Regulation of cytochrome c oxidase contributes to health and optimal life

The generation of cellular energy in the form of ATP occurs mainly in mitochondria by oxidative phosphorylation.Cytochrome c oxidase(CytOx),the oxygen accepting and rate-limiting step of the respiratory chain,regulates the supply of variable ATP demands in cells by“allosteric ATP-inhibition of CytOx.”This mechanism is based on inhibition of oxygen uptake of CytOx at high ATP/ADP ratios and low ferrocytochrome c concentrations in the mitochondrial matrix via cooperative interaction of the two substrate binding sites in dimeric CytOx.The mechanism keeps mitochondrial membrane potentialΔΨm and reactive oxygen species(ROS)formation at low healthy values.Stress signals increase cytosolic calcium leading to Ca^2+-dependent dephosphorylation of CytOx subunit I at the cytosolic side accompanied by switching off the allosteric ATPinhibition and monomerization of CytOx.This is followed by increase ofΔΨm and formation of ROS.A hypothesis is presented suggesting a dynamic change of binding of NDUFA4,originally identified as a subunit of complex I,between monomeric CytOx(active state with highΔΨm,high ROS and low efficiency)and complex I(resting state with lowΔΨm,low ROS and high efficiency).

Hippocampal mitochondrial cytochrome C oxidase activity and gene expression in a rat model of chronic cerebral ischemia

The present study established a rat model of chronic cerebral ischemia using bilateral common carotid artery permanent ligation to analyze cytochrome C oxidase activity and mRNA expression in hippocampal mitochondria. Results showed significantly decreased cytochrome C oxidase activity and cytochrome C oxidase II mRNA expression with prolonged ischemia time. Further analysis revealed five mitochondrial cytochrome C oxidase II gene mutations, two newly generated mutations, and four absent mutational sites at 1 month after cerebral ischemia, as well as three mitochondrial cytochrome C oxidase III gene mutations, including two newly generating mutations, and one disappeared mutational site at 1 month after cerebral ischemia. Results demonstrated that decreased cytochrome C oxidase gene expression and mutations, as well as decreased cytochrome C oxidase activity, resulting in energy dysmetabolism, which has been shown to be involved in the DatholoQical Process of ischemic brain iniurv.

Construction of Cox7a2 fluorescent vector and its effect on cytochrome C oxidase activity in mouse Sertoli cell line TM4

Objective To construct Cox7a2 fluorescent vector and study its effect on cytochrome C oxidase ( COX) activity in mouse Sertoli cell line TM4. Methods The coding region of CoxTa2 was amplified from mouse Sertoli cell line TM4 by RT-PCR. PCR product was

Tumor necrosis factor receptor-associated protein 1 regulates hypoxia-induced apoptosis through a mitochondria-dependent pathway mediated by cytochrome c oxidase subunit Ⅱ

Background:Tumor necrosis factor receptor-associated protein 1(TRAP1)plays a protective effect in hypoxic cardiomyocytes,but the precise mechanisms are not well clarified.The study is aimed to identify the mechanism of TRAP1 on hypoxic damage in cardiomyocytes.Methods:In this study,the effects of TRAP1 and cytochrome c oxidase subunit Ⅱ(COXⅡ)on apoptosis in hypoxia-induced cardiomyocytes were explored using overexpression and knockdown methods separately.Results:Hypoxia induced cardiomyocyte apoptosis,and TRAP1 overexpression notably inhibited apoptosis induced by hypoxia.Conversely,TRAP1 silencing promoted apoptosis in hypoxic cardiomyocytes.Further investigation revealed that the proapoptotic effects caused by the silencing of TRAP1 were prevented by COXⅡ overexpression,whereas COXⅡ knockdown reduced the antiapoptotic function induced by TRAP1 overexpression.Additionally,changes in the release of cytochrome c from mitochondria into the cytosol and the caspase-3 activity in the cytoplasm,as well as reactive oxygen species production,were found to be correlated with the changes in apoptosis.Conclusions:The current study uncovered that TRAP1 regulates hypoxia-induced cardiomyocyte apoptosis through a mitochondria-dependent apoptotic pathway mediated by COXⅡ,in which reactive oxygen species presents as an important component.

Synthesis, Characterization of Heterodinuclear Co-Cu Complex and Its Electrocatalytic Activity towards O_2 Reduction: Implications for Cytochrome c Oxidase Active Site Modeling

A new dinucleating ligand consisting of a tetraphenylporphyrin derivative covalently linked with tris(2-benzimidazylmethyl)-amine and its homodinuclear Co-Co and heterodinuclear Co-Cu complexes were synthesized and spectroscopically characterized. The heterobimetallic cobalt-copper complex bearing three benzimidazole ligands for copper, as cytochrome c oxidase active site model, was applied to the surface of glassy carbon electrode to show electrocatalytic activity for O 2 reduction in aqueous solution at an acidity level close to physiological pH value. The kinetic parameters of this electrocatalytic process were obtained.

Evolution for extreme living:variation in mitochondrial cytochrome c oxidase genes correlated with elevation in pikas(genus Ochotona)

The genus Ochotona(pikas)is a clade of cold-tolerant lagomorphs that includes many high-elevation species.Pikas offer a unique opportunity to study adaptations and potential limitations of an ecologically important mammal to high-elevation hypoxia.We analyzed the evolution of 3 mitochondrial genes encoding the catalytic core of cytochrome c oxidase(COX)in 10 pika species occupying elevations from sea level to 5000 m.COX is an enzyme highly reliant on oxygen and essential for cell function.One amino acid property,the equilibrium constant(ionization of COOH),was found to be under selection in the overall protein complex.We observed a strong relationship between the net value change in this property and the elevation each species occupies,with higher-elevation species having potentially more efficient proteins.We also found evidence of selection in low-elevation species for potentially less efficient COX,perhaps trading efficiency for heat production in the absence of hypoxia.Our results suggest that different pika species may have evolved elevation-specific COX proteins,specialization that may indicate limitations in their ability to shift their elevational ranges in response to future climate change.

A Comparative Study of Human and Animal Hairs:Microscopic Hair Comparison and Cytochrome c Oxidase I Species Identification

Human and animal hairs have been used in forensic investigations for over a century.Hair is stable under adverse natural conditions;hence,it is often recovered at the crime scene,and it is necessary to determine whether the hair is of human or animal origin.Morphological and genetic characteristics are useful to differentiate human hair from animal hair.In the present study,we analyzed the distinguishing characteristics of hair of various species.In addition,we explore species identification by cytochrome c oxidase I mitochondrial gene analysis.We confirm that both the microscopic and molecular analyses of hairs are useful in forensic investigations.

Cytochrome c Oxidase Subunit 1-based Human RNA Quantification to Enhance mRNA Profiling in Forensic Biology

RNA analysis offers many potential applications in forensic science,and molecular identification of body fluids by analysis of cell‑specific RNA markers represents a new technique for use in forensic cases.However,due to the nature of forensic materials that often admixed with nonhuman cellular components,human‑specific RNA quantification is required for the forensic RNA assays.Quantification assay for human RNA has been developed in the present study with respect to body fluid samples in forensic biology.The quantitative assay is based on real‑time reverse transcription‑polymerase chain reaction of mitochondrial RNA cytochrome c oxidase subunit I and capable of RNA quantification with high reproducibility and a wide dynamic range.The human RNA quantification improves the quality of mRNA profiling in the identification of body fluids of saliva and semen because the quantification assay can exclude the influence of nonhuman components and reduce the adverse affection from degraded RNA fragments.

Benthodytes tetrapapillata sp.nov.,a new elasipodid sea cucumber(Elasipodida:Psychropotidae)from a seamount in the Western Pacific Ocean

Benthodytes tetrapapillata sp.nov.was collected from a seamount located on the Caroline Ridge at a depth of 2289 m,during the cruise of R/V Kexue in June 2019.We provided detailed descriptions of external and deposits morphology.The phylogenetic analyses based on cytochrome c oxidase I(COI)and a concatenated dataset of 16S and COI genes showed that the new species belonged to Benthodytes that is not monophyletic.Both details of morphological comparisons and molecular analyses confirmed that Benthodytes tetrapapillata sp.nov.is a new psychropotid species.A state of main morphological characters in valid species of Benthodytes is also provided in this study.

DNA barcoding of fishes from Zhoushan coastal waters using mitochondrial COI and 12S rRNA genes

Accurate species identification is a key component of biodiversity research.DNA barcoding is an effective molecular method used for fish species identification.We aimed to study the DNA barcoding of fish in Zhoushan coastal waters,explore the differences and applicability of two gene fragments(12S rRNA and COI)of DNA barcoding in fish species identification,and established a comprehensive fish barcoding reference database.Two hundred and eighty-seven captured fish samples from Zhoushan coastal waters were identified using morphological characteristics and DNA barcoding.A total of 26412S rRNA sequences(belonging to eight orders,31 families,55 genera,and 66 species)and 188 COI sequences(belonging to seven orders,30 families,48 genera,and 58 species)were obtained.The lengths of the 12S rRNA sequences ranged from 165 to 178 bp,and the guanine-cytosine(GC)content was 45.37%.The average 12S rRNA interspecific and intraspecific genetic distances(K2P)were 0.10%and 26.66%,respectively.The length of the COI sequence ranged 574–655 bp,and the content of GC was 45.97%.The average 12S rRNA interspecific and intraspecific genetic distances(K2P)were 0.16%and 27.45%,respectively.The minimum interspecific genetic distances of 12S rRNA and COI(1.23%and 1.86%)were both greater than their maximum intraspecific genetic distances(2.42%and 8.66%).Three molecular analyses(NJ tree,ABGD,and GMYC)were performed to accurately identify and delineate species.Clustering errors occurred when the 12S rRNA sequences were delimited using the NJ tree method,and the delimitation results of ABGD and GMYC are consistent with the final species identification results.Our results demonstrate that DNA barcoding based on 12S rRNA and COI can be used as an effective tool for fish species identification,and 12S rRNA has good application prospects in the environmental DNA(eDNA)metabarcoding of marine fish.

Genetic diversity and population structure of the sea star Asterias amurensis in the northern coast of China

The sea star Asterias amurensis is widely viewed as a severe“marine pest”because of its broad feeding habits.Over the past few decades,A.amurensis undergoes massive and sporadic population outbreaks worldwide,causing extensive economic and ecological losses to the local aquaculture industry and marine ecosystem.Understanding the genetic diversity and population structure of A.amurensis can provide vital information for resource management.By analyzing the polymorphism of the mitochondrial cytochrome C oxidase subunit I(COI)gene and ten simple sequence repeat(SSR)microsatellites markers,the genetic diversity and population structure of A.amurensis of four populations along the northern coast of China was uncovered.A total of 36 haplotypes were identified,and a main haplotype was found in four populations.The Qingdao(QD)population displayed the highest genetic diversity among all the populations.The AMOVA and pairwise F_(st)showed that there was small but statistically significant population differentiation among the four populations,especially between QD and Weihai(WH).Moreover,the principal component analysis(PCA)and admixture analysis showed that several individuals in Yantai(YT)and Dalian(DL)had little genetic association with other individuals.Overall,this study provided useful information of the genetic diversity and population structure of A.amurensis and will contribute to the resource management of A.amurensis in China.

MitochondrialtRNA^(leu(UUR)) Gene Mutation and the DecreasedActivity of Cytochrom e c Oxidase in Preeclam psia

To explore the roles of mitochondria tRNA leu(UUR) gene mutation at nucleotide 3243 and the activity of cytochrome c oxidase in pathogenesis of preeclampsia, 57 patients with preeclampsia and 60 normotension pregnancy women were screened for tRNA leu(UUR) nt3243 A→G mutation with the method of polymers chain reaction (PCR) and restriction fragment length polymorphism. Cytochrome c oxidase activity was determined by measuring the rate of cyanide sensitive oxidation of reduced cytochrome c using luminosity photographer. The results showed that cytochrome c oxidase activity was significantly lower in the preeclampsia group (0.30±0.39/min, n = 32) than that in the controls (0.73±0.54/min, n = 26, P <0.01). The mitochondria DNA mutation at position 3243 was not found in our series. The results suggested that the decreased activity of cytochrome c oxidase might impair the energy production, leading to the mitochondria dysfunction and placenta dysfunction in preeclampsia patients. Mitochondria dysfunction may be involved in the pathogenesis of preeclampsia. The mutation of mitochondria DNA may not be the common contributor of preeclampsia in our series.

Mitochondrial protective and anti-apoptotic effects of Rhodiola crenulata extract on hippocampal neurons in a rat model of Alzheimer's disease

In our previous study, we found that the edible alcohol extract of the root of the medicinal plant Rhodiola crenulata（RCE） improved spatial cognition in a rat model of Alzheimer＇s disease. Another study from our laboratory showed that RCE enhanced neural cell proliferation in the dentate gyrus of the hippocampus and prevented damage to hippocampal neurons in a rat model of chronic stress-induced depression. However, the mechanisms underlying the neuroprotective effects of RCE are unclear. In the present study, we investigated the anti-apoptotic effect of RCE and its neuroprotective mechanism of action in a rat model of Alzheimer＇s disease established by intracerebroventricular injection of streptozotocin. The rats were pre-administered RCE at doses of 1.5, 3.0 or 6.0 g/kg for 21 days before model establishment. ATP and cytochrome c oxidase levels were significantly decreased in rats with Alzheimer＇s disease. Furthermore, neuronal injury was obvious in the hippocampus, with the presence of a large number of apoptotic neurons. In comparison, in rats given RCE pretreatment, ATP and cytochrome c oxidase levels were markedly increased, the number of apoptotic neurons was reduced, and mitochondrial injury was mitigated. The 3.0 g/kg dose of RCE had the optimal effect. These findings suggest that pretreatment with RCE prevents mitochondrial dysfunction and protects hippocampal neurons from apoptosis in rats with Alzheimer＇s disease.

Seasonal phenotypic flexibility in body mass,basal thermogenesis,and tissue oxidative capacity in the male Silky Starling(Sturnus sericeus)

Background: Acclimatization to winter conditions is an essential prerequisite for the survival of small birds in the northern temperate zone.Changes in photoperiod,ambient temperature and food availability trigger seasonal physiological and behavioral acclimatization in many passerines.Seasonal trends in metabolic parameters are well known in avian populations from temperate environments;however,the physiological and biochemical mechanisms underlying these trends are incompletely understood.In this study,we used an integrative approach to measure variation in the thermogenic properties of the male Silky Starling(Sturnus sericeus) at different levels or organization,from the whole organism to the biochemical.We measured body mass(Mb),basal metabolic rate(BMR),energy budget,the mass of selected internal organs,state 4 respiration and cytochrome c oxidase(COX) activity in the heart,liver and muscle.Methods: Oxygen consumption was measured using an open-circuit respirometry system.The energy intake of the birds were then determined using an oxygen bomb calorimeter.Mitochondrial state 4 respiration and COX activity in heart,liver and pectoral muscle were measured with a Clark electrode.Results: The results suggest that acclimatization to winter conditions caused significant change in each of the measured variables,specifically,increases in Mb,organ mass,BMR,energy intake and cellular enzyme activity.Furthermore,BMR was positively correlated with body mass,energy intake,the mass of selected internal organs,state 4 respiration in the heart,liver and muscle,and COX activity in the heart and muscle.Conclusions: These results suggest that the male Silky Starling's enhanced basal thermogenesis under winter conditions is achieved by making a suite of adjustments from the whole organism to the biochemical level,and provide further evidence to support the notion that small birds have high phenotypic plasticity with respect to seasonal changes.

First record of the little fire ant,Wasmannia auropunctata(Hymenoptera:Formicidae),in Chinese mainland

In January 2022,we received ant specimens collected from three field colonies from Shantou City,Guangdong Province,China.They were identified as the little fire ant,Wasmannia auropunctata,through morphological and molecular analyses.Wasmannia auropunctata is listed as one of the 100 most dangerous invasive species by the International Union for Conservation of Nature(IUCN)and has spread from its native range in South America to every continent except Antarctica.DNA analysis of mitochondrial cytochrome c oxidase subunit I(COI)in nine specimens of W.auropunctata found that they had a close genetic relationship with specimens from Argentina.This study represents the first formal record of the establishment of W.auropunctata outdoor in Chinese mainland.However,the invasion stage and occurrence degree of W.auropunctata in China are not clear to date.The implementation of quarantine measures,investigation of the occurrence and distribution,and development of monitoring and control strategies are needed to actively respond to the threat posed by this highly invasive ant.

Effects of sustained cold and heat stress on energy intake, growth and mitochondrial function of broiler chickens

To study the correlation of broiler chickens with energy intake, growth and mitochondrial function which exposed to sustained cold and heat stress and to find out the comfortable temperature, 288 broiler chickens（21-day with（748±26） g, 144 males and 144 females） were divided randomly into six temperature-controlled chambers. Each chamber contained six cages including eight AA broilers per cage, each cage as a repeat. After acclimation for one week（temperature, 21℃; relative humidity, 60%）, the temperature of each chamber was adjusted（finished within 1 h） respectively to 10, 14, 18, 22, 26, or 30℃（RH, 60%） for a 14-day experimental period. After treatment, gross energy intake（GEI）, metabolizable energy intake（MEI）, the ratio of MEI/BW, metabolizability, average daily gain（ADG）, the concentration of liver mitochondria protein and cytochrome c oxidase（CCO） were measured respectively. Our results confirmed that when the temperature over 26℃ for 14 days, GEI, MEI and CCO activities were decreased significantly（P〈0.05）, but the concentration of liver mitochondria protein was increased and metabolizability of broilers was not influenced（P〉0.05）. Compared with treatment for 14 days, the ratio of MEI/body weight（BW） were also decreased when the temperature over 26℃ after temperature stress for 7 days（P〈0.05）, meanwhile mitochondrial protein concentration was increased at 10℃ and CCO activity was not affected（P〉0.05）. Additionally at 22℃, the ADG reached the maximal value. When kept in uncomfortable temperatures for a long time, the ADG and CCO activities of broiler were reduced, which was accompanied by mitochondrial hyperplasia. In summary, our study focused on the performance of broilers during sustained cold and heat environmental temperatures ranging from 10 to 30℃. From the point of view of energy utilization, moreover, 22 to 26℃ is comfortable for 28–42 day s broilers. And these could provide the theoretical basis on the high efficient production.

The role of temperature as a driver of metabolic flexibility in the Red-billed Leiothrix(Leiothrix lutea)

Background:The thermoregulatory ability of animals is strongly influenced by the temperature of their environment.Acclimation to cold requires a range of physiological and morphological adjustments.In this study,we tested the hypothesis that a small passerine,the Red-billed Leiothrix(Leiothrix lutea),can maintain homeothermy in cold conditions by adjusting the physiology and biochemistry of its tissue and organs and return to its former physiological and biochemical state when moved to a warm temperature.Methods:Phenotypic variation in thermogenic activity of the Red-billed Leiothrixs(Leiothrix lutea)was investigated under warm(35℃),normal(25℃)or cold(15℃)ambient temperature conditions.Oxygen consumption was measured using an open-circuit respirometry system.Mitochondrial state-4 respiration and cytochrome-c oxidase(COX)activity in liver,kidney heart and pectoral muscle were measured with a Clark electrode.Results:Birds acclimated to an ambient temperature of 15℃ for 4 weeks significantly increased their basal metabolic rate(BMR)compared to a control group kept at 25℃.Birds acclimated to 35℃ decreased their BMR,gross energy intake(GEI)and digestible energy intake(DEI).Furthermore,birds acclimated to 15℃ increased state-4 respiration in their pectoral muscles and cytochrome-c oxidase(COX)activity in their liver and pectoral muscle,compared to the 25℃ control group.Birds acclimated to 35℃ also displayed lower state-4 respiration and COX activity in the liver,heart and pectoral muscles,compared to those kept at 25℃.There was a positive correlation between BMR and state-4 respiration,and between BMR and COX activity,in all of the above organs except the liver and heart.Conclusions:Our study illustrates that the morphological,physiological,and enzymatic changes are associated with temperature acclimation in the Red-billed Leiothrix,and supports the notion that the primary means by which small birds meet the energetic challenges of cold conditions is through metabolic adjustments.

Morphological and molecular characterization of the rice root-knot nematode, Meloidogyne graminicola, Golden and Birchfeild, 1965 occurring in Zhejiang, China

The rice root-knot nematode Meloidogyne graminicola is a severe pest of rice. In China, it was first reported from Hainan Province, and later from several other provinces. In the present study, a rice root-knot nematode population found from the rice cultivation areas of Zhejiang Province, China is characterized via molecular analysis using internal transcribed spacer(ITS) and cytochrome c oxidase subunit Ⅱ(coxⅡ)-16 S rRNA genes and scanning electron microscopy(SEM) observations of males and the second-stage juveniles. Morphometric data and molecular sequence comparisons for all M. graminicola populations occurring in China are also provided. The overall morphology of M. graminicola found in Zhejiang match well with the original description, though males have a slightly longer body and stylet, and a shorter tail, while the second-stage juvenile is also slightly longer than in the original description. This is the first report of M. graminicola from Zhejiang. Phylogenetic studies based on coxⅡ suggest that all the Chinese populations belong to Type B. This study expands knowledge of the increasing distribution and phylogenetic relationships of M. graminicola that occur in China.

Prevalence and genotype distribution of Enterobius vermicularis among kindergarteners in Shiraz and Khorramabad cities, Iran

Objective: To study the prevalence and genotype of Enterobius(E.)vermicularis from adhesive tape samples in the cities of Shiraz and Khorramabad, Iran. Methods: A total of 1 000 adhesive tape samples from kindergartens in Shiraz(500 samples) and Khorramabad(500 samples) were collected and tested using a microscope to find E. vermicularis egg/s. A questionnaire was filled out for each sample. In order to characterize the genotype of E. vermicularis, the PCR-sequencing method of the mitochondrial cytochrome C oxidase subunit 1(cox1) gene was used. Genomic DNA was extracted from the positive scotch tape samples of E. vermicularis. The cox1 gene was amplified by the polymerase chain reaction and sequenced. The sequence data were aligned using the BioEdit software and compared with the published sequences in GenBank. Phylogenetic analysis was performed using the maximum likelihood method. Results: The parasitological method showed that 15 out of the 500 samples from Shiraz(3.00%) and 12 out of the 500 samples from Khorramabad(2.40%) were infected with E. vermicularis eggs. BLAST analysis indicated that the sequenced isolates belonged to E. vermicularis genotype B while three different haplotypes were also identified. Conclusions: This is the first study on genotyping E. vermicularis in the cities of Shiraz and Khorramabad. Considering the public health importance of the disease, further studies are necessary to characterize the genotype of E. vermicularis in human populations from other regions of Iran.

Genetic variation and phylogenetic relationship of Hypoderaeum conoideum(Bloch, 1782) Dietz, 1909(Trematoda: Echinostomatidae) inferred from nuclear and mitochondrial DNA sequences

Objective:To explore genetic variations of Hypoderaeum conoideum collected from domestic ducks from 12 different localities in Thailand and Lao PDR,as well as their phylogenetic relationship with American and European isolates.Methods:The nucleotide sequences of their nuclear ribosomal DNA(ITS),mitochondrial cytochrome c oxidase subunit 1(CO1),and NADH dehydrogenase subunit 1(ND1)were used to analyze genetic diversity indices.Results:We found relatively high levels of nucleotide polymorphism in ND1(4.02%),whereas moderate and low levels were observed in CO1(2.11%)and ITS(0.96%),respectively.Based on these polymorphisms,the 20 ND1,12 CO1,and 18 ITS haplotypes were classified,and several common haplotypes were observed in all samples.At least three major lineages,namely American,European and Asian lineages,have been classified by phylogenetic analyses based on ND1 sequences.Conclusions:Our report demonstrates that the ND1 gene is the most suitable genetic marker to explore genetic variation and phylogenetic relationship of Hypoderaeum conoideum.However,a combination of all loci for ND1,CO1 and ITS would be of great value toward further genetic investigation of this endemic worldwide parasite.Thus,comprehensive molecular genetic analyses of Hypoderaeum conoideum from its worldwide distribution is needed to further understanding of the evolutionary and systematic relationships of this parasite.