Enhanced oxidative stress is a hallmark of cisplatin nephrotoxicity, and inhibition of poly(ADP-ribose) polymerase 1 (PARP1) attenuates oxidative stress during cisplatin nephrotoxicity; however, the precise mechanisms behind its action remain elusive. Here, using an in vitro model of cisplatin-induced injury to human kidney proximal tubular cells, we demonstrated that the protective effect of PARP1 inhibition on oxidative stress is associated with sirtuin 3 (SIRT3) activation. Exposure to 400 µM cisplatin for 8 hours in cells decreased activity and expression of manganese superoxide dismutase (MnSOD), catalase, glutathione peroxidase (GPX), and SIRT3, while it increased their lysine acetylation. However, treatment with 1 µM PJ34 hydrochloride, a potent PARP1 inhibitor, restored activity and/or expression in those antioxidant enzymes, decreased lysine acetylation of those enzymes, and improved SIRT3 expression and activity in the cisplatin-injured cells. Using transfection with SIRT3 double nickase plasmids, SIRT3-deficient cells given cisplatin did not show the ameliorable effect of PARP1 inhibition on lysine acetylation and activity of antioxidant enzymes, including MnSOD, catalase and GPX. Furthermore, SIRT3 deficiency in cisplatin-injured cells prevented PARP1 inhibition-induced increase in forkhead box O3a transcriptional activity, and upregulation of MnSOD and catalase. Finally, loss of SIRT3 in cisplatin-exposed cells removed the protective effect of PARP1 inhibition against oxidative stress, represented by the concentration of lipid hydroperoxide and 8-hydroxy-2'-deoxyguanosine; and necrotic cell death represented by a percentage of propidium iodide–positively stained cells. Taken together, these results indicate that PARP1 inhibition protects kidney proximal tubular cells against oxidative stress through SIRT3 activation during cisplatin nephrotoxicity.
Acetylation ; Catalase ; Cell Death ; Cisplatin* ; Deoxyribonuclease I ; Down-Regulation* ; Glutathione Peroxidase ; Humans ; In Vitro Techniques ; Kidney ; Lipid Peroxides ; Lysine ; Oxidative Stress* ; Plasmids ; Poly Adenosine Diphosphate Ribose* ; Poly(ADP-ribose) Polymerases* ; Propidium ; Sirtuin 3* ; Superoxide Dismutase ; Transfection ; Up-Regulation

Kidney ischemia and reperfusion injury (IRI) is associated with a high mortality rate, which is attributed to tubular oxidative and nitrative stresses; however, an effective approach to limit IRI remains elusive. Spermidine, a naturally occurring polyamine, protects yeast cells against aging through the inhibition of oxidative stress and necrosis. In the present study, spermidine supplementation markedly attenuated histological damage and kidney dysfunction during IRI. In addition, exogenous spermidine potently inhibited poly(ADP-ribose) polymerase 1 (PARP1) activation and DNA nitrative/oxidative stress following IRI. Conversely, inhibition of ornithine decarboxylase (ODC) via siRNA transfection in vivo significantly enhanced DNA nitration, PARP1 activation, and functional damage during IRI. Finally, in ODC knockdown kidneys, PARP1 inhibition attenuated histological and functional damage induced by IRI, but not DNA nitrative stress. In conclusion, these data suggest that spermidine protects kidneys against IRI through blocking DNA nitration and PARP1 activation and this finding provides a novel target for prevention of acute kidney injury including IRI.
Acute Kidney Injury ; Aging ; DNA* ; Ischemia* ; Kidney* ; Mortality ; Necrosis ; Ornithine Decarboxylase ; Oxidative Stress ; Poly(ADP-ribose) Polymerases ; Reperfusion Injury* ; Reperfusion* ; RNA, Small Interfering ; Spermidine* ; Transfection ; Yeasts

Treatment with cisplatin for cancer therapy has a major side effect such as nephrotoxicity; however, the role of poly (ADP-ribose) polymerase 1 (PARP1) in necrosis in response to cisplatin nephrotoxicity remains to be defined. Here we report that cisplatin induces primary necrosis through PARP1 activation in kidney proximal tubular cells derived from human, pig and mouse. Treatment with high dose of cisplatin for 4 and 8 hours induced primary necrosis, as represented by the percentage of propidium iodide-positive cells and lactate dehydrogenase release. The primary necrosis was correlated with PARP1 activation during cisplatin injury. Treatment with PJ34, a potent PARP1 inhibitor, at 2 hours after injury attenuated primary necrosis after 8 hours of cisplatin injury as well as PARP1 activation. PARP1 inhibition also reduced the release of lactate dehydrogenase and high mobility group box protein 1 from kidney proximal tubular cells at 8 hours after cisplatin injury. Oxidative stress was increased by treatment with cisplatin for 8 hours as shown by 8-hydroxy-2'-deoxyguanosine and lipid hydroperoxide assays, but PARP1 inhibition at 2 hours after injury reduced the oxidative damage. These data demonstrate that cisplatin-induced PARP1 activation contributes to primary necrosis through oxidative stress in kidney proximal tubular cells, resulting in the induction of cisplatin nephrotoxicity and inflammation.
Animals ; Cisplatin* ; Humans ; Inflammation ; Kidney* ; L-Lactate Dehydrogenase ; Lipid Peroxides ; Mice ; Necrosis* ; Oxidative Stress ; Poly(ADP-ribose) Polymerases* ; Propidium

Although the sternalis muscle has been well known to anatomists, it is quite unfamiliar to clinicians. During routine educational dissection, we came across a well-defined bilateral double sternalis muscle innervated by the intercostal nerve, respectively. The right sternalis muscle 1) became tendinous to insert into the sternum and 2) crossed midline and then intermingled with the left pectoralis major muscle, which could be classified into a double with single cross based on Snosek et al.'s criteria. The left sternalis muscle was composed of two bellies, which were combined at the midway, and became tendinous to insert into the contralateral manubrium, which could be classified into a bicipital diverging with double cross based on Snosek et al.'s criteria. The detailed knowledge on the sternalisis is important for clinicians as well as for anatomists, since the clinical importance of the sternalis muscle has been highlighted in recent years.
Anatomists ; Cadaver* ; Humans ; Intercostal Nerves ; Manubrium ; Sternum

The hallmark of cisplatin-induced acute kidney injury is the necrotic cell death in the kidney proximal tubules. However, an effective approach to limit cisplatin nephrotoxicity remains unknown. Spermidine is a polyamine that protects against oxidative stress and necrosis in aged yeasts, and the present study found that exogenous spermidine markedly attenuated tubular necrosis and kidney dysfunction, but not apoptosis, during cisplatin nephrotoxicity. In addition, exogenous spermidine potently inhibited oxidative/nitrative DNA damage, poly(ADP-ribose) polymerase 1 (PARP1) activation and ATP depletion after cisplatin injection. Conversely, inhibition of ornithine decarboxylase (ODC) via siRNA transfection in vivo significantly increased DNA damage, PARP1 activation and ATP depletion, resulting in acceleration of tubular necrosis and kidney dysfunction. Finally, exogenous spermidine removed severe cisplatin injury induced by ODC inhibition. In conclusion, these data suggest that spermidine protects kidneys against cisplatin injury through DNA damage and tubular necrosis, and this finding provides a novel target to prevent acute kidney injury including nephrotoxicity.
Acceleration ; Acute Kidney Injury ; Adenosine Triphosphate ; Apoptosis ; Cell Death ; Cisplatin* ; DNA Damage ; Kidney ; Lipid Peroxidation ; Necrosis* ; Ornithine Decarboxylase ; Oxidative Stress ; Poly(ADP-ribose) Polymerases ; RNA, Small Interfering ; Spermidine* ; Transfection ; Yeasts

PURPOSE: To evaluate the change of accommodation-convergence parameters after implantation of Artisan phakic intraocular lens (PIOL). METHODS: Prospective study for the patients with the Artisan PIOL implantation was performed. A total of 37 patients (3 males and 34 females) enrolled the study. Preoperatively, convergence amplitude, the stimulus accommodative convergence per unit of accommodation (AC/A) ratio and the near point of convergence (NPC) were evaluated. After the Artisan PIOL implantation, the identical evaluations were repeated at 1 week, 1, 3, and 6 months after the surgery. RESULTS: Mean age was 24.3 +/- 4.8 years old, and preoperative refractive error was -8.92 +/- 4.13 diopters (D). After the implantation, mean refractive errors significantly decreased to within +/-1.00 D, and noticeable complications were not found. The convergence amplitude and the stimulus AC/A ratio increased 1 month after the surgery, but progressively stabilized afterward to near preoperative values. NPC didn't show any significant change over follow-up period up to 6 months. CONCLUSIONS: These results regarding implantation of the Artisan PIOL revealed the increase of accommodation-convergence relationship within first 1 month after the surgery, but progressive stabilization was noted during follow-up periods.
*Accommodation, Ocular ; Adult ; *Convergence, Ocular ; Female ; Follow-Up Studies ; Humans ; Lens Implantation, Intraocular/*methods ; Male ; Myopia/*diagnosis/*surgery ; *Phakic Intraocular Lenses ; Postoperative Period ; Prospective Studies ; Refractive Surgical Procedures/methods ; Treatment Outcome ; Visual Acuity ; Young Adult

Dual left anterior interventricular coronary artery (also called left anterior descending artery, hereafter referred as LAD) is a rare congenital coronary artery anomaly. Notably, type IV dual LAD has never been reported in Koreans. During a routine dissection, a new variant of type IV dual LAD was found in a 57-year-old Korean male cadaver, whose cause of death was unknown. One LAD originated from the right coronary artery, coursed through the anterior interventricular sulcus, but did not reach the apex cordis. The other LAD arose from the left coronary artery, never entered the anterior interventricular sulcus through its course, but reached the apex cordis, where it met the posterior interventricular branch of the right coronary artery. This is the first report on a new variant of type IV dual LAD in Koreans, which is of clinical importance during procedures containing the coronary artery.
Arteries ; Cadaver ; Cause of Death ; Coronary Vessel Anomalies ; Coronary Vessels* ; Humans ; Korea ; Male ; Middle Aged

After renal injury, selective damage occurs in the proximal tubules as a result of inhibition of glycolysis. The molecular mechanism of damage is not known. Poly(ADP-ribose) polymerase (PARP) activation plays a critical role of proximal tubular cell death in several renal disorders. Here, we studied the role of PARP on glycolytic flux in pig kidney proximal tubule epithelial LLC-PK1 cells using XFp extracellular flux analysis. Poly(ADP-ribosyl)ation by PARP activation was increased approximately 2-fold by incubation of the cells in 10 mM glucose for 30 minutes, but treatment with the PARP inhibitor 3-aminobenzamide (3-AB) does-dependently prevented the glucose-induced PARP activation (approximately 14.4% decrease in 0.1 mM 3-AB-treated group and 36.7% decrease in 1 mM 3-AB-treated group). Treatment with 1 mM 3-AB significantly enhanced the glucose-mediated increase in the extracellular acidification rate (61.1±4.3 mpH/min vs. 126.8±6.2 mpH/min or approximately 2-fold) compared with treatment with vehicle, indicating that PARP inhibition increases only glycolytic activity during glycolytic flux including basal glycolysis, glycolytic activity, and glycolytic capacity in kidney proximal tubule epithelial cells. Glucose increased the activities of glycolytic enzymes including hexokinase, phosphoglucose isomerase, phosphofructokinase-1, glyceraldehyde-3-phosphate dehydrogenase, enolase, and pyruvate kinase in LLC-PK1 cells. Furthermore, PARP inhibition selectively augmented the activities of hexokinase (approximately 1.4-fold over vehicle group), phosphofructokinase-1 (approximately 1.6-fold over vehicle group), and glyceraldehyde-3-phosphate dehydrogenase (approximately 2.2-fold over vehicle group). In conclusion, these data suggest that PARP activation may regulate glycolytic activity via poly(ADP-ribosyl)ation of hexokinase, phosphofructokinase-1, and glyceraldehyde-3-phosphate dehydrogenase in kidney proximal tubule epithelial cells.
Animals ; Cell Death ; Epithelial Cells* ; Glucose ; Glucose-6-Phosphate Isomerase ; Glycolysis ; Hexokinase ; Kidney* ; LLC-PK1 Cells ; Oxidoreductases ; Phosphofructokinase-1 ; Phosphopyruvate Hydratase ; Poly Adenosine Diphosphate Ribose* ; Poly(ADP-ribose) Polymerases* ; Pyruvate Kinase ; Swine

Cyclosporin A (CsA) does not only exert a toxic effect on kidney parenchymal cells, but also protects them against necrotic cell death by inhibiting opening of mitochondrial permeability transition pore. However, whether CsA plays a role in hydrogen peroxide-induced kidney proximal tubular cell death is currently unclear. In the present study, treatment with CsA further increased apoptosis and necrosis in HK-2 human kidney proximal tubule epithelial cells during exposure to hydrogen peroxide. In addition, hydrogen peroxide-induced p53 activation and BH3 interacting-domain death agonist (BID) expression were higher in CsA-treated cells than those in non-treated cells, whereas hydrogen peroxide-induced activation of mitogen-activated protein kinases including p38, c-Jun N-terminal kinase, and extracellular signal-regulated kinase and activation of protein kinase B were not significantly altered by treatment with CsA. In oxidant-antioxidant system, reactive oxygen species (ROS) production induced by hydrogen peroxide was further enhanced by treatment with CsA. However, expression levels of antioxidant enzymes including manganese superoxide dismutase, copper/zinc superoxide dismutase, and catalase were not altered by treatment with hydrogen peroxide or CsA. Treatment with CsA further enhanced mitochondrial membrane potential induced by exposure to hydrogen peroxide, although it did not alter endoplasmic reticulum stress based on expression of glucose-regulated protein 78 and 94. Taken together, these data suggest that CsA can aggravate hydrogen peroxide-induced cell death through p53 activation, BID expression, and ROS production.
Apoptosis ; Catalase ; Cell Death ; Cyclosporine ; Endoplasmic Reticulum Stress ; Epithelial Cells ; Humans ; Hydrogen Peroxide ; Hydrogen ; JNK Mitogen-Activated Protein Kinases ; Kidney ; Membrane Potential, Mitochondrial ; Mitogen-Activated Protein Kinases ; Necrosis ; Permeability ; Phosphotransferases ; Proto-Oncogene Proteins c-akt ; Reactive Oxygen Species ; Superoxide Dismutase

Adult stem cells possess the characteristics of self-renewal, multipotent, plasticity as well as slow cycling rate. We investigated a location of slow-cycling cells, that is, adult stem-like cells, in various organs in the 8 week-old mice which administered bromodeoxyuridine (BrdU) at neonatal phase. BrdU-retaining cells (slow-cycling cells) were observed in speramtogonia at the edge of seminiferous tubules in testes, hair root cells surrounding hair follicles, the cells in the inner nuclear layer of the retina, the myocytes in the hearts, the cells in the parenchyma and the Glisson's capsule of liver, the cells in the epithelial layer of bronchioles, and the tubular epithelial cells in the kidneys. In conclusion, various organs of adult mouse expressed slow-cycling cells, indicating that the cells may associate with normal cell turnover and repair after damages.
Adult Stem Cells* ; Adult* ; Animals ; Bromodeoxyuridine ; Bronchioles ; Epithelial Cells ; Hair ; Hair Follicle ; Heart ; Humans ; Kidney ; Liver ; Mice* ; Muscle Cells ; Plastics ; Retina ; Seminiferous Tubules ; Testis