Higher Adenosine Triphosphate Levels Research Articles

RipE expression correlates with high ATP levels in Ehrlichia, which confers resistance during the extracellular stage to facilitate a new cycle of infection.

Ehrlichiosis is a potentially life-threatening disease caused by infection with the obligatory intracellular bacteria Ehrlichia species. Ehrlichia japonica infection of mice provides an animal model of ehrlichiosis as it recapitulates full-spectrum and lethal ehrlichiosis in humans. The E. japonica transposon mutant of EHF0962, which encodes a previously uncharacterized hypothetical protein, is attenuated in both infection and virulence in mice. EHF0962 was hence named here as resistance-inducing protein of Ehrlichia (RipE). Using this ΔripE mutant, we studied how RipE protein contributes to Ehrlichia pathogenesis. Ehrlichia species have an intracellular developmental cycle and a brief extracellular stage to initiate a new cycle of infection. Majority of RipE proteins were expressed on the surface of the smaller infectious dense-core stage of bacteria. Extracellular ΔripE E. japonica contained significantly less adenosine triphosphate (ATP) and lost infectivity more rapidly in culture compared with wild-type (WT) E. japonica. Genetic complementation in the ΔripE mutant or overexpression of ripE in WT E. japonica significantly increased bacterial ATP levels, and RipE-overexpressing E. japonica was more virulent in mice than WT E. japonica. RipE is conserved among Ehrlichia species. Immunization of mice with recombinant RipE induced an in vitro infection-neutralizing antibody, significantly prolonged survival time after a lethal dose of E. japonica challenge, and cross-protected mice from infection by Ehrlichia chaffeensis, the agent of human monocytic ehrlichiosis. Our findings shed light on the extracellular stage of Ehrlichia, highlighting the importance of RipE and ATP levels in Ehrlichia for extracellular resistance and the next cycle of infection. Thus, RipE is a critical Ehrlichia protein for infection as such can be a potential vaccine target for ehrlichiosis.

Tumor microenvironment activation amplify oxidative stress promoting tumor energy remodeling for mild photothermal therapy and cuproptosis

Tumor metabolic reprogramming requires high levels of adenosine triphosphate (ATP) to maintain treatment resistance, which poses major challenges to chemotherapy and photothermal therapy. Especially, high levels of ATP promote copper ion efflux for limiting the curative effect of cuproptosis. Here, an H2S-responsive mesoporous Cu2Cl(OH)3-loading chemotherapeutic cisplatin (CDDP) was synthesized, and the final nanoparticle, CDDP@Cu2Cl(OH)3-CDs (CDCuCDs), was encapsulated by electrostatic action with carbon dots (CDs). CDCuCDs reacted with overproduction H2S in colon tumor to produce photothermic copper sulfide for photothermal therapy. CDDP was released by lysis to achieve chemotherapeutic effects. Importantly, CDDP elevated H2O2 levels in cells through a cascade reaction and continuously transforms H2O2 into highly cytotoxic •OH through chemodynamic therapy between H2O2 and Cu+, which enables nanoparticles to generate •OH and improve the chemotherapeutic efficacy. Highly toxic •OH disrupts mitochondrial homeostasis, prohibiting it from performing normal energy-supplying functions. Down-regulated ATP inhibits heat shock protein expression, which promotes the therapeutic effect of mild photothermal therapy and reduces the efflux of intracellular copper ions, thus improving the therapeutic effect of cuproptosis. Our research provides a potential therapeutic strategy using overproduction H2S responses in tumors, allowing tumor microenvironment-activated •OH nanogenerators to promote tumor energy remodeling for cancer treatment.

Detrusor Overactivity After Partial Bladder Outlet Obstruction Is Associated With High Urinary Adenosine Triphosphate Levels in Female Wistar Rats.

Bladder outlet obstruction (BOO) commonly causes detrusor overactivity (DO). In this study, a post hoc analysis of previous obtained data, we investigate if DO occurring in initial phases of BOO is associated with changes in urinary adenosine triphosphate (ATP) levels. Adult female Wistar rats were submitted to partial BOO (pBOO) or to sham obstruction. Cystometry was performed at 3 or 15 days after pBOO and saline voided was collected for ATP determination. Normality was tested using Shapiro-Wilk test. The mean frequency of voiding contractions (VCs) of the sham-operated animals at 15 days after surgery, plus or minus 3 standard deviations, was used to represent the normal range. Statistical analyses were performed using the chi-square and Mann-Whitney tests. DO was indicated by a VC frequency greater than or equal to 0.9 VCs/min. DO was observed in 63% of animals at 3 days and in 33% at 15 days following pBOO. ATP levels were significantly higher in rats with DO compared to those without DO. The DO phenotype, occurring in the initial phases of BOO, is associated with comparatively high urinary ATP levels.

Applying a multi-faceted infection control strategy to improve hospital environmental cleaning quality

BackgroundAlong with existing infection control policies, repeated education and training of environmental service workers (ESWs) improves their compliance and ultimately reduces hospital-associated infection (HAI) rates. However, only limited studies have explored the health behavioral determinants of ESWs regarding their cleaning performance after implementing an educational intervention with multi-faceted infection control strategy. ObjectiveTo determine whether an educational intervention with multi-faceted infection control strategy improves the health behavioral determinants associated with ESWs’ cleaning performance. MethodsTwenty-eight ESWs who received an educational intervention with multi-faceted hospital infection control strategy were included. ESWs’ knowledge, perceived benefits and barriers, self-efficacy, health literacy, and cleaning performance were evaluated at pre-intervention, post-intervention, and 3-month follow-up. ResultsHAI-related adenosine triphosphate (ATP) levels decreased significantly at post-intervention and 3-month follow-up compared with pre-intervention levels (all p < 0.05). All post-intervention ATP levels met the standard criterion after the 2nd environmental cleaning, with a median score of 267 (range, 71–386). High baseline ATP levels (odds ratio [OR] = 4.195, 95%CI 2.500–7.042, p < 0.05) were positively associated with qualified post-intervention ATP levels, while high education (OR = 0.480, 95%CI 0.276–0.833, p < 0.05) and high baseline knowledge scores (OR = 0.481, 95%CI 0.257–0.903, p = 0.023) were negatively associated with qualified post-intervention ATP levels. ConclusionEducational intervention using a multi-faceted infection control strategy improves health behavioral determinants (baseline education, knowledge scores and ATP levels) associated with ESWs’ hospital cleaning performance. Receiving an educational intervention may increase HAI knowledge of environmental cleaning among ESWs with high education or low baseline HAI knowledge.

Acidic electrolyzed oxidizing water delayed the breakdown occurrence in pulp of fresh longan by regulating the metabolisms of respiratory and energy

The efficacy of acidic electrolyzed oxidizing water (AEOW) treatment on the respiratory and energy metabolism during storage of harvested longan and its relationship with pulp breakdown were investigated. Results confirmed that the AEOW treatment exerted a significant inhibitory effect on longan pulp breakdown, with 15.5 % lower pulp breakdown index than the control at 6 d. Compared to the control fruit, the AEOW treatment displayed lower activities of polyphenol oxidase (PPO), ascorbic acid oxidase (AAO), alternate oxidase (AOX), cytochrome C oxidase (CCO), succinate dehydrogenase (SDH), pulp phosphohexose isomerase (PGI), and higher levels of nicotinamide adenine dinucleotide kinase (NADK), glucose-6-phosphate dehydrogenase (G-6-PDH) and glucose-6-phosphate dehydrogenase (6-PGDH). Furthermore, the AEOW treatment increased the amount of NADPH and NADP while decreasing that of NADH and NAD. In terms of energy metabolism, the AEOW treatment maintained higher levels of adenosine triphosphate (ATP), adenosine diphosphate (ADP), energy charge, H+-ATPase, Ca2+-ATPase, and Mg2+-ATPase, but lower content of adenosine monophosphate (AMP). Therefore, the AEOW treatment inhibited respiratory metabolism by decreasing the terminal oxidase activities, tricarboxylic acid cycle (TCA cycle) and glycolytic pathway (EMP), and increasing the pentose phosphate pathway (PPP), which helped to maintain a higher energy status and further led to attenuation of longan pulp breakdown.

Gas-Mediated Tumor Energy Remodeling for Sensitizing Mild Photothermal Therapy.

The metabolic reprogramming of tumors requires ATP to maintain therapeutic resistance, posing a major challenge for photothermal therapy (PTT). Although raising the temperature can help in tumor ablation, it frequently leads to severe side effects such as chronic inflammation and hyper-immunogenicity. Therefore, improving therapeutic responsiveness and promoting wound healing are crucial considerations in PTT development. In this context, we proposed a gas-mediated energy remodeling strategy to enhance the efficacy of mild PTT at moderate temperatures while minimizing side effects. In the proof-of-concept study, an FDA-approved drug-based hydrogen sulfide (H2S) donor was developed to provide a sustained supply of H2S to tumor sites, serving as an adjuvant to mild PTT. This approach proved to be highly effective in disrupting the mitochondrial respiratory chain, inhibiting ATP generation, and reducing the overexpression of heat shock protein 90 (HSP90), which ultimately amplified the therapeutic outcome. With the ability to reverse tumor thermotolerance, this strategy delivered a greatly potent antitumor response, achieving complete tumor ablation in a single treatment while minimizing harm to healthy tissues. Thus, it holds great promise to overcome the limitations of PTT and may serve as a valuable paradigm for the future clinical translation of photothermal nanoplatforms.

Gas‐Mediated Tumor Energy Remodeling for Sensitizing Mild Photothermal Therapy

AbstractThe metabolic reprogramming of tumors requires high levels of adenosine triphosphate (ATP) to maintain therapeutic resistance, posing a major challenge for photothermal therapy (PTT). Although raising the temperature helps in tumor ablation, it frequently leads to severe side effects. Therefore, improving the therapeutic response and promoting healing are critical considerations in the development of PTT. Here, we proposed a gas‐mediated energy remodeling strategy to improve mild PTT efficacy while minimizing side effects. In the proof‐of‐concept study, a Food and Drug Administration (FDA)‐approved drug‐based hydrogen sulfide (H2S) donor was developed to provide a sustained supply of H2S to tumor sites, serving as an adjuvant to PTT. This approach proved to be highly effective in disrupting the mitochondrial respiratory chain, inhibiting ATP generation, and reducing the overexpression of heat shock protein 90 (HSP90), which ultimately amplified the therapeutic outcome. With the ability to reverse tumor thermotolerance, this strategy delivered a greatly potent antitumor response, achieving complete tumor ablation in a single treatment while minimizing harm to healthy tissues. Thus, it holds great promise to be a universal solution for overcoming the limitations of PTT and may serve as a valuable paradigm for the future clinical translation of photothermal nanoagents.

Creatine metabolism at the uterine-placental interface throughout gestation in sheep†.

The placenta requires high levels of adenosine triphosphate to maintain a metabolically active state throughout gestation. The creatine-creatine kinase-phosphocreatine system is known to buffer adenosine triphosphate levels; however, the role(s) creatine-creatine kinase-phosphocreatine system plays in uterine and placental metabolism throughout gestation is poorly understood. In this study, Suffolk ewes were ovariohysterectomized on Days 30, 50, 70, 90, 110 and 125 of gestation (n = 3-5 ewes/per day, except n = 2 on Day 50) and uterine and placental tissues subjected to analyses to measure metabolites, mRNAs, and proteins related to the creatine-creatine kinase-phosphocreatine system. Day of gestation affected concentrations and total amounts of guanidinoacetate and creatine in maternal plasma, amniotic fluid and allantoic fluid (P < 0.05). Expression of mRNAs for arginine:glycine amidinotransferase, guanidinoacetate methyltransferase, creatine kinase B, and solute carrier 16A12 in endometria and for arginine:glycine amidinotransferase and creatine kinase B in placentomes changed significantly across days of gestation (P < 0.05). The arginine:glycine amidinotransferase protein was more abundant in uterine luminal epithelium on Days 90 and 125 compared to Days 30 and 50 (P < 0.01). The chorionic epithelium of placentomes expressed guanidinoacetate methyltransferase and solute carrier 6A13 throughout gestation. Creatine transporter (solute carrier 6A8) was expressed by the uterine luminal epithelium and trophectoderm of placentomes throughout gestation. Creatine kinase (creatine kinase B and CKMT1) proteins were localized primarily to the uterine luminal epithelium and to the placental chorionic epithelium of placentomes throughout gestation. Collectively, these results demonstrate cell-specific and temporal regulation of components of the creatine-creatine kinase-phosphocreatine system that likely influence energy homeostasis for fetal-placental development.

P2X7 purinergic receptor: A potential target in heart diseases (Review).

The P2X7 purinergic receptor (P2X7R) is a non‑selective cation channel activated by high levels of adenosine triphosphate that are commonly present in serious conditions. Activation of this purinergic receptor is closely related to the development of various disease states including inflammatory and neurodegenerative disorders, orthopedic diseases and types of cancer. Accumulating evidence has shown that the P2X7R plays a crucial role in the development of various heart diseases. For example, activation of P2X7Rs may alleviate myocardial ischemia‑reperfusion injury by releasing endogenous cardiac protective substances. In contrast to these findings, activation of P2X7Rs can promote the development of acute myocardial infarction and myocarditis by inducing inflammatory responses. Activation of these receptors can also contribute to the development of different types of cardiomyopathies including diabetic cardiomyopathy, dilated cardiomyopathy and hypertrophic cardiomyopathy by inducing cardiac hypertrophy, fibrosis and apoptosis. Notably, inhibition of P2X7Rs can improve cardiac structure and function abnormalities following acute myocardial infarction, reduction of inflammatory responses following myocarditis and attenuation of the cardiomyopathy process. Furthermore, recent evidence has demonstrated that P2X7Rs are highly active in patients infected with coronavirus disease‑2019 (COVID‑19). Hyperactivation of P2X7Rs in COVID‑19 may induce severe myocardial injury through the activation of several signaling pathways. The present study reviewed the important role of the P2X7R in cardiac dysfunctions and discusses its use as a possible new therapeutic approach for the prevention and treatment of several myocardial diseases.

Soluble sugars, organic acids and energy metabolism involved in the wound healing of muskmelons elicited by benzothiadiazole

Benzothiadiazole (BTH) is an analogue of salicylic acid, which can reduce the disease severity in many harvested horticultural products via inducing resistance. In this study, muskmelons (cv. Perfect) were used as the materials to investigate BTH dipping on the sugars, organic acids and energy metabolism during healing. Results demonstrated that BTH increased fructose, glucose and sucrose contents at wounds of muskmelons in the early stage of healing. Further, BTH enhanced the contents of pyruvic acid, malic acid, fumaric acid, α-ketoglutaric acid and succinic acid, but decreased citric acid content. BTH also increased nicotinamide adenine dinucleotide (NAD) and its reduced form (NADH) contents. Moreover, BTH up-regulated ATP synthase subunit β and CmAAC expressions, but down-regulated CmAOX1D, CmUCP and CmPM-ATPase expressions, thereby maintaining the higher levels of adenosine triphosphate (ATP) and energy charge (EC). Taken together, BTH could promote the levels of soluble sugars, organic acids and energy at muskmelon wounds, which provide carbon skeletons and energy required for synthesis, transportation and cross linking of precursors of suberin and lignin, contributing to the formation of healing tissue.

Differential Immunomodulatory Effects of Head and Neck Cancer-Derived Exosomes on B Cells in the Presence of ATP.

Head and neck squamous cell carcinoma (HNSCC) is an aggressive malignancy. Tumor-derived exosomes (TEX) have immunoregulatory properties. Adenosine triphosphate (ATP) and its immunosuppressive precursor adenosine (ADO) have been found in cancerous tissue. We investigated the effect of TEX on B cells in the presence of ATP. TEX were isolated from human HNSCC cell line (PCI-13) cultures and co-cultured with peripheral blood B cells of healthy donors, with or without TEX in different concentrations and with or without a low (20 µM) or high (2000 µM) ATP dose. We were able to demonstrate that TEX inhibit B-cell proliferation. The addition of TEX to either ATP concentration showed a decreasing trend in CD39 expression on B cells in a dose-dependent manner. High ATP levels (2000 µM) increased apoptosis and necrosis, and analysis of apoptosis-associated proteins revealed dose-dependent effects of ATP, which were modified by TEX. Altogether, TEX exhibited dual immunomodulatory effects on B cells. TEX were immunosuppressive by inhibiting B-cell proliferation; they were immunostimulatory by downregulating CD39 expression. Furthermore, TEX were able to modulate the expression of pro- and anti-apoptotic proteins. In conclusion, our data indicate that TEX play an important, but complex, role in the tumor microenvironment.

High and stable ATP levels prevent aberrant intracellular protein aggregation in yeast.

Adenosine triphosphate (ATP) at millimolar levels has recently been implicated in the solubilization of cellular proteins. However, the significance of this high ATP level under physiological conditions and the mechanisms that maintain ATP remain unclear. We herein demonstrated that AMP-activated protein kinase (AMPK) and adenylate kinase (ADK) cooperated to maintain cellular ATP levels regardless of glucose levels. Single-cell imaging of ATP-reduced yeast mutants revealed that ATP levels in these mutants underwent stochastic and transient depletion, which promoted the cytotoxic aggregation of endogenous proteins and pathogenic proteins, such as huntingtin and α-synuclein. Moreover, pharmacological elevations in ATP levels in an ATP-reduced mutant prevented the accumulation of α-synuclein aggregates and its cytotoxicity. The present study demonstrates that cellular ATP homeostasis ensures proteostasis and revealed that suppressing the high volatility of cellular ATP levels prevented cytotoxic protein aggregation, implying that AMPK and ADK are important factors that prevent proteinopathies, such as neurodegenerative diseases.

Hygienic monitoring in long-term care facilities using ATP, crAssphage, and human noroviruses to direct environmental surface cleaning

BackgroundNorovirus and C. difficile are associated with diarrheal illnesses and deaths in long-term care (LTC) facilities and can be transmitted by contaminated environmental surfaces. Hygienic monitoring tools such as adenosine triphosphate (ATP) bioluminescence and indicators of fecal contamination can help to identify LTC facility surfaces with cleaning deficiencies. MethodsHigh-touch surfaces in 11 LTC facilities were swabbed and tested for contamination by norovirus, a fecal indicator virus, crAssphage, and ATP which detects organic debris. High levels of contamination were defined as log ATP relative light unit values or crAssphage log genomic copy values in the 75th percentile of values obtained from each facility. ResultsOver 90% of surfaces tested positive for crAssphage or gave failing ATP scores. Norovirus contamination was not detected. Handrails, equipment controls, and patient beds were 4 times more likely than other surfaces or locations to have high levels of crAssphage. Patient bed handrails and tables and chairs in patient lounges had high levels of both ATP and crAssphage. ConclusionsSurfaces with high levels of ATP and crAssphage were identified. Quantifying levels of contamination longitudinally and before and after cleaning might enhance infection prevention and control procedures for reducing diarrheal illnesses in LTC facilities.

Carvacrol delays Phomopsis stem-end rot development in pummelo fruit in relation to maintaining energy status and antioxidant system

Pummelo fruit rapidly depreciate in commodity value due to postharvest fungal decay and fruit quality deterioration. Here, we used carvacrol (CVR) to control Phomopsis stem-end rot (SER) caused by Diaporthe citri in pummelo fruit stored at 25 °C. Antifungal activity of CVR inhibited D. citri growth and Phomopsis SER development. Harvested pummelo fruit treated with CVR delayed firmness loss and lowered electrolyte leakage, and retarded hydrogen peroxide (H2O2) and malondialdehyde (MDA) accumulation. Unlike the control fruit, the CVR-treated fruit maintained higher levels of adenosine triphosphate and energy charge, and increased ATPase, succinate dehydrogenase (SDH), malate dehydrogenase (MDH), and cytochrome C oxidase (CCO) activities, along with up-regulated expression levels of the respective genes. CVR improved the antioxidant capacity, as evidenced by higher non-enzymatic antioxidants amounts, higher activities of superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), ascorbate peroxidase (APX) and glutathione reductase (GR), and up-regulated expression levels of ROS-scavenging-related genes. Collectively, CVR treatment maintained the energy status and antioxidant capacity in D. citri-infected pummelo fruit, which revealed antifungal mechanisms critical for controlling postharvest fungal diseases.

A new cell-sized support for 3D cell cultures based on recombinant spider silk fibers.

It is now generally accepted that 2D cultures cannot accurately replicate the rich environment and complex tissue architecture that exists in vivo, and that classically cultured cells tend to lose their original function. Growth of spheroids as opposed to 2D cultures on plastic has now been hailed as an efficient method to produce quantities of high-quality cells for cancer research, drug discovery, neuroscience, and regenerative medicine. We have developed a new recombinant protein that mimics dragline spidersilk and that self-assembles into cell-sized coils. These have high thermal and shelf-life stability and can be readily sterilized and stored for an extended period of time. The fibers are flexible, elastic, and biocompatible and can serve as cell-sized scaffold for the formation of 3D cell spheroids. As a proof of concept, recombinant spidersilk was integrated as a scaffold in spheroids of three cell types: primary rat hepatocytes, human mesenchymal stem cells, and mouse L929 cells. The scaffolds significantly reduced spheroid shrinkage and unlike scaffold-free spheroids, spheroids did not disintegrate over the course of long-term culture. Cells in recombinant spidersilk spheroids showed increased viability, and the cell lines continued to proliferate for longer than control cultures without spidersilk. The spidersilk also supported biological functions. Recombinant spidersilk primary hepatocyte spheroids exhibited 2.7-fold higher levels of adenosine triphosphate (ATP) continued to express and secrete albumin and exhibited significantly higher basal and induced CYP3A activity for at least 6 weeks in culture, while control spheroids without fibers stopped producing albumin after 27 days and CPY3A activity was barely detectable after 44 days. These results indicate that recombinant spidersilk can serve as a useful tool for long-term cell culture of 3D cell spheroids and specifically that primary hepatocytes can remain active in culture for an extended period of time which could be of great use in toxicology testing.

Development-related mitochondrial properties of retinal pigment epithelium cells derived from hEROs.

To explore the temporal mitochondrial characteristics of retinal pigment epithelium (RPE) cells obtained from human embryonic stem cells (hESC)-derived retinal organoids (hEROs-RPE), to verify the optimal period for using hEROs-RPE as donor cells from the aspect of mitochondria and to optimize RPE cell-based therapeutic strategies for age-related macular degeneration (AMD). RPE cells were obtained from hEROs and from spontaneous differentiation (SD-RPE). The mitochondrial characteristics were analyzed every 20d from day 60 to 160. Mitochondrial quantity was measured by MitoTracker Green staining. Transmission electron microscopy (TEM) was adopted to assess the morphological features of the mitochondria, including their distribution, length, and cristae. Mitochondrial membrane potentials (MMPs) were determined by JC-1 staining and evaluated by flow cytometry, reactive oxygen species (ROS) levels were evaluated by flow cytometry, and adenosine triphosphate (ATP) levels were measured by a luminometer. Differences between two groups were analyzed by the independent-samples t-test, and comparisons among multiple groups were made using one-way ANOVA or Kruskal-Wallis H test when equal variance was not assumed. hEROs-RPE and SD-RPE cells from day 60 to 160 were successfully differentiated from hESCs and expressed RPE markers (Pax6, MITF, Bestrophin-1, RPE65, Cralbp). RPE features, including a cobblestone-like morphology with tight junctions (ZO-1), pigments and microvilli, were also observed in both hEROs-RPE and SD-RPE cells. The mitochondrial quantities of hEROs-RPE and SD-RPE cells both peaked at day 80. However, the cristae of hEROs-RPE mitochondria were less mature and abundant than those of SD-RPE mitochondria at day 80, with hEROs-RPE mitochondria becoming mature at day 100. Both hEROs-RPE and SD-RPE cells showed low ROS levels from day 100 to 140 and maintained a normal MMP during this period. However, hEROs-RPE mitochondria maintained a longer time to produce high levels of ATP (from day 120 to 140) than SD-RPE cells (only day 120). hEROs-RPE mitochondria develop more slowly and maintain a longer time to supply high-level energy than SD-RPE mitochondria. From the mitochondrial perspective, hEROs-RPE cells from day 100 to 140 are an optimal cell source for treating AMD.

Double‐Lock Nanomedicines Enable Tumor‐Microenvironment‐Responsive Selective Antitumor Therapy

AbstractA hybrid nanocarrier for reducing the off‐target adverse effects of chemotherapy via selective drug delivery to the tumor cells is reported. The model active agent, combretastatin A4 (CA4) phosphate is deposited onto the magnetite (Fe3O4) nanoparticles as the core, followed by lipid coating as the shell. Upon nanocarrier administration and biodistribution to the tumor site, the high level of adenosine triphosphate in the extracellular space of tumor induces the cargo release via phosphate displacement. Then, the CA4 phosphate is dephosphorylated by the alkaline phosphatase that is overexpressed at the plasma membrane of certain tumor cells, resulting in enhanced intracellular uptake of hydrophobic CA4. These sequential two‐step unlocking processes enable the preferable accumulation of CA4 within tumor cells. Such approach is applicable to a wide range of chemotherapeutics and is promising for efficacy enhancement and side‐effect reduction of antitumor chemotherapy.

A Balance between the Activities of Chloroplasts and Mitochondria Is Crucial for Optimal Plant Growth.

Energy metabolism in plant cells requires a balance between the activities of chloroplasts and mitochondria, as they are the producers and consumers of carbohydrates and reducing equivalents, respectively. Recently, we showed that the overexpression of Arabidopsis thaliana purple acid phosphatase 2 (AtPAP2), a phosphatase dually anchored on the outer membranes of chloroplasts and mitochondria, can boost the plant growth and seed yield of Arabidopsis thaliana by coordinating the activities of both organelles. However, when AtPAP2 is solely overexpressed in chloroplasts, the growth-promoting effects are less optimal, indicating that active mitochondria are required for dissipating excess reducing equivalents from chloroplasts to maintain the optimal growth of plants. It is even more detrimental to plant productivity when AtPAP2 is solely overexpressed in mitochondria. Although these lines contain high level of adenosine triphosphate (ATP), they exhibit low leaf sucrose, low seed yield, and early senescence. These transgenic lines can be useful tools for studying how hyperactive chloroplasts or mitochondria affect the physiology of their counterparts and how they modify cellular metabolism and plant physiology.

Hydrolysis of Extracellular ATP by Vascular Smooth Muscle Cells Transdifferentiated into Chondrocytes Generates Pi but Not PPi.

(1) Background: Tissue non-specific alkaline phosphatase (TNAP) is suspected to induce atherosclerosis plaque calcification. TNAP, during physiological mineralization, hydrolyzes the mineralization inhibitor inorganic pyrophosphate (PPi). Since atherosclerosis plaques are characterized by the presence of necrotic cells that probably release supraphysiological concentrations of ATP, we explored whether this extracellular adenosine triphosphate (ATP) is hydrolyzed into the mineralization inhibitor PPi or the mineralization stimulator inorganic phosphate (Pi), and whether TNAP is involved. (2) Methods: Murine aortic smooth muscle cell line (MOVAS cells) were transdifferentiated into chondrocyte-like cells in calcifying medium, containing ascorbic acid and β-glycerophosphate. ATP hydrolysis rates were determined in extracellular medium extracted from MOVAS cultures during their transdifferentiation, using 31P-NMR and IR spectroscopy. (3) Results: ATP and PPi hydrolysis by MOVAS cells increased during transdifferentiation. ATP hydrolysis was sequential, yielding adenosine diphosphate (ADP), adenosine monophosphate (AMP), and adenosine without any detectable PPi. The addition of levamisole partially inhibited ATP hydrolysis, indicating that TNAP and other types of ectonucleoside triphoshatediphosphohydrolases contributed to ATP hydrolysis. (4) Conclusions: Our findings suggest that high ATP levels released by cells in proximity to vascular smooth muscle cells (VSMCs) in atherosclerosis plaques generate Pi and not PPi, which may exacerbate plaque calcification.

Near-saturated relative humidity alleviates chilling injury in zucchini fruit through its regulation of antioxidant response and energy metabolism

To investigate the effect of relative humidity (RH) on chilling injury (CI), zucchini fruit were stored in cold rooms (4 ± 0.4 ℃) with different RHs (near-saturated RH [NSH] with 96–100% and normal RH with 72–76% served as control). Storage in NSH delayed weight loss and CI, maintained firmness and skin color. Higher antioxidant enzyme activities and greater scavenging capacities of free radicals were found in NSH-fruit than in the control fruit. The decrease of the unsaturated fatty acids was delayed in NSH-fruit due to lower activities of related membrane lipid degrading enzymes as compared to the control fruit. NSH-fruit also maintained higher activities of energy metabolism-associated enzymes than control fruit, leading to high levels of adenosine triphosphate (ATP). Taken together, we attributed the alleviation of CI by NSH storage to its enhancement of antioxidant capacities and its effect on maintaining higher energy status in zucchini fruit.