P1B-type heavy metal ATPases(HMAs)are transmembrane metal-transporting proteins that play a key role in metal homeostasis.We here reported the characterization of rice OsHMA6,a member of the P1B-type ATPase family.Phy...P1B-type heavy metal ATPases(HMAs)are transmembrane metal-transporting proteins that play a key role in metal homeostasis.We here reported the characterization of rice OsHMA6,a member of the P1B-type ATPase family.Phylogenetic tree analysis showed that OsHMA6 belonged to the Cu/Ag subgroup of the HMA family and had a close evolutionary relationship with OsHMA9.Amino acid sequence alignment showed 82.78%consistency between OsHMA6 and OsHMA9.OsHMA6 expressed in all organs at different growth stages,including spikelet,and abundant in leaf blades,however,OsHMA9 most strongly expressed in roots,but very low in spikelet.Excessive Cu^2+can up-regulate the expression of OsHMA6 and OsHMA9 in rice seedlings.The heterologous expression in yeast showed that OsHMA6 can significantly rescue the growth of yeast strain CM52 when supplied with 3 or 6 mmol/L Cu^2+.Compared with the empty vector pYES2,the Cu concentration in OsHMA6-pYES2 decreased by 23.4%and 30.3%under 3 or 6 mmol/L Cu2+,respectively.Subcellular localization revealed that OsHMA6 was located in the plasma membrane.These results suggested that OsHMA6,similar to OsHMA9,is likely a copper efflux protein located in the plasma membrane.展开更多
Platinum(Pt)-based antitumor agents are effective in the treatment of many solid malignancies. However, their efficacy is limited by toxicity and drug resistance. Reduced intracellular Pt accumulation has been consist...Platinum(Pt)-based antitumor agents are effective in the treatment of many solid malignancies. However, their efficacy is limited by toxicity and drug resistance. Reduced intracellular Pt accumulation has been consistently shown to correlate with resistance in tumors. Proteins involved in copper homeostasis have been identified as Pt transporters. In particular, copper transporter receptor 1(CTR1), the major copper influx transporter, has been shown to play a significant role in Pt resistance. Clinical studies demonstrated that expression of CTR1 correlated with intratumoral Pt concentration and outcomes following Pt-based therapy. Other CTRs such as CTR2, ATP7 A and ATP7 B, may also play a role in Pt resistance. Recent clinical studies attempting to modulate CTR1 to overcome Pt resistance may provide novel strategies. This review discusses the role of CTR1 as a potential predictive biomarker of Pt sensitivity and a therapeutic target for overcoming Pt resistance.展开更多
The experiment was conducted to as- sess the effects of dietary supplementation of Cu on the growth performance, digestive enzymes, tissue minerals and absorptive transporters in small intestinal mucosa of weanling pi...The experiment was conducted to as- sess the effects of dietary supplementation of Cu on the growth performance, digestive enzymes, tissue minerals and absorptive transporters in small intestinal mucosa of weanling pigs. One hundred crossbred pigs weaned at 28 ＋ 2 d of age were assigned randomly to one of the following diets with 5 replicates：corn-soy- bean basal diet with 10,100,175,250 mg/kg of Cu as CuSO4·5H20. The results showed that 250 mg/kg Cu had a positive effect （ P 〈 0.05） on average daily gain, daily feed intake and ratio of gain/feed. Com- pared to 10 mg/kg Cu, higher Cu had significant effect on the apparent digestibility of protein and fat （P 〈 0.05 ）. The supplementing of Cu improved am- ylase and lipase activity in jejunum content and lipase in pancreas （ P 〈 0.05） and had no effect on intestinalmorphology. The liver Cu elevated approximately 4- fold in pigs fed diet with 250 mg/kg Cu compared with pigs fed diet with 10 mg/kg Cu, no increases were observed in pigs receiving the lower level of Cu （100 and 175 mg/kg）. Both Fe and Zn contents in kidney and liver were not affected by Cu supplemen- tation. There was no positive effect （ P 〉0.05） of Cu supplementation on PepT1 （peptide transporter 1 ） and SGLT1 （sodium/glucose cotransporter） mRNA abundance in intestinal mucosa. However, higher sup- plementing level （250 mg/kg） significantly elevated the DMT1 （divalent metal transporter） mRNA abun- dance in duodenum mucosa. These results suggested that dietary supplementation with 250 mg/kg Cu could improve growth performance, nutrient digestibil- ity and intestinal enzyme activities of weanling pigs.展开更多
Cisplatin belongs to platinum-based drugs and is widely used in cancer chemotherapy.Ototoxicity is one of the major dose limiting side-effects of cisplatin.For toxicity to occur cisplatin must first be transported fro...Cisplatin belongs to platinum-based drugs and is widely used in cancer chemotherapy.Ototoxicity is one of the major dose limiting side-effects of cisplatin.For toxicity to occur cisplatin must first be transported from the bloodstream into cochlear cells.Three copper transporters are considered pathways for regulating the uptake and translocation of cisplatin into cells:Ctr1,ATP7A and ATP7B.Our recent study with cochlear organotypic cultures shows that cochlear hair cells can be destroyed by cisplatin at low concentrations from 10μm to 100μn.However,high doses of cisplatin cannot damage hair cells,maybe due to intrinsic feedback reactions that increase export of platinum by ATP7B when the platinum concentration is high in extracellular space.Cimitidine is a specific copper transporter inhibitor that can block the entrance of copper and platinum,and may prevent cisplatin-induced cochlear hair cell injury.To evaluate this hypothesis,we treated cochlear organotypic cultures with cisplatin (10 μm or 50 μm) alone,or cisplatin combined with cimitidine at concentrations ranging from 10-2000 μm for 48 hours.cisplatin at 10 μm damaged about 20% hair cells.In contrast,when cimitidine (10 μm,100 μm and 2000 μm) was added to the culture,near 100% cochlear hair cell survived.At higher concentration (50 μm),cisplatin destroyed about 80% of cochlear hair cells.However,100 μmcimitidine rescued about 50% hair cells from cisplatin damage,and 2000μm cimitidine protected about 80% hair cells.The data of western blot showed that CTR1 and ATP7B expressions were increased in cisplatin treated cochlear tissue,but cimitidine significantly reduced CTR1 and ATP7B.In addition,ATP7A expression was depressed a little after cisplatin treatment.Considering that Ctr1 is involved in copper and platinum influx,but the ATP7A and ATP7B are copper export transporters,the results suggest that cimitidine can effectively block the entrance by copper transporters and stop the influx of cisplatin.展开更多
Based on the momentum and mass conservation equations,a comprehensive model of heap bioleaching process is developed to investigate the interaction between chemical reactions,solution flow,gas flow,and solute transpor...Based on the momentum and mass conservation equations,a comprehensive model of heap bioleaching process is developed to investigate the interaction between chemical reactions,solution flow,gas flow,and solute transport within the leaching system.The governing equations are solved numerically using the COMSOL Multiphysics software for the coupled reactive flow and solute transport at micro-scale,meso-scale and macro-scale levels.At or near the surface of ore particle,the acid concentration is relatively higher than that in the central area,while the concentration gradient decreases after 72 d of leaching.The flow simulation between ore particles by combining X-ray CT technology shows that the highest velocity in narrow pore reaches 0.375 m/s.The air velocity within the dump shows that the velocity near the top and side surface is relatively high,which leads to the high oxygen concentration in that area.The coupled heat transfer and liquid flow process shows that the solution can act as an effective remover from the heap,dropping the highest temperature from 60 to 38 °C.The reagent transfer coupled with solution flow is also analyzed.The results obtained allow us to obtain a better understanding of the fundamental physical phenomenon of the bioleaching process.展开更多
AIM: To study the effect of copper transporting P-type ATPase in copper metabolism of hepatocyte and pathogenesis of Wilson disease (WD).METHODS: WD copper transporting properties in some organelles of the cultu...AIM: To study the effect of copper transporting P-type ATPase in copper metabolism of hepatocyte and pathogenesis of Wilson disease (WD).METHODS: WD copper transporting properties in some organelles of the cultured hepatocytes were studied from WD patients and normal controls. These cultured hepatocytes were incubated in the media of copper 15mg.L-1 only, copper 15 mg. L-1 with vincristine (agonist of P-type ArPase) 0.5mg. L-1, or copper 15 mg. L-1 withvanadate (antagonist of P-type ATPase) 18.39 mg. L-1separately. Microsome (endoplasmic reticulum and Golgi apparatus), lysosome, mitochondria, and cytosol were isolated by differential centrifugation. Copper contents in these organelles were measured with atomic absorption spectrophotometer, and the influence in copper transportion of these organelles by vanadate and vincristine were comparatively analyzed between WD patients and controls.WD copper transporting P-type ATPase was detected by SDS-PAGE in conjunction with Western blot in liver samples of WD patients and controls.RESULTS: The specific WD proteins (Mr155 000 lanes) were expressed in human hepatocytes, including the control and WD patients. After incubation with medium containing copper for 2 h or 24 h, the microsome copper concentration in WD patients was obviously lower than that of controls,and the addtion of vanadate or vincristine would change the copper transporting of microsomes obviously. When incubated with vincristine, levels of copper in microsome were significantly increased, while incubated with vanadate,the copper concentrations in microsome were obviously decreased. The results indicated that there were Wdproteins, the copper transportion P-type ATPase in the microsome of hepatocytes. WD patients possessed abnormal copper transporting function of WD protein in the microsome, and the agonist might correct the defect of copper transportion by promoting the activity of copper transportion P-type ATPase.CONCLUSION: Copper transportion P-type ATPase plays an important role in hepatocytic copper metabolism.Dysfunction of hepatocytic WD protein copper transportion might be one of the most important factors for WD.展开更多
Platelets have essential roles in both health and disease. Normal platelet function is required for hemostasis.Inhibition of platelet function in disease or by pharmacological treatment results in bleeding disorders.O...Platelets have essential roles in both health and disease. Normal platelet function is required for hemostasis.Inhibition of platelet function in disease or by pharmacological treatment results in bleeding disorders.On the other hand,hyperactive platelets lead to heart attack and stroke.Calcium is a major second messenger in platelet activation,and elevated intracellular calcium leads to hyperactive platelets.Elevated platelet calcium has been documented in hypertension and diabetes;both conditions increase the likelihood of heart attack and stroke. Thus,proper regulation of calcium metabolism in the platelet is extremely important.Plasma membrane Ca2+-ATPase(PMCA)is a major player in platelet calcium metabolism since it provides the only significant route for calcium efflux.In keeping with the important role of calcium in platelet function,PMCA is a highly regulated transporter.In human platelets,PMCA is activated by Ca2+/calmodulin,by cAMP-dependent phosphorylation and by calpain-dependent removal of the inhibitory peptide.It is inhibited by tyrosine phosphorylation and calpain-dependent proteolysis.In addition,the cellular location of PMCA is regulated by a PDZ-domain-dependent interaction with the cytoskeleton during platelet activation.Rapid regulation by phosphorylation results in changes in the rate of platelet activation,whereas calpain-dependent proteolysis and interaction with the cytoskeleton appears to regulate later events such as clot retraction.In hypertension and diabetes,PMCA expression is upregulated while activity is decreased, presumably due to tyrosine phosphorylation.Clearly,a more complete understanding of PMCA function in human platelets could result in the identification of new ways to control platelet function in disease states.展开更多
Metal–metal battery bears great potential for next-generation large-scale energy storage system because of its simple manufacture process and low production cost.However,the cross-over of metal cations from the catho...Metal–metal battery bears great potential for next-generation large-scale energy storage system because of its simple manufacture process and low production cost.However,the cross-over of metal cations from the cathode to the anode causes a loss in capacity and influences battery stability.Herein,a coating of poly(ionic liquid)(PIL)with poly(diallyldimethylammonium bis(trifluoromethanesulfonyl)imide)(PDADMA^(+)TFSI^(−))on a commercial polypropylene(PP)separator serves as an anion exchange membrane for a 3.3 V copper–lithium battery.The PIL has a positively charged polymer backbone that can block the migration of copper ions,thus improving Coulombic efficiency,long-term cycling stability and inhibiting self-discharge of the battery.It can also facilitate the conduction of anions through the membrane and reduce polarization,especially for fast charging/discharging.Bruce-Vincent method gives the transport number in the electrolyte to be 0.25 and 0.04 for PP separator without and with PIL coating,respectively.This suggests that the PIL layer reduces the contribution of the internal current due to cation transport.The use of PIL as a coating layer for commercial PP separator is a cost-effective way to improve overall electrochemical performance of copper–lithium batteries.Compared to PP and polyacrylic acid(PAA)/PP separators,the PIL/PP membrane raises the Coulombic efficiency to 99%and decreases the average discharge voltage drop to about 0.09 V when the current density is increased from 0.1 to 1 mA cm^(−2).展开更多
基金the Agricultural Science and Technology Innovation Program Cooperation and Innovation Mission(Grant No.CAAS-XTCX2016001)Shenzhen Science and Technology Projects(Grant No.JSGG20160608160725473)+1 种基金China Postdoctoral Science Foundation(Grant No.2018M641558)Fundamental Research Funds for Science,Technology and Innovation Commission of Shenzhen Municipality(Grant No.JCYJ20160530191619099).
文摘P1B-type heavy metal ATPases(HMAs)are transmembrane metal-transporting proteins that play a key role in metal homeostasis.We here reported the characterization of rice OsHMA6,a member of the P1B-type ATPase family.Phylogenetic tree analysis showed that OsHMA6 belonged to the Cu/Ag subgroup of the HMA family and had a close evolutionary relationship with OsHMA9.Amino acid sequence alignment showed 82.78%consistency between OsHMA6 and OsHMA9.OsHMA6 expressed in all organs at different growth stages,including spikelet,and abundant in leaf blades,however,OsHMA9 most strongly expressed in roots,but very low in spikelet.Excessive Cu^2+can up-regulate the expression of OsHMA6 and OsHMA9 in rice seedlings.The heterologous expression in yeast showed that OsHMA6 can significantly rescue the growth of yeast strain CM52 when supplied with 3 or 6 mmol/L Cu^2+.Compared with the empty vector pYES2,the Cu concentration in OsHMA6-pYES2 decreased by 23.4%and 30.3%under 3 or 6 mmol/L Cu2+,respectively.Subcellular localization revealed that OsHMA6 was located in the plasma membrane.These results suggested that OsHMA6,similar to OsHMA9,is likely a copper efflux protein located in the plasma membrane.
文摘Platinum(Pt)-based antitumor agents are effective in the treatment of many solid malignancies. However, their efficacy is limited by toxicity and drug resistance. Reduced intracellular Pt accumulation has been consistently shown to correlate with resistance in tumors. Proteins involved in copper homeostasis have been identified as Pt transporters. In particular, copper transporter receptor 1(CTR1), the major copper influx transporter, has been shown to play a significant role in Pt resistance. Clinical studies demonstrated that expression of CTR1 correlated with intratumoral Pt concentration and outcomes following Pt-based therapy. Other CTRs such as CTR2, ATP7 A and ATP7 B, may also play a role in Pt resistance. Recent clinical studies attempting to modulate CTR1 to overcome Pt resistance may provide novel strategies. This review discusses the role of CTR1 as a potential predictive biomarker of Pt sensitivity and a therapeutic target for overcoming Pt resistance.
基金supported by Program for Changjiang Scholars and In-novative Research Team in University with grant No. IRTO555-5,China Ministry of Education
文摘The experiment was conducted to as- sess the effects of dietary supplementation of Cu on the growth performance, digestive enzymes, tissue minerals and absorptive transporters in small intestinal mucosa of weanling pigs. One hundred crossbred pigs weaned at 28 ＋ 2 d of age were assigned randomly to one of the following diets with 5 replicates：corn-soy- bean basal diet with 10,100,175,250 mg/kg of Cu as CuSO4·5H20. The results showed that 250 mg/kg Cu had a positive effect （ P 〈 0.05） on average daily gain, daily feed intake and ratio of gain/feed. Com- pared to 10 mg/kg Cu, higher Cu had significant effect on the apparent digestibility of protein and fat （P 〈 0.05 ）. The supplementing of Cu improved am- ylase and lipase activity in jejunum content and lipase in pancreas （ P 〈 0.05） and had no effect on intestinalmorphology. The liver Cu elevated approximately 4- fold in pigs fed diet with 250 mg/kg Cu compared with pigs fed diet with 10 mg/kg Cu, no increases were observed in pigs receiving the lower level of Cu （100 and 175 mg/kg）. Both Fe and Zn contents in kidney and liver were not affected by Cu supplemen- tation. There was no positive effect （ P 〉0.05） of Cu supplementation on PepT1 （peptide transporter 1 ） and SGLT1 （sodium/glucose cotransporter） mRNA abundance in intestinal mucosa. However, higher sup- plementing level （250 mg/kg） significantly elevated the DMT1 （divalent metal transporter） mRNA abun- dance in duodenum mucosa. These results suggested that dietary supplementation with 250 mg/kg Cu could improve growth performance, nutrient digestibil- ity and intestinal enzyme activities of weanling pigs.
文摘Cisplatin belongs to platinum-based drugs and is widely used in cancer chemotherapy.Ototoxicity is one of the major dose limiting side-effects of cisplatin.For toxicity to occur cisplatin must first be transported from the bloodstream into cochlear cells.Three copper transporters are considered pathways for regulating the uptake and translocation of cisplatin into cells:Ctr1,ATP7A and ATP7B.Our recent study with cochlear organotypic cultures shows that cochlear hair cells can be destroyed by cisplatin at low concentrations from 10μm to 100μn.However,high doses of cisplatin cannot damage hair cells,maybe due to intrinsic feedback reactions that increase export of platinum by ATP7B when the platinum concentration is high in extracellular space.Cimitidine is a specific copper transporter inhibitor that can block the entrance of copper and platinum,and may prevent cisplatin-induced cochlear hair cell injury.To evaluate this hypothesis,we treated cochlear organotypic cultures with cisplatin (10 μm or 50 μm) alone,or cisplatin combined with cimitidine at concentrations ranging from 10-2000 μm for 48 hours.cisplatin at 10 μm damaged about 20% hair cells.In contrast,when cimitidine (10 μm,100 μm and 2000 μm) was added to the culture,near 100% cochlear hair cell survived.At higher concentration (50 μm),cisplatin destroyed about 80% of cochlear hair cells.However,100 μmcimitidine rescued about 50% hair cells from cisplatin damage,and 2000μm cimitidine protected about 80% hair cells.The data of western blot showed that CTR1 and ATP7B expressions were increased in cisplatin treated cochlear tissue,but cimitidine significantly reduced CTR1 and ATP7B.In addition,ATP7A expression was depressed a little after cisplatin treatment.Considering that Ctr1 is involved in copper and platinum influx,but the ATP7A and ATP7B are copper export transporters,the results suggest that cimitidine can effectively block the entrance by copper transporters and stop the influx of cisplatin.
基金Projects(50934002,51104011) supported by the National Natural Science Foundation of ChinaProject(IRT0950) supported by Program for Changjiang Scholars and Innovative Research Team in Chinese UniversityProject(20100480200) supported by China Postdoctoral Science Foundation
文摘Based on the momentum and mass conservation equations,a comprehensive model of heap bioleaching process is developed to investigate the interaction between chemical reactions,solution flow,gas flow,and solute transport within the leaching system.The governing equations are solved numerically using the COMSOL Multiphysics software for the coupled reactive flow and solute transport at micro-scale,meso-scale and macro-scale levels.At or near the surface of ore particle,the acid concentration is relatively higher than that in the central area,while the concentration gradient decreases after 72 d of leaching.The flow simulation between ore particles by combining X-ray CT technology shows that the highest velocity in narrow pore reaches 0.375 m/s.The air velocity within the dump shows that the velocity near the top and side surface is relatively high,which leads to the high oxygen concentration in that area.The coupled heat transfer and liquid flow process shows that the solution can act as an effective remover from the heap,dropping the highest temperature from 60 to 38 °C.The reagent transfer coupled with solution flow is also analyzed.The results obtained allow us to obtain a better understanding of the fundamental physical phenomenon of the bioleaching process.
基金Supported by Key Clinical Program of Ministry of Ministry of Health(No.37091)"211 Project"of SUMS sponsored by Ministry of Health and Guangdong Provincial Natural Science Foundation,No.990064
文摘AIM: To study the effect of copper transporting P-type ATPase in copper metabolism of hepatocyte and pathogenesis of Wilson disease (WD).METHODS: WD copper transporting properties in some organelles of the cultured hepatocytes were studied from WD patients and normal controls. These cultured hepatocytes were incubated in the media of copper 15mg.L-1 only, copper 15 mg. L-1 with vincristine (agonist of P-type ArPase) 0.5mg. L-1, or copper 15 mg. L-1 withvanadate (antagonist of P-type ATPase) 18.39 mg. L-1separately. Microsome (endoplasmic reticulum and Golgi apparatus), lysosome, mitochondria, and cytosol were isolated by differential centrifugation. Copper contents in these organelles were measured with atomic absorption spectrophotometer, and the influence in copper transportion of these organelles by vanadate and vincristine were comparatively analyzed between WD patients and controls.WD copper transporting P-type ATPase was detected by SDS-PAGE in conjunction with Western blot in liver samples of WD patients and controls.RESULTS: The specific WD proteins (Mr155 000 lanes) were expressed in human hepatocytes, including the control and WD patients. After incubation with medium containing copper for 2 h or 24 h, the microsome copper concentration in WD patients was obviously lower than that of controls,and the addtion of vanadate or vincristine would change the copper transporting of microsomes obviously. When incubated with vincristine, levels of copper in microsome were significantly increased, while incubated with vanadate,the copper concentrations in microsome were obviously decreased. The results indicated that there were Wdproteins, the copper transportion P-type ATPase in the microsome of hepatocytes. WD patients possessed abnormal copper transporting function of WD protein in the microsome, and the agonist might correct the defect of copper transportion by promoting the activity of copper transportion P-type ATPase.CONCLUSION: Copper transportion P-type ATPase plays an important role in hepatocytic copper metabolism.Dysfunction of hepatocytic WD protein copper transportion might be one of the most important factors for WD.
文摘Platelets have essential roles in both health and disease. Normal platelet function is required for hemostasis.Inhibition of platelet function in disease or by pharmacological treatment results in bleeding disorders.On the other hand,hyperactive platelets lead to heart attack and stroke.Calcium is a major second messenger in platelet activation,and elevated intracellular calcium leads to hyperactive platelets.Elevated platelet calcium has been documented in hypertension and diabetes;both conditions increase the likelihood of heart attack and stroke. Thus,proper regulation of calcium metabolism in the platelet is extremely important.Plasma membrane Ca2+-ATPase(PMCA)is a major player in platelet calcium metabolism since it provides the only significant route for calcium efflux.In keeping with the important role of calcium in platelet function,PMCA is a highly regulated transporter.In human platelets,PMCA is activated by Ca2+/calmodulin,by cAMP-dependent phosphorylation and by calpain-dependent removal of the inhibitory peptide.It is inhibited by tyrosine phosphorylation and calpain-dependent proteolysis.In addition,the cellular location of PMCA is regulated by a PDZ-domain-dependent interaction with the cytoskeleton during platelet activation.Rapid regulation by phosphorylation results in changes in the rate of platelet activation,whereas calpain-dependent proteolysis and interaction with the cytoskeleton appears to regulate later events such as clot retraction.In hypertension and diabetes,PMCA expression is upregulated while activity is decreased, presumably due to tyrosine phosphorylation.Clearly,a more complete understanding of PMCA function in human platelets could result in the identification of new ways to control platelet function in disease states.
基金supported by grant from the Research Grants Council(City U 11305220)of the Hong Kong Special Administrative Region,China
文摘Metal–metal battery bears great potential for next-generation large-scale energy storage system because of its simple manufacture process and low production cost.However,the cross-over of metal cations from the cathode to the anode causes a loss in capacity and influences battery stability.Herein,a coating of poly(ionic liquid)(PIL)with poly(diallyldimethylammonium bis(trifluoromethanesulfonyl)imide)(PDADMA^(+)TFSI^(−))on a commercial polypropylene(PP)separator serves as an anion exchange membrane for a 3.3 V copper–lithium battery.The PIL has a positively charged polymer backbone that can block the migration of copper ions,thus improving Coulombic efficiency,long-term cycling stability and inhibiting self-discharge of the battery.It can also facilitate the conduction of anions through the membrane and reduce polarization,especially for fast charging/discharging.Bruce-Vincent method gives the transport number in the electrolyte to be 0.25 and 0.04 for PP separator without and with PIL coating,respectively.This suggests that the PIL layer reduces the contribution of the internal current due to cation transport.The use of PIL as a coating layer for commercial PP separator is a cost-effective way to improve overall electrochemical performance of copper–lithium batteries.Compared to PP and polyacrylic acid(PAA)/PP separators,the PIL/PP membrane raises the Coulombic efficiency to 99%and decreases the average discharge voltage drop to about 0.09 V when the current density is increased from 0.1 to 1 mA cm^(−2).
