 13C-pyruvate, at baseline and 24 h after treatment, to evaluate dynamic changes in pyruvate-lactate exchange. Furthermore, ex vivo 13C-MRS steady state metabolic tracing experiments were performed after U-13C-glucose or 5-13C-glutamine injection, 24 h after treatment. The HP 13C-lactate-to-pyruvate ratio was not modified in response to BRAF/MEK inhibition, whereas the production of 13C-lactate from 13C-glucose was significantly reduced 24 h after treatment with vemurafenib, trametinib, or with the combined inhibitors. Conversely, 13C-glutamine metabolism was not modified in response to BRAF/MEK inhibition. In conclusion, we identified 13C-glucose fluxomic as a potential marker of response to BRAF/MEK inhibition in YUMM1.7 melanoma xenografts.
miR-142-3p affects cell size, invasion and contractility in St-T1b cells. (A) MiR-142-3p reduces stromal cell size, 320-fold magnification. (B) Quantification of the effect of miR-142-3p on stromal cell size (n = 376, Kruskal-Wallis test). (C) Migration assay through fibronectin (an ITGAV ligand)-coated chamber. Scale bar, 250 µm. (D) MiR-142-3p reduces the migratory potential of stromal cells (n = 10, t-test). (E) Collagen contraction assay after 48, 96, and 144 h in St-T1b and ectopic stromal cells (ESCs), Scale bar, ~5 mm. (F,G) MiR-142-3p reduces contractility of both eutopic St-T1b and ectopic ESCs cells (n = 3, multiple t-tests, data are representative of two independent repeats). Data in the whole figure panel represent mean ± s.d. For all figures in this panel * p < 0.05; ** p < 0.01; *** p < 0.001, **** p < 0.0001, and n.s. p > 0.05.
Downregulated microRNA-142-3p signaling contributes to the pathogenesis of endometriosis, an invasive disease where the lining of the uterus grows at ectopic locations, by yet incompletely understood mechanisms. Using bioinformatics and in vitro assays, this study identifies cytoskeletal regulation and integrin signaling as two relevant categories of miR-142-3p targets. qPCR revealed that miR-142-3p upregulation in St-T1b cells downregulates Rho-associated protein kinase 2 (ROCK2), cofilin 2 (CFL2), Ras-related C3 botulinum toxin substrate 1 (RAC1), neural Wiskott-Aldrich syndrome protein (WASL), and integrin α-V (ITGAV). qPCR and Western-blotting showed miR-142-3p effect on WASL and ITGAV was significant also in primary endometriotic stroma cells. Luciferase reporter assays in ST-T1b cells then confirmed direct regulation of ITGAV and WASL. On the functional side, miR-142-3p upregulation significantly reduced ST-T1b cell size, the size of vinculin plaques, migration through fibronectin-coated transwell filters, and the ability of ST-T1b and primary endometriotic stroma cells to contract collagen I gels. These results suggest that miR-142-3p has a strong mechanoregulatory effect on endometrial stroma cells and its external administration reduces the invasive endometrial phenotype. Within the limits of an in vitro investigation, our study provides new mechanistic insights into the pathogenesis of endometriosis and provides a perspective for the development of miR-142-3p based drugs for inhibiting invasive growth of endometriotic cells.
Selection flow of the final study population. Figure 1 represents the flow diagram for patient inclusion/exclusion path. (flow diagram was drawn by Lucidchart©, Lucid Software Inc., 2021;; access date: 31 May 2021).
Characteristics of hypertensive patients at baseline (T0) and after 6-month antihypertensive therapy (T1).
Characteristics of hypertensive patients at baseline (T0) and after 6-month antihypertensive therapy (T1) according to the treatment assigned.
Two-by-two comparisons by treatment.
miR profile could be associated to CV risk, and also to prognosis/outcome in response to therapeutic approach. We aimed to evaluate if anti-hypertensive drugs enalapril, losartan or olmesartan have effects on monocyte miR profile in essential hypertensives without target organ involvement. For this purpose, 82 hypertensives and 49 controls were included; we evaluated SBP/DBP, lipid profile, glucose, CRP, fibrinogen, arterial stiffness indices (PWV; AIx), and cIMT at baseline (T0) and after 24 weeks of treatment (T1). Subjects with LDL-C ≥ 160 mg/dL, TG ≥ 200 mg/dL, BMI ≥ 30, and other additional CV risk factors were excluded. Patients who were prescribed to receive once-a-day enalapril 20 mg, losartan 100 mg or olmesartan 20 mg were eligible for the study. At T1, we found a significant improvement of SBP (−18.5%), DBP (−18%), HDL-C and LDL-C (+3% and −5.42%), glucose (−2.15%), BMI (−3.23%), fibrinogen (−11%), CRP (−17.5%,), AIx (−49.1%) PWV (−32.2%), and monocyte miR expression (miR-221: −28.4%; miR-222: −36%; miR-145: +41.7%) with respect to baseline. miR profile was compared to control subjects at baseline and at T1. We found some little difference in the behaviour of the three treatments on some variables: olmesartan was the most effective in reducing fibrinogen, DBP, CRP, and AIx (−13.1%, −19.3%, −21.4%, and −56.8%, respectively). Enalapril was the drug more significantly increasing the expression of miR-145. In conclusion, enalapril, losartan and olmesartan are effective in improving mechanical and humoral factors associated to AS and atherogenesis. These drugs appear to be able to modify miRs 221/222 and miR-145 expression in drug-naïve hypertensives, making it closer to that of control subjects; additionally, this provides a good blood pressure compensation, contributing to slow the progression of vascular damage.
The difference of microRNA between remission and non-remission in patients with major depressive disorder: (a) let-7e; (b) miR-21-5p; (c) miR-223 (d) miR-145; (e) miR-146a; (f) miR-155. Before antidepressants treatment, let-7e, miR-21-5p, miR-145, and miR-146a were significantly lower than non-remission group but were not different from health controls. Antidepressants significantly increased let-7e, miR-21-5p, miR-223, miR-145, miR-146a and miR-155 expression levels.* p < 0.05; ** p < 0.01.
ROC analysis of ability of microRNAs to discriminate remission from non-remission. (a) let-7e; (b) miR-21-5p; (c) miR-223 (d) miR-145; (e) miR-146a; (f) miR-155. (a,d,e) let-7e, miR-145, and miR-146a showed acceptable discrimination (0.7 ≥ AUC > 0.6) between remission and non-remission (b,f) miR-21-5p and miR-155 showed poor discrimination (0.6 ≥ AUC > 0.5). (c) miR-223 showed no discrimination.
Primers for qRT-PCR.
Demographic and clinical characteristics of patients with major depressive disorder and health control.
Difference of exosomal microRNA expression between remission and non-remission at baseline.
The intracellular microRNAs that negatively regulate Toll-like receptor 4 signaling pathways in peripheral blood mononuclear cells are associated with major depressive disorder (MDD). However, that the distribution of these microRNAs in exosomes could be a biomarker of central nervous system diseases is just beginning to be explored. In the present study, we isolated serum exosomes from patients with MDD and healthy controls to explore the levels of exosomal microRNAs, including let-7e, miR-21-5p, miR-223, miR-145, miR-146a, and miR-155. We also investigated the changes of these exosomal microRNAs after antidepressant treatment and their association with clinical changes in scores on the Hamilton Depression Rating Scale. An ANCOVA adjusted by age, sex, BMI, and smoking showed higher expression levels of miR-146a (p = 0.006) in patients with MDD compared to controls. Patients who achieved remission showed significantly lower let-7e, miR-21-5p, miR-145, miR-146a, and miR-155 levels before treatment and increased levels after antidepressant treatment compared with the non-remission group. Through receiver operating characteristic (ROC) analysis, let-7e, miR-145, and miR-146a showed acceptable discrimination between the remission and non-remission groups, whereas miR-21-5p and miR-155 showed poor discrimination. These findings demonstrate that exosomal microRNAs may play essential roles in predicting antidepressants response.
Primers sequences for RT-PCR.
The constant dialogue between the plant world and the animal world (including man among them) has been known since the time of Adam and Eve, where an apple was the origin of the evils of the world. Apart from Snow White—who might have something to object to when it comes to the use of apples—fruits, plants, and natural extracts have been known for millennia as remedies for human health-related ailments. In the light of such evidence, the aim of the present work was to investigate from a biological point of view the potential role of apple exosomes in inflammatory processes on human cells. To this end we isolated and characterized apple exosomes and treated human cells such as macrophages and NCTC L929 as cancer cells in order to evaluate the tumorigenic and anti-inflammatory effect of apple exomes. Microscopic and molecular biology analyses were conducted to characterize exosomes and to assess cell proliferation, death, and miRNA line, as well as gene expression and the uptake of exosomes by cells. The results confirm the absolute biological safety of exosomes and their anti-inflammatory effect, mediated mainly by miRNA146 production by M2 macrophages.
Patients with locally advanced rectal cancer (LARC) who achieve a pathological complete response (pCR) to neoadjuvant chemoradiotherapy (NACRT) have an excellent prognosis, but only approximately 30% of patients achieve pCR. Therefore, identifying predictors of pCR is imperative. We employed a microRNA (miRNA) microarray to compare the miRNA profiles of patients with LARC who achieved pCR (pCR group, n = 5) with those who did not (non-pCR group, n = 5). The validation set confirmed that miRNA-148a was overexpressed in the pCR group (n = 11) compared with the non-pCR group (n = 40). Cell proliferation and clonogenic assays revealed that miRNA-148a overexpression radio-sensitized cancer cells and inhibited cellular proliferation, before and after irradiation (p < 0.01). Apoptosis assays demonstrated that miRNA-148a enhanced apoptosis before and after irradiation. Reporter assays revealed that c-Met was the direct target gene of miRNA-148a. An in vivo study indicated that miRNA-148a enhanced the irradiation-induced suppression of xenograft tumor growth (p < 0.01). miRNA-148a may be a biomarker of pCR following NACRT and can promote apoptosis and inhibit proliferation in CRC cells by directly targeting c-Met in vitro and enhancing tumor response to irradiation in vivo.
Background: Radioimmunotherapy utilize a targeting antibody coupled to a therapeutic isotope to target and treat a tumor or disease. In this study we examine the synthesis and cell binding of a polymer scaffold containing a radiotherapeutic isotope and a targeting antibody. Methods: The multistep synthesis of a fluorescent or (149)Promethium-labeled Trastuzumab-polyethyleneimine (PEI), Trastuzumab, or PEI is described. In vitro uptake, internalization and/or the binding affinity to the Her2/neu expressing human breast adenocarcinoma SKBr3 cells was investigated with the labeled compounds. Results: Fluorescent-labeled Trastuzumab-PEI was internalized more into cells at 2 and 18 h than fluorescent-labeled Trastuzumab or PEI. The fluorescent-labeled Trastuzumab was concentrated on the cell surface at 2 and 18 h and the labeled PEI had minimal uptake. DOTA-PEI was prepared and contained an average of 16 chelates per PEI; the compound was radio-labeled with (149)Promethium and conjugated to Trastuzumab. The purified (149)Pm-DOTA-PEI-Trastuzumab had a radiochemical purity of 96.7% and a specific activity of 0.118 TBq/g. The compound demonstrated a dissociation constant for the Her2/neu receptor of 20.30 ± 6.91 nM. Conclusion: The results indicate the DOTA-PEI-Trastuzumab compound has potential as a targeted therapeutic carrier, and future in vivo studies should be performed.
Photos of patients 8 (A), 10 (B) and 11 (C) with specific facial phenotypes-a thin nasal back with a pointed tip, a small pointed chin, thin lips, almond-shaped eyes, hypoplasia of the middle third of the face and large protruding auricles.
X-ray of patients 8 (A), 11 (B) and 12 (C). We see the absence of hypertrophic callus at the fracture site in patient 8, the presence of hypertrophic callus in patient 11 with the classic triad of signs and the absence of ossification of the interosseous membrane in patient 12.
Demographic characteristics of patients with OI type V.
Radiographic characteristics of patients with OI type V.
Comparative analysis of the clinical characteristics of the studied patients versus litera- ture data.
Osteogenesis imperfecta (OI) is a large group of genetically heterogeneous diseases resulting from decreased bone density and an abnormal microarchitecture, which are clinically manifested by abnormal bone fractures. A distinctive clinical feature of this group of diseases is the presence of spontaneous fractures and skeletal deformities. However, the clinical manifestations of different types of OI are characterized by marked polymorphism with variable severity of skeletal and extra-skeletal features. Previous studies have shown that a mutation (c.-14C>T) in the IFITM5 gene is responsible for autosomal dominant OI type V. However, the mutation has a variable expression pattern and marked clinical heterogeneity. In this study, a clinical and genetic analysis of 12 cases with molecularly confirmed OI type V from 12 unrelated families was performed. Significant clinical heterogeneity of the disease with the same molecular defect was detected. In six subjects (50%), there were no classic signs of OI type V (formation of a hyperplastic bone callus, calcification of the interosseous membrane and dislocation of the radial head). In all cases, the mutation occurred de novo.
Angiotensin-converting enzyme 2 (ACE2) is a regulator in the renin-angiotensin system. ACE2 expression was analysed immunohistochemically in 15,306 samples from 119 tumour types and in 608 samples of 76 normal tissue types. In normal tissue, ACE2 was most abundant in testis and corpus luteum, kidney, small intestine and capillaries of selected organs. At least an occasional weak ACE2 positivity of tumour cells was seen in 83 of 119 (70%) tumour types. ACE2 tumour cell positivity was particularly frequent in papillary (94%) and clear cell (86%) renal cell carcinoma, colorectal adenocarcinoma (81%), mucinous ovarian cancer (61%), cholangiocarcinoma (58%), hepatocellular carcinoma (56%), and in adenocarcinomas of the stomach (47%), pancreas (42%), and the lung (35%). ACE2-positive capillaries were found in 409/12,644 (3%) of analysable tumours, most frequently in tumours with endocrine/neuroendocrine activity. Presence of ACE2-positive capillaries was linked to low stage in papillary thyroid cancer and low grade in neuroendocrine neoplasms. In conclusion, ACE2 expression can occur both in tumour cells and tumour-associated capillaries in a broad variety of different tumour types at highly variable frequencies.
Several recent studies have shown that both strands of certain miRNAs derived from miRNA duplexes are involved in cancer pathogenesis. Our own recent studies revealed that both strands of the miR-150 duplex act as tumor-suppressive miRNAs in lung adenocarcinoma (LUAD) through the targeting of several oncogenes. The aim of the study here was to further investigate the tumor-suppressive roles of miR-150-3p (the passenger strand) in lung squamous cell carcinoma (LUSQ) and its control of cancer-promoting genes in LUSQ cells. The downregulation of miR-150-3p in LUSQ tissues was confirmed by data in The Cancer Genome Atlas (TCGA). The ectopic expression of miR-150-3p attenuated cancer cell aggressive features, e.g., cell cycle arrest, migration and invasive abilities. Our target search strategy successfully identified a total of 49 putative targets that were listed as subjects of miR-150-3p regulation in LUSQ cells. Interestingly, among these targets, 17 genes were categorized as related to the “cell cycle” based on Gene Ontology (GO) classification, namely CENPA, CIT, CCNE1, CCNE2, TIMELESS, BUB1, MCM4, HELLS, SKA3, CDCA2, FANCD2, NUF2, E2F2, SUV39H2, CASC5, ZWILCH and CKAP2). Moreover, we show that the expression of HELLS (helicase, lymphoid specific) is directly controlled by miR-150-3p, and its expression promotes the malignant phenotype of LUSQ cells.
Diabetic retinopathy (DR) is a chronic complication associated with diabetes and the number one cause of blindness in working adults in the US. More than 90% of diabetic patients have obesity-associated type 2 diabetes (T2D), and 60% of T2D patients will develop DR. Photoreceptors undergo apoptosis shortly after the onset of diabetes, which contributes to the retinal dysfunction and microvascular complications leading to vision impairment. However, how diabetic insults cause photoreceptor apoptosis remains unclear. In this study, obesity-associated T2D mice and cultured photoreceptors were used to investigate how decreased microRNA-150 (miR-150) and its downstream target were involved in photoreceptor apoptosis. In the T2D retina, miR-150 was decreased with its target ETS-domain transcription factor (ELK1) and phosphorylated ELK1 at threonine 417 (pELK1T417) upregulated. In cultured photoreceptors, treatments with palmitic acid (PA), to mimic a high-fat environment, decreased miR-150 but upregulated ELK1, pELK1T417, and the translocation of pELK1T417 from the cytoplasm to the cell nucleus. Deletion of miR-150 (miR-150−/−) exacerbates T2D- or PA-induced photoreceptor apoptosis. Blocking the expression of ELK1 with small interfering RNA (siRNA) for Elk1 did not rescue PA-induced photoreceptor apoptosis. Translocation of pELK1T417 from cytoplasm-to-nucleus appears to be the key step of diabetic insult-elicited photoreceptor apoptosis.
Timeline of the brain presentation in rats after complete calvariectomy: normal brain before superior mesenteric artery ligation (healthy, h), brain swelling immediately after occlusion of the superior mesenteric artery (a), and after BPC 157 therapy, a gradual decrease of the swelling, immediately after therapy (b), and at 5 min (c) and 15 min (d) thereafter. The camera was attached to a VMS-004 Discovery Deluxe USB microscope (Veho, Denver, CO, USA). Proportional change of the brain surface area assessed for the brain-swelling recording, as % of volume in healthy means ± SD, mm), brain, abdominal aorta, inferior mesenteric artery, inferior caval vein, and superior mesenteric artery relative to the volume at the time of the occlusion (SMA occlusion before medication) (white bars) and at 1, 5, 15, and 30 min after medication application (10 µ g/kg BPC 157 (light gray bars), 10 ng/kg BPC 157 (dark gray bars), or 5 mL/kg saline (white bars) (SMA occlusion after medication). # p ˂ 0.05, at least, vs. healthy values (100%); * p ˂ 0.05, at least, vs. corresponding control.
The neuropathologic scores.
Gastric pentadecapeptide BPC 157 therapy counteracts multiple organ dysfunction syndrome in rats, which have permanent occlusion of the superior mesenteric artery close to the abdominal aorta. Previously, when confronted with major vessel occlusion, its effect would rapidly activate collateral vessel pathways and resolve major venous occlusion syndromes (Pringle maneuver ischemia, reperfusion, Budd–Chiari syndrome) in rats. This would overwhelm superior mesenteric artery permanent occlusion, and result in local, peripheral, and central disturbances. Methods: Assessments, for 30 min (gross recording, angiography, ECG, pressure, microscopy, biochemistry, and oxidative stress), included the portal hypertension, caval hypertension, and aortal hypotension, and centrally, the superior sagittal sinus hypertension; systemic arterial and venous thrombosis; ECG disturbances; MDA-tissue increase; and multiple organ lesions and disturbances, including the heart, lung, liver, kidney, and gastrointestinal tract, in particular, as well as brain (cortex (cerebral, cerebellar), hypothalamus/thalamus, hippocampus). BPC 157 therapy (/kg, abdominal bath) (10 µg, 10 ng) was given for a 1-min ligation time. Results: BPC 157 rapidly recruits collateral vessels (inferior anterior pancreaticoduodenal artery and inferior mesenteric artery) that circumvent occlusion and ascertains blood flow distant from the occlusion in the superior mesenteric artery. Portal and caval hypertension, aortal hypotension, and, centrally, superior sagittal sinus hypertension were attenuated or eliminated, and ECG disturbances markedly mitigated. BPC 157 therapy almost annihilated venous and arterial thrombosis. Multiple organ lesions and disturbances (i.e., heart, lung, liver, and gastrointestinal tract, in particular, as well as brain) were largely attenuated. Conclusions: Rats with superior mesenteric artery occlusion may additionally undergo BPC 157 therapy as full counteraction of vascular occlusion-induced multiple organ dysfunction syndrome.
Due to endothelial impairment, high-dose lithium may produce an occlusive-like syndrome, comparable to permanent occlusion of major vessel-induced syndromes in rats; intracranial, portal, and caval hypertension, and aortal hypotension; multi-organ dysfunction syndrome; brain, heart, lung, liver, kidney, and gastrointestinal lesions; arterial and venous thrombosis; and tissue oxidative stress. Stable gastric pentadecapeptide BPC 157 may be a means of therapy via activating loops (bypassing vessel occlusion) and counteracting major occlusion syndromes. Recently, BPC 157 counteracted the lithium sulfate regimen in rats (500 mg/kg/day, ip, for 3 days, with assessment at 210 min after each administration of lithium) and its severe syndrome (muscular weakness and prostration, reduced muscle fibers, myocardial infarction, and edema of various brain areas). Subsequently, BPC 157 also counteracted the lithium-induced occlusive-like syndrome; rapidly counteracted brain swelling and intracranial (superior sagittal sinus) hypertension, portal hypertension, and aortal hypotension, which otherwise would persist; counteracted vessel failure; abrogated congestion of the inferior caval and superior mesenteric veins; reversed azygos vein failure; and mitigated thrombosis (superior mesenteric vein and artery), congestion of the stomach, and major hemorrhagic lesions. Both regimens of BPC 157 administration also counteracted the previously described muscular weakness and prostration (as shown in microscopic and ECG recordings), myocardial congestion and infarction, in addition to edema and lesions in various brain areas; marked dilatation and central venous congestion in the liver; large areas of congestion and hemorrhage in the lung; and degeneration of proximal and distal tubules with cytoplasmic vacuolization in the kidney, attenuating oxidative stress. Thus, BPC 157 therapy overwhelmed high-dose lithium intoxication in rats.
Gross presentation of rats with vesicovaginal fistulas. Bladder presentation and lesion (defect) (a), urinary leaking through the vagina (b), vesical stones (c) (full arrows) always noted in all control rats. These disturbances were absent in rats treated with BPC 157 regimens presenting with the preserved bladder presentation (d) (dashed arrow).
Adhesion formation, scored 0-7, Min/Med/Max, time line. Therapy regimens starting at the day 14 after fistula creation (therapy day 0), included BPC 157 given once daily intraperitoneally, last application at 24 h before sacrifice, 10 µg/kg (light gray bars) or 10 ng/kg (dark gray bars) or given perorally, in drinking water 10 µg/kg/day (dashed light gray bars) or 10 ng/kg (dashed dark gray bars) until the sacrifice. Controls received simultaneously an equal volume of saline (5 mL/kg) intraperitoneally (white bars) or drinking water (12 mL/rat/day) (black bars). * p < 0.05, relative to control.
Bladder defect diameter, mm, means ± SD, time line. Therapy regimens starting at the day 14 after fistula creation (therapy day 0), included BPC 157 given once daily intraperitoneally, last application at 24 h before sacrifice, 10 µg/kg (light gray bars) or 10 ng/kg (dark gray bars) or given perorally, in drinking water 10 µg/kg/day (dashed light gray bars) or 10 ng/kg (dashed dark gray bars) until the sacrifice. Controls received simultaneously an equal volume of saline (5 mL/kg) intraperitoneally (white bars) or drinking water (12 mL/rat/day) (black bars). * p < 0.05, relative to control.
With the stable gastric pentadecapeptide BPC 157 therapy known to heal various both external and internal rat fistulas, we attempt to approach vesicovaginal fistula, continuous urine leaking through vagina, bladder stones, and a possible therapy solution among rats with well-formed 2 week-fistulas (vaginal/vesical 4 mm large defects) started with delayed therapy. Subsequent control fistula course (the subsequent 1, 2, 4, and 6 weeks) since beginning revealed the failed healing, fistula leaking, adhesions, urinary leaking through vagina, failed epithelization, collagenization, granulation tissue and neovascularization, increased inflammation, and necrosis. Thereby, the later intervals revealed the persistent inability to sustain even minimal volume, vesical, and vaginal defects and stone formation at the end of the experiment (fistula-time day 56). BPC 157 therapy (10 µg/kg, 10 ng/kg, intraperitoneally once time daily or perorally in drinking water until sacrifice) was initiated with a considerable delay (at 2 weeks after fistula formation). Already within 1 week therapy, all BPC 157 regimens stopped urinary leaking through vagina, reversed the otherwise resistant poor healing course to the increased epithelization, collagenization, granulation tissue and neovascularization, decreased inflammation, and decreased necrosis. Thereby, at later intervals, all BPC 157 rats exhibited a five times larger volume that can be sustained before leaking as in healthy, vesical, and vaginal defects completely closed and no stone formation. Thus, macro/microscopic and functional recovery, and counteracted stone formation. Concluding, BPC 157 therapy’s beneficial effects resulted in healing and no stone formation, with µg- and ng-regimens, either given daily perorally in drinking water or intraperitoneally.
Top-cited authors
László Vecsei
  • University of Szeged
Masaru Tanaka
  • ELKH-SZTE Neuroscience Research Group, Eötvös Loránd Research Network, University of Szeged (ELKH-SZTE)
Dario Didona
  • Philipps University of Marburg
Giovanni Paolino
  • San Raffaele Scientific Institute
Alexander Orekhov
  • Institute of General Pathology and Pathophysiology