ABSTRACT
Microplastics (MPs) and pharmaceuticals and personal care products (PPCPs) are two types of emerging contaminants widely present in the global aquatic ecosystem. The ecological risks associated with the coexistence of these two contaminants have garnered increasing attention from researchers. In this study, we selected 15 typical hydrophilic PPCPs, including Sulfacetamide (SA), Thiamphenicol, Florfenicol, Chloramphenicol (CHL), Ampicillin, Cephalexin, Ofloxacin, Fluorouracil, Phenytoin, Theophylline, Cimetidine, Methylparaben, Diethyltoluamide, Benzophenone-2 (BP-2), and Benzophenone-4, as adsorbates. We evaluated the adsorption potential of five traditional plastics (TPs), namely Polyamide 6 (PA6), Polystyrene (PS), Polyethylene terephthalate (PET), Polyvinyl chloride (PVC), and Polyurethane (TPU), as well as three biodegradable plastics (BDPs), including Polylactic acid (PLA), Polybutylene succinate (PBS), and Poly (ε-caprolactone) (PCL), for these adsorbates. Out of the 120 combinations of MPs and PPCPs tested, only 24 exhibited significant adsorption behavior. Notably, the adsorption performance of the three BDPs was stronger than that of the three typical TPs (PS, PET, and PVC). Based on their adsorption potential, PA6, BDPs, phenytoin, and BP-2 were identified as potential sources of high ecological risk. To further explore the adsorption mechanism, we investigated the adsorption behaviors of SA, BP-2, and CHL on PA6. The conclusions were as follows: SA, BP-2, and CHL all reached adsorption equilibrium within 24 h, with the partition coefficient (Kd) following this order: BP-2 (8.051) ⫠SA (0.052) > CHL (0.018). The primary forces of adsorption were electrostatic interactions, intermolecular hydrogen bonding, and hydrophobic interaction, respectively. Additionally, weak electrostatic effects were observed in the adsorption of CHL and BP-2. The effects of pH, ionic strength, and fulvic acid on adsorption capacity varied. These results highlight a complex adsorption mechanism between MPs and hydrophilic contaminants in the aquatic environment. This study provides a basis for further evaluating the ecological risks of MPs and PPCPs combined pollution.
Subject(s)
Cosmetics , Water Pollutants, Chemical , Plastics , Microplastics , Adsorption , Ecosystem , Phenytoin , Polystyrenes/chemistry , Hydrophobic and Hydrophilic Interactions , Pharmaceutical Preparations , Water Pollutants, Chemical/analysisABSTRACT
Highly precise and controllable liner implosions driven by a pulsed power facility have extensive applications in exploration of advanced hydrodynamics at the extremes of pressure and material velocity. In this paper, we describe a new pulsed power facility developed in China named FP-2 (a series of facilities for Fluid Physics investigations-the second generation) for liner implosions. Benefiting from the reliable and stable operation of 48 rail gap switches, the FP-2 facility can steadily transmit a current of 10.5 MA to a dummy load of 10 nH in the case of a charging voltage of ±40 kV. The first quarter cycle is 5.5 µs, and the percentage shot-to-shot deviation of the current history is less than 1%. When the aluminum liners of 60 mm in height and 0.6 mm in thickness are adopted, the maximum velocity of 4.5 and 7.5 km/s has been achieved with the liner diameter of 90 and 60 mm, respectively, at the diameter of 10 mm. Experimental results show that the percentage shot-to-shot deviation of the liner velocity history is less than 1%. As impact on the target, the maximum of the impact time deviation measured from four perpendicular fiber pins is less than 20 ns. Due to the modular design of FP-2, it is convenient for a future upgrade. The confirmation of high-quality implosion on FP-2, such as high repeatability, high reliability, and high symmetry, makes it a bright prospect to explore the advanced hydrodynamic problems at extremes of pressure and material velocity in the future.
ABSTRACT
Objective We have previously demonstrated benefits of kidney preservation utilizing an oxygenated subnormothermic ex vivo perfusion platform. Herein, we aim to compare pulsatile versus centrifugal (steady and uniform flow) perfusion with the goal of optimizing renal preservation with these devices. Materials and methods: Pig kidneys were procured following 30 min of warm ischemia by cross-clamping both renal arteries. Paired kidneys were cannulated and underwent either: oxygenated pulsatile or centrifugal perfusion using a hemoglobin oxygen carrier at room temperature with our ex vivo machine perfusion platform for 4 hr. Kidneys were reperfused with whole blood for 4 hr at 37° C. Renal function, pathology and evidence of inflammation were assessed post-perfusion. Results: Both pump systems performed equally well with organs exhibiting similar renal blood flow, and function post-reperfusion. Histologic evidence of renal damage using apoptosis staining and acute tubular necrosis scores was similar between groups. This was corroborated with urinary assessment of renal damage (NGAL 1) and inflammation (IL-6), as levels were similar between groups. Conclusion: In our porcine model with added warm ischemia simulating the effects of reperfusion after transplantation, pulsatile perfusion yielded similar renal protection compared with centrifugal perfusion kidney preservation. Both methods of perfusion can be used in ex vivo kidney perfusion systems.
Subject(s)
Kidney Transplantation , Kidney , Organ Preservation , Animals , Perfusion , Pulsatile Flow , SwineABSTRACT
BACKGROUND: The optimal method of oxygen delivery to donor kidneys during ex vivo machine perfusion has not been established. We have recently reported the beneficial effects of subnormothermic (22°C) blood perfusion in the preservation of porcine donation after circulatory death kidneys. Since using blood as a clinical perfusate has limitations, including matching availability and potential presence of pathogen, we sought to assess hemoglobin-based oxygen carrier (HBOC-201) in oxygen delivery to the kidney for renal protection. METHODS: Pig kidneys (n = 5) were procured after 30 minutes of warm in situ ischemia by cross-clamping the renal arteries. Organs were flushed with histidine tryptophan ketoglutarate solution and subjected to static cold storage or pulsatile perfusion with an RM3 pump at 22°C for 4 hours with HBOC-201 and blood. Thereafter, kidneys were reperfused with normothermic (37°C) oxygenated blood for 4 hours. Blood and urine were subjected to biochemical analysis. Total urine output, urinary protein, albumin/creatinine ratio, flow rate, resistance were measured. Acute tubular necrosis, apoptosis, urinary kidney damage markers, neutrophil gelatinase-associated lipocalin 1, and interleukin 6 were also assessed. RESULTS: HBOC-201 achieved tissues oxygen saturation equivalent to blood. Furthermore, upon reperfusion, HBOC-201 treated kidneys had similar renal blood flow and function compared with blood-treated kidneys. Histologically, HBOC-201 and blood-perfused kidneys had vastly reduced acute tubular necrosis scores and degrees of terminal deoxynucleotidyl transferase 2'-deoxyuridine, 5'-triphosphate nick end labeling staining versus kidneys treated with cold storage. Urinary damage markers and IL6 levels were similarly reduced by both blood and HBOC-201. CONCLUSIONS: HBOC-201 is an excellent alternative to blood as an oxygen-carrying molecule in an ex vivo subnormothermic machine perfusion platform in kidneys.
Subject(s)
Kidney Transplantation/adverse effects , Organ Preservation Solutions/administration & dosage , Organ Preservation/methods , Perfusion/methods , Reperfusion Injury/prevention & control , Animals , Blood Substitutes/administration & dosage , Blood Substitutes/chemistry , Disease Models, Animal , Hemoglobins/administration & dosage , Hemoglobins/chemistry , Humans , Organ Preservation/instrumentation , Organ Preservation Solutions/chemistry , Oxygen/analysis , Oxygen/metabolism , Perfusion/instrumentation , Reperfusion Injury/etiology , Sus scrofa , Warm Ischemia/adverse effectsABSTRACT
INTRODUCTION: The current methods of preserving donor kidneys in nonoxygenated cold conditions minimally protect the kidney against ischemia-reperfusion injury (IRI), a major source of complications in clinical transplantation. However, preserving kidneys with oxygenated perfusion is not currently feasible due to the lack of an ideal perfusion mechanism that facilitates perfusion with blood at warm temperature. Here, we have designed an innovative renal pump circuit system that can perfuse blood or acellular oxygen carrier under flexible temperatures, pressures, and oxygenation. We have tested this apparatus to study optimal conditions of storage of our porcine model of donation after cardiac death (DCD) kidneys. METHODS: Porcine kidneys were retrieved after 30 minutes of cross-clamping renal pedicles in situ. Cessation of blood mimics postcardiac death in humans and simulates DCD warm ischemic injury. Procured kidneys were flushed and subjected to static cold storage (SCS) for 4 hours. For warm perfusion, kidneys were cannulated for pulsatile oxygenated perfusion with blood:PlasmaLyte for 4 hours at 15 °C, 22 °C, and 37 °C. To mimic posttransplant scenario, all kidneys were reperfused with blood for an additional 4 hours at 37 °C. RESULTS: Compared with all other groups, 22 °C perfusion resulted in significant reduction of acute tubular necrosis (ATN), apoptosis, kidney damage markers, Toll-like receptor signaling, and cytokine production. It was associated with maximal renal blood flow and urine output. Kidneys stored at 15 °C thrombosed within 2 hours under this condition. Martius Scarlet Blue staining confirmed that 22 °C was the optimal temperature to minimize hemorrhage and blood clots. CONCLUSION: Our novel study shows that oxygenated perfusion at near-room-temperature provides optimal donor kidney storage conditions.
ABSTRACT
PARP2 belongs to a family of proteins involved in cell differentiation, DNA damage repair, cellular energy expenditure, and chromatin modeling. In addition to these overlapping functions with PARP1, PARP2 participates in spermatogenesis, T-cell maturation, extra-embryonic endoderm formation, adipogenesis, lipid metabolism, and cholesterol homeostasis. Knowledge of the functions of PARP2 is far from complete, and the mechanism(s) by which the gene and protein are regulated are unknown. In this study, we found that two different mechanisms are used in vitro to regulate PARP2 levels. In the presence of serum, PARP2 is degraded through the ubiquitin-proteasome pathway; however, when serum is removed or dialyzed with a 3.5 kDa molecular cut membrane, PARP2 rapidly becomes sodium dodecyl sulphate- and urea-insoluble. Despite the presence of a putative serum response element in the PARP2 gene, transcription is not affected by serum deprivation, and PARP2 levels are restored when serum is replaced. The loss of PARP2 affects cell differentiation and gene expression linked to cholesterol and lipid metabolism. These observations highlight the critical roles that PARP2 plays under different physiological conditions, and reveal that PARP2 is tightly regulated by distinct pathways.
Subject(s)
Poly(ADP-ribose) Polymerases/metabolism , Animals , Cells, Cultured , Humans , Mice , Poly(ADP-ribose) Polymerases/blood , Poly(ADP-ribose) Polymerases/geneticsABSTRACT
BACKGROUND: Carbon monoxide (CO) inhalation protects organ by reducing inflammation and cell death during transplantation processes in animal model. However, using CO in clinical transplantation is difficult due to its delivery in a controlled manner. A manganese-containing CO releasing molecules (CORM)-401 has recently been synthesized which can efficiently deliver 3 molar equivalents of CO. We report the ability of this anti-inflammatory CORM-401 to reduce ischemia reperfusion injury associated with prolonged cold storage of renal allografts obtained from donation after circulatory death in a porcine model of transplantation. METHODS: To stimulate donation after circulatory death condition, kidneys from large male Landrace pig were retrieved after 1 hour warm ischemia in situ by cross-clamping the renal pedicle. Procured kidneys, after a brief flushing with histidine-tryptophan-ketoglutarate solution were subjected to pulsatile perfusion at 4°C with University of Wisconsin solution for 4 hours and both kidneys were treated with either 200 µM CORM-401 or inactive CORM-401, respectively. Kidneys were then reperfused with normothermic isogeneic porcine blood through oxygenated pulsatile perfusion for 10 hours. Urine was collected, vascular flow was assessed during reperfusion and histopathology was assessed after 10 hours of reperfusion. RESULTS: We have found that CORM-401 administration reduced urinary protein excretion, attenuated kidney damage markers (kidney damage marker-1 and neutrophil gelatinase-associated lipocalin), and reduced ATN and dUTP nick end labeling staining in histopathologic sections. CORM-401 also prevented intrarenal hemorrhage and vascular clotting during reperfusion. Mechanistically, CORM-401 appeared to exert anti-inflammatory actions by suppressing Toll-like receptors 2, 4, and 6. CONCLUSIONS: Carbon monoxide releasing molecules-401 provides renal protection after cold storage of kidneys and provides a novel clinically relevant ex vivo organ preservation strategy.
Subject(s)
Carbon Monoxide/pharmacology , Kidney Transplantation/adverse effects , Manganese/chemistry , Organ Preservation/methods , Reperfusion Injury/prevention & control , Adenosine/chemistry , Allografts/pathology , Allopurinol/chemistry , Animals , Carbon Monoxide/metabolism , Cold Ischemia/adverse effects , Glutathione/chemistry , Insulin/chemistry , Kidney/pathology , Male , Models, Animal , Organ Preservation/instrumentation , Organ Preservation Solutions/chemistry , Raffinose/chemistry , Reperfusion Injury/etiology , Reperfusion Injury/pathology , Sus scrofaABSTRACT
A 633 nm laser interferometer has been designed based on a novel concept, which, without the acousto-optic modulator or the demodulator circuit, adopts the fibers to connect all elements except photodetectors and oscilloscope in this system to make it more compact, portable, and efficient. The noteworthy feature is to mathematically compare the two divided interference signals, which have the same phase-shift caused by the electron density but possess the different initial phase and low angular frequencies. It is possible to read the plasma density directly on the oscilloscope by our original mathematic demodulation method without a camera. Based on the Abel inversion algorithm, the radial electron density profiles versus time can be obtained by using the multi-chord system. The designed measurable phase shift ranges from 0 to 2π rad corresponding to the maximum line integral of electron density less than 3.5 × 10(17) cm(-2), and the phase accuracy is about 0.017 rad corresponding to the line integral of electron density accuracy of 1 × 10(15) cm(-2). After the construction of eight-chord interferometer, it will provide the detailed time resolved information of the spatial distribution of the electron density in the field-reversed configuration (FRC) plasma target produced by the "Yingguang-1" programmed-discharge device, which is being constructed in the Key Laboratory of Pulsed Power, China Academy of Engineering Physics.
ABSTRACT
STUDY DESIGN: A biomechanical study of cervical artificial disk replacement (CADR). OBJECTIVE: This study aimed to investigate the range of motion (ROM) of the treated segment, the ROM of the adjacent segments, the global ROM in the sagittal plane, and the total neck ROM in the 3 cardinal planes after single-level Discover CADR. SUMMARY OF BACKGROUND DATA: CADR could theoretically preserve the motion function of the treated segment without affecting the adjacent segments significantly. Although previous studies have reported excellent clinical outcomes and ROM of the treated segment after CADR, few studies have focused on the ROM of the adjacent segments, the global ROM, and the total neck motion. METHODS: C5/6 Discover CADR was performed in 58 patients (37 male and 21 female) between September 2008 and September 2010. Anteroposterior, lateral, and flexion-extension lateral radiographies were performed before operation and at the 1-year follow-up. Clinical parameters, including the Japanese orthopedic association score, the neck disability index, and the visual analogue scale, were evaluated. The ROM of the treated segment (C5/6) and the adjacent segments (C4/5 and C6/7) and the global ROM (C2/7) were measured by radiography. To evaluate the total neck ROM, the cervical ROM device was advocated. Preoperative and postoperative data were compared using the paired t test. RESULTS: The Japanese orthopedic association score was 14.3 at the 1-year follow-up as compared with the preoperative score of 8.7. Other scoring systems had improved postoperatively, including the neck disability index from 85.1 to 68.6 and the visual analogue scale from 7.8 to 3.3. Compared with the preoperative ROM, the postoperative ROM increased by 3.0 degrees (27.0%) in C5/6, 1.3 degrees (13.7%) in C4/5, and 1.8 degrees (17.6%) in C6/7. The postoperative global ROM also increased by 6.7 degrees (15.2%) compared with preoperative global data. Compared with the preoperative total neck motion, the postoperative total neck motion increased by 8.3 degrees (9.3%) in the sagittal plane and 6.1 degrees (7.7%) in the coronal plane. There was an insignificant increase of 0.8 degrees (0.6%) in the horizontal plane. CONCLUSIONS: This study demonstrated that the single-level Discover CADR increased the ROM of the treated segment and the adjacent segments. There was also an increase in the global ROM and the total neck motion in the sagittal and the coronal planes, although there was no significant difference in the horizontal plane before and after operation.
Subject(s)
Cervical Vertebrae/physiology , Cervical Vertebrae/surgery , Range of Motion, Articular/physiology , Total Disc Replacement/methods , Biomechanical Phenomena/physiology , Female , Follow-Up Studies , Humans , Male , Treatment OutcomeABSTRACT
Platelet-rich plasma (PRP) has a pool of multiple growth factors efficient at inducing the proliferation and osteogenic differentiation of human adipose-derived stem cells (hADSCs). Bone morphogenetic protein (BMP)-2 is a strong stimulator for the osteogenic differentiation of hADSCs. The purpose of this study was to verify the effect of PRP-released growth factors and microsphere-encapsulated BMP-2 on the proliferation and osteoblastic differentiation of hADSCs and to construct a novel tissue-engineered bone. The BMP-2-loaded microspheres and hADSCs were embedded in activated PRP gel. Another 5 composites (hADSCs/platelet-poor plasma [PPP]; hADSCs/PRP; hADSCs/BMP-2/PPP; hADSCs/BMP-2/PRP; and hADSCs/BMP-2+microspheres/PPP) were also constructed. The DNA content, alkaline phosphatase activity, mRNA expression of alkaline phosphatase, osteopontin, osteocalcin, and mineralization of hADSCs in each composite were compared. The DNA content was higher in all PRP-containing composites, meaning that PRP-released growth factors stimulated proliferation of hADSCs. Alkaline phosphatase increased in BMP-2/PRP and BMP-2+microspheres/PRP composites in the first 7 days, meaning that BMP-2 had a synergistic effect with PRP in the early differentiation of hADSCs. Osteopontin, osteocalcin, and mineralization assays were higher in BMP-2+microspheres/PRP composite than in the BMP-2/PRP composite up to 21 days, meaning that a continuous delivery of BMP-2 stimulates osteoblastic differentiation of hADSCs at the early stage and the final maturation stage. These results suggest that sustained delivery of BMP-2 in combination with PRP is better than a single administration of PRP or BMP-2 in the osteogenic differentiation of hADSCs.
Subject(s)
Adipose Tissue/cytology , Adipose Tissue/physiology , Bone Morphogenetic Protein 2/therapeutic use , Osteogenesis/physiology , Platelet-Rich Plasma , Stem Cells/cytology , Stem Cells/physiology , Adipose Tissue/drug effects , Cells, Cultured , Delayed-Action Preparations/administration & dosage , Humans , Intercellular Signaling Peptides and Proteins/administration & dosage , Osteogenesis/drug effects , Stem Cells/drug effects , Treatment OutcomeABSTRACT
Spontaneous discitis is unusual and typically affects children. Hematogenous delivery of an infectious organism is the likely main cause. Common treatment method including conservative and surgical treatments, which also needs prolonged antimicrobial therapy based on an effective inhibitory concentration, can be achieved on the local disc space. Intradiscal antibiotic concentration was measured after the disc was harvested after preventive administration of antibiotics in previous studies. On the one hand the disc cannot simulate the infection situation when the inflammation leads to end plate destruction, vascular invasion and increase of permeability. On the other hand antibiotic concentrations were measured in vitro which cannot tell the actual situation in vivo. It is necessary to find a reliable evaluation method to decide whether the antibiotic can penetrate and make an effective inhibitory concentration in the local disc at the beginning of the treatment in vivo. Systemic antibiotics like nutrients enter and leave the disc by the only way of passive diffusion. The postcontrast MRI has been widely used as a noninvasive method of studying transport into the disc. The enhancement following contrast administration can be measured in T1 sagittal MR images by placing suitable cursors and evaluating the signal intensity (SI) of the region. Therefore we hypothesise that serial postcontrast MRI can be used to measure antibiotic concentration in the infected intervertebral disc in vivo. If the hypothesis is verified, we can better determine the choice of antibiotics and antibiotic treatment regime at the beginning of the treatment to improve the treatment success rate.
Subject(s)
Anti-Bacterial Agents/pharmacokinetics , Discitis/drug therapy , Anti-Bacterial Agents/therapeutic use , Humans , Reproducibility of ResultsABSTRACT
Some controversy still exists over the optimal treatment time and the surgical approach for cervical myelopathy due to ossification of the posterior longitudinal ligament (OPLL). The aim of the current study was first to analyze the effect of intramedullary spinal cord changes in signal intensity (hyperintensity on T2-weighted imaging and hypointensity on T1-weighted imaging) on magnetic resonance imaging (MRI) on surgical opportunity and approach for cervical myelopathy due to OPLL. This was a prospective randomized controlled study. Fifty-six patients with cervical myelopathy due to OPLL were enrolled and assigned to either group A (receiving anterior decompression and fusion, n = 27) or group P (receiving posterior laminectomy, n = 29). All the patients were followed up for an average 20.3 months (12-34 months). The clinical outcomes were assessed by the average operative time, blood loss, Japanese Orthopedic Association (JOA) score, improvement rate (IR) and complication. To determine the relevant statistics, we made two factorial designs and regrouped the data of all patients to group H (with hyperintensity on MRI, n = 31), group L (with hypointensity on MRI, n = 19) and group N (no signal on MRI, n = 25), and then to further six subgroups as well: AH (with hyperintensity on MRI from group A, n = 15), PH (with hyperintensity on MRI from group P, n = 16), AL (with hypointensity on MRI from group A, n = 10), PL (with hypointensity on MRI from group P, n = 9), AN (no signal intensity on MRI from group A, n = 12) and PN (no signal intensity on MRI from group P, n = 13). Both hyperintensity on T2-weighted imaging and hypointensity on T1-weighted imaging had a close relationship with the JOA score and IR. The pre- and postoperative JOA score and postoperative IR of either group H or group L was significantly lower than that of group N (P < 0.05), regardless of whether the patients had received anterior or posterior surgery. On the other hand, both the JOA score and IR of subgroup AH were higher than those of subgroup PH at 1 week, 6 and 12 months postoperatively (P < 0.05), as well as between subgroup AL and PL; but in group N, there was no difference between the subgroup AN and PN (P > 0.05). In conclusion, regardless of hyperintensity on T2-weighted imaging or hypointensity on T1-weighted imaging in patients with OPLL, severe damage to the spinal cord is indicated. Surgical treatment should be provided before the advent of intramedullary spinal cord changes in signal intensity on MRI. The anterior approach is more effective than posterior approach for treating cervical myelopathy due to OPLL characterized by intramedullary spinal cord changes in signal intensity on MRI.
Subject(s)
Cervical Vertebrae/pathology , Ossification of Posterior Longitudinal Ligament/pathology , Spinal Cord Diseases/pathology , Spinal Cord/pathology , Aged , Cervical Vertebrae/surgery , Decompression, Surgical/methods , Female , Humans , Laminectomy , Magnetic Resonance Imaging , Male , Middle Aged , Ossification of Posterior Longitudinal Ligament/surgery , Postoperative Period , Prospective Studies , Spinal Cord/surgery , Spinal Cord Diseases/surgery , Spinal Fusion , Treatment OutcomeABSTRACT
CASK is a member of the membrane-associated guanylate kinase family. In mammals it is an essential protein, as CASK knockout mice die after birth and its deletion in humans has developmental consequences. CASK plays a role in the transcription of genes required for forebrain development, and in the nervous systems of Drosophila and C. elegans, it participates in receptor localization at the plasma membrane. This role in organizing supramolecular protein complexes to appropriate subcellular regions is shared in mammals and is regulated by phosphorylation. CASK is a kinase and regulator of cell proliferation and adhesion, which adds to an expanding list of roles. In this study we report for the first time that CASK is degraded in a characteristic fashion in mammalian cells. We found that CASK is a long-lived protein despite the fact that it contains three putative PEST sequences. Finally, we provide detailed evidence that CASK degradation is mediated through a ubiquitin-proteasome pathway and this is phosphorylation-dependent. Together, these results provide evidence that post-translational modifications to CASK are major regulatory steps leading to its proteasomal degradation. This regulation not only has important implications on how CASK participates in its many disparate roles, but highlights how altering this regulation may contribute to the pathogenesis of human disease.
Subject(s)
Guanylate Kinases/metabolism , Proteasome Endopeptidase Complex/metabolism , Protein Processing, Post-Translational , Amino Acid Sequence , Animals , Cell Line , Enzyme Stability , Guanylate Kinases/chemistry , Humans , Molecular Sequence Data , Phosphorylation , Protein Transport , Sequence Analysis, Protein , Ubiquitin/metabolism , Ubiquitinated Proteins/metabolism , UbiquitinationABSTRACT
Mouse F9 cells differentiate into primitive endoderm when treated with retinoic acid (RA) and into parietal endoderm in response to RA and dibutyryl (db-) cAMP. G protein signaling either blocks or mimics RA-induced differentiation, the latter signaling through the Wnt-beta-catenin pathway. In our study, we found that a constitutively active Galpha13 mutant induces F9 cells to differentiate into parietal endoderm in the absence of exogenous agents. Galpha13 expression and subsequent differentiation are accompanied by beta-catenin translocation to the nucleus. Differentiation and changes in cell morphology are supported by rearrangements to the F-actin cytoskeleton. ERM (ezrin-radixin-moesin) proteins, known to link F-actin to transmembrane receptors, are also redistributed during differentiation. Furthermore, morpholino antisense and shRNA approaches show that moesin expression is essential since its knockdown leads to altered F-actin distribution and subsequent apoptosis. Moesin-depleted cells, however, remain attached to the substrate when Galpha13 is constitutively expressed, but they do not differentiate into extraembryonic endoderm. Our study demonstrates a link between Galpha13 signaling that regulates differentiation of F9 cells through primitive to parietal endoderm and a moesin requirement for cell survival.
Subject(s)
Apoptosis , Cell Differentiation , Endoderm/cytology , GTP-Binding Protein alpha Subunits, G12-G13/metabolism , Microfilament Proteins/metabolism , Actins/metabolism , Active Transport, Cell Nucleus , Animals , Cell Line, Tumor , Cell Nucleus/metabolism , Cell Survival , Lithium Chloride/pharmacology , Mice , Microfilament Proteins/deficiency , Microfilament Proteins/genetics , Protein Transport/drug effects , RNA, Antisense/genetics , Signal Transduction , Transfection , beta Catenin/metabolismABSTRACT
OBJECTIVE: To study the clinical and pathological characteristics of patients with microscopic polyangiitis (MPA) with medium artery involvement. METHODS: Hospitalized patients with MPA in recent two years were retrospectively studied. Their clinical and pathological features were compared between patients with and without renal medium artery involvement. RESULTS: Thirty-nine patients had renal pathology confirmed MPA. Nine cases were with medium artery involvement. For the 30 patients without medium artery involvement, 24/30 had crescentic glomerulonephritis and 11/30 also had focal segmental glomerular fibrinoid necrosis; clinically, 21/30 patients were pANCA/MPO-ANCA positive, 26/30 had acute renal failure with an average duration of 14 weeks before MPA was diagnosed, eight cases achieved complete remission after intensive immunosuppressive therapy. Nine MPA patients had medium artery involvement, manifested as segmental fibrinoid necrosis of major branch of arcuate artery, glomerulus ischemia was predominant, but crescentic lesions were mild, none of them reached crescentic glomerulonephritis. Five of the nine were pANCA/MPO-ANCA positive, eight out of the nine patients had acute renal failure with an average duration of eight weeks before diagnosed, seven of the nine achieved complete remission after intensive immunosuppressive therapy. In comparison, MPA with medium artery involvement had a shorter duration (p < 0.05), less crescentic glomerulonephritis in patients with acute renal failure (p < 0.01) and more patients achieved complete remission after treatment (p < 0.05). CONCLUSION: In present study, about 23% MPA patients had medium artery involvement and their impaired renal function is mainly due to extensive glomerular ischemia. These patients progressed to acute renal failure quicker and responded to therapy better.